Abstract Book

ABI1 Protein Phosphatase 2C inhibitor designed to improve crop yield.

Maciej Janicki, Agata Cieśla & Agnieszka Ludwików

Adam Mickiewicz University in Poznan, Poland

Reversible protein phosphorylation is a key protein modification involved in the regulation of many cellular processes. Phosphorylation/dephosphorylation processes are catalysed by two groups of enzymes: protein kinases and protein phosphatases. There are more than 1000 genes encoding protein kinases and protein phosphatases in Arabidopsis. The protein phosphatase ABI1 plays a crucial role in plant growth, development and response to abiotic stresses. Type 2C group A protein phosphatases are known effectors of the ABA signalling pathway. Understanding ABA perception and signalling has been a major goal of plant research for decades. In this study, we present the crystal structure of the ABI1 protein obtained by X-ray crystallography. According to the literature, the structure of ABI1 has only been solved in complex with the active site bound ABA receptor, and it is unclear whether the conformation changes significantly upon substrate or inhibitor binding. The novel crystal structure of the free-standing ABI1 phosphatase provides an opportunity as a new target for the search for molecules as PP2C inhibitors. We present here an approach for the identification of effective inhibitors of ABI1-like PP2C. Overall, the crystal structure of the ABI1 protein provides important insights into its organisation and functional implications. This knowledge paves the way for a comprehensive understanding of the role of the ABI1 protein in plant physiology and for plant improvement strategies aimed at enhancing abiotic stress tolerance.

The role of biodiversity in mitigating climate change – soil fauna and regenerative agriculture

Carolyn-Monika Görres

Hochschule Geisenheim University, Germany

Anthropogenic climate change is a major global crisis threatening the livelihood of people around the globe. Agriculture is part of the problem contributing about 20 % to annual global greenhouse gas (GHG) emissions. These are mainly the result of soil carbon (C) losses due to land-use conversion and soil-degrading land management practices, nitrous oxide (N2O) emissions tied to fertilization, and methane (CH4) emissions from livestock and rice production. But agriculture is and needs to be part of the solution to mitigating climate change. Many countries try to reduce agricultural GHG emissions and improve soil C sequestration on agricultural land through improved land management practices. One promising way forward is the implementation of regenerative agriculture. There is no agreed consensus definition, but it usually comprises management techniques such as reduced or no till, cover cropping, crop rotation, reduced use of synthetic inputs (fertilizer and pesticides), use of organic amendments, increased use of perennials and agroforestry, crop- livestock rotation and managed grazing. Many of these techniques were and still are traditionally used by smallholder farmers and are partly rediscovered for a wider application since they are not only aimed at increasing soil organic C and soil health, but also at increasing biodiversity. Biodiversity loss is the other major global crisis of our times and industrial agriculture is once again one of the main contributors. Climate change and biodiversity loss are tightly interconnected and the challenge is to identify on a regional and farm-level regenerative agricultural practices that provide a win-win situation for climate change mitigation and biodiversity while sustaining yields. However, while this has been recognized in theory, in practice the climate change community and the biodiversity community – both scientists and practitioners – are too often not working together resulting in serious knowledge gaps which prevents the timely identification of win-win situations. This is especially the case for soil fauna. Soil animals are direct producers of GHG emissions via their respiratory and metabolic activities and can indirectly change soil C and nitrogen (N) cycles by changing physical, chemical and biological soil properties, e.g. through bioturbation, defecation, herbivory, and litter fragmentation and redistribution. In addition, they can create microhabitats which offer favorable conditions to microorganisms. Thus, soil fauna is able to substantially effect the spatial and temporal variability of GHG fluxes in ecosystems. However, emissions of carbon dioxide (CO2), CH4 and N2O from and associated with soil fauna remain poorly quantified and have been limited to only a few regions, species and land use types. Since 2010, only ~200 original GHG emission studies addressing soil fauna have been published, with the vast majority dedicated to earthworms. Two huge barriers preventing the quantification of soil faunal GHG emissions is the lack of suitable non-invasive methods for spatial and temporal soil fauna monitoring and a lack of ecological knowledge on species groups. In this presentation, I will make the case that the conference theme “Bridging disciplines, building connections” is essential and has a huge potential in further illuminating the “black box” soil. New methods need to be developed to allow non-invasive field monitoring of soil biodiversity, e.g. based on ecoacoustics, and new soil health indices are needed focusing on different soil faunal groups and soil food webs for inclusion in GHG flux models. However, since the ongoing biodiversity crisis also affects belowground communities, time is of the essence: We might already be losing soil fauna species before we are able to understand their role in mitigating climate change in agroecosystems!

Sustainable Agriculture for Food Security in Changing Climate

Muhammad Farooq

Department of Plant Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al-Khoud 123, Oman

Email: farooqcp@squ.edu.om

An increase in global food demand, declining natural resources, deteriorating natural ecosystems, and changing climate are threatening the sustainability of agriculture and food security. The major challenges to agriculture and food security, due to climatic changes, include the occurrence of un-predicted temperature fluctuations, heat waves, drought episodes, floods, and epidemics of biotic stresses, etc., The climatic changes and intensive tillage have also increased soil erosion causing a decline in soil organic matter, deteriorating soil structure, and giving rise to soil salinity resulting in a loss of overall soil fertility and productivity. In addition to environmental pollution, pesticide use has resulted in the development of resistant pest biotypes. Sustainable agriculture, the system of harvesting better crop yields with minimal adverse environmental impact and without bringing non-agricultural land under cultivation, is the pragmatic option to ensure food security for future generations in the changing climate. With a focus on environmental sustainability through an ecosystem approach, sustainable agriculture aims to maximize options for crop production through the management of biodiversity and ecosystem services.

Dehydration of Plant-Based Ingredients: Enhancing Supply Chain Resilience and Ensuring Food Security

Dr. Muhammad Tuseef Asghar1*, Dr. Muhammad Azhar Ali1, Dr. Sadaf Shakoor2, Sadaf Fatima3, Hafiz Azhar Rasool4, Muhammad Usman Afzal5, Muhammad Mohsin Ali6

University of Agriculture, Faisalabad, Department of Food
University of Agriculture, Faisalabad Sub Campus Burewala
TruECaP Seasonings, Burewala,
NAMAL University
PMAS Arid Agriculture University
PARC-National Agricultural Research Centre (NARC),

Email: mtuseef@uaf.edu.pk

Seasonal variations in agricultural produce often lead to market uncertainties, posing challenges for both producers and consumers. This study explores the potential of dehydrating key culinary ingredients such as onions, garlic, ginger, green chilies, and tomatoes to mitigate these uncertainties and enhance food security. We aim to preserve these produce items' nutritional value and flavor by employing a low-temperature dehydration technique. The resulting dehydrated products are not only easy to transport and versatile for various culinary uses but also have an extended shelf life. This process benefits smallholding producers by reducing post-harvest losses and ensuring a consistent supply of produce. Additionally, the value-added dehydrated products open new market opportunities for producers. These dehydrated ingredients offer end consumers several advantages, including convenience, space-saving storage, and reduced cooking time and energy consumption. The proposed project emphasizes a holistic approach from farm to fork, highlighting its economic and environmental benefits. This initiative aligns with the UN's Sustainable Development Goals (SDGs) 2, 3, 8, and 9, contributing to global food security and sustainability.

Rapid Introgression of SBE mutant genes into Pakistani Wheat through Marker-Assisted Speed Breeding

Muhammad Sajjad*a, Saima Mir Arainb, Zahid Mahmoodc, , Dongcheng Liud*

Nuclear Institute of Agriculture, NIA, Tandojam, 70050, Pakistan
Crop Sciences Institute, National Agricultural Research Centre (NARC),

Park Road, Islamabad, 44000, Pakistan

State Key Laboratory of North China Crop Improvement and Regulation, College of Agronomy, Hebei Agriculture University,

Baoding 071000, Hebei, China

Email : muhammad.sajjad@comsats.edu.pk; liudongcheng@hebau.edu.cn

Wheat, serving as a staple for billions worldwide, is becoming more important with increasing rate of diabetes. High amylose wheat (HAW) is considered as a potential remedy against type 2 diabetes and obesity. Trait introgression (TI) through traditional methods is laborious, sensitive, resource intensive and time consuming. Here, we report a rapid introgression of high amylose trait from 'Patwin' SBEII quintuple mutant to a Pakistani wheat variety through Marker-Assisted Speed Breeding (MASB). The integration of KASP genotyping with speed breeding method facilitated precise trait selection and accelerated breeding cycles. The resultant SBEII mutant line exhibited substantial increase (73%) in amylose content in comparison to the wild-type control. The increase in resistant starch (RS) was more than 20 folds. The MASB with KASP expedited trait introgression, shortening the time frame from 6-7 years to 2.5 years. This study exemplifies the acceleration of trait introgression into our elite cultivars through MASB.

Analyzing current agronomic management for sustainable wheat production enhancing nitrogen use efficiency in changing climate

Abdul Wakeel

Institute of Soil and Environmental Sciences, University of Agriculture, Faisalabad-Pakistan

Email: abdulwakeel77@gmail.com

Increasing wheat production and gross margins (GMs) of farms also lies in optimization of agronomic management settings which includes key factors like sowing date (SD) and nitrogen (N) fertilizer use. While maximizing wheat productivity, N use efficiencies (NUE) (agronomic/economic) may also be impacted due to changed N rates. To examine these questions, a Monte Carlo analysis was conducted at two study sites, Faisalabad, and Bahawalpur, in arid and semi-arid regions, using the process-based cropping system model APSIM (previously robustly calibrated/validated for the study sites). A broad range of combinations of N rate (67-804 kg N ha-1) and SD (30th October, 10th, 20th, 30th November and 10th December) were used. APSIM simulations were run for 32 years of historical climate data from 1991 to 2022. Wheat production, GMs, agronomic and economic NUE were calculated and reported for each crop cycle and averaged 32 years of simulations. Our results indicate that maximum wheat production and GMs were achieved when the current recommended N rate was multiplied by a factor of 2.0x and 1.5x (corresponding to 268 and 201 kg N ha-1) at both sites. The optimum SDs to maximize production and GMs were 20th and 30th of November. Even for late SD, farmers can maximize their production and GMs by incrementally increasing N fertilizer rate. Agronomic and economic NUE showed a declining trend when the N fertilizer rate was increased, keeping production and GMs as key targets. This is likely due to increased environmental N-losses at higher N-application rates, and we have suggested further work to explore additional strategies of N fertilizer timing and splitting aimed at ameliorating these increased losses. It concludes that there are tradeoffs between higher production, GMs and NUE, at increased N fertilizer rates keeping production and GMs as key targets compromising NUE. Our analysis also suggests that robustly calibrated/validated crop models like APSIM are useful tools to optimize agronomic management settings globally, where farmers are still not able to find ideal combination of these settings. In recent local studies we come up with few more agronomic solutions to enhance NUE in wheat-maize cropping system

Potential Role of Epigenetics in Crop Improvement

Abdul Rehman Khan, Zahid Ahmad Khan, Shahid Masood Shah

Department of Biotechnology, COMSATS University Islamabad Abbottabad Campus

Epigenetics refers to the changes in the gene activities that can be inherited mitotically and/or meiotically but it does not involve changes in DNA sequence. There are three main mechanisms included in epigenetics: DNA methylation, histone modification and RNA mediated gene regulation. For the last two decades, the study of these mechanisms in response to various environmental factors and their inheritance have been the center of interest. A large part of these changes is transmitted only during the lifetime of an individual and are reset in the next generations. But a significant amount of these changes are being transmitted to the multiple subsequent generations and provides and additional source of genetic variability (epigenetic variability). Crop breeders/scientists are always in search of new genetic variations. This variability at the DNA level enables us to use it as the basis for crop improvement. Therefore, any technique that can increase the genetic diversity is of interest. In this regard, epigenetics can be very useful, as it involves an additional layer of information with the ability to regulate the gene activities. There are various reports confirming the transgenerational inheritance of DNA methylation variations (an epigenetic modification) induced by different sources. This part of genetic diversity (epigenetic diversity) for crop improvement is largely unexplored. Currently, different approaches are in use for the induction of epigenetic diversity and its use in crop improvement. One such approach includes development of epiRIL lines where the mutants with silenced methyltransferases are produced and the epigenetic variability created due to this is being evaluated and used. Another technique chemically induce epigenetic diversity through demethylating agents like 5-Azacytidine, check their transgenerational heritability and use this transgenerationally heritable epigenetic diversity for crop improvement. Both these techniques have shown promising results in different crops and have opened a new and unexplored level that has the potential that must be utilize for crop improvement in this era of population growth and climate change.

Genome-based Breeding for Development of Climate Resilient Crops in Rapid Time

Muhammad Ramzan Khan*, Nazia Rehman, Muhammad Kashif Naeem, Amna Muhammad, Muhammad Uzair, Aamir Riaz, Wajya Ajmal, Fara Muqaddas, Maryam Murtaza, Bilal Saleem, Bushra Bibi, Muhammad Aqeel

Functional Genomics and Bioinformatics Program, National Institute for Genomics and Advanced Biotechnology, National Agricultural Research Centre, Park Road, Islamabad, Pakistan.
Wheat program, Crop Sciences Institute, National Agricultural Research Centre, Park Road, Islamabad,

Email: mrkhan@parc.gov.pk

According to food security analysts the global agricultural production must double by 2050 to meet the demands of increasing world human population, but this challenge is further exacerbated by unpredictable climate change. The availability of new climate-resilient crop varieties in rapid time is an uphill task to feeding the global population. Lack of application of new breeding technologies (NBTs) is the major bottleneck for speedy development of the climate resilient varieties. For rapid increase in yield, the utilization of NBTs including implementation of UAV-based high throughput phenomics, speed cloning, genome–based breeding by design, genomic selection, and Express- Editing galvanized by speed breeding are the desirable strategies to meet the rising food demand. Speed cloning can reduce the linkage drag to a minimum level -imminent during the Marker Assisted Selection. Pre-breeding in the form of the discovery of new genes and association of markers with traits through Genome Wide Association Studies (GWAS) has added to traits-specific breeding. Advancements in NGS have revolutionized genome-based breeding by determining the genomic constitution of parents and offspring for a particular target environment (TE). In this regard, new genomic platforms, PCR- based KASP markers, and options for indigenous PCR-based NGS library preparation kits have facilitated the use of genomic selection based on genetic variants existing in indigenous cultivars. A decade of CRISPR/Cas technology has brought improved nutritional value, disease resistance, and improvement and expansion in crops. i.e., Genomics to phenomics along with integrated applications of all these technologies - if supplemented with speed breeding, the desirable crops can be achieved in less than half time. Hence, the implementation of genome-based speed breading technologies has a great potential for accelerating the pace of development of new high-yielding climate-smart varieties.

Hydrological assessment for the flash flood management with eco-system base adaptation (EBA) in Punjab Pakistan

(A case of Soon Valley)

Muhammad Imran Azam, Umer Bin Khalid, Masood Arshad, Sohail Ali Naqvi Hamza Mukhtar, Muhammad Ahamd

*Freshwater Programe WWF-Pakistan

The hydrological assessment was performed for the flash flood management with eco-system base adaptation (EBA) structures of the Soon Valley (Ramsar Site), Punjab, Pakistan. The project area is one of the most dramatic physio- graphic features of the Punjab province. The frequent natural disaster in this region that devastates the inhabitants, the economy, and the infrastructure is the flash flood. The impacts of flash floods in Soon Valley were investigated using the Geographic Information System (GIS), Remote Sensing (RS), Water Modelling system (WMS) and Hydrologic Modelling System (HEC-HMS). Utilizing the SCS curve number method and HEC-HMS model, the hydrological analysis includes the identification of suitable sites for eco-system base adaptation (EbA) interventions. As per assessment Eco-system Base Adaptation Structures (EbA) i.e. retention ponds, delay action reservoirs, slope stabilization and Ablution Water Reuse System (AWRS) were designed and constructed to manage the flash flood of this area. By the construction of aforementioned interventions the flood intensity is minimized upto 70% and also control the erosion of this area. The results of this project highlight the significance of adaptive infrastructure for flash flood management, climate resilience infrastructure (NBS) to promote the smart agriculture of this area. The project aims to provide a sustainable framework and wayforward for the communities, experts and government departments for sustainable agriculture, water management of the Soon Valley.

Green synthesis and characterization of Zinc Oxide Nanoparticles using Thymus linearis and evaluation of different biological activities

Javed Iqbal1*, Farishta Zarshan1, Banzeer Ahsan Abbasi2, Zakir Ullah3, Tariq Mahmood3

Department of Botany, Bacha Khan University, Charsadda, 24420, Khyber Pakhtunkhwa,
Department of Botany, Rawalpindi Women University, 6th Road, Satellite Town, Rawalpindi
Department of Plant Sciences, Faculty of Biological Sciences, Quaid-i-Azam University Islamabad

Email: javed89qau@gmail.com

In the present research study, ZnONPs have been prepared using Thymus linearis (TL) aqueous extract as both reducing and stabilizing agent. Several analytical tools including ultraviolet-visible spectroscopy, SEM, EDX, XRD, DLS and FT-IR were employed to validate the synthesis of as-prepared ZnONPs. FTIR analysis was done to determine the presence of different bioactive functional groups adsorbed on the surface of TL-ZnONPs. SEM analysis was performed to determine the shape and morphology of the TL-ZnONPs. Further, XRD analysis was done to understand about the crystalline nature of biogenic ZnO nanoparticles and the size was calculated as 30 nm. To determine the elemental composition, EDX analysis was performed which revealed the successful formation of TL-ZnONPs. Furthermore, ZnONPs were examined for multifaceted biological activities. The antibacterial properties were analyzed using different bacterial strains revealing significant antibacterial potential. The maximum resistance was observed in case of Enterococcus faecalis (MIC: 125 µg/ml), however, Staphylococcus saprophyticus (MIC: 31.25 µg/ml) was found to be the most sensitive among all strains. The fungicidal properties were demonstrated using different fungal strains. The maximum resistance against all doses was observed in case of Aspergillus flavus (MIC: 125 µg/ml). However, Aspergillus niger (MIC: 31.25 µg/ml) was found to be the most sensitive among all strains. The TL-ZnONPs were evaluated for antioxidant potentials and the highest DPPH scavenging potential was reported 85.03%, total reducing power (81.4%) and total antioxidant capacity (79 %) were observed at highest of 1050 µg/ml. Further, our TL-ZnONPs revealed excellent cytotoxic potential against nascent brine shrimp’s larva’s (IC50 34.16 µg/ml). The anticancer property of TL-ZnONPs were investigated and have shown strong potential against HepG-2 cell lines with IC50: 24.85 µg/ml.

Biochar Production and Utilization for Metal Removal from Agricultural Soils: A Decade-Long Journey in Soil Amendment and Environmental Protection

Muhammad Farooq Qayyum

Department of Soil Science, Faculty of Agricultural Sciences & Technology,

Bahauddin Zakariya University Multan

farooq.qayyum@bzu.edu.pk

We have been working on various aspects of biochar research over the past decade in Pakistan. The journey started from 2012 with a funding from Higher Education Commission Pakistan, to work on biochar preparation, characterization and use as amendment in alkaline soils. A variety of feedstock undergone pyrolysis to prepare biochars. Pot and field experiments were conducted using funds from this project. In 2015, we received another funding from the International Foundation for Science, Sweden and focused on utilization of biochar for mitigating cadmium uptake by the field crops. The results of these experiments were published in different peer-reviewed journals. We received more fundings from the Higher Education Commission Pakistan in 2015, and 2018 which further enhanced our motivation in the field of biochar research. In most of the studies, we have worked on remediation of potentially toxic elements (also termed as heavy metals) in sewage irrigated peri-urban lands. Heavy metals, a significant problem in Pakistani soils, hinder nutrient absorption by plants, leading to reduced crop growth, development, and yields, contributing to food scarcity. We have used a variety of biochar which were different based on the nature of feedstock. We studied sorption of single metals or combined in solution forms on various biochars. Moreover, the research was extended over different agricultural fields while growing different cereals. The effect of different biochars on soil mineralogical properties were also studied while analyzing soil samples from the incubation and field experiments. More recently, use of green-synthesized nanoparticles for modification of biochar was done and used as amendments in the plant growth trials.

Impact of Green-Synthesized Silver Nanoparticles (AgNPs) on Wheat Carbohydrate Metabolism and Protein Quantification under Heat Stress in Pakistan's Changing Climate

Naveed Iqbal Raja

Department of Botany, PMAS Arid Agriculture University Rawalpindi, Rawalpindi, Pakistan

Email: drnaveedraja@uaar.edu.pk

In the present investigation, the role of GS-AgNP treatment in wheat plants was carried out in reducing heat stress with the aim of facilitating scientists on this topic. The effect of GS-AgNPs against heat stress has rarely been deliberated in wheat plants, and only a few studies have been established earlier in this scenario. This work illustrated the effect of GS-AgNPs on the regulation of carbohydrates metabolism, SOD, proteins, crude fibers, and minerals changes in wheat plants. Data were analysed using PCA analysis, correlation parameters, and normal probability distribution in PAST 3 software. The results indicated that heat stress alone caused severe changes in carbohydrates metabolism, SOD, proteins, crude fibers, and minerals immediately so that plants could not recover without foreign stabilizers such as GS-AgNPs. The application of GS-AgNPs increases the flux of carbohydrates metabolism, SOD, and proteins, including HSPs, crude fibers, and minerals, in wheat plants to reduce the effect of heat stress. The 50 mg/l concentration of GS-AgNPs has shown an increase in carbohydrates metabolism and SOD activity, while crude fibres have shown a significant enhancement at 100 mg/l of GS-AgNPs. The crude and true proteins were also shown pronounced increase in treatment to a concentration of 50 mg/l of GS-AgNPs. GS-AgNPs stimulated HSP production; most importantly, smHSP production was observed in the present results with other HSPs in wheat plants treated with a 50 mg/l concentration of GS-AgNPs. The mineral distribution was also regulated by the respective treatment of GS-AgNPs, and the highest amounts of Ca, P and Fe were found to be highest in wheat under heat stress. In general, we computed the expected model based on GS-AgNPs on the genes/factors that respond to heat stress and their potential role in mitigating heat stress in wheat. In addition, we discussed the prospective signalling pathway triggered by GS-AgNPs in wheat against heat stress. In the future, this work might be helpful in distinguishing the genetic variation due to GS-AgNPs in promoting tolerance in wheat against heat stress.

Indigenous Nature-based solutions and Ecosystem based approaches in addressing climate change in Pakistan

Ahmad Mahmood, Amar Matloob, Muqarrab Ali, Tanveer ul Haq

Department of Climate Change, MNS-University of Agriculture, Multan, Pakistan.

Email: ahmad.mahmood@mnsuam.edu.pk

Pakistan, ranked 8th among the most climate-vulnerable countries, faces multiple environmental challenges, including floods, heatwaves, droughts, glacial lake outburst floods (GLOF), biodiversity loss, shifting rainfall patterns, and economic instability. These issues also contribute to food insecurity and mass migration. In response, indigenous Nature-Based Solutions (NbSs) and Ecosystem-Based Adaptation (EbA) practices are being employed to address climate change. EbA emphasizes ecosystem conservation, sustainable management, and restoration, while NbSs adopt a broader, multifunctional approach that integrates nature into sustainable development and climate resilience. Indigenous knowledge plays a vital role in climate adaptation. Traditional practices such as animal grazing for improved crop yields, rainwater harvesting, agroforestry, and integrated farming systems have been used for centuries. These practices include community-level water management, crop diversification, and sustainable livestock management. Furthermore, nomadic and semi-nomadic livestock systems, terracing, soil conservation, and spate irrigation are long-standing methods that enhance biodiversity and ecosystem health. The perspective highlights that solutions to Pakistan’s climate challenges already exist through indigenous practices that promote regenerative, sustainable, and organic agriculture. To mitigate the impacts of climate change, Pakistan must prioritize and expand these time- tested ecosystem-based and nature-driven approaches.

Indigenous Nature-based solutions and Ecosystem based approaches in addressing climate change in Pakistan

Ahmad Mahmood, Amar Matloob, Muqarrab Ali, Tanveer ul Haq

Department of Climate Change, MNS-University of Agriculture, Multan, Pakistan.

Email: ahmad.mahmood@mnsuam.edu.pk

Enhancing Caralluma tuberculata’s Antioxidant and Hypoglycemic Properties: The Role of Light Regime and Selenium Nanoparticles

Amir Ali, Zia-ur-Rehman Mashwani*

Department of Botany, PMAS Arid Agriculture University, Rawalpindi

Email: mashwani@uaar.edu.pk

In vitro plant cultures have emerged as a viable source of biomass enriched with potent secondary metabolites, holding significant promise for medicinal applications. This study aimed to explore the antioxidative and hypoglycemic potential of selenium nanoparticles (SeNPs) and light stressed mediated in vitro cultures of Caralluma tuberculata callus extract. The morphological and physicochemical features of biogenic selenium nanoparticles were evaluated by UV–Visible spectrophotometry, Scanning Electron Microscopy, Energy Dispersive X-rays, Fourier-transform infrared spectrometry, and Zeta Potential. The development of optimal callus was achieved through a two-week incubation in a dark environment and shifted to normal condition by using Murashige and Skoog (MS) media supplemented with 100 µg/L of Selenium NPs, along with 0.5 mg/L of 2,4-Dichlorophenoxyacetic acid and 0.5 mg/L of 6- Benzyladenine, resulting in maximum biomass production at 56 days. This well- developed callus, rich in secondary metabolites, was selected to evaluate its antioxidative and hypoglycemic properties. The antioxidative potential of the callus extract was assessed through various tests, including ABTS+ scavenging, hydrogen peroxide inhibition, hydroxyl radical scavenging, and reducing power. Likewise, the hypoglycemic capacity of the callus extract was probed through assays involving alpha-amylase, alpha-glucosidase, glucose absorption by yeast cells, anti-sucrase, and anti-lipase activities. Notably, concentrations of 200 and 800 µg/ml exhibited pronounced efficacy in mitigating radical species. To delve into the hypoglycemic effects of the callus extract, assessments involving alpha amylase, alpha-glucosidase, and glucose absorption by yeast cells were performed across varying glucose concentrations (5 mmol/L, 10 mmol/L, and 25 mmol/L). The outcomes consistently demonstrated a dose-dependent relationship, with higher concentrations of the callus extract exerting potent inhibitory effects on the catalytic sites of the enzymes. The findings of this study propose a mechanism wherein selenium nanoparticles and light stressed mediated in vitro callus cultures of C. tuberculata potentially interact with charged species, operating as competitive inhibitors. This interaction retards the enzyme- substrate reaction, thereby impeding enzymatic degradation. These noteworthy outcomes hold promising implications for future medical applications of extract derived from callus cultures under the nanoparticles and light stressed condition.

ASSESSMENT OF WATER UTILIZATION DYNAMICS OF MINI DAMS: A CASE STUDY POTOHAR PAKISTAN

Usman, M.,1,2*, Umair .Y.C1, Saira A.1, Usman, K3., Cheema M.J.M.1,4,

Bashir, A.5, and Hussain, Z.6, Email: us.usman791@uaar.edu.pk

Water scarcity due to climate change is a major crisis spreading globally and it’s threatening all sectors. The agriculture sector is facing alarming challenges due to inadequate availability of water during critical stages. Rainfed agriculture is practiced in almost all hydroclimatic zones, and globally, more than 80 percent of the cropped area depends on rainfall alone. In Pakistan, about 23 Mha is cultivable crop land, and nearly 25 percent is designated as rainfed agriculture. The land and water productivity in rainfed regions is very low due to climate change, uneven rainfall pattern, absence of irrigation network, and multifarious hydrogeological dynamics. The government has introduced various projects in the Potohar region, e.g., construction of small dams, mini dams, water ponds, and fruit valleys. These water reservoirs are multifunctional and constructed by different government organizations under different schemes. Although some department level studies have been conducted to examine the impact of water reservoirs, no comprehensive research has been reported in the scientific literature. The present research has been conducted to identify the spatial spread of water reservoirs and to examine water storage and utilization dynamics in the Potohar region. A questionnaire was designed to gather information about water utilization dynamics for irrigation, poultry, dairy, fisheries, and other purposes under the command of mini dams. Various performance indexes were defined to evaluate the water utilization for poultry, dairy, irrigation, fishery, and overall performance index. Moreover, the research work has established knowledge for the stakeholders, which helps them to introduce more effective schemes and projects to enhance land and water productivity. In view of the surveyed and secondary data from different government organizations, it has been estimated that only 10 percent farmers are using water management techniques. Construction of water reservoirs in the region has promoted irrigated agriculture, but mostly farmers are practicing traditional flood irrigation or open pipe irrigation. The mini dam owners have adopted fish farming as an easy and economical approach for revenue generation from stored water. It is therefore recommended that an integrated farming approach be promoted in the region to enhance land and water productivity. It is suggested that benchmarks should be established for water reservoir performance assessment to track socio-economic, environmental and eco-hydrological benefits.

Climate smart agriculture - a way forward for climate change adaptation

Dr. Muhammad Sanaullah

Institute of Soil & Environmental Sciences, University of Agriculture Faisalabad, Pakistan

Climate change and unsustainable soil management practices have significantly depleted soil organic matter (SOM), contributing to the release of greenhouse gases (GHGs) and compromising soil health and productivity. Poor plant growth, limited SOM input, and accelerated decomposition under changing climatic conditions further hinder efforts to enhance SOM levels. Consequently, soil capital and sustainability are under threat, with more than 40% of agricultural soils worldwide classified as degraded, and salinity levels rising due to these climatic impacts. Intensive conventional farming practices that strip SOM have accelerated soil degradation, leading to the loss of vital soil resources. Understanding the impact of climate change on sustainable soil functioning is crucial for developing effective management strategies. Climate-smart agriculture (CSA) offers a promising approach to restore SOM, improve soil structure and fertility, and mitigate atmospheric GHGs emissions. By integrating sustainable soil management practices, such as cover cropping, reduced tillage, organic amendments, and diversified cropping systems, CSA aims to enhance soil carbon sequestration while promoting resilience against climate variability. However, the wide variation in estimates of soil organic carbon content and sequestration potential underscores the need for comprehensive assessments of soil properties. This knowledge is essential to inform policies and practices that prioritize SOM restoration as a key component of climate-smart agriculture. Effective soil management focused on SOM enhancement can not only improve soil health and agricultural productivity but also play a critical role in combating climate change and ensuring long-term sustainability.

Enhancing Plant Immunity Against Tan spot disease by Targeting Brassinosteroid-Regulated Fatty Acid Biosynthesis to Inhibit ToxA Activity

Dr Muhammad Junaid Yousaf Abdul Wali Khan University Mardan junaidyousaf44@gmail.com

Plant-pathogen interactions are intricate processes with profound implications for agriculture and food security. This study investigates the interplay between brassinosteroids, fatty acid biosynthesis, and the pathogen Pyrenospora tritici-repentis (Ptr) to enhance plant defenses by inhibiting ToxA, a known Ptr virulence factor. To achieve this, 11-day-old Triticum aestivum seedlings were infected with Ptr. Five days post-inoculation, we assessed growth parameters, photosynthetic efficiency, secondary metabolites, and ROS scavenging activities. As expected, Ptr infection reduced growth and photosynthesis due to lower secondary metabolite levels and subsequent ROS scavenging compared to the control. These effects were more pronounced when brassinosteroids (bL) were blocked before infection. Surprisingly, applying bL before infection improved growth and photosynthesis by increasing secondary metabolites and ROS scavenging. Further analysis of SRA profiles (SRR8816261, SRR8816277, SRR8816282, SRR8816278) from the NCBI SRA repository and GC-MS identified multiple metabolites in bL-Ptr treatment. Molecular docking and simulation using Gromacs revealed four fatty acid molecules with strong binding to ToxA, reducing its interaction with host Tsn1 proteins and diminishing its virulence.

Investigating the Protective role of cellulose and lignin-derived smoke cues in seed germination under heavy metal stress Chromium (Cr) Kishwer Naseer

ARID university Rawalpindi

Email: kishwernaseer03@gmail.com

Seed dormancy is an important characteristic for many plant species. One well-known tool for encouraging plant growth and development is smoke created from plants. When it comes to species with varying levels of seed coat permeability, heat shock and smoke-induced seed dormancy breaking which promotes germination are significant complimentary variables. In this circumstance, seeds cannot sprout even in the presence of ideal environmental factors. Smoke and its active components induce seed germination, increase seedling growth and mitigation of abiotic stresses. Contamination of heavy metal, particularly with chromium (Cr), shows an important environmental stressor that severely affects growth of plant, development, and productivity. Agriculturally significant crops such as wheat this is of particular concern for them which are foundational to global food security. Plants exposed to chromium may get variety of negative consequences, such as oxidative stress, altered metabolic processes, and decreased germination rates which eventually compromise crop quality and production. Research on smoke-derived cues from cellulose and lignin has shown promise in the search for novel ways to increase plant resilience against such abiotic stresses. Recent studies have shown that some substances found in smoke, which are mostly produced when cellulose and lignin burn, can cause seeds to germinate and drive plant growth responses. These results imply that cues obtained from smoke may be able to lessen the detrimental effects of heavy metal stress on plants. As common organic polymers, lignin and cellulose burn to produce a complex mixture of chemicals, some of which are known to have physiologically active characteristics that can have a variety of effects on plant physiology. Among these, the ability to mitigate the harmful impacts of heavy metals like chromium presents a strong opportunity for agricultural innovation. The purpose of this study proposal is to examine how smoke cues originating from cellulose and lignin contribute to seed germination and plant growth in the presence of chromium stress. Through an investigation of the fundamental mechanisms by which smoke cues give resistance to heavy metal toxicity, this work aims to open new avenues for augmenting the environmental stress- resilience of important agricultural crops. In the face of mounting environmental concerns, these techniques could use smoke-derived cues as a sustainable way to enhance crop growth in heavy metal-contaminated areas. This would promote agricultural production and food security.

Application of machine learning to avoid detrimental effects of using whole genome markers in additive and epistatic prediction models

Faisal Ramzan1, Saif-ur-Rehman1, Waseem Abbass1, M. Mahboob Ali Hamid1, Felix Heinrich2, Armin

Schmidt2, Mehmet Gultas3
Institute of Animal and Dairy Sciences, University of Agriculture Faisalabad, Jail Road, 38000 Faisalabad, Pakistan
Breeding Informatics Group, Department of Animal Sciences, Georg-August University, Margarethe von Wrangell-Weg 7, 37075 Göttingen, Germany
Faculty of Agriculture, South Westphalia University of Applied Sciences, 59494 Soest, Germany

drfaisal.ramzan@uaf.edu.pk

The increasing availability of high-density genomic markers, such as single nucleotide polymorphisms (SNPs), has established genomic prediction as a crucial tool in animal and plant breeding. This process involves predicting phenotypes solely based on genomic data. However, the large number of SNPs compared to the limited number of individuals can result in overfitting and reduced prediction accuracy. Artificial intelligence-based feature selection can help mitigate this by filtering out irrelevant SNPs and enhancing model performance. Feature selection (FS) methods identify a subset of predictor variables most relevant to the response variable, thereby preventing overfitting in high-dimensional datasets and reducing computation time and resources. Another challenge in genomic prediction is the often-overlooked interaction between SNPs, known as epistasis, which significantly contributes to the genetic architecture of quantitative traits. Epistasis is frequently ignored in genomic selection approaches using parametric models due to the high computational load, especially when interactions among a large number of markers are involved. In this study, we investigated an incremental feature selection method that ranks individual SNPs and epistatic SNP pairs based on their relative importance in trait manifestation. This ranking was then integrated with a random forest prediction model. We applied our approach to a simulated as well as a real goat dataset to test its performance. Our method resulted in a significant increase in prediction accuracy when using only a subset of available SNPs. Models using all available SNPs showed the lowest performance. Importantly, including epistasis in the prediction improved accuracy compared to models considering only additive effects. Our study demonstrates that the incremental feature selection approach can substantially enhance prediction accuracy. Furthermore, incorporating epistasis into genomic prediction further improves prediction performance.

Safer Biosolid Based Organic Fertilizer: Co-Composting of Sludge to Reduce Antibiotic Resistance Genes

Rabeea Zafar1, 2,* and Muhammad Arshad1

Institute of Environmental Sciences and Engineering, School of Civil and Environmental Engineering, National University of Sciences and Technology (NUST),

Sector H-12, Islamabad, 44000, Pakistan

Department of Nutritional Sciences and Environmental Design, Faculty of Sciences,

Allama Iqbal Open University, Sector H-8, Islamabad, 44000, Pakistan

rabeea.zafar@aiou.edu.pk

Antibiotic resistance is becoming even stronger and more widespread with the passing of time and is expected to increase the human death rate by a huge number in the upcoming years. It has been reported that in 2013, the USA alone had 200,000 people infected with antibiotic resistant bacteria, of which 23,000 died. The World Health Organization has thus declared it to be one of the top three major threats to public health in the 21st century. It can also prove to be an economic threat considering the level to which extra money would be spent on public health care and in general, to combat this increasingly spreading antibiotic resistance. Antibiotic resistance genes (ARGs) have been detected in various types of organic solid wastes such as livestock waste, agricultural waste, municipal waste, wastewater treatment plant sludge, leakage of sewerage in water bodies, and livestock farming. These microorganisms transfer readily between organisms and the environment and thus the resistance proliferates quickly. Given that the root cause of this problem is the excessive exposure of antibiotics to living organisms, implementing techniques related to organic waste management could offer potential solutions. In this study sludge containing resistant genes was co- composted with rice straw. Sludge from the wastewater treatment plant and rice straw from the rice mill was collected and the experiment was performed for 60 days. The C/N ratio was 29.85 and moisture content was 67%. During the experiment, the C/N ratio decreased towards the end and reached 18 at the end of the experiment. The sludge before and after the experiment was sent for metagenomic analysis after DNA extraction. The results show that there was more than 80% reduction in resistant genes and 99% of the resistant bacterial species were killed due to high temperature >50°C during initial phases of the experiment. The co-composting experiment yielded highly promising results, achieving a reduction of over 80% in antibiotic resistance genes (ARGs).

Development of Dual Purpose Bio-based Polymer Coated Urea Fertilizer: Controlled Nitrogen Release and Tailored Zinc Supply

Tooba Khan, Muhammad Ansar Farooq*, Aamir Alaud Din

Institute of Environmental Sciences and Engineering (IESE), School of Civil and Environmental Engineering (SCEE), National University of Sciences and Technology (NUST), Islamabad, Pakistan

Email: ansar@iese.nust.edu.pk

The increasing global demand for food has exerted significant pressure on agricultural lands to boost yields, leading to the excessive use of urea fertilizer. However, this practice has contributed to various environmental issues due to the low Nitrogen Use Efficiency (NUE). Additionally, zinc (Zn) deficiency in soils has emerged as a concurrent challenge for both plant and human health. To address both issues, this study aimed to develop a Zn-enriched, slow-release urea fertilizer. A dual-coated urea fertilizer was formulated, featuring an inner layer of bio-polyurethane and an outer layer composed of ZnO-loaded mesoporous silica nanopartilces (Zn@MSN) embedded within a hydrogel matrix. Various characterization analyses, including Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), and Brunauer-Emmett-Teller (BET) surface area analysis, were conducted on both MSN and Zn@MSN. The SEM results for MSN revealed a successful achievement of particle sizes of around 70 nm. The FTIR spectra of Zn@MSN confirmed the incorporation of ZnO within the silica matrix, with characteristics bands observed at 464 cm-1 for Zn-O-Si vibrations, 803 cm-1 for Si-O-Si bending vibrations, 1632 cm-1 for Si-OH vibrations and 3436 cm-1 for OH stretching. XRD analysis showed the characteristic peak of silica between 10° and 15° 2θ. BET surface area analysis further validated the successful loading of ZnO into the MSN pores, with MSN exhibiting a surface area of 184.9786 m²/g and a BJH adsorption pore volume reduced to 0.562717 cm³/g, indicating effective integration of ZnO within the mesoporous structure. After successful formulation of Zn@MSN, Liquified Cotton Stalk (LCS) was synthesized to form the inner layer of biopolyurethane. For the outer layer, a hydrogel matrix was created using sodium alginate and copper sulfate within which previously synthesized Zn@MSN were dispersed. Subsquently, the urea granules were dually coated with the hydrogel matrix using the dip-coating technique. To evaluate the slow-release properties of the final product, water and sand column release tests were carried out. The samples were collected at regular intervals over the period of 30 days. The Kjeldahl Method (TKN) was employed for quantifying Nitrogen and Atomic Absorption Spectroscopy (AAS) was performed to quantify Zn. To further validate the results, plant assays and field testing would be carried out as well.

Assessment of indoor microbiological air quality at different sites of he althcare center: A Cross-Sectional Study

Shabana Bano, Imran Hashmi

School of Civil and Environmental Engineering (SCEE), Institute of Environmental Sciences and Engineering (IESE), National University of Sciences and Technology (NUST), Sector H-12, Islamabad, 44000, Pakistan.

Email: imranhashmi@iese.nust.edu.pk

Humans are at risk of exposure to airborne microorganisms in both homes and hospitals. In hospitals, these microorganisms can spread easily due to crowded spaces, poor infection control, and overcrowding. Aim of this study was to evaluate bacterial load in indoor air of a healthcare center located in Islamabad. A passive sampling technique was employed to measure bacterial load, using gravitational settling plate method for data collection. A total of 60 indoor air samples were collected over three months from 10 sampling sites. The mean bacterial concentration ranged from (67-160 CFU/m3) after 20 minutes exposed sampling plate. The mean bacterial concentration recorded in the current study was well above standard provided by WHO (<100 CFU/m3) for healthcare facilities. Gram-positive bacteria were the most prevalent among most indoor environment samples. The temperature, relative humidity and carbon dioxide levels in the rooms were also above WHO guidelines, creating favorable conditions for bacterial growth and multiplication. The high concentration of bacteria in the indoor air may be attributed to inadequate ventilation, high temperatures, overcrowding, and improper waste management. It is imperative to regularly monitor indoor microbial loads and implement effective infection prevention and control measures.

Phenotypic and genotypic characteristics of a live-attenuated genotype I Vaccine of Japanese enceph alitis virus

Muhammad Naveed Anwar, Sourat Mudassar, Sidra Iqbal, Muhammad Umar Zafar Khan, Muhammad Shahid Ma hmood2, Sultan Ali

Institute of Microbiology, Faculty of Veterinary Science, University of Agriculture, Faisalabad, Punjab 38000 Pakistan

Email: naveed.anwar@uaf.edu.pk;dr.naveed903@gmail.com

The phenotypic and genotypic characteristics of a live-attenuated genotype I (GI) strain (SD12-F120) of Japanese encephalitis virus (JEV) were compared with its virulent parental SD12 strain to gain an insight into the genetic changes acquired during the attenuation process. SD12-F120 formed smaller plaque on BHK-21 cells and showed reduced replication in mouse brains compared with SD12. Mice inoculated with SD12-F120 via either intraperitoneal or intracerebral route showed no clinical symptoms, indicating a highly attenuated phenotype in terms of both neuroinvasiveness and neurovirulence. SD12-F120 harbored 29 nucleotide variations compared with SD12, of which 20 were considered silent nucleotide mutations, while nine resulted in eight amino acid substitutions. Comparison of the amino acid variations of SD12- F120 vs. SD12 pair with those from other four isogenic pairs of the attenuated and their virulent parental strains revealed that the variations at E138 and E176 positions of E protein were identified in four and three pairs, respectively, while the remaining amino acid variations were almost unique to their respective strain pairs. These observations suggest that the genetic changes acquired during the attenuation process were likely to be strain-specific and that the mechanisms associated with JEV attenuation/virulence are complicated.

PHA Nanocomposites for Packaging Applications

Farha Masood

Department of Biosciences, COMSATS University, Islamabad, Punjab, 45550 Pakistan

Email: farha.masood@yahoo.com

Green packaging materials with good mechanical and thermal properties is a desirable choice for future environmental protection. Nanocomposites based on polymer and surface engineered nanofiber can overcome the shortcomings of synthetic polymers used in packaging industries. Here, we report the use of surface functionalized sepiolite for preparation of nanocomposites that exhibit excellent good mechanical, thermal properties and biodegradability. The surface functionalization of the hydrophilic sepiolite was done with different silane molecules to achieve their uniform dispersion within the hydrophobic components of poly-3-hydroxyalkanoates (PHA). The silane molecules were used as compatibilizing agent. The nanocomposites were developed by melt blending technique. The characterization studies were carried out by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and thermogravimetric analysis (TGA). Good interfacial interaction was found between polymer matrix and surface functionalized sepiolite in scanning electron micrographs. FTIR analysis showed the formation of crosslinks in nanocomposites. Additionally, the effect of different concentration of surface functionalized sepiolite on mechanical, thermal and biodegradation properties of PHA was investigated. Mechanical properties of the resulting nanocomposites were 5 folds increased as compared to pristine polymer. Thermal stability was also remarkably enhanced. A significant weight loss of these clay polymer nanocomposites was evident after soil burial indicating their biodegradability. Thus, these eco-friendly clay polymer nanocomposites can be used for different packaging applications.

Impact of Integrated Livestock Farming on Food Security and Agricultural Sustainability

Muhammad Mahboob Ali Hamid, Muhammad Riaz, Waseem Abbas, M. Saim Arif

Institute of Animal and Dairy Sciences, Faculty of Animal Husbandry, University of Agriculture Faisalabad, Punjab 38000 Pakistan

Email: dr.mmahboob@uaf.edu.pk

The most pressing challenge in agriculture today is food security, climate change, growing human population and land shrinkage with deteriorating soil health. Global food security faces two primary challenges in today’s world. The first challenge is that one in nine people do not get enough energy and protein in their diet with small holder subsistence farmers being 50% and landless families in low- and medium-income countries accounting for 20% of them. The other challenge is specialized intensive agricultural practices often lead to soil and environmental degradation. Integrated livestock farming (ILF) emerges as one of the most cost-effective and efficient strategies to these issues. By integrating livestock and crop production, this system enhances economic yield per unit area and time for small and marginal farmers. Furthermore, it effectively mitigates environmental pollution by recycling of waste materials through a linked approach. ILF effectively complete the nutrient and energy cycles as livestock convert plant residues and by-products into both high- quality protein and manure, with manure serves as an organic fertilizer thereby reducing the need for mineral fertilizers. By integrating livestock with crop production and producing meat, milk and eggs, the ILF ensures food security, nutritional security and stable income for farmers thus emphasizing the urgent need for eco-friendly, sustainable and economically viable solution. In this manner, an ILF offers more benefits than monoculture and specialized farming by generating extra income and year-round employment.

Yield Predication of Canola by using AQUA CROP MODEL as an AI tool under rainfed conditions in Pakistan

Muhammad Yousaf Raza1, Mukhtar Ahmed1, Abdul Manaf1, Mahmood ul Hassan2

1.Department of Agronomy, PMAS-Arid Agriculture University Rawalpindi, Pakistan 46300, 2.Department of Plant Breeding and Genetics, PMAS-Arid Agriculture University Rawalpindi,

46300 Pakistan

Email: yousafraza9060@gmail.com

Context: Canola, a major oilseed crop, is highly sensitive to climate change. Rising temperatures, shifting precipitation patterns, and increased frequency and intensity of extreme weather events pose significant threats to canola production, particularly in rainfed environments. Understanding the impact of climate change on canola phenology, yield, and evapotranspiration under rainfed conditions is crucial for developing effective adaptation strategies and ensuring sustainable canola production using artificial intelligence.

Objectives: Calibration and evaluation of the AQUA-CROP model for Canola crops under rainfed conditions. Evaluation of the impact of sowing date and plant population on phenology and yield of Canola cultivar.To employ 4 socio- economic scenarios (SSP1-2.6, SSP2-4.5, SSP3-7.0, and SSP5-8.5) for early (2030s), mid (2050s), and late (2090s) periods in the 21st century to assess the impacts of increased atmospheric CO2 and temperature on Canola phenology, yield, evapotranspiration, and water use efficiency under rainfed conditions compared with baseline.

Method: Climate Data: Collection of historical and future climate data for the study region from reliable sources, such as Global Climate Models (GCMs).Crop Modeling: Employ a crop simulation model, such as the Aqua Crop Model developed by the Food and Agriculture Organization (FAO), to simulate canola growth and yield under various climate scenarios. Climate Change Scenarios: Simulate canola phenology, yield, and evapotranspiration under different climate change scenarios representing a range of temperature and precipitation changes.

Therapeutic Potential of a Novel Membrane Permeable MCU Inhibitor in Skeletal Muscle Injury

Ameer Hamza, Laiba Tariq Abbasi, Shehzad Ali1, Saima Barki, Noor Ul Ain, Eisha-Tur-Razia, Muhammad Rizwan Alam

Department of Biochemistry, Quaid-i-Azam University, 45320 Islamabad Pakistan.

Email: mralam@qau.edu.pk

Peripheral artery disease (PAD) is a highly prevalent pathology that affects more than 200 million people worldwide. Critical limb ischemia (CLI) is a serious complication that arises when PAD is left untreated. It can lead to limb amputation in 33% of cases and death in 25%. Skeletal muscle ischemia reperfusion injury (IRI) is a pathological condition that occurs when hindlimb is reperfused after prolonged ischemia. Mitochondrial dysfunction plays key role in IRI injury resulting in cell death and inflammation. Mitochondrial calcium overload is one of major causes of mitochondrial dysfunction during IRI. It occurs due to activity of Mitochondrial calcium uniporter (MCU) complex that is present in the mitochondrial inner membrane. Thus, inhibiting MCU complex could be potential therapy for IRI. In our research we have used a recently developed MCU inhibitor, Ru265 because of its high cell permeability, low toxicity, low IC50 value and a high mitochondrial uptake. Here, we aimed to evaluate the potential of Ru265 to alleviate skeletal muscle IRI. Acute hindlimb IRI was induced in bALB/c albino mice and verified by analysis of overall body weight, limb functionality, muscle weight, infarct size, and by markers of cellular damage such as serum LDH levels. Ru265 (10µm) was injected intraperitoneally at the onset of reperfusion. Our data showed that Ru265 application has no effect on decline in body weight as well as limb functionality due to IRI. Moreover, its application demonstrated no positive effect on damaged limb muscles; instead, it increased infarct area as verified by TTC staining. In addition, rise in serum LDH levels showed that Ru265 application further aggravated hindlimb skeletal muscle IRI. Thus, our findings reveal that Ru265 application has no positive effect on treatment of hindlimb skeletal muscle IRI in mice.

In-Vitro Evaluation for Synergistic Activity of Indigenously Isolated Phage with Antibiotics Against Multi-Drug-Resistant Staphylococcus Aureus clinical isolates Syed Damin Abbas Hamdani

Shifa College of Pharmaceutical Sciences, Shifa Tameer-e-Millat University, Jaffar Khan Jammali Road, Sector

H-8/4, 44000 Islamabad, Pakistan.

Email: Daman.abbas99@gmail.com

Multidrug-resistant Staphylococcus aureus (MDRSA) is a superbug against which antibiotic resistance cases have dramatically increased over the past decade. It is among the high-priority pathogens against which there is an urgent need for the development of new strategies, as per the World Health Organization. The use of bacteriophages (phages) is one of the potential strategies that can help combat such pathogens. Combining antibiotics with phages can result in the synergistic antibacterial activity of both, termed phage-antibiotic synergism (PAS). In our study, we evaluated the phage- antibiotic combinations to establish PAS activity against clinical isolates of MDRSA. An indigenously isolated novel phage named SDAH-01 was isolated, characterized, and tested for antibacterial activity against the MDRSA strain. Once the anti-MDRSA activity of SDAH-01 was established, it was evaluated for its potential synergistic activity with antibiotics selected from various pharmacological classes. Eighty-four different combinations of each antibiotic and phage were tested to determine the optimal combination showing maximum synergistic activity. As compared to monotherapies, combinations of SDAH-01 with amikacin, vancomycin and erythromycin significantly inhibited bacterial growth indicating synergistic activity. This study demonstrates the potential of PAS in the management of MDRSA infections, suggesting its clinical use.

Aspergillus; an abundant truffle in research Samira Khaliq, Ammara Aftab, Shahtaj Baloch Institute of Bichemistry, University of Balochistan, Quetta, Pakistan

Email: samkhaliq@gmail.com

Aspergillus was first identified in 1729 and is an asexual spore-forming fungi, sometimes switching to sexual stages. Most Aspergillus species are of commercial importance but some are also known to cause infections. They are commercially used in fermentation processes to produce alcoholic beverages as well as natural products such as anthraquinone (antibacterial and antifungal compound), citric acid (1.4 million tons/year) and enzymes (glucose oxidase, lysozyme, and lactase). In our first project we studied the in vitro inhibition of acetylcholine esterase (AChE) extracted from A.niger versus AChE extracted from healthy human blood, by n-hexadecylphosphocholine (HePC). It was observed that AChE from A.niger was inhibited competitively whereas AChE from human blood was inhibited non-reversibly by HePC. In our second project we isolated A.oryzae from soil samples collected from Quetta city. After an initial screening on the plates containing calcium phytate, it was found that this strain produced phytase in high concentrations. Hence, this strain was characterized for its phytase production in submerged fermentation. It was then mutated through γ-irradiation and Phytase production kinetics were studied in wild versus mutated strains. The results showed that although the mutated strain could grow at temperatures of up to 55°C however, the optimum temperature for Phytase production was 35°C at an acidic pH of 5.7. The kinetic parameters show that sucrose and CSL were good inducers of Phytase. In our third and most recent project we isolated A.sydowii from sea water samples and characterized its growth parameters on various Carbon and Nitrogen sources in submerged fermentation. We then fractionated the extracellular components present in the media via dissolving and separating in different solvents and partially purifying the samples via TLC. Each sample was then further analyzed by HPLC, FTIR and NMR. We are currently analyzing the NMR data to elucidate the structure of the compounds present in each sample and hence identify them.

NMR-based metabolic fingerprinting of grapevine cultivars

Kashif Ali

Department of Biosciences, Faculty of Life Sciences, SZABIST University, Karachi, 75600, Pakistan

Email: kashif.ali@szabist.edu.pk

Metabolic fingerprinting is a tool to perform metabolic comparison of different samples. This technique is widely applied to highlight metabolic differences at different stages of development, disease of healthy state, or taxonomical differences, etc. This tool is now widely applied to the agriculture to achieve various goals, including quality control, disease and pest management, and assistance to breeding or genetic modification programs. Chemical characterization of economically important crops, i.e., chemotaxonomy, is one of the major objectives accomplished by the metabolic fingerprinting studies. Not only different types, samples and crops are studied, analytical platforms used for such studies also varied considerably. Liquid and gas chromatography coupled with mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy are the most widely used among the others.Choosing the analytical platform is mainly depends upon the aim of the study, target metabolites, and the availability.Nuclear magnetic resonance, although less sensitive as compare to mass spectroscopy-based techniques, is an optimum choice offering discrete advantages of simple sample preparation, shorter measurement time, and the use of spectroscopic data for quantification without any calibration curve. Current study used NMR in combination with various multivariate data analysis methods, including principal component analysis (PCA), partial least square discriminant analysis (PLS-DA), hierarchical clustering analysis (HCA), to perform chemical classification or chemotaxonomy of resistant and susceptible grapevine cultivars. Total twelve cultivars were subjected to metabolic fingerprinting, out of which eight are resistant and four are susceptible. The differentiating metabolites are identified as phenylpropanoids and flavonoids, whose concentrations are higher in the resistant cultivars as compared to susceptible cultivars. The overlapping of NMR resonances of different metabolites is the major problem for compound identification which is usually overcome by the use of different 2D techniques. In this study, 1H–1H correlated spectroscopy (COSY) and heteronuclear multiple bonds coherence (HMBC) spectra were recorded to identify the metabolites.

Enhancing Air Quality and Livestock Nutrition: The Socio-Environmental Benefits of Urea-Treated Rice Straw in Pakistan

Waseem Abbas*, Faisal Ramzan, Muhamad Mahboob Ali Hamid and Muhammad Sharif

Institute of Animal and Dairy Sciences, University of Agriculture, Faisalabad, Pakistan

Email: waseem.abbas@uaf.edu.pk

In Pakistan, we are facing severe smog in Lahore, Faisalabad, Gujranwala, and Gujrat division. Many factors contribute to air pollution, such as dust particles, vehicle emissions, and factory smoke, but an important reason is the burning of rice straw by farmers before wheat cultivation. The leftover rice straw from mechanical harvesting presents a significant challenge for Pakistani farmers on multiple fronts. These residues interfere with the cultivation operations needed to prepare the field for the subsequent wheat planting season. While tilling the land to incorporate the straw is an option, the high cost of fuel makes this approach economically unviable for many farmers. Unfortunately, the most common solution adopted by farmers is burning the rice straw. This practice, while seemingly quick and cost-effective, creates a serious environmental issue. Smoke generated from burning rice straw significantly contributes to air pollution, leading to smog formation that negatively impacts air quality in the immediate vicinity and surrounding areas. Therefore, finding an alternative use for rice straw benefits the environment and helps farmers feed their animals. Rice straw presents a viable alternative to wheat straw, given its abundance in regions where rice is cultivated extensively. However, rice straw is less palatable and has a lower nutritional value compared to wheat straw, making it less desirable for animal feed. Enhancing the nutritional quality and palatability of rice straw is essential to make it preferable for animal feed. Urea treatment of rice straw offers a potential solution to this problem. This treatment involves adding urea, a nitrogenous compound, to rice straw, initiating a chemical reaction that makes the straw easier for ruminants to digest and increases its nutrient content. Prior research has demonstrated the effectiveness of urea treatment in enhancing the nutritional value and digestibility of rice straw for ruminants.

Prevalence of quinolone resistance in Escherichia coli and Pseudomonas aeruginosa isolated from poultry in South Punjab

Israr Maqbool1, Syed Qaswar Ali Shah1*, Muhammad Waqar Aziz1, Sidra Rasheed1 1Department of Zoology, Cholistan University of Veterinary and Animal Sciences Bahawalpur, Pakistan Email: israrmaqbool916@gmail.com

The rise of antibiotic resistance in Escherichia coli and Pseudomonas aeruginosa poses a significant threat to public health. Since antibiotics used in veterinary medicine are also employed to treat human infections, resistance to drugs such as quinolones is a major concern. This study investigates quinolone resistance patterns in E. coli and P. aeruginosa associated with poultry infections. We collected 300 samples from domestic and commercial chickens across various poultry farms, butcher shops, and shopping malls. The isolated microbes were identified using standard microbiological protocols, and quinolone susceptibility was tested using the Kirby-Bauer disk diffusion method. The results revealed quinolone resistance in 19% of E. coli isolates and 22% of P. aeruginosa isolates. Phenotypically resistant isolates were further analyzed for quinolone resistance genes using PCR, and plasmid- mediated resistance genes qnrA, qnrB, and qnrS were identified. Gene sequencing confirmed the presence of these resistance genes. The preliminary findings highlight a concerning trend of emerging resistance genes in poultry- borne microbes, which necessitates urgent action to reduce the risk of zoonotic transmission. There is a critical need for stringent regulatory and legislative measures regarding the sale and use of antibiotics in food animals like poultry.

Enhancing linezolid activity in the treatment of oral biofilms using novel chitosan microneedles with iontophoretic control

Saman Zafar, Sadia Jafar Rana, Elshaimaa Sayed, Tahir Ali Chohan, Israfil Kucuk, Kazem Nazari, Muhammad Sohail Arshad, Zeeshan Ahmad

Bahauddin Zakariya University, Bosan Road, Multan, Punjab, Pakistan

Email: sohailarshad@bzu.edu.pk

This study aimed to prepare and assess active microneedle (MN) patches based on a novel biomaterial and their effective coupled (physical and electrical) transdermal delivery of a model drug (Linezoid). Modified MN patches (e.g. fabricated from Linezoid, boronated chitosan, polyvinyl alcohol and D-sorbitol) were engineered using a vacuum micromoulding method. Physicochemical, FTIR (Fourier transform infrared), in-silico, structural and thermal analysis of prepared formulations were conducted to ascertain MN quality, composition and integrity. In- vitro mechanical tests, membrane toxicity, drug release, antibiofilm, ex-vivo mucoadhesion, insertion and in-vivo antibiofilm studies were performed to further validate viability of the coupled system. Optimized MN patch formulation (CSHP3 - comprising of 3 % w/v boronated chitosan, 3.5 % w/v PVA and 10 % w/w D-sorbitol) exhibited sharp-tipped, equi-distant and uniform-surfaced micron-scaled projections with conforming physicochemical features. FTIR analysis confirmed modification (i.e., boronation) of chitosan and compatibility as well as interaction between CSHP3 constituents. In-silico analysis indicated non-covalent interactions between all formulation constituents. Moreover, boronated chitosan-mucin glycoprotein complex showed a stronger bonding (~1.86 times higher CScore) as compared to linezolid-mucin counterpart. Thermal analysis indicated amorphous nature of CSHP3. A ~ 1.42 times higher tensile strength was displayed by CSHP3 as compared to control (i.e., pure chitosan, polyvinyl alcohol and D-sorbitol-based MN patch). Membrane toxicity study indicated non-toxic and physiological compatible nature of CSHP3. Within 90 min, 91.99 ± 2.3 % linezolid was released from CSHP3. During release study on agarose gel, CSHP3-iontophoresis treatment resulted in a ~ 1.78 and ~ 1.20 times higher methylene blue-covered area and optical density, respectively, within 60 min as compared to CSHP3 treatment alone. Staphylococcus aureus biofilms treated with CSHP3 exhibited 65 ± 4.2 % reduction in their mass. CSHP3 MN patches remained adhered to the rabbit oral mucosa for 6 ± 0.15 h. Mucosa treated with CSHP3 and CSHP3-iontophoresis combination showed a generation of pathways in the epithelium layers without any damage to the underlying lamina propria. Eradication of Staphylococcus aureus from oral mucosal wounds and complete tissue regeneration was recorded following 7-day treatment using CSHP3- iontophoresis coupled approach.

Assessment of Biopesticidal Potential of Rosmarinic acid against fruitfly, Bactrocera zonata

Anum Imtiaz1, Saira Khan1, 2*, Mohammad Maroof Shah1, Ihsan ul Haq3, Muhammad Asif4

Department of Biotechnology, COMSATS University Islamabad, Abbottabad Campus, University Road, 22060, Abbottabad, Pakistan
Department of Medical Lab Technology, University of Haripur, Haripur, Pakistan
Insect pest Management Program, National Agricultural Research Center, Islamabad, Pakistan 4. Department of Management Sciences, COMSATS University Islamabad, Abbottabad Campus, University Road, 22060, Abbottabad, Pakistan

Email: sairakhan@uoh.edu.pk

For cleaner production, plant based pesticides have long been recognized as safer alternatives to synthetic pesticides for their high target specificity, less non-target impacts, low environmental persistence and by posing fewer risks to humans and the environment. In nature, plants produce secondary metabolites during their interaction with insects and these metabolites can act as toxicants, antifeedants, anti-oviposition agents and deterrents towards the insects. In our previous publications, bioactive pesticidal compound; rosmarinic acid (RA) was isolated from the plant, Isodon rugosus and was identified by using various analytical techniques. This study evaluated insecticidal activities of RA against Bactrocera zonata. For male toxicity bioassay, two methodologies were followed; male annihilation technique (MAT) and bait application technique (BAT) and for female, feeding toxicity bioassay was performed. Two different solvent based dilutions of rosmarinic acid (methanolic and aqueous) were assayed along with synthetic insecticide and neem based biopesticide. The toxicity data was recorded after every 24 hour till 72 hours in all the treatments. RA aqueous solution exhibited 80% mortality against male B. zonata in MAT bioassay and 70% in BAT bioassay while 80% mortality in female toxicity bioassay. All these results encouraging studying further about RA as a botanical insecticide and its mode of action. As a potential botanical insecticide RA may fit well in IPM programs to control Bactrocera zonata.

Next-Generation CAR T Cells Using CRISPR-Cas9 Gene Editing for Improved Cancer Immunotherapy

Tariq Nadeem1, Saira Khan2, Muhammad Adeel Rana3

1Center of Excellence in Molecular Biology, University of The Punjab, Lahore, Pakistan. 2Deparment of Medical Laboratory Technology, University of Haripur, Haripur, Pakistan.

3Department of Microbiology, Quaid-e-Azam University, Islamabad, Pakistan.

Email: tariqbiotech27@g mail.com

Chimeric Antigen Receptor (CAR) T cell therapy has emerged as a promising immunotherapy for hematological malignancies. However challenges such as limited persistence, off-target effects, reduced efficacy in solid tumors, T cell exhaustion, cytokine release syndrome, and resistance to therapy hinder its broader applicability. Gene editing technologies, especially CRISPR-Cas9, are powerful tools to address these limitations. We will explore the potential of gene editing to optimize CAR T cell function and safety. We will also discuss the strategies to improve CAR T cell persistence, proliferation, and effector function by targeting co-inhibitory receptors or enhancing signaling pathways. Additionally, we will discuss the engineering of CAR T cells to recognize multiple tumor antigens to overcome tumor heterogeneity and minimize the risk of antigen escape. Furthermore, we will address the challenges of genetic defects in CAR T cells and how gene editing can be used to correct these defects. Safety considerations will also be discussed, including off-target effects and ethical implications. Although gene editing holds great promise, it presents potential risks, including off-target effects and unintended genetic alterations. We will also discuss the strategies to minimize these risks, including improved CRISPR-Cas9 specificity and rigorous safety assessments. Ethical considerations and regulatory challenges will also be discussed to ensure this technology's safe and responsible application in clinical settings. In conclusion, the future of gene-edited CAR T cells holds the potential to revolutionize cancer treatment, offering innovative and effective therapeutic modalities.

Simulation of Source Sink Partitioning in Wheat under Varying Nitrogen Regimes using DSSAT-CERES-Wheat model

Mukhtar Ahmed1,2*, Muhammad Bilal1, Shakeel Ahmad3

Department of Agronomy, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46300, Pakistan
Swedish University of Agricultural Sciences Umea Sweden 3.Department of Agronomy, Bahauddin Zakariya University Multan-60800, Pakistan

Email: ahmadmukhtar@uaar.edu.pk

Grain yields in wheat can be limited by the assimilate supply (source) or by the carbohydrate demand of the grains (sink). Recently, there have been questions regarding the capability of crop models to simulate the physiology of source- sink interactions in crops; however, crop models scarcely tested with source- sink partitioning. DSSAT_CERES_Wheat model was used with details of field experimental data having treatments of manipulated source (i.e., assimilate supply), sink (i.e., kernel number). The aim of the present study was to assess the impact of different levels of nitrogen and source-sink manipulation on wheat crop and to model source-sink partitioning in wheat under varying N- Regimes and climatic conditions. CERES-Wheat model was calibrated using 2015-16 observed data while model was evaluated using two-year field collected data of two sites i.e. Islamabad and Chakwal. The model was able to simulate treatments impacts on phenology (R2, RMSE and d-index values of 0.89, 2.80 days and 0.97 respectively at Islamabad while at Chakwal R2 = 0.89, RMSE = 2.65 days and d-index = 0.94), leaf area index (R2 = 0.94, 0.94, RMSE = 0.51, 0.38 and d-index = 0.98 and 0.92 at Islamabad and Chakwal respectively), grain yield (R2 = 0.97,0.96, RMSE = 0.17, 0.2 t ha-1, and d-index = 0.95 and 0.93 at Islamabad and Chakwal respectively), harvest index, soil nitrogen, crop nitrogen and grain nitrogen with good accuracy. The observed range for dry matter water use efficiency (DM_WUE) was 34.1 to 14.5 kg ha-1 mm-1) while grain WUE remained in the range of 10.3 to 3.7 kg ha-1 mm-1. Crop response to modified radiation use efficiency (RUE) was variable among sites which could be critical for studying crop environment interactions, improving WUE, estimating genetically and atmospheric CO2-related increased RUE, analyzing impact of solar dimming and source manipulations under biotic stress.

Evaluating Antibiotic Knowledge and Self-Medication Practices among University Students in Islamabad: A Cross-Sectional Study

Fizza Maryam

National University of Medical Sciences, Islamabad Pakistan

Email: fizza.maryam272000@gmail.com

Background: There has been a dramatic increase in antibiotic resistance in recent years. Antibiotic resistance results from the indiscriminate use of antibiotics due to people's lack of knowledge about them. This study aims to determine whether students' knowledge about antibiotics influences their behavioral beliefs about antibiotics.

Methodology: We carried out a descriptive cross-sectional study using an online survey targeted at students from the National University of Medical Sciences, Islamabad. Participants consented by completing a questionnaire divided into three sections: demographics, general knowledge about antimicrobial usage, and understanding of antimicrobial resistance. We collected 197 valid responses from verified university students from different programs. The study categorized participants into two groups based on their completion of a formal microbiology course.

Results: In our study, 76.1% of respondents reported antibiotic use within the past six months, with 65% experiencing gastrointestinal issues following antibiotic therapy. Notably, 86.3% of individuals did not utilize probiotic supplements during their antibiotic course. Analysis of the data revealed significant differences in antibiotic-related knowledge and practices across different study programs. Specifically, adherence to medical prescriptions varied (p=0.030), with some programs showing higher compliance than others. Self-medication behaviors differed significantly among programs (p=0.001), indicating varying practices in self-treating with antibiotics. Additionally, perceptions of self-medication as a cause of antimicrobial resistance (AMR) were significantly different (p=0.002), highlighting discrepancies in understanding the role of self medication in AMR. The perceived impact of AMR on public health also showed significant variation (p=0.000), reflecting differences in the recognition of AMR's health burden. Finally, general awareness of AMR varied significantly among programs (p=0.000), underscoring diverse levels of knowledge about AMR. Conclusions: There are significant variations in knowledge and practices regarding antibiotics and AMR among students from different study programs. Targeted educational interventions are necessary to improve awareness and proper antibiotic use, especially in non-health-related faculties.

Isolation and Screening of Drought Tolerant Plant Growth Promoting Rhizobacteria (PGPR) In a Semi-Arid Agro ecosystem Associated with Aloe Barbadensis

Falak Sajjad

University of Management and Technology Daska road Sialkot

Email: falak.sajjad@skt.umt.edu.pk

Introduction: Drought stress is the leading cause of reducing the crop yield in arid and semiarid regions of the world thus, classified as a main agricultural drawback. Regrettably the strategies to enhance the drought tolerant ability of plants are highly technical and labor-intensive, and consequently difficult to be done practically. However, the ability of Plant Growth Promoting Rhizobacteria (PGPR) in improving the plant growth under drought stress thought to have great economic importance in agricultural systems.

Objectives: The present study was conducted to analyze the effects of PGPRs against water stress and to isolate and characterize drought tolerance PGPR strains from rhizospheric region of Aloe Barbadensis, find out the morphological characterization of selected isolates and evaluate the drought tolerant efficiency of PGPR obtained from the rhizosphere of Aloe Barbadensis.

Methodology: Rhizosphere soil samples were collected from Aloe barbadensis plants in a semi arid region. PGPR were isolated using standard microbiological techniques and screened for drought tolerance via polyethylene glycol (PEG) induced osmotic stress assays. Selected strains were characterized for plant growth-promoting traits including production of indole-3-acetic acid (IAA), phosphate solubilization, and HCN producing activity. The most promising isolates were further tested for their ability to enhance growth and drought tolerance in model plants under controlled conditions.

Programmable Prodrug Nanomedicine for Normalization Cancer Immunotherapy

Dr. Madiha Saeed

Interdisciplinary Research Center in Biomedical Materials, COMSATS University Islamabad Lahore Campus, Lahore 54000, Pakistan

Email: madihasaeed@cuilahore.edu.pk

The tumor immune microenvironment (TIME) is comprised of a complex milieu that contributes to stunting antitumor immune responses by restricting T cells to accumulate in the vicinity of the tumor. Nanomedicine-based strategies are being proposed as a salvage effort to reinvigorate antitumor immunity. Various strategies, however, often fail to unleash the antitumor immune response. The effective nano/biomaterial-based delivery and TIME normalization approaches that promote T cell-mediated antitumor immune response will be discussed, with a focus on tumor immune microenvironment responsive and highly sophisticated cyclodextrin-based host-guest supramolecular prodrugs. Based on currently available evidence, it seems as if the ultimate success of cancer immunotherapy and nanomedicine hinges on the capacity to normalize the TIME. Here, how nanomedicines target immunosuppressive cells and signaling pathways to broaden the impact of cancer immunotherapy are explored. Acquisition of the urgently needed knowledge of nanomedicine-mediated immune normalization will guide researchers and scientists towards applications of cancer immunotherapy.

Exploring the Potential of Bacteriophages and Biochar to Improve Nitrogen Use Efficiency

Sadaf Khan1, Muhammad Faraz Bhatti1, Fazal Adnan2, Farhan Younas3, Ghulam Haider1*

Agricultural Sciences and Technology, National University of Sciences and Technology, Islamabad Department of Microbiology and Biotechnology, National University of Sciences and

Technology, Islamabad, Pakistan

3.Sulaiman Bin Abdullah Aba AlKhail-Centre for Interdisciplinary Research in Basic Sciences, IIU, Islamabad, Pakistan

Email: sadafkhani340@gmail.com

The extensive use of nitrogen fertilizers to meet the food demands of rapidly growing populations has contributed to global warming through the emission of greenhouse gases (GHGs). Lower nitrogen use efficiency (NUE) has led to the loss of nearly half of the applied nitrogen into the environment via ammonia volatilization, nitrate leaching, and nitrous oxide (N₂O) emission during nitrification and denitrification. Nitrous oxide is a potent greenhouse gas with a higher global warming potential (GWP) compared to CO2 and CH4, demands effective mitigation strategies. Various approaches are employed globally to improve NUE, including synthetic nitrification inhibitors (SNI), biological nitrification inhibitors (BNI), and biochar-based slow-release nitrogen fertilizers. Here we aimed utilizing biocontrol agents like bacteriophages targeting nitrifying and denitrifying bacteria to slow down the nitrification and denitrification processes. Additionally, a comparative analysis was conducted between biochar-based slow-release nitrogen fertilizers and organic extracts doped biochar-based slow-release nitrogen fertilizers in maize (Zea mays) under controlled conditions. Active agricultural soil was obtained for the isolation of nitrifying bacteria. The phylogenetic analysis of isolated bacteria confirmed their similarity to Pseudomonas spp., and a bacteriophage against this bacterium was isolated with positive results from the spot assay. Further assays will be conducted in the next phase of study. The results of the biochar maize study revealed a 98.54% increase in shoot biomass with 100 kg/ha nitrogen-treated biochar and a 14.58% to 24.01% increase in plant height with 150 kg/ha nitrogen-treated biochar, compared to the control. While the sorghum extracts-treated nitrogen-loaded biochar enhanced plant photosynthetic activity and total dry biomass yield compared to Nitrogen-treated biochar. Organic extracts treated nutrient-doped biochar can be further investigated to develop commercial fertilizers.

Synergistic prospect of bioaugmentation in agricultural lands towards heavy metal remediation stress for sustainable agriculture

Nimra Qureshi1, Bisma Ahmad1, Rabia Amir*

1Department of Agriculture Sciences and Technology, National University of Science and Technology, Islamabad

Email: nimra.a.s.qureshi77@gmail.com

Pakistan has the highest urbanization rate in South Asia. Being an agricultural country, the water demands of Pakistan are very high leading to the utilization of untreated effluents from the cities are being for urban and peri-urban irrigation purposes causing increase in the exposure of living organisms to heavy metals ultimately causing biological toxicity. Sunflower (Helianthus annuus L.) is the fourth leading resource of edible vegetable oil worldwide. Edible oil imports provide most of the oil for consumption while only 30% of the total domestic needs are met through local production with sunflower contributing to about 29.12%. Major domestic sunflower cultivation comes from Punjab and Sindh where sunflower is used as an important intercropping species as sustainable agricultural practice. The rise of industrialization possesses greater soil contamination risks to these agricultural zones thus impacting the productivity of major crops including sunflower. The use of biofertilizers in developing environmentally friendly agriculture is a sustainable approach for improving the yield of oilseed crops in terms of quality and quantity. Phytoremediation has proven to be a suitable method for reducing cadmium toxicity from soil. Plant-microbe-assisted biological reclamation of soil is a beneficial and innovative strategy for mitigating cadmium toxicity in soil. The present study aims to analyze the morphological and physiochemical responses of Helianthus annuus under cadmium stress and devise strategies for mitigating cadmium stress in soil while enhancing the crop's natural biological properties. Bacterial inoculation in soil improves the resilience of sunflower grown under cadmium stressed soil by regulating its biochemical responses and improving its morphological properties. By leveraging the natural bioremediation potential of sunflower in conjunction with PGPB, this study contributes to the development of integrated strategies for improving soil health, ensuring food security, and promoting environmental sustainability in regions vulnerable to heavy metal pollution.

Ubiquitin specific protease 1 is involved in grain size regulation in wheat

Usman Aslam1, 3, Rao Sohail Ahmad Khan2, Rana Muhammad Atif1

Seed Genetics Lab, Department of Plant Breeding and Genetics, University of Agriculture Faisalabad, Pakistan
Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture Faisalabad, Pakistan
Shoni Dharti Research Laboratory, Sohni Dharti Seeds International, Sahiwal, Pakistan

Email: usmanaslam@uaf.edu.pk

Wheat is Pakistan’s major food crop that fulfils approximately 70% of our daily caloric demands. Current yield of Pakistani wheat is considerably lower than the world average. Bringing substantial yield increases per-unit-area through innovative approaches is crucial to achieve national food security. Grain size is a key agronomic component of yield. Translating the effects of grain size related genes (GSRGs) from Arabidopsis to wheat could be an efficient fast- lane strategy to increase yields. Present research was aimed to find and structurally characterize gene orthologs that can influence grain size in local wheat varieties. Ubiquitin specific proteases (USP) have been known to regulate endoreduplication and organ size especially seed size in Arabidopsis thaliana and rice. Hence, USP was selected for its role in grain size control in wheat and named as grain size related gene-1 (GSRG1). The wheat transcriptomic data revealed that GSRG1 predominantly expressed in grains particularly, embryo, endosperm, and aleurone layer. This tissue-specific expression of GSRG1 was also substantially greater in these tissues compared to that of TaGS3, TaGS5, TaGW2 and TaGW7 indicating that each gene drives discrete mechanics to control final grain size. We later screened one small seeded (SS) and one large seeded (LS) wheat genotypes which were then tested for possible SNPs in genomic sequence of GSRG1 by PCR and sequencing. We found SNPs in 8th intron and 11th exon of GSRG1A in LS compared to SS. The cellular examination of SSG and LSG seeds also showed significant differences in nuclear contents and cell size in seed coat. Thus, these findings suggest that allelic variations in GSRG1A contribute to grain size phenotypes in wheat making it a potential candidate to increase wheat yields through targeted mutagenesis approaches.

Molecular Identification of Bacterial and Antimicrobial Resistance in Drinking Water Sources

Junaid Ahmed Kori1, Rasool Bux Mahar2, and Munazza Raza Mirza1*

Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan.
S.-Pakistan Center for Advanced Studies in Water, Mehran University of Engineering and Technology, Jamshoro 76062, Pakistan.

Email: munzihyder@yahoo.com

Plant Growth-Promoting Rhizobacteria (PGPR) Reduce Adverse Effects of Salinity and Drought Stresses by Regulating Nutritional Profile of Barley

Sania Zaib,1,2,* Akmal Zubair, 1 Safdar Abbas, 1 Javed Hussain,3 Ishaq Ahmad, 3 and Samina N. Shakeel1

1 Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan

2 Department of Biological Sciences, Faculty of Sciences, International Islamic University, Islamabad, Pakistan

3 Experimental Physics Department, National Centre for Physics, Islamabad, Pakistan

Email: saniazaibsz@gmail.com

For molecular quality assessment, 42 surface drinking water samples from Hyderabad and Karachi, Pakistan, were collected. The physicochemical quality of all drinking water samples was found to be in the range as per standard of EPA and WHO guidelines. For molecular identification of bacteria and antibiotic resistance genes, 16S rDNA based amplicon sequencing of 42 samples and shotgun metagenomics of 9 samples were carried out. In QIIME2 analysis of 16S rDNA amplicons, on average, 26.1% of the high quality reads were further processed for bacterial taxonomic analysis and results showed high bacterial prevalence in all samples. The bacterial classification at phylum and genus levels showed 40 bacterial phyla and more than 500 bacterial genera in Hyderabad and Karachi drinking water samples. Whereas, PICRUSt functional predictions of the representative reads of these genera were contributing to 421 KEGG pathways. In shotgun metagenomics of 9 samples, HL-01A sample harbored the most antibiotic resistance genes as per NCBI (19 genes), CARD (16 genes), MEGARes (18 genes), ARG-ANNOT (18 genes), and ResFinder (16 genes) databases, which could contribute to resistance against sulfonamide, beta-lactam, streptomycin, quinolones, tetracycline, macrolide, and cephalosporin. In conclusion, plate count, 16S rDNA amplicon sequencing, and shotgun metagenomics showed higher prevalence of bacteria that also harbored several bacterial antibiotic resistance genes, which could lead to several waterborne diseases, and also signifies that water sources should be monitored to avoid any future bacterial infections as their increasing antibiotic resistance pattern is alarming. PGPR applications depicted reduced Cl− contents in 200 mM salt stressed barley roots (KT2440 = 7.7 mg/kg and SBW25 = 6.3 mg/kg) and stems (KT2440 = 406.4 mg/kg and SBW25 = 365.5 mg/kg) as compared to controls (roots = 8.9 and stems = 469.5), while they displayed a significant increase in the barley leaves (KT2440 = 405 mg/kg and SBW25 = 416.4 mg/kg) when compared to control (110.6 mg/kg) under the same stress condition. In 1000 mM salt stress, a significant reduction in the Cl− content was observed in PGPR-applied barley roots (KT2440 = 7.6 mg/kg), stems (KT2440 = 1205.8 mg/kg and SBW25 = 1008.3 mg/kg), and leaves (KT2440 = 967.8 mg/kg and SBW25 = 530.8 mg/kg) when compared to controls (roots = 15.2 mg/kg, stems = 1605.2 mg/kg, and leaves = 1165.2 mg/kg). On the other hand, a significant increase in the Cl− content was noticed in PGPR applied barley roots (KT2440 = 29.5 mg/kg and SBW25 = 25.8 mg/kg), stems (KT2440 = 1023.8 mg/kg and SBW25 = 894.9 mg/kg), and leaves (KT2440 = 369.2 mg/kg and SBW25 = 409.8 mg/kg) when compared to controls (roots = 13.5 mg/kg, stems = 505.3 mg/kg, and leaves = 219.9 mg/kg) under drought stress condition. PGPR application was also found to be effective for enhancing the uptake of micronutrients (Mn, Fe, Co, Ni, Cu, and Zn) in barley plant parts under control and also under stressed conditions. Overall, our findings revealed an improvement in the uptake of macro- and micronutrients for the enhancement of salinity and drought stress tolerance. Conclusively, these PGPR species are an effective source of plant stress tolerance and elevated growth of barley and related plants under stress conditions.

Insulin-gene-expression and Resistance in Different Models of Diet-Induced Obesity, Hepatomegaly and Metabolic Syndrome

Ramadori GP,Göttingen,Germany,

Emial: gulianoramadori@gmail.com

Background. The role of insulin-gene-expression in the pancreas and of the liver in insulin clearance, in development of hyperinsulinemia and hyperglycemia as the main markers of diabetes type II ,in obese persons with metabolic syndrome, remains still ill defined.

Methods. Three groups of adult rats (160 g b.w.) were fed a low-fat-diet (LFD,chow 13% kcal fat) or high-fat-diet (HFD, 35% fat), or HFD+ 30% ethanol+ 30% fructose (HF-EFr,22% fat). After overnight-fast animal weight was determined, blood was drown at different times when animals were culled (at one, four and eight weeks) and plasma was stored together with pancreatic and hepatic tissues, which were shock-frozen with liquid nitrogen and used for extraction of total RNA. Plasma was used for clinical chemistry and RNA for PCR-analysis. Before storing livers were weighted. mRNAs were isolated for subsequent RT-PCR analysis.

Results. After eight weeks, body weight increased three-fold in the LFD group, 2.8-fold in the HFD group, and 2.4-fold in the HF-EFr (p < 0.01) group, while HF- EFr-fed rats, which had consumed less food during weeks 48(p<0.05) had the greatest liver weight. At Week 8, HOMA-IR values, fasting serum glucose, C- peptide, and triglycerides levels were significantly increased in LFD-fed rats compared to that at earlier time points. The greatest plasma levels of glucose, triglycerides and leptin (insulin?) were observed in the HF-EFr at week 8. Gene expression of pancreatic-insulin was significantly greaterin the HFD and HF-EFr groups versus the LFD. insulin: Cpeptide ratio and HOMA-IR values were higher in HF-EFr. Hepatic gene-expression of insulin-receptor-substrate-1/2 was downregulated in the HF-EFr,while the expression of phospho-ERK-1/2 and inflammatory-mediators were greatest in the liver of HF-EFr-fed rats. Chronic intake of both LFD and HFD induced obesity, MetS (hypergycemia, hyperinsulinemia, hypertriglyceridemia), and intrahepatic-fat accumulation.

Pakistan Society of Nephrology (PSN) Research Task Force initiatives to tackle the global burden of Chronic Kidney Disease

Professor Khaja H Mujtaba Quadri MD

Inaugural Chair Research, Pakistan Society of Nephrology

Head of Nephrology, Maroof International Hospital, Islamabad, Pakistan

Abstract:

10 percent of all individuals (850 million) are globally afflicted by Chronic Kidney disease ( Ref 1):The Pakistan Society of Nephrology undertook several initiatives between 2022 and 2024 inclusive of setting up a Research Taskforce, to address the gap in capacity to perform high quality clinical and translational research within the country. The taskforce conducted five onsite /blended workshops in all provincial and federal capitals and combined this with 11 monthly journal clubs and research fora, culminating in the formation of four working groups and initiation of a National multicenter Randomized controlled trial and a chronic Kidney disease registry. The entire process and proposed roadmap forward was documented (Ref 2) A PSN Research Committee has now taken the effort forward. It is anticipated that local generation of valid data shall inform public health policy to address this major National non communicable disease challenge.

Hsp90 Inhibiton and Induction of Immune Response in Cancer Cells

Yusuf Tutar

Division of Medicinal Biochemistry, Department of Basic Medical Sciences, Faculty of Medicine, Recep Tayyip Erdogan University, Rize, Türkiye

Email: yusuf.tutar@erdogan.edu.tr

Abstract

Cancer treatment has been a challenge since suppressing aggressive tumor mechanism over a single target often lead failure. Therefore, blocking cancer cell signaling through Heat shock protein (Hsp) inhibition may improve survival rates. Hsp90 is at the center of these signaling and antiapoptotic pathways. Therefore, this study employed thiazolyl benzodiazepine (TB) which has a promising activity against human breast epithelial carcinoma (MCF-7) as hydrazone is an important group for ROS-activated anticancer pro-drugs. This compound exerts anticancer activity through Hsp90 inhibition on MCF-7 cell lines and induces immune system via ERK/MAPK. TB reverses immune suppression at MCF-7 cells by TRAF6, TRIF and MYD88 and may be used as monotherapies or combined with other treatments. TB suppression dependent rewiring of cancer cells self-destruct and provide a unique anti-cancer treatment.

Penicillium chrysogenum as a cell factory for the biosynthesis of complex peptides

Dr. Hazrat Ali

Senior Scientist at National Institute for Biotechnology and Genetic Engineering, Faisalabad

In this presentation, I will explain the genome mining strategies that I adopted for the discovery of three new secondary metabolite biosynthetic pathways in Penicillium chrysogenum. These pathways synthesize more than fifty secondary metabolites and having diverse biological activities. The remarkable feature of all three biosynthetic pathways is the large number of intermediates and/or products that is the result of a distinct un-specificity of the adenylation domains in Non ribosomal peptide synthetase (NRPS) and/or modifying enzymes involved. The aim of this thesis is to discover, identify and ultimately assign secondary metabolites to their corresponding NRPS genes and gene clusters in P. chrysogenum. Herein a combination of genome mining strategies and functional analysis is employed exploiting an analytical pipeline that allows an un-biased detection of the impact of genetic modifications on a level of the extracellular metabolites. As the products and intermediates of secondary metabolite gene clusters show complex chemical structures and are mostly present only at low concentrations, an advanced mass spectrometry based pipeline was developed which enables the identification of structural complex metabolites at low concentrations directly from a biological matrix. Through the construction of individual gene inactivation mutants and metabolic profiling of the growth medium, final products and intermediates were identified to construct the biosynthetic pathway/s for the metabolites isolated. Furthermore, we also observed that by the deletion of the biosynthetic roquefortine/meleagrine pathway resulted in increase in production of chrysogen and related metabolites and vice versa even though the expression of the genes involved remained unaltered. This phenomenon is likely due to a re-direction of the metabolic (nitrogen) flux yielding increased levels of particular metabolites. This implies that for an optimal P. chyrsogenum strain, it is desirable to inactivate all unnecessary secondary metabolite genes. Remarkably, this is a phenomenon that to some extent already happened during the classical strain improvement that resulted in the silencing or mutational inactivation of various secondary metabolite genes.

Applied Biotechnology and Biosciences in times of COVID-19”: Pakistan Experience

Dr. Muhammad Tariq

Country Director, Chemonics International, Pakistan

Email: muhammad.tariq@smme.nust.edu.pk

During those challenging times of COVID-19 since February 2020, both Pakistan and the world needed urgent biotechnological and bioscientific advancements to quickly detect, prevent and treat increasing morbidity and mortality. Chemonics, USAID and the Government of Pakistan quickly sourced, procured and distributed 200 ventilators (trained 539 health workers), Personal Protective Equipment (PPE) for healthcare workers, one million Rapid Antigen Tests (411 borders point of entry staff trained), therapeutic Oxygen commodities distributed to 163 tertiary and secondary level facilities for managing lung diseases, and mobile testing increased to an additional 4000 PCR/day) throughout in areas deployed with four mobile Biosafety Level-2 labs for NIH & provincial governments. We also developed COVID-19 PPE calculator, which went into the world and the Netherlands 1+ solutions academy developed it into a course. In addition, Tariq et al., supported the Government in developing digital health products during COVID-19 times. The COVID-Management Information System (MIS) was developed which founded the current Pakistan Infectious Diseases Management Information System (IDMIS) for Integrated Disease Surveillance. Tariq et al., in collaboration with School of Mechanical and Manufacturing Engineering (SMME) and Ataur Rehman School of Applied Biosciences (ASAB) made a number of scientific developments including the integration of IDMIS with the Integrated Disease Surveillance and Response (IDSR) system, Vaccines Logistics Management Information System (LMIS), Electronic Medical Record (EMR), COVID-MIS, lab information systems, and Global Flood Awareness System, thus making a comprehensive digital platform which is ready for today as well as for times of next pandemic.

Safe, efficient and sustainable alternatives to conventional Microbiome transplants.

Dr. Raees Khan

National University of Medical Sciences, Rawalpindi Pakistan

The human intestine harbors a huge number of diverse microorganisms where a variety of complex interactions take place between the microbes as well as the host and gut microbiota. Significant long-term variations in the gut microbiota (dysbiosis) have been associated with a variety of health conditions including inflammatory bowel disease (IBD). Conventional fecal microbiota transplantations (FMT) have been utilized to treat IBD and have been proved promising. However, various limitations such as transient results, pathogen transfer, storage and, reproducibility render conventional FMT less safe and less sustainable. Defined Synthetic microbial communities (SynCom) have been used to dissect the host-microbiota-associated functions using gnotobiotic animals or in vitro cell models. This talk will focus on the limitations of the conventionally used microbiome transplants and how synthetic and minimal microbiomes can be a better, safer and sustainable alternative to conventionally used microbiome transplants.

Anaerobic Digestion coupled with Microbial Electrolysis Systems, Second Generation Anerobic Digestion: A Pathway to Biogas Upgradation and Enhanced Methane Production

Muhammad Adil Nawaz Khana, Malik Badshah*

Department of Microbiology, Quaid-i-Azam University Islamabad, 45320, Pakistan.

Email: malikbadshah@qau.edu.pk.

Microbial electrolysis systems (MES) present an innovative method for biogas upgradation, utilizing bioelectrochemical processes to boost methane content. In MES, electroactive microorganisms at the cathode facilitate the conversion of carbon dioxide (CO₂) and hydrogen (H₂) into methane (CH₄) when an external voltage is applied. This approach enhances the overall energy yield and purity of biogas, making it more suitable for various applications such as grid injection and vehicle fuel. Key factors influencing the efficiency of MES include the applied voltage, choice of electrode materials, and the composition of the microbial community. By integrating MES with anaerobic digestion processes, it is possible to achieve in-situ biogas enhancement, offering a cost-effective and sustainable solution for improving the quality of biogas as a renewable energy source.

Assessment of Antioxidant Capacity and LC-MS Chemical Profiling in MRQ74 & MRQ76 Rice Varieties

Norhaslinda Ridzwan1, Mohd Adzim Khalili Rohin*1, Norhayati Abd Hadi1,

Mohd Fadzelly Abu Bakar2, Norazlina Saad3, & Ruziana Ishak4

Nanomaterials Based Morphologically Distinct Interfaces Improving the Food Safety Measures

Mian Hasnain Nawaz

Interdisciplinary Research Center in Biomedical Materials (IRCBM), COMSATS University Islamabad, Lahore Campus.

Email: mhnawaz@cuilahore.edu.pk

Smart packaging of food is an emerging and challenging field of research that monitor the food quality and conditions in real-time and enable consumers to recognize freshness of food. This will lead to lessen health, food loss and environmental concerns. A number of smart packaging materials based upon different detection systems are available. Out of these, fluorescent and Colorimetric based indicators have gained center of attention owing to on- time detection based on color change. Here, smart packaging material as colorimetric and fluorescent indicators for chicken meat spoilage are fabricated and their response was analyzed. Novel nanomaterials corroborated fluorescence probing of spoilage indicators via monitoring their Fluorescence emission spectra. Moreover, the composite material based on biopolymer and metal oxide nanocomposite spin coated on a glass slide acted as colorimetric indicator. The colorimetric response of the nanocomposite towards chicken meat spoilage was monitored by visual color change and RGB color model. The colorimetric composite formed was selective for volatile amines specifically released by spoiling chicken meat. The optimization of affecting parameters further facilitated the selectivity and analytic performance of the protocol and proved that the fabricated material can be used on commercial scale to indicate the freshness of the packed meat.

AI for Antibiotic Discovery

Prof. Cesar de la Fuente

University of Pennsylvania

Website: https://delafuentelab.seas.upenn.edu/

Abstract

Computers can be programmed for superhuman pattern recognition of images and text; however, their application in biology and medicine is still in its infancy. In this talk, I will discuss our advances over the past decade, which are accelerating discoveries in the crucial and underinvested area of antibiotic discovery. We have pioneered antibiotics designed by computers with proven efficacy in preclinical animal models, demonstrating that machines and artificial intelligence (AI) can be used to design therapeutic molecules. For the first time, we successfully mined the human proteome for antibiotics. Recently, we expanded our mining efforts to explore extinct species. Using AI, my lab discovered the first therapeutic molecules in extinct organisms, including Neanderthals and the whoolly mammoth, launching the field of molecular de-extinction. Collectively, our efforts have dramatically accelerated antibiotic discovery, reducing the time needed to identify preclinical candidates from years to hours. I believe we are on the cusp of a new era in science where advances enabled by AI will help control antibiotic resistance, infectious disease outbreaks, and pandemics.

Smart Probiotics: Use of AI to Revolutionize Microbiome Health and Nutrition

Dr. Shakira Ghazanfar

Pakistan Agricultural Research Council (PARC)-Islamabad

The integration of artificial intelligence (AI) with probiotics brings with it the biggest paradigm shifts in microbiology a breakthrough for optimizing health and nutrition across sectors. So far, AI is the most technologically advanced means of isolating, discovering, formulating, and applying probiotics by implementing advanced machine learning algorithms, data analytics, and predictive modeling. Analysis of metagenomics and microbiome datasets accelerates identification and selection of probiotic strains to enable the formulation of well-defined probiotic products for specific health outcomes. Valued to advance productivity in livestock, gut health of the animals, and higher yields of milk from them to reduce the global agricultural challenge, animal nutrition is one of its great potential applications in several domains. Overall, AI capacities for real-time monitoring and data analysis allow a much deeper understanding of the underlying microbial dynamics and hence create innovative probiotic interventions in human health, disease prevention, and therapeutic treatments. This article will illustrate the convergence of AI and probiotics, discussing such technological advancement in detail, case studies, and applications. In this context, it will look into the emerging new avenues and present the directions for the future course of AI-powered probiotics in the biotech industry to pave the way for innovative health and nutrition strategies globally. AI technologies changes the process of design, use, and effectiveness of probiotics. Now, through the potential implementation of AI, researchers can more fully understand the very intricate interactions that exist within the gut microbiome, ultimately leading to the design of the "smart probiotics" tailored to the unique health-related needs of an individual. It will include the intersection of AI and microbiome science- an overview from the perspective of how smart probiotics are enhancing gut health, supporting immune function, and improving nutrition across human and animal species. It underlines the prospects of AI in the process of redesigning and consumptive application of probiotics both in the therapeutic and agricultural fields.

Unveiling Eye Color with DNA: Exploring the Potential of a “Biological Eyewitness”

Murad Ali Rahat, Sumaia Saif, Akhtar Rasool, Muhammad Israr

Department of Forensic Sciences, University of Swat

Forensic DNA analysis has traditionally focused on individual identification, offering little insight into physical appearance. However, recent advancements in DNA-based prediction of externally visible characteristics (EVCs) like eye color present a new frontier. This approach aims to create a “biological eyewitness” by analyzing DNA to infer physical traits. This presentation delves into the potential of DNA phenotyping for eye color prediction, using a specific example: the Pakhtun population of Malakand Division, Pakistan. The study investigated the link between Single Nucleotide Polymorphisms (SNPs) and eye color variations. Researchers analyzed DNA samples and eye color data from 893 participants. The findings not only revealed brown as the dominant eye color but also identified interesting gender differences. Females exhibited a higher prevalence of brown eyes compared to males, who showed a greater tendency towards intermediate and blue hues. Further analysis established statistically significant correlations between specific SNP genotypes and eye color. Individuals with brown eyes primarily possessed CT and TT genotypes, while blue eyes were exclusively linked to the CC genotype. The analysis of intermediate eye color genotypes presented a more nuanced picture, with both CT and CC genotypes present.

The Genetic Architecture of the People of Khyber Pakhtunkhwa

Muhammad Ilyas

Centre for Omic Sciences, Islamia College University Peshawar – Pakistan.

Email: milyas@icp.edu.pk

We have been involved in population genetics projects like the first Pashtun’s genome project and first Pakistan Ancient DNA project. We are using some innovative approaches to detect ethnic and even tribal-specific mutations that may be the key to rare and common diseases with higher prevalence in the population under study. We worked on genome-wide data analysis of human populations in Pakistan and their genetic relationship with other human populations in order to evaluate the genetic background of these groups. We aim to provide insights into the processes that have modeled the extant genetic diversity in these populations (such as migrations, bottlenecks and admixture) as well as the consequences of the diversity observed, including their relation to disease susceptibility. Our goal is to pursue this mission by emphasizing individual and collaborative faculty investigations in confluence with this genetic research cycle paradigm.

The Influence of Gut Microbial Strains and Genomic Variants on Immunotherapy Efficacy in NSCLC Patients

Muhammad Faheem Raziq1*

Department of Sciences, School of Interdisciplinary Engineering & Sciences (SINES), National University of Sciences & Technology (NUST), Srinagar Highway, Sector H-12,

Islamabad, Pakistan

faheemraziq1999@gmail.com

Abstract Cancer treatment has been a challenge since suppressing aggressive tumor mechanism over a single target often lead failure. Therefore, blocking cancer cell signaling through Heat shock protein (Hsp) inhibition may improve survival rates. Hsp90 is at the center of these signaling and antiapoptotic pathways. Therefore, this study employed thiazolyl benzodiazepine (TB) which has a promising activity against human breast epithelial carcinoma (MCF-7) as hydrazone is an important group for ROS-activated anticancer pro-drugs. This compound exerts anticancer activity through Hsp90 inhibition on MCF-7 cell lines and induces immune system via ERK/MAPK. TB reverses immune suppression at MCF-7 cells by TRAF6, TRIF and MYD88 and may be used as monotherapies or combined with other treatments. TB suppression dependent rewiring of cancer cells self-destruct and provide a unique anti-cancer treatment.

Biochemical and cytotoxic Analysis of Green Synthesized Silver Nanoparticles on Cal-51 Breast Cancer Cells.

Aamna Dilshad

Email: aamnadilshad8@gmail.com

The use of green synthesis in producing nanoparticles has demonstrated potential in nanomedicine, particularly in medical applications. This study explores the use of Moringa oleifera leaf extract for the green synthesis of silver nanoparticles (AgNPs). An aqueous extract with a 6.2% yield was utilized to reduce silver salt, producing AgNPs with attributes such as biocompatibility, eco-friendliness, and cost-effectiveness. These green-synthesized AgNPs exhibited enhanced biocompatibility, reduced toxicity, and specific targeting of Cal-51 breast cancer cells, making them suitable for cancer theranostics. The characterization of green-synthesized AgNPs was carried out using various techniques, including UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), Dynamic Light scattering (DLS), X-ray diffraction (XRD), and Particle Imaging. Additionally, qualitative phytochemical screening, thin- layer chromatography (TLC), and antioxidant assays were performed. The green Ag-NPs demonstrated significant inhibition of free radical scavenging in DPPH (97% (IC50=25.09μg/ml)) and H2O2 (92% (IC50=6.372μg/ ml) assays at 1000 μg/ml concentration. The percentage hemolysis of Green Ag-NPs was found to be less than 40% at lower concentrations, indicating their safe usage. Furthermore, the green Ag-NPs exhibited notable uptake efficiency in breast cancer cells and demonstrated promising cytotoxicity against triple-negative breast cancer cell lines (CAL-51) with nearly 60% inhibition with (IC50= 4.09μg/ml) of Green Ag-NPs compared to 20% (IC50= 38μg/ml) for Mo aq. leaf extract. In silico studies evaluated the potential of phytochemicals from Mo aq. leaf extract showing favorable interactions with integral proteins of Cal-51 Cancer cells. This research underscores the advantages of green synthesis, highlighting its role in facilitating sustainable and safe nanotechnological advancements for medical applications.

Synthesis of Micro-fabricated Bacterial Cellulose and Fish Collagen Peptide Scaffold to accelerate skin wound healing and protect against infection

Tayyaba Bibi

National University of Medical Sciences Rawalpindi

Email: tayyababibi97@gmail.com

The most important problem in biomedicine is always coming up with novel and creative methods to treat skin wounds. Skin wound treatment and management are very challenging across the world, mostly in diabetic patients due to the conversion of simple minor wounds into chronic wounds. Biological dressings are promising candidates for wound healing due to their characteristics such as biocompatibility, nontoxicity and non-immunogenicity. Bacterial cellulose and Fish collagen peptides are considered as important wound healing materials. Collagens can directly change the wound microenvironment, act as a scaffold for cell attachment and provide biologically active principles or antimicrobials to help wounds heal. This study describes the use of bio-fabricated bacterial cellulose (BC), fish collagen peptides(FCP) and Staphylococcus epidermidis hybrid model and evaluating its skin wound healing potential through in vitro and in vivo applications in a mouse model. The study consisted of the preparation of BC and FCP based scaffold, and its characterization via scanning electron microscopy(SEM), Fourier transform infrared spectroscopy (FTIR) and X-Ray diffraction analysis. In vitro experiments revealed about inhibition capability of this biological dressing against infection causing pathogenic strain Staphylococcus aureus. The wound healing potential was investigated in vivo on female albino mice. The invivo experiments showed that mice treated with BC-FCP and Staphylococcus epidermidis based scaffold exhibited faster wound healing as compared to BC-FCP treated and non-treated groups. The results of our study concluded that the innovative scaffold based on BC-FCP has a high potential for wound healing and may be used in the future for wound healing applications.

Fabrication Of Biodegradable Bioplastic Using Corn Starch

Haniah Iqbal Naseem1, Tayyaba Wasi1, Abdul wadood1, M.Farooq Usmani 1, M.Ebad Khan1, Saima Kashif1

Department of Biomedical Engineering at NED University of Engineering and Technology Karachi, Pakistan

Email: Haniahiqbal54@gmail.com

Abstract:

Plastics have emerged as a widespread environmental challenge due to their persistent nature and harmful effects on ecosystems and human health. This study explores the development of biodegradable plastics using corn starch as a sustainable alternative to conventional plastics. The process involves blending corn starch with vinegar, glycerol, and water, followed by heating and molding to form a bioplastic composite. The surface properties of the bioplastic samples were evaluated using contact angle testing. Biodegradability tests under different environmental conditions including soil, water and air confirmed the material's eco-friendly attributes. Overall, this study underscores the potential of starch-based bioplastics as a promising solution to mitigate plastic pollution while meeting diverse industrial needs.

Comprehensive biochemical analysis of formulated essential oil and In-silico docking evaluation of its bioactive compounds with skin receptors for hair fall prevention

Swaira Rafique1, Muhamad Aqmal Bin Othman2, Muhammad Qasim Hayat1,

*Muhammad Faraz Bhatti1

1Department of agriculture science and technology Atta-ur-Rahman School of Applied Biosciences, NUST, Islamabad, Pakistan; 2Department of chemistry, Universiti Malaya, Kuala Lumpur, Malaysia

Correspondence: mfbhatti@asab.nust.edu.pk

Abstract:

Hair fall is prevalent global concern affecting individuals. This research study will be focused on investigating the effects of formulated essential oil in preventing hair fall and comparing its efficacy with commercially available essential oils. Comprehensive analysis of bioactive compounds in formulated essential oil will be analyzed through Gas Chromatography Mass Spectrometry (GC-MS). Antifungal and antibacterial assays of formulated essential oil will be performed to confirm its potential role in inhibiting fungal and bacterial infections that are responsible for hair fall and will be compared to the commercially available essential oils. Phytochemical analysis will be performed for assessing the active chemical compounds having direct role in prevention of hair fall. Furthermore, in-silico docking studies will be performed to analyze how these bioactive compounds interact with human hair skin receptors and its effect on skin cells as well.

The current study will be focused on how these compounds intercept specific hair fall phases (Anagen, Catagen, Telogen) and also determining which pathway will be regulated. Molecular docking studies will further reveal binding affinities and partial mechanism of action of formulated oil on hair cells. This multidisciplinary approach will provide a scientific validation of a novel formulated oil in comparison to chemically available products in market.

Enhancement of methane contents by recirculation of Biogas along with External Hydrogen in Methanogenic Reactor Coupled with Microbial Electrolysis Cell

Asfa Zafar (asfazafar2019@gmail.com)

Quaid-e-Azam University Islamabad

Agrarian countries like Pakistan can easily overcome electricity crises with the use of biogas as transport fuel. But burning efficiency of biogas is less than natural gas. Biogas has considerable amount of CO₂ i.e.: 40-50 percent that can be upgraded to methane. For this, bio-gas upgradation is done. In-situ biogas upgradation was done by recirculation of H2 gas through a peristaltic pump allowing mixing of CH₄ to CO₂ leading to improved methane contents up to 99%. Similar results were noted when hydrogen gas recirculation for 2hrs and then stopping recirculation for half an hour. Hydrogen gas recirculation for longer periods with little breaks allow great percentage of biogas upgradation while avoiding disruption of hydrolytic and fermentative reactions and also preventing accumulation of Volatile Fatty Acids. Four reactors were set for the experiment. R1 acted as a control with no recirculation of gases, R2 was control with recirculation of hydrogen gas with no electrodes while R3 had single set of electrodes (SSE) with applied voltage and R4 had double set of electrodes (DSE) with applied voltage of 0.5 V. As R4 had double set of electrodes, it had a greater surface area for exo electrogens and hydro genotropic methanogens to grow and allowed biogas upgradation with methane content up to 95 %. R4 with 8-hour hydrogen gas recirculation and half an hour interval of break after every 2hr showed maximum biogas upgradation rates of 99 %.

High-Accuracy Machine Learning Models Reveal Gut Microbiome-Metabolite Signatures in Breast Cancer

Haseeb Manzoor 1* (haseebmanzoor667@gmail.com)

1Department of Sciences, School of Interdisciplinary Engineering & Sciences (SINES), National University of Sciences & Technology (NUST), Srinagar Highway, Sector H-12, Islamabad, Pakistan

Breast cancer remains the leading cause of cancer-related mortalities among women, underscoring the need for novel mechanistic insights into its cancer biology. Recent evidence has implicated the gut microbiome and its associated metabolites in the pathogenesis of breast cancer. This study employed comprehensive metagenomic analyses to profile the gut microbiome and its metabolite signatures in breast cancer patients. Our investigation identified 40 key metabolites from 471 microbial species that may play a crucial role in the metabolic landscape associated with breast cancer. Multivariate association analysis revealed significant positive associations (p-value < 0.05) between the presence of E. coli, siderophore, and thiopeptide production and breast cancer incidence. Moreover, we could distinguish microbiome-metabolite signatures among pre- and postmenopausal breast cancer cases and controls utilizing a custom ensemble machine learning classifier, with an accuracy of >90% and AUC exceeding 95%. This model outperformed traditional algorithms, and the high-ranking features i.e., E. coli, siderophore, and thiopeptide validated the statistical findings. Based on our results, we propose a mechanistic model in which E. coli secretes siderophores under iron-limiting conditions, leading to iron sequestration from the host and potentially promoting angiogenesis and tumor progression. This hypothesis, supported by statistical analysis and existing literature, suggests that microbial iron acquisition mechanisms could be critical in breast cancer pathophysiology. These findings warrant further functional analyses to validate the proposed mechanisms and explore their therapeutic potential. Our study highlights the gut microbiome and its metabolites as promising targets for the development of innovative breast cancer therapies, offering new avenues for research and clinical intervention.

Genetic Diversity of Shiga Toxin-Producing E. coli (STEC) in Cattle and Buffaloes from Islamabad

Zarrin Basharat1, Hamid Irshad2, Aitezaz Ahsan 2, Arfan Yousaf 3, Naowarat Kanchanakhan 4, Tepanata Pumpaibol 4, Wattasit Siriwong 4, Pinidphon Prombutara 5, Ibrar Ahmed 1,11*, Mudussar Nawaz 3,Abd Ullah 7,8,Humaira Amin1,12,Audrey Thevenon 9 ,Ijaz Khan 10,Muhammad Usman Zaheer 11, Sangeeta Rao 11 and Mo Salman 11

1Alpha Genomics Private Limited, 45710, Islamabad, Pakistan.

Email:alphagenomics.co@gmail.com

Shiga toxin-producing E. coli (STEC) are considered important zoonotic pathogens of great economic significance, associated with diarrhea, hemolytic uremic syndrome (HUS), hemorrhagic colitis (HC), and death in humans. The study aimed to investigate the distribution of various STEC virulence gene markers and antimicrobial susceptibility (AST) profiles associated within E. coli isolates from recto-anal mucosal swabs (RAMS) of slaughtered cattle and buffaloes in Islamabad, Pakistan. The RAMS (n=200) were analyzed using multiplex PCR for the presence of stx1, stx2, eae, and ehxA genes. The samples positive for one or more of the virulence genes were inoculated onto Sorbitol MacConkey agar (SMAC) for isolation of STEC. The isolates were further analyzed for the presence of virulence genes using multiplex PCR. Of the 200 RAMS, 118 (59%) were positive for one or more virulence genes. E. coli isolates (n=18) with one or more virulence genes were recovered from the 118 positive samples. The DNA of the isolates positive for one or more virulent gene was extracted and subjected to whole-genome sequencing using Illumina. Analysis of the WGS data indicated that the E. coli isolates could be differentiated into 11 serotypes. Most E. coli isolates (13/18; 72.2%) carried five genes (stx1, stx2, Iha, iss, and IpfA) in various combinations. In addition to these five genes, other virulence genes identified in these isolates were espI, ireA, espP, exhA, epeA, mcmA, mch, ast, celB, eilA, katP, and capU. The AST was performed using Kirby-Bauer disc diffusion test. The study indicated that all the isolates were resistant to rifampicin and a significant proportion of the isolates were MDR. A wide range of antimicrobial resistance genes (ARGs) were detected among the isolates, reflecting the complex nature of resistance mechanisms. The study results indicate that cattle and buffaloes slaughtered in Islamabad might be the carriers of antimicrobial resistant STEC of zoonotic significance, thus representing a source of human infection.

Functional assessment of the of MgO and MnO nanoparticles supplementation in an acute lung injury rat model Saman Saleem1, Nimra Razzaq2, Dr Arslan Shaukat3

Department of Physiology ,Government College University Faisalabad (GCUF), Pakistan

Email: samirana392@gmail.com

Acute lung injury (ALI) and the more severe acute respiratory distress syndrome (ARDS) are the pulmonary manifestation of an acute systemic inflammatory process characterized clinically by pulmonary illnesses, hypoxemia and edema. The combination of Magnesium and Manganese oxide nanoparticle supplements have been effective for treatment of acute lung injury. The nanoparticle form of an active ingredient or drug has a greater physiological effect, lesser toxicity higher absorption rates. Concerning, it has been previously reported that Magnesium oxide and Manganese oxide nanoparticles can be used for lung injury. The use of the nanoparticle from MgO& MnO in ameliorating the effect of lung injury has not been undertaken in a designed experimental setup. The rats were divided into five groups’ positive control, negative control, standard control, magnesium oxide & manganese oxide as a combination in low dose, and high dose groups. The lung injury be induced by passive exposure to cigarette smoke in twenty- eight number of Albino rats (4 cigarettes per day). Positive control (PC) and standard control (SC) low dose treatment (LDT), high dose treatment (HDT), each consisting of seven rats, were given an oral gavage containing a combination of MgO &MnO at a dose of 150 mg/kg BW for low dose treatment 300 mg/kg BW for high dose treatment. Seven more healthy rats were used as a negative control group; they were not exposed to smoking (NC). Every rat received a standard chow maintenance and water ad libitum. At days 28th rats were decapitated via the cervical region in order to harvest the lungs and blood. For hematological analysis, a full blood profile was performed. The body weight and relative organ weight was calculated. In terms of statistics, was used to examine all the data. Total oxidant status, and malondialdehyde level all significantly increased in the PC group. Body weight, total antioxidant capacity, total protein, albumin, globulin, and platelet count levels were all significantly lower in PC, while they were all significantly higher in the treatment group. In the PC group exposed to cigarette smoke, microscopic inspection of lung sections revealed damaged histological features indicating alveolar degradation. These findings thus point to a critical role for combination of MgO & MnO nanoparticles in the rat model of lung injury caused by cigarette smoke.

Immunomodulatory and antioxidant effects of MgO and MnO nanoparticle combination supplementation in Inflammatory Bowel Disease (IBD) in male albino rats.

Nimra Razzaq1, Saman Saleem2, Arslan Shaukat3

Department of Physiology, Government College University Faisalabad (GCUF), Pakistan.

Email: nimrarazzaq178@gmail.com

Inflammatory Bowel Disease (IBD) is characterized by excessive oxidative stress, chronic inflammation, and dysregulated gene expression, all of which contribute to the relentless progression of the disease and exacerbating tissue damage. This study explored the therapeutic potential of Magnesium Oxide (MgO) and Manganese Oxide (MnO) nanoparticles, designed to disrupt the deleterious cycle of oxidative stress, inflammation, and aberrant gene expression in IBD. MgO and MnO nanoparticles, with enhanced bioavailability and targeted delivery capabilities, were synthesized to modulate immune responses, reduce oxidative damage, and normalize the expression of key inflammatory genes. Healthy Albino rats (n=35) were kept in five separated groups i.e.; group A (negative control), group B (Positive control), group C (Standard control; IBD rats given Sulfasalazine treatment), group D (Low dose group; IBD rats given MgO and MnO combination supplementation at 150mgkg/body), group E (High dose group; IBD rats given MgO and MnO combination nanoparticle supplementation at 300mg/kg/body) each group contained (n=7) rats. Administered orally, the MgO and MnO nanoparticles accumulated in the inflamed colon, where they demonstrated a protective effect by improving gut barrier integrity, mitigating colonic injury, and modulating gene expression such as YAP and EZH-1. This nanoparticle-based approach offers a promising and targeted therapeutic strategy for managing IBD, potentially overcoming the limitations of conventional treatments and reducing the risk of adverse side effects. After 28 days, the animals were euthanized. Blood, fecal and colon samples were collected for hematological and biochemical analyses. Organ samples were subjected to histological examination to assess the anti-inflammatory and anti- oxidative potential of the MgO and MnO nanoparticle combination. Serum biochemical assays, performed using spectrophotometry, revealed significant effects (p≤0.05) on total antioxidant capacity, total oxidative stress (TOS), malondialdehyde (MDA), lipid profile, and liver enzymes, including Aspartate Amino Transferase (AST) and Alanine Transaminase (ALT). Hematological biomarkers also showed significant improvements with both high and low dose nanoparticle supplementation (p≤0.05). Histological analysis of the colon confirmed the ameliorative effects of MgO and MnO nanoparticle supplementation on structural derangement in IBD groups. In conclusion, MgO and MnO nanoparticle combinatorial supplementation at doses of 150 mg/kg and 300 mg/kg body weight demonstrated superior anti-inflammatory, anti-ulcerative, and anti- oxidative effects, offering a promising therapeutic strategy for managing IBD.

Integrative Multi-Omics Strategies for Precision Biomarker Discovery and Targeted Therapies in Neurological Disorders

Dr. Marwa Zafarullah

marwazafar@gmail.com

Neurological disorders, including neurodegenerative, neurodevelopmental and neuromuscular disorders present significant challenges due to their complex nature, often leading to insufficient diagnostic and therapeutic approaches. To address these challenges, integrative multi-omics strategies, which combine transcriptomics, proteomics, and metabolomics, offer a comprehensive framework for understanding and treating these conditions. By integrating diverse layers of biological data, these strategies can uncover novel biomarkers and identify targeted therapeutic interventions, thereby enhancing diagnostic precision and supporting personalized treatment approaches. This holistic methodology promises to advance the field of neurology, leading to more effective management and improved outcomes for patients with neurological disorders.

Discovery of novel small molecule kinase inhibitor to potentiate sorafenib's antiangiogenic effects on hepatocellular carcinoma

Ishaq N. Khan

Background: Hepatocellular carcinoma (HCC) is a type of liver cancer that heavily relies on the formation of new blood vessels, a process mediated by the VEGFR2 protein. This protein plays a crucial role in the development and progression of HCC. Current targeted therapies, such as sorafenib and lenvatinib, inhibit VEGFR2 but have shown limited effectiveness in improving patient outcomes.

New Development: In an effort to develop more effective treatments, we have developed 20 new small molecule kinase inhibitors. Among these, two inhibitors demonstrated particularly strong inhibitory effects on VEGFR2. These compounds were found to be much more potent than FDA- approved sorafenib, with SMI1 and SMI2 showing inhibitory concentrations (IC50) of 0.72-1.3 nM compared to sorafenib's IC50 of 68 nM.

Testing: In an in-vitro experiments, SMI1 was subjected to various laboratory tests to evaluate its efficacy. The compound significantly inhibited both basal and VEGF-induced tube formation, which is a critical step in the formation of new blood vessels that supply tumors. In an in-vivo setups, to further assess the efficacy of SMI1, we conducted experiments on 40 SCID/Beige mice. The mice were divided into four groups, with six mice from each group injected subcutaneously with HepG2WT cells and four mice with HepG2Luc cells for live imaging. Two weeks post-injection, when the tumors had reached approximately 200 mm², drug treatments were initiated. Tumor volumes were measured weekly using the ellipsoid formula (V = 1/2 (Length x Width²)), and live imaging was conducted using an IVIS system following luciferin injection.

Results: SMI1 demonstrated a significant reduction in VEGFR2 expression (p < 0.001) compared to sorafenib. Both SMI1 and sorafenib significantly inhibited the growth of HepG2WT HCC tumors in mice. SMI1 reduced tumor growth by 69.2%, while sorafenib achieved a 72.5% reduction. However, the combination of SMI1 and sorafenib was the most effective, suppressing tumor growth by 41.8% compared to the negative control. Immunohistochemical analysis supported these findings, showing a lower mitotic index in the SMI1-treated group (MI=8) and the combination treatment group (MI=3) compared to both negative (MI=11) and positive controls (MI=11). Importantly, no significant toxicity was observed in the treated mice, and in vivo imaging confirmed the absence of metastasis.

Conclusion: The novel compound SMI1 exhibits strong potential as an effective therapeutic agent for HCC. It can be used alone or in combination with sorafenib to enhance anti-angiogenic effects, leading to better suppression of tumor growth. These promising results provide a compelling rationale for advancing SMI1 to clinical trials in HCC patients.

DNA origami vaccine (DoriVac) for cancer and infectious disease

Zeng, Y.C.1,3,4, Young, O.J.1,3,4,7, Rajwar, A., Dembele, H3, Mulligan, K.1, Si, L.3, Ku, M.W.3, Wintersinger, C.H.1,4, MacDonald, J.I.3, Isinelli, G.3,8, Zhang, T.9, Anastassacos, F.M.3,4, Dellacherie, M.O.5, Sobral, M.5, Bai, H.3, Graveline, A.R.3, Vernet, A.3, Sanchez, M.3, Choi. Y.2, Ferrante, T.H.3, Keskin, D.B.6, Fell, G.G.10, Neuberg, D.10, Goyal, G3, Wu. C.J.6, Mooney. D.J.3,5, Ingber, D.3, Kwon, I.C.1,2, Ryu, J.H.1,2, Shih. W.M.1,3,4

1Department of Cancer Biology, Dana-Farber Cancer Institute, 2Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology (KIST), 3Wyss Institute for Biologically Inspired Engineering at Harvard University, 4Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 5Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, 6Department of Medical Oncology, Dana-Farber Cancer Institute, 7Harvard-Massachusetts Institute of Technology (MIT) Division of Health Sciences and Technology, Massachusetts Institute of Technology, 8Department of Drug and Health Sciences, University of Catania, Catania, Italy, 9Department of Cell Biology, Harvard medical school, 10Department of Biostatistics and Computational Biology, Dana-Farber Cancer Institute

DoriVac aims to leverage DNA nanotechnology and immune activators to stimulate stronger and long-lasting immune responses, targeting both cancer and various infectious diseases. For cancer, the development of improved therapies for patients whose tumors are resistant to immune checkpoint blockade is an unmet need. Our results reveal that square block (SQB) DoriVac with model tumor antigen fabricated enhances antigen cross-presentation by dendritic cells, and that uniform 3.5 nm spacing of CpG on SQBs differentially induces Th1 polarization. Our recent preliminary studies have indicated that Barrel DoriVac appears to be a promising strategy of capturing multiple neoantigens to better realize personalized vaccine formulation. Both formulations have been validated in aggressive mouse melanoma models where they showed durable tumor control in combination with immune checkpoint inhibitor anti-PD-L1. For infectious diseases, the induction of strong cellular immune responses has been limited. SQB DoriVac fabricated with infectious-disease (SARS-CoV-2, HIV and Ebola) associated peptides induced robust humoral and cellular immune responses with significant improvement compared to bolus vaccine formulation verified both on mouse and human lymph node organ chip.

BST-2 Facilitates Circulatory Survival and Enhances Pulmonary Metastatic Seeding of Breast Cancer Cells

Wasifa Naushad

Department of Pathology, Microbiology and Immunology, New York Medical College, Valhalla, NY, USA

Email:wnaushad@nymc.edu

Bone marrow stromal antigen 2 (BST-2) plays a pivotal role in driving various aspects of cancer progression and metastasis. In this study, we demonstrate that elevated BST-2 expression correlates with poor survival among invasive breast cancer patients, underscoring its association with disease severity. However, the precise mechanisms underlying BST-2's pro-metastatic effects remain incompletely elucidated. Our investigation involved analyzing BST-2 expression levels in human tissue samples to assess its correlation with tumor aggressiveness. We employed migration, invasion, and competitive experimental metastasis assays to evaluate cellular responses following BST-2 knockdown. Utilizing a mouse model of breast cancer, we further established that BST-2 promotes metastasis independently of primary tumor growth. Additionally, we found that suppressing BST-2 enhances sensitivity to anoikis in non-adherent cancer cells. In experiments using a basement membrane matrix, silencing BST-2 inhibited invadopodia formation, extracellular matrix degradation, and subsequent cell invasion. Competitive experimental pulmonary metastasis assays demonstrated that BST-2 depletion reduced viable circulating tumor cell (CTC) numbers and impaired lung colonization efficiency. Our data uncover novel functions of BST-2 in promoting i) invadopodia formation, ii) extracellular matrix degradation, and iii) protection of CTCs against hemodynamic stress. We propose that physical (tractional forces) and biochemical (extracellular matrix type/composition) cues may regulate BST-2's roles in cell survival and invadopodia development. Together, these findings underscore BST-2 as a critical factor enabling cancer cells to invade, survive in circulation, and establish metastatic colonies.

Pakistan Society of Nephrology (PSN) Research Task Force initiatives to tackle the global burden of Chronic Kidney Disease

Professor Khaja H Mujtaba Quadri MD

Inaugural Chair Research, Pakistan Society of Nephrology

Head of Nephrology, Maroof International Hospital, Islamabad, Pakistan

Clinical characteristics, Treatment Response, and Genome Sequences of SARS-CoV-2 infected Patients in Pakistan

Shah Jahan

Institute of Allied Health Sciences, University of Health Sciences, Lahore Pakistan

Pakistan experienced five distinct COVID-19 waves, resulting in 335,728 infections and 9,849 deaths. The Alpha variant of SARS-CoV-2 was dominant during the third wave, causing over 70% of infections and leading to mild to severe symptoms and fatalities due to limited understanding of the virus’s pathogenesis, which persists today. A study of 452 patients, ranging from symptomatic to severe, aimed to understand local epidemiology, clinical characteristics, and treatment responses. The average patient age was 58 years, with a male-to-female ratio of 61.7% to 38.2%. Of these patients, 27.3% had known exposure, 11.9% had travel history, and 60% had unknown infection sources. The remdesivir group showed significant improvements in oxygen saturation (58%), ferritin levels (58.2%), chest X-ray results (67.8%), and discharge rates (66.7%) compared to the untreated group. Remdesivir was especially effective for patients with moderate COVID-19. This study highlights that remdesivir can enhance outcomes in moderate cases and underscores the need for targeted research to inform health strategies in varied epidemiological contexts. Genomic analysis of 102 samples from seven hospitals, all with a PCR cutoff value under 25 cycles, revealed that the B.1.1.7 (Alpha variant) was predominant (97.9%), while B.1.351 (Beta variant) was present in only 2% of cases. This analysis showed multiple introductions and local establishment of B.1.1.7, with eight samples matching European genomes, indicating recent transmission. Other genomes displayed signs of evolution, suggesting ongoing transmission either within Pakistan or in regions with sparse genome sequencing. Vaccines remain effective against B.1.1.7, but the presence of B.1.351—against which some vaccines are less effective highlights the need for continued genomic surveillance. Samples collected from major Pakistani cities during the four pandemic waves (May 2020 to December 2022) underwent whole genome sequencing. Analysis using in silico and machine learning tools revealed an increase in mutation frequency from 12.19 in the first wave to 41.22 in the fourth. Strains B.1.36, B.1.160, and B.1.255 were most common in the first wave; B.1.36 and B.1.247 in the second; B.1.1.7 and B.1.36 in the third; and B.1.617.2 (Delta) in the fourth. Notably, B.1.36 was present in all waves, indicating its adaptability. Phylogenetic analysis identified similar genomes and revealed that C>T mutations were most common, while T>A substitutions were rare. Four types of mutations significantly impacted viral characteristics, suggesting high genetic variability in third-wave strains. This study provided valuable insights into SARS-CoV-2 evolution and transmission in Pakistan, aiding in the development of effective preventive measures.

Miracles of Genome

Shaukat Iqbal Malik

Capital University of Science and Technology

The human genome is important because the entirety of a person or organism's genetic informations is contained in their genome, sometimes known as the "book of life". Scientists can learn more about gene mutations that cause diseases like cancer by mapping a person's DNA. If all the DNA from a single human cell was stretched out end-to-end, it would make a six-foot- long strand comprised of a six billion letter code. It’s hard to imagine how that much DNA can be packed into a cell’s nucleus, which is so small it can only be seen with a specialized microscope. The secret lies in the highly structured and tightly packed nature of the genome. The research could eventually lead to the prescription of individualized precision medicine by medical professionals. Because of a genome, it can search for genes linked to different disorders. comprehend hereditary diseases and how they are treated. examine historical patterns of human migration. The nuclear genome comprises approximately 3 200 000 000 nucleotides of DNA, that encode the genetic instructions for all human cells divided into 24 linear molecules, the shortest 50 000 000 nucleotides in length and the longest 260 000 000 nucleotides, each contained in a different chromosome. These 24 chromosomes consist of 22 autosomes and the two sex chromosomes, X and The mitochondrial genome is a circular DNA molecule of 16 569 nucleotides, multiple copies of which are located in the energy-generating organelles called mitochondria. Each of the approximately 1013 cells in the adult human body has its own copy or copies of the genome. These are called somatic cells, in contrast to sex cells or gametes, which are haploid and have just 23 chromosomes, comprising one of each autosome and one sex chromosome. Both types of cell have about 8000 copies of the mitochondrial genome, 10 or so in each mitochondrion. The genome allows for advancements in personalized medicine, where treatments and medical care can be tailored to an individual's genetic profile. This can lead to more effective and less harmful treatments. The study of genomes across different species—helps us understand human evolution and how we are related to other organisms. It provides insight into the genetic changes that have shaped human development and adaptation. The knowledge of the human genome facilitates drug discovery and development by identifying new drug targets and understanding how individuals might respond differently to medications. Genomic information is used in forensic science to identify individuals, establish family relationships, and solve crimes by comparing DNA evidence. Genome raises important ethical and social questions about privacy, genetic discrimination, and the potential for genetic modification. It’s important to address these issues as genomic technologies advance. Overall, the human genome is central to many fields of science and medicine, driving progress and offering profound insights into what makes us human.

INVESTIGATION OF POTENTIAL RISK VARIANTS ASSOCIATED WITH SCHIZOPHRENIA IN LOCAL ETHNIC GROUPS

Umme Habiba1, Muhammad Mazhar1, Rafaqat Ishaq1, Muhammad Ilyas1,2, Muhammad Raza Memon3, Asad Tammezud Din Nazami4, Aafia Malik5, Uzma Abdullah1, Sadia Saeed1, Ghazala Kaukab Raja1, Pakeeza Arzoo Shaiq1* 1.University Institute of Biochemistry and Biotechnology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan.

Riphah International University Malakand Campus, Khyber Pakhtunkhwa, Pakistan 3.Department of Psychiatry, Liaquat University of Medical and Health Sciences, Jamshoro 76090, Pakistan
Institute of Psychiatry, WHO Collaborating Center for Mental Health, Benazir Bhutto Hospital, Murree, Road, Rawalpindi 23000, Pakistan
Department of Psychiatry, Jinnah Hospital Usmani Road, Quaid-i-Azam Campus, Lahore 54550, Pakistan

Email: pakeezaarzoo@uaar.edu.pk

Introduction: Schizophrenia is a polygenic disorder, however, the specific risk genetic variants of schizophrenia are still largely unknown. Single nucleotide polymorphism (SNP) is Important genetic factor for the susceptibility of schizophrenia. Investigating individual candidate gene contributing to disease risk remains important.

Methods: In a case control study, four SNPs which are reported as associated variants, including rs 3738401, rs821616,rs6675281 and rs1468412were genotyped in 200 schizophrenia patients and 200 healthy controls to investigate possible association between loci and schizophrenia in Pakistani population. The associated variant was further subjected to Insilco functional analysis, and the all the variants which were in linkage disequilibrium with the associated SNP were investigated. Phenome wide association analysis across UK Biobank was also done to investigate the association of SNPs in Genome Wide Association Studies.

Results: Our result showed significant association between rs821616 and schizophrenia (p = 0.0073, OR = 0.35, 95%, CI = 0.16- 0.75) in allele A to T, with MAF 0.33 in the studies sample(MAF cases = 0.12,MAF Controls=0.21). The rs821616, a missense variant, results into amino acid substitution from Serine to Cysteine at position 704 in exon number 11Current study analysed S704C polymorphic region rather than large translocation due to the fact that common polymorphic region needed smaller sample size. Phenome wide analysis of 821616 indicated that this SNP is reported to have associations with a large number of disorders as well, the most significant being with psychiatric, immunological, neurological and cognitive disorders (p=0.04868, p=0.02122, p=0.03654, p=0.0312403).

Conclusions:

The findings indicate that rs821616 in DISC1, might be involved in the pathogenesis of schizophrenia and, the allele T is risk posing variant for schizophrenia in not only Pakistani population but also other ethnic groups, since it causes a change in the structure of DISC 1 protein that interferes with the interaction of DISC1 with others.

DEVELOPING DRUGGABLE GENE EDITING TOOLS FOR Β- THALASSEMIA THERAPY

Afsar Ali Mian1*, Irfan Hussain1, Fawad Ur Rehman1, Hammad Hassan1, Ambrin Fatima1, Zehra Fadoo1, Dilawar Khan2, Oliver Ottmann3 and Salman Kirmani1

1: Aga Khan University, Karachi, Pakistan,

2: National University of Sciences and Technology, Islamabad, Pakistan 3: University of Cardiff, Cardiff, United Kingdom

β-thalassemia, a severe genetic disorder caused by mutations in the beta- globin (HBB) gene, leading to anemia that requires regular blood transfusions. With a carrier frequency exceeding 7%, the country has over 100,000 registered patients. Approximately 9,000 children are born annually with β- Thalassemia Major and remains a significant health burden, especially in Pakistan. Conventional treatment is challenging to sustain with < 1% of patients surviving beyond age of 20. The human BCL11A gene plays a crucial role in suppressing fetal globin (HBG1/2) gene expression. Disrupting BCL11A gene could potentially increase fetal hemoglobin (HbF) production, which is beneficial for patients with β-thalassemia. Our computational research identified the ZnF4 domain of multiple zinc finger (ZnF) as a key regulatory region that interacts with HBG1/2. Targeting this domain could be a promising therapeutic strategy in treating β-thalassemia. Hematopoietic stem cells (HSCs) isolated from β-thalassemia patients were edited using CRISPR-Cas9 to disrupt the ZnF4 domain of BCL11A gene. The impact of BCL11A deletion on fetal hemoglobin production and hematopoietic differentiation was analyzed. Induced pluripotent stem cells (iPSCs) generated from the same patients were differentiated into erythroid progenitors to compare the effects of ZnF4 editing. Our results demonstrate that CRISPR-Cas9-mediated disruption of BCL11A's ZnF4 domain in HSCs significantly increased HbF levels (p<0.001) while sparing other erythroid target genes. The edited HSCs maintained their ability to form erythroid colonies with increased γ-globin expression. Comparative analysis showed similar responses to ZnF4 editing in both iPSC- derived and patient-isolated HSCs. After validating the CRISPR-Cas9 construct for HbF induction, we designed a targeting peptide specific to CD34+ cells for druggable delivery. Liposome/Exosomes decorated with this peptide were successfully validated in in-vitro and in-vivo models, ensuring targeted delivery of payload to specific cells. This study highlights the critical role of BCL11A's ZnF4 domain in fetal globin repression and demonstrates the potential of targeted gene editing in HSCs. This study represents a significant step forward in the quest for effective therapies for β-thalassemia, combining innovative gene-editing techniques with targeted delivery systems to potentially transform patient care.

Artificial Intelligence (AI) based identification, sorting and transplantation of stem-cells for personalized

diagnostics and therapeutics.

Dr. Ahmad Ahsan Nawaz

Facility Head / Senior Scientist at Max-Planck-Institut für die Physik des Lichts, Erlangen, Germany

In the realm of medical research and personalized healthcare, the ability to harness artificial intelligence (AI) and deep neural networks (DNNs) for identifying and sorting cells without the need for labeling holds immense promise. This innovative approach can revolutionize both research and clinical applications, particularly those focused on personalized diagnosis and treatment. Traditionally, much emphasis has been placed on molecular markers to distinguish various cell types, but this method often involves time- consuming and expensive staining processes. Not only can these processes alter cellular properties, but they can also introduce contamination, limiting the utility of the samples. A compelling alternative to molecular markers is AI- powered label-free cell identification through the observation of mechanical and morphological characteristics. Introducing a cutting-edge microfluidic device equipped with real-time image processing and DNNs, we have transformed this concept into reality. Our microfluidic chip utilizes a surface acoustic wave generator to actively sort up to 10000 cells per second, based on their bright-field images, with the assistance of DNNs. This novel approach has proven successful in enriching lymphocytes, granule-monocytes, and red blood cells from human blood. Additionally, our innovation incorporates lasers and a fluorescence detection unit, which allows us to assign a fluorescence signal to each captured bright-field image. Harnessingthis AI technology, along with traditional molecular staining, to compile a labeled dataset featuring thousands of images of various blood cells. Using this dataset, we trained a DNNs with an impressive latency of under 1 millisecond. The ability to sort unstained neutrophils from human blood with a remarkable target concentration of 90%. This pioneering method of employing AI and deep learning for image-based cell sorting opens up a vast range of potential applications. We have already demonstrated its potential in studying conditions like long COVID-19 disease progression, transplantation of human eye cells and even enabling the label- free enrichment of stem cells for transplantation for blood cancer patients.

The Mitochondrion - From Power Generator to Living Drug

Muhammad Rizwan Alam

Department of Biochemistry, Quaid-i-Azam University, Islamabad

Email: mralam@qau.edu.pk

Mitochondria, traditionally known as the bioenergetic factories, contribute to a plethora of cell functions including calcium homeostasis, signal transduction, redox balance and cell death. Besides regulating the physiology of diverse cell types, these double-membrane organelles havealso been associated with the pathogenesis of several diseases including myocardial infraction(MI) and diabetes. Intriguingly, the healthy and living mitochondria isolated from different tissues have also been employed as therapeutic modalities (living drugs) to alleviate the cell damage under the settings of MI and brain stroke along with various other pathologies. Our research work highlights a fundamental involvement of mitochondria in the physiology of pancreatic β- cells and circulating platelets. Under disease settings, we have unveiled the crucial role of mitochondria in the ischemia-reperfusion injury of heart. On the similar lines, our research group has recently uncovered, a so far unreported, impact of the cell-free mitochondrial DNA in the overactivation of platelets in patients with type II diabetes mellitus. Along with exploring the mechanistic significance of mitochondria in health and disease processes, our recent data also reveals the use of mitochondrial transplantation as a promising therapeutic regime in peripheral artery disease. Altogether, our research discoveries establish a diversified role of mitochondria in the (patho)physiology of different cells along with demonstrating the application of mitochondrial transplantation in regenerative medicine.

m6A and its regulators at the crossroads of cancer and male infertility

Dr. Muhammad Jadoon Khan

Department of Biosciences, COMSATS University Islamabad (CUI), Pakistan

N6-methyladenosine (m6A) is the most prevalent internal modification in eukaryotic mRNA, playing a role in RNA metabolism e.g., its splicing, stability, nuclear export, translation, and decay thus regulating gene expression. This epitranscriptomic modification has emerged as a key player in stem cell biology, influencing pluripotency, differentiation, and self-renewal capabilities. In the context of cancer, m6A methylation is reported with dual role of promoting or suppressing tumorigenesis. Recent studies have also highlighted a significant role of m6A methylation in reproductive biology, implicating it in various aspects of fertility and embryonic development. In this talk, molecular mechanisms underlying m6A methylation, its dynamic regulation and how its dysregulation can contribute to cancer progression and male infertility, will be explained. Further, differential RNA binding proteome with respect to m6A would be presented in knockout model of key enzymes catalyzing the RNA methylation, in mouse embryonic stem cell. Also, some preliminary data related to the role of m6A in cancer onset and progression, and male infertility would be discussed.

Expression Profiler: A Comprehensive Pipeline of Integrative Genomics and Functional Annotation using Transcriptomics data.

Dr. Farhan Haq

COMSATS, Islamabad

Differentially expressed genes (DEGs) show variable expressions in different conditions. Identifying DEGs helps in understanding the molecular mechanisms underlying various biological processes, diseases and responses to treatments. Biological Insights: Reveal gene function, regulatory mechanisms and biological pathways Disease Research: Identify disease biomarkers and understand disease mechanisms Therapeutic Targets: Discover potential targets for new treatments. In conclusion, Expression Profiler is an interactive, visually appealing tool created with Python that offers insights into gene pathways, enrichment, GO annotations, protein interactions, drug targets, and informing therapeutic strategies.

Stimuli-Responsive Nano-Drug Delivery System for Colitis Therapy

Dr. Muhammad Naeem

Department of Biological Sciences, National University of Medical Sciences, Rawalpindi, Punjab, Pakistan

Email: m.n aeem@numspak.edu.pk

Introduction: Ulcerative colitis is a chronic persistent idiopathic inflammatory condition of the large intestine that affects millions of people worldwide. Conventional therapeutic dosage forms are available to treat colitis but most of them have limited efficacy because of problems related to reduced drug solubility, less drug availability at inflamed sites, and systemic toxicity. Nanotechnology-based drug delivery systems have the potential to accumulate and gather in the colon tissues experiencing inflammation due to a phenomenon called as “epithelial enhanced permeability and retention effect” ultimately leading to improved therapeutic efficacy and localized and targeted therapy, which increases the local bioavailability and reduces systemic toxicity. Methods: Budesonide-loaded dual-sensitive nanoparticles were prepared using enzymesensitive azo-polyurethane and pH-sensitive methacrylate copolymer for the treatment of colitis. The therapeutic potential of the enzyme/pH dual-sensitive nanoparticles was evaluated using a rat colitis model and compared to single pH-triggered nanoparticles. Results: Clinical activity scores, colon/body weight ratios, myeloperoxidase activity, and proinflammatory cytokine levels were markedly decreased by dual-sensitive nanoparticles compared to single pH-triggered nanoparticles and budesonide solution. Moreover, dual-sensitive nanoparticles accumulated selectively in inflamed segments of the colon. In addition, dualsensitive nanoparticle plasma concentrations were lower than single pH-triggered nanoparticles, and no noticeable in vitro or in vivo toxicity was observed. Conclusion: Our results demonstrate that enzyme/pH dual-sensitive nanoparticles are an effective and safe colon-targeted delivery system for colitis therapy. Keywords: azo- polyurethane, methacrylate copolymer, budesonide, nanoparticles, colitis.

THERAPEUTIC PROSPECTS AND CHALLENGES OF PLANT BASED NATURAL PRODUCTS IN DRUG DISCOVERY

Dr. Yamin Bibi

Department of Botany, Rawalpindi Women University, Rawalpindi

Email: yamin.bibi @f.rwu.edu.pk

Green natural products refers to chemical substances produced during plant metabolism and play a crucial role in different signaling cascades, defense mechanism against pathogen attack, insect attack, mechanical injury, and other types of biotic and abiotic stresses. These natural products present a vast and largely untapped reservoir of chemical diversity that can serve as the foundation for new drug development. These bioactive compounds offer promising therapeutic prospects ranging from antiseptics to anticancer particularly in the context of addressing emerging drug resistance and targeting complex diseases such as cancer, cardiovascular disorders, oxidative stress and neurodegenerative diseases. Spectroscopic methods sorted out the structures of novel components through GC/LC-MS, IR and NMR. Advancements in technology, such as high-throughput screening and bioinformatics, have facilitated the identification and characterization of these natural products, accelerating the drug discovery process. Furthermore, the integration of ethnobotanical knowledge with modern pharmacology has provided a synergistic approach to discovering new drugs, leading to several plant-derived compounds being successfully developed into clinically approved medications. Despite these advancements, challenges such as sustainability, standardization, and the complexity of plant matrices remain. In conclusion, plant-based natural products hold significant promise for the future of drug discovery, offering a diverse and potent source of novel therapeutics that could address current and emerging health challenges. The integration of traditional knowledge with cutting-edge research methodologies will be crucial in unlocking the full potential of these natural resources as well as continuous availability and resupply of natural medicinal products

Decoding the Multifaceted Regulatory Roles of CircRNAs: A Focus on the CircRNA–miRNA–mRNA Axis

Faryal Mehwish Awan*1 , Burton B. Yang2, 3, 4, Ayesha Obaid1 , Sadiq Noor Khan1 , Saira Khan1 , Anam Naz5 , Rabea Basri1

Department of Medical Lab Technology, The University of Haripur, Haripur, Khyber Pakhtunkhwa, Pakistan
Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto, Canada 3.Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada
Institutes of Medical Sciences, University of Toronto, Toronto, Canada
Institute of Molecular Biology and Biotechnology (IMBB), The University of Lahore (UOL), Lahore, Pakistan

E-mail: faryal.mehwish@uoh.edu.pk

The study of Non-coding RNAs (ncRNAs) is still a largely enigmatic field, with the boundaries between their beneficial and detrimental effects yet to be clearly defined. These molecules can affect RNA stability, control the flux of genetic information and play crucial roles in mediating epigenetic modifications. A large number of studies have highlighted the potential roles of ncRNAs in infections, transmission and therapeutics. Our study holistically mapped the regulatory networks driven by circRNAs, miRNAs and mRNAs to uncover plausible interactions and interplay amongst them in order to explore their diverse functions and possible therapeutic options. We found that circRNAs including hsa_circ_0004214, hsa_circ_0017836 and hsa_circ_0007813 can modulate miRNA function and have the potential to be therapeutic targets as well as diagnostic and prognostic indicators. Our results from RNA datasets, clinical tissue samples and cell lines led us to conclude that circRNAs could play a crucial role in the development of different diseases. Our results have added an additional layer to the understanding of the mutual regulation of various classes of ncRNAs. We concluded that the circRNA based therapeutics will be a promising direction in the field of biomedicine. By leveraging AI's computational power, researchers can uncover the complexities of circRNA functions and their implications in health and disease, paving the way for innovative diagnostic and therapeutic approaches.

Identification of potential microRNA diagnostic panels in HCV induced HCC

Aqsa Ikram1, Bisma Rauff2

1Institute of Molecular Biology and Biotechnology (IMBB), University of Lahore (UOL), Lahore, Pakistan

2 Department of Biomedical Engineering, UET Lahore, 51600, Narowal campus, Narowal, Pakistan

MiRNAs are non-coding RNAs, and they play a crucial role in gene regulation. Differential expression patterns of miRNA have been observed in different malignancies, such as HCC caused by HCV and HBV infection. In the current study, utilizing multiple bioinformatics tools, six serum-based miRNA biomarkers (miR-1, miR-200b, miR-320d, miR-346, and miR-451a) were identified for HCV induced HCC diagnosis. Furthermore, RT-PCR was used to check their and immune-related biomarkers (IL-6, and NF-κB) expression levels in study groups (G1= control, G2= HCV, G3= HCV- HCC, and G4 =HCC). It was found that miR-1, miR-346, and miR-451 expression were significantly low in G2, G3, and groups, as compared with G1 group. However, the expression of miR-200b, miR-320d, NF-κB and IL-6, expression was significantly upregulated in G3 as compared with G1 group. miR-451a(r = 0.5941, p< 0.0171) was positively correlated with NF-κB, and miR-1 (r = -0.6896, p <0.0043) was negatively correlated with IL-6 in the G3 group. A panel of miRNAs combining biomarkers improved the sensitivity and specificity (87.10% and 0.5565%, respectively) of G3 as compared to G1 group. The current study data reveal that miRNA panel (miR-1, miR-200b, miR-320d, miR- 346, and miR-451a) may serve as therapeutic targets and diagnostic biomarkers for HCV-HCC.

Fabrication of green composite hand knitted silk mesh reinforced with silk hydrogel

Natasha Bokhari a,b , Asif Ali a,c , Abida Yasmeen b , Hamad Khalid a , Sher Zaman Safi d , Faiza Sharif a,*

A Interdisciplinary Research Centre in Biomedical Materials, COMSATS University Islamabad, Lahore Campus, Lahore 54000, Pakistan b Department of Chemistry, Lahore College for Women University, Lahore 54000, Pakistan c Research Unit Plasma Technology (RUPT), Department of Applied Physics, Faculty of Engineering and Architecture, Ghent University, Ghent 9000, Belgium d Faculty of Medicine, Bioscience & Nursing MAHSA University, Jenjarom 42610, Selangor, Malaysia.

Soft tissue defects like hernia and post-surgical fistula formation can be resolved with modern biomaterials in the form of meshes without post- operative complications. In the present study hand knitted silk meshes were surface coated with regenerated silk fibroin hydrogel and pure natural extracts. Two phytochemicals (Licorice extract (LE) and Bearberry extract (BE)) and the two honeybee products (royal jelly (RJ) and honey (HE)) were incorporated separately to induce antibacterial, anti-inflammatory, and wound healing ability to the silk hydrogel coated knitted silk meshes. Meshes were dip coated with a blend of 4 % silk hydrogel (w/v) and 5 % extracts. Dried modified meshes were characterized using SEM, DMA, GC–MS and FTIR. Antimicrobial testing, in vitro cytotoxicity, in-vitro wound healing and Q-RT-PCR were also performed. The presence of coating on the mesh improved the mechanical strength/Young's modulus by 1602.8 %, UTS by 451.7 % and reduced the % strain by 51.12 %. Cytocompatibility testing and gene expression results suggest that out of four variables MHRJ presented best cell viability, % wound closure and expression of wound healing marker genes. In-vivo analyses in rat hernia model were carried out using only MHRJ variant, which also confirmed the non- toxic nature and wound healing characteristics of the modified mesh. The improved cell proliferation and activated wound healing in vitro and in vivo suggested that MHRJ could be a valuable candidate to promote cell infiltration and activate soft tissue and hernia repair as a biomedical implant.

Islets of Langerhans in primates and rodents: Variations of cytoarchitecture and metabolic signatures

Dr. Shahzad Irfan

Physiology, G.C. University Faisalabad

Email: irfan.shahzad@daad-alumni.de

Islets of Langerhans, a unique composite of multiple endocrine cells, are considered the pinnacle for glycemic control. Diabetes mellitus is the consequence of even slight subnormality in the cellular interplay of these endocrine cells. Islets of Langerhans are Hypothalamic-Pituitary independent yet strictly endocrine, employing all forms of cell signaling (Autocrine/intracrine, paracrine, juxtacrine). The interplay of different cells within islets, through hormones, and the subsequent glycemic control in different physiological states makes islets of Langerhans a unique and royal enterprise in the clinical aspect of metabolic health. Diabetes mellitus is fundamentally a human disease and is non-existent in the animal kingdom. A wealth of data is being generated every year by using rodent models of hyperglycemia and diabetes by using beta cell-selective chemical toxicants (STZ, Alloxan). Rodents are convenient, inexpensive, and easy to manage animal models. However, the casual extrapolation of rodent data toward humans in understanding pathophysiology or treatment options needs to be critically reviewed from a cellular and physiological perspective. One key indicator for the caution lies in the fact that the success rate of new drug application (NDA) from phase 1 to approval is only 4.8% for diabetes, while for all the other diseases it is close to 10%. The current talk will highlight the key and fundamental differences at cellular and physiological levels between rodent and primate islets of Langerhans.

Unveiling a Novel Signature Biomarker for Early Diagnostic and Prognostic Evaluation of Pancreatic Ductal Adenocarcinoma through Integrative Computational Analysis with In-vitro Validation in Pakistani Cohort.

Nimra Munir

Quaid-e-Azam University, Islamabad

Email: nimramunir1111@gmail.com

Pancreatic ductal adenocarcinoma (PDAC) is ranked third highest cause of cancer-related mortalities death due to its poor prognosis. The primary issue in PDAC management is the delayed diagnosis, which frequently occurs at advanced stages, severely limiting curative treatment options. Mucins, a class of specialized O-glycoproteins, demonstrate significant potential as diagnostic biomarkers due to their significant over-expression in PDAC. A comprehensive analysis was performed to evaluate the expression profile of Membrane- Associated Mucins (MAMs) gene for its utilization as an early diagnostic and prognostic biomarker. The upregulated key MUC genes were then compared to assess their overexpression in PDAC versus normal pancreatic tissue. The expression profiling of key MUC gene was investigated in correlation with socio-behavioral and demographic factors along with the cancer stage specific analysis. Furthermore, disease-free survival analysis was conducted to evaluate prognostic significance. With the aim of validating the signature biomarker (MUC3A/MUC4/MUC13/MUC16) as a potent early diagnostic and prognostic marker a comprehensive analysis of multi-omics data of Pakistani cohort was performed utilizing the integrated whole exome sequencing, transcriptome analysis and RT-qPCR. The key MUC genes represent an early- stage overexpression as well as the signature biomarker positively correlates with survival analysis with poor prognosis. The in-vitro validation of signature biomarker in Pakistani cohort reveals the pathogenic effect of mutations in the key MUC genes and its profound effect on its protein structure and functionality. In transcriptome analysis, MUC4 was the only one with statistically significant differential expression. In conclusion, our findings suggest MUC3A/MUC4/MUC13/MUC16 as robust diagnostics and prognostic biomarker and liquid biopsy as non-invasive approach to be integrated into daily clinical practice. This analysis can also be extended to other malignancies.

AN IN VIVO EXPERIMENTAL VALIDATION OF THE PREDICTED T-CELL EPITOPES FOR HUMAN RESPIRATORY SYNCYTIAL VIRUS (HRSV)

Aiman Fatima, Tahira Qousain, Kausar Hussain Shah and Syed Aun Muhammad Institute of Molecular Biology and Biotechnology (IMBB), Bahauddin Zakariya University Multan, 60800-Multan, Pakistan

Email: eimyshah19@gmail.com

Respiratory Syncytial Virus (RSV) is a major cause of lower respiratory tract infections (LRTIs) in young children and infants. In Pakistan annually ~0.125 million children of age <5 years die because of LRTIs. Against RSV infections, only prophylactic treatment (Palivizumab) is in practice as there is no proper medication and licensed vaccine available so far. This study was designed to investigate potential vaccine candidates against Pakistani serotypes of RSV by reverse vaccinology approach. For this purpose, RSV related most antigenic localized peptide sequences were identified from the available genomic and proteomic databases. The most antigenic T-cell epitopes (3RSV-MHC-I: FSSKFWPYF and 6RSV-MHC-II: FWPYFTLIH) for RSV were predicted (from SH protein of hRSV) and synthesized. The host-pathogen interaction was investigated. In vivo experimental validation of the synthesized epitope peptides was done in rats(Sprague Dawley) and the immune response was investigated through Hematology (WBCs, LYM, NEUT), IFN-γ, Granzyme B and IgG specific assays. It was observed that the doses of Peptides along with IFA and/or CFA resulted in relatively lower levels of WBCs, LYM, NEUT, IFN, IgG and Granzyme-B as compared to the doses of peptides alone. The synthetic peptide 3RSV-MHC-I induced significantly higher levels of WBCs, LYM, NEUT, IFN, IgG and Granzyme-B when administered as 1st booster dose without any adjuvant. The levels of IgG and Granzyme-B were relatively higher with 6RSV- MHC-II peptide (2.97 g/L, 613 pg/ml respectively) than that with 3RSV-MHC-I peptide. The results indicated that both synthetic peptides could be effective vaccine candidates against hRSV. This study helped in understating the RSV pathobiology and lead to the discovery of new vaccine candidates against RSV.

Transplantation of Hydrogel-Entrapped Mitochondria as a Novel Therapeutic Regime for Skeletal Muscle Ischemia Reperfusion Injury

Laiba Tariq Abbasi1, Saima Barki1, Ameer Hamza1, Fazal Wahid1, Muhammad Rizwan Alam1,2

Department of Biochemistry, Quaid-I-Azam University, Islamabad.

Email: mralam@qau.edu.pk

Skeletal muscle ischemia-reperfusion injury (IRI) is a global health concern that potentially causes severe limb damage, organ failure, and even limb amputation. Mitochondrial dysfunction during IRI triggers excessive oxidative stress and cell death. The present study was designed to explore the effectiveness of mitochondria entrapped in hydrogel Pluronic F127 (PF127) to protect mitochondria from TE injuries and thus MT as an effective therapy to alleviate skeletal muscle IRI. Hindlimb IRI was induced in bALB/c male albino mice and then verified by analyzing its impact on muscle weight, infarct size, and the markers of cellular damage and inflammation. Mitochondria were obtained from liver of healthy mice, quantified and functionally characterized. Likewise, PF127 was characterized for biocompatibility, and temperature- sensitive gelation parameters prior to its in vivo use. PF127-entrapped mitochondria were injected into the hindlimb muscles post ischemia at the onset of reperfusion. Data showed that liver-derived mitochondria were viable and were sensitive to extracellular calcium. 15% PF127 demonstrates biocompatibility and effective temperature-sensitive gelation properties. The in vivo experiments revealed a successful development of IRI as indicated by increased infarct size, muscle weight, fibrosis and elevated serum LDH level, all of which were mitigated upon transplantation of naked mitochondria. However, PF127 coating did not significantly improve MT except for muscle fibrosis probably by causing systemic inflammation as highlighted by elevated serum C5a levels. In conclusion, the data confirms that MT ameliorates IRI in skeletal muscles while the mitochondria entrapment limited the efficiency of MT therapy by causing an increase in systemic inflammation.

Ru265 Modulates Agonist-Induced Platelet Responses Ajwa Ashfaq1, Durre Shehwar1, Manahil Zubair1, Khadeeja Ahsan1, Muhammad Rizwan Alam1,2

Department of Biochemistry, Quaid-I-Azam University,

Email: mralam@qau.edu.pk

Activated platelets are essential for hemostasis and thrombosis. Agonists, such as ADP, adrenaline, collagen and thrombin trigger intracellular signaling cascades that result in calcium mobilization from intracellular stores, ultimately leading to activation and aggregation of platelets. Elevation of both cytosolic and mitochondrial calcium is involved in activation and aggregation of platelets. Mitochondrial calcium uniporter (MCU) is a key transporter that regulates uptake of calcium by mitochondria. Ru265, a derivative of Ru360, is a novel inhibitor of MCU with its implications in research and therapy. Our study was aimed at finding the role of Ru265 in agonists-induced platelet aggregation, cell free DNA (cfDNA) release and clot retraction. MCU was blocked by Ru265 for a short (15 minutes) and long (30 minutes) time to determine whether or not it has any time-dependent effects. Platelet aggregation profiling was performed by using a time-lapsed assay on spectrophotometer. Interestingly, our data reveals a time-dependent effect of Ru265 on ADP-induced aggregation. Ru265 significantly decreased ADP- induced platelet aggregation after shorter incubation time while after a prolonged incubation time, it significantly increased ADP-induced aggregation. However, in case of adrenaline and thrombin, both shorter and prolonged incubation significantly increased the platelet aggregation. Measurement of cfDNA release by Sytox Green, yielded no observable effect of Ru265 with all three. Furthermore, clot retraction assay revealed that Ru265 significantly attenuated the clot retraction process in the absence of thrombin and significantly increased clot retraction in the presence of thrombin. Altogether, the present study revealed a significant involvement of Ru265-dependent mitochondrial calcium signaling in agonist-induced platelet responses.

Harnessing AI and Bioinformatics for Precision: A Novel Methodology for Personalized Cancer Vaccine Development Targeting Neoantigens

Rehman

Department of Biological Sciences, International Islamic University, Islamabad1

Email: arisharehman83@gmail.com

Traditional cancer vaccines often face limitations in efficacy because they struggle to distinguish between tumor-specific and normal body antigens, which can lead to suboptimal targeting and potential adverse effects. To address these challenges, we propose a novel methodology that integrates artificial intelligence (AI) and bioinformatics technologies to improve neoantigen identification and personalize vaccine development. Our approach begins with a biopsy of tumor tissue from cancer patients, followed by comprehensive genome sequencing to identify genetic mutations unique to the tumor. These mutations are analyzed to predict neoantigens—peptides specific to cancer cells due to these genetic alterations. Next, we apply Weighted Correlation Network Analysis (WGCNA) to the gene expression data from the tumor. WGCNA helps identify clusters of co-expressed genes and relevant pathways, refining the selection of neoantigens associated with tumorigenesis. To enhance precision further, we propose developing an AI- powered bioinformatics tool designed to differentiate between neoantigens, normal body antigens, and other cancer-associated antigens. This tool will integrate data from genomic sequencing and WGCNA to accurately classify and select the most promising neoantigens for vaccine targeting.

PCR Based Assay for the Screening of Repeat Expansion in FMR1 Gene

Muhammad Mehtab, Alina Murtaza, Dr. Muhammad Ansar* Department of Biochemistry, Quaid-I-Azam University, Islamabad

Email: ansar@qau.edu.pk

Fragile X Syndrome (FXS) is the most common form of inherited intellectual disability and a significant X-linked neurodevelopmental disorder. This condition arises from an insertional mutation involving a CGG trinucleotide repeat in the 5' untranslated region (UTR) of the FMR1gene on the X chromosome. The abnormal expansion of this triplet sequence leads to hypermethylation and subsequent silencing of the FMR1 gene, resulting in the absence of the encoded protein, Fragile X Mental Retardation Protein (FMRP). The study focused on the development and validation of a PCR-based assay for the screening of repeat expansions in the FMR1 gene, specifically targeting CGG trinucleotide repeats associated with Fragile X syndrome. The research aimed to enhance the efficiency and accuracy of identifying these repeat expansions using a rapid and cost-effective method. The study draws on recent advancements in PCR-based methodologies to streamline the screening process and improve diagnostic capabilities for Fragile X syndrome. Through a comprehensive evaluation of the PCR assay's performance parameters, this research contributes to the ongoing efforts to enhance screening strategies for genetic disorders linked to repeat expansions. For this purpose, normal individuals from different regions of Pakistan were recruited. Blood samples were collected from both male and female normal healthy individuals. After DNA extraction from these blood samples, PCR was performed, and PCR products were resolved on 2% agarose gel to check the size of CGG repeats. The average number of CGG repeats in healthy individuals was found to be between 5-44 reported by various studies. This study will have significant impacts on a better knowledge of FXS, its diagnosis, and prognosis in the Pakistani population. However, further studies will be required to determine the frequency of CGG repeat expansion in FXS.

In Silico Characterization Of Hypothetical Pathogenicproteins Of Leishmania (L. Infantum And L. Donovani)

Nabiha Naeem

University of Management and Technology (UMT) Lahore

Email: nabiha.naeem@umt.edu.pk

Leishmania is the protozoan parasite that causes one of the most fatal forms of parasitic disease known as visceral Leishmaniasis, transmitted by the bite of infected female phlebotomine sandflies, characterized by weight loss, anemia, irregular bouts of fever, and enlargement of the liver and spleen. Annually, 50,000 to 90,000 cases of visceral leishmaniasis occur worldwide, with only between 25 to 45% reported to WHO. This parasite pathogen contains several virulence factors, including metalloprotease, and lipophosphoglycan, however many of its proteins are still hypothetical; and uncharacterized. The epidemic of this pathogen needs the characterization of all of its proteins to know the virulence factors and their effects on human health. The current study used simple and robust in silico approaches to characterize structures and functions of selected 25 hypothetical proteins of Leishmania (L. infantum, L. donovani). The selected 48 proteins were first filtered through homology analysis for template accessibility. After filtering, 25 hypothetical proteins were selected further for the conserved domain analysis (CDD), Pfam, InterProscan, and Scanprosite, subcellular localization via CELLO v.2.5, and signalIP 5.0, physicochemical characterization via ExPASy ProtParam, and structural modeling via I-TASSER and SAVES server (v.6.0). The VirulentPred tool was used for pathogenicity assessment indicating that 18 out of 25 proteins acted as potential virulent factors, functional annotation was evaluated via CATH v.4.3 to assign the probable function to hypothetical proteins respectively. All this information related to functions and the pathogenic involvement of these proteins may help exploit the molecule to act as a potential therapeutic target for Leishmania.

Integrating Computational Method For Druggable Beta-Catenin Inhibitory Phytocompound From Glycine Max L. Seeds To Treat Ovarian Cancer

Samra Hafeez, Asma Ahmed*, Maryam Mureed, Hamna shahid, Afaq Akram Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore, Punjab, Islamic Republic of Pakistan

Email: theafaqshah@gmail.com

Ovarian Cancer is the most lethal gynecological malignancy and is ranked as the seventh leading cause of cancer deaths in females. Wnt signaling is an evolutionarily conserved regulatory pathway that governs numerous normal cellular and developmental processes such as cell fate determination, cell proliferation and migration. However, aberrant Wnt signaling has also been identified as a key mechanism in cancer biology. ß -catenin is the key mediator of the Canonical Wnt pathway. The main components of this complex include AXIN, adenomatous polyposis coli (APC), casein kinase 1 (CK1), and glycogen synthase kinase 3ß (GSK3ß), as well as the E3 ligase, ßTrCP. Protein phosphatase 2A (PP2A) is also associated with the ß-catenin destruction complex. AXIN is a scaffolding protein that has interaction sites for multiple proteins including PP2A, APC, GSK3ß, and CK1. The most common genetic alteration in the Wnt/ß-catenin pathway involved in EOC is in the ß-catenin gene, CTNNB1. Soybeans (Glycine max L.), a functional food widely consumed in Asia, has been reported as the main source of isoflavones. Phytoestrogen properties of soy isoflavones showed their activity as ligands for estrogen receptors and exhibited the estrogenic potency as reported in the previous in vitro and in vivo studies. The major functional components include Carbohydrates, fats, Proteins, Saponins, Lecithin, Linolenic acid, Linoleic acid, Phytosterols and Isoflavones. Due to these valuable constituents, it possesses multiple therapeutic activities. Current in-silico study showed that Daidzein from G. max L. has maximum binding affinity with beta catenin, which may be ultimately inhibited followed by the inactivation of Wnt/β -Catenin pathway. Moreover, it has been shown to relive sleep disorders, may help managing diabetes, prevents osteoporosis, improves blood circulation and provide good care of pregnancy.

Title: In Silico Saturation Mutagenesis Analysis of VEGF-A Binding to VEGFR2: Insights into Binding Affinity and Therapeutic Potential Muhammad Rehan Athar, Syeda Yumna Farrukh*

University of Lahore

Email: yumna.farrukh@imbb.uol.edu.pk

Angiogenesis, the formation of new blood vessels, is critical in both physiological and pathological conditions, with Vascular Endothelial Growth Factor (VEGF) and its receptor VEGFR2 playing pivotal roles. While the VEGF- VEGFR2 interaction is a key target for therapies in diseases like cancer, where angiogenesis is often dysregulated, the impact of specific amino acid mutations on this interaction remains incompletely understood. This study employs in silico saturation mutagenesis to comprehensively explore the effects of mutations on the VEGF-A binding site with VEGFR2. Using the 3v2a model from RCSB, we generated and analyzed 95 mutated VEGF models at residues 17, 21, 25, 63, and 64, alongside the wild-type, using Modeller and HADDOCK3 for docking simulations. Our results reveal significant variability in binding affinities across mutants, with some exhibiting enhanced binding compared to the wild-type. Notably, mutations at positions of 25 showed the most pronounced effects, with Arginine substitution resulting in the most favorable disruption in binding affinity of -10.0 kcal/mol and Asparagine substitution resulting in the most enhanced binding affinity of -14.6 kcal/mol. DOCKQ scores remained relatively high, with some outliners. These findings provide crucial insights into the molecular determinants of VEGF-VEGFR2 binding, offering potential avenues for the rational design of angiogenesis modulators and highlighting specific mutations that could inform the development of targeted therapies for angiogenesis-related disorders.

Identification and In-Silico Analysis of Deleterious Non-Synonymous SNPs in EHBP1 Gene

Fajar baig, Naveed khan

Department of Biotechnology, Abdul Wali Khan University Mardan, KPK, Pakistan

fajarbaigdec@gmail.com

Prostate cancer is one of the frequently reported cancer in men worldwide while it is third most prevalent urinogenital cancer among males in Pakistani population. Variations in several oncogenes and tumor suppressor genes have been studied that have role in prostate cancer. GWAS has identified SNPs in EHBP1 gene to be associated with prostate cancer. EHBP1 gene encodes an adaptor protein regulating vesicular trafficking of endocytes thereby recruiting EPs15-homology containing proteins 1 / 2 (EHD1/2) and Rab8 family members. In this study, we used in- silico approaches for identifying the most damaging non-synonymous SNPs (nsSNPs), playing a significant structural and functional role in EHBP1 protein. Data of Non-synonymous SNPs were recruited from ensemble. Deleterious nsSNPs were identified using Sift, Poly-Phen2, Phd-SNP, SNP & GO and Fathmm. Structural, functional, stability analysis and conservation profile of nsSNPs were verified using Mut-Pred, I-mutant, Mu-pro and Consurf web server respectively. STRING was used for protein-protein interaction. Our study observed that there are 32 most damaging nsSNPs found in EHBP1 gene. Structural and functional analysis of these nsSNPs manifest that they have deleterious effect on protein structure and function. Stability analysis showed that 31 of these nsSNPs decreases protein stability and are located in highly conserved regions. Gene-gene interactions showed relation of EHBP1 with other genes delineate its importance in several pathways and co‐expressions. In future, these 32 SNPs are needed to be explored through population based studies in diseases associated with EHBP1 gene such as prostate cancer consequently investigating novel biomarkers.

In silico Characterization and expression of OmpC gene in Nicotiana tabacum for the development of vaccine against colon cancer

Fatima Ijaz, Mohammad Tahir Waheed

Department of Biochemistry, Quaid-i-Azam University, Islamabad 45320, Pakistan

Email: tahirwaheed@qau.edu.pk, fatima.ijaz.a@gmail.com

Colon cancer is world's 3rd most frequently diagnosed cancer worldwide. 20% of global cancer burden is attributable to infectious agents including bacteria. Salmonella a gram negative bacteria responsible for salmonellosis is among the most common food borne pathogens, and prolonged salmonellosis leads to development of colon cancer. Porins found in the outer membrane of Salmonella can be used to elicit immune response. The production of plant- based vaccine against colon cancer using outer membrane proteins can be helpful for the people of developing countries. In this study, firstly full length Outer Membrane Protein C (OmpC) containing vaccine construct was subjected to in silico analysis. The 3D structure was generated, refined and validated, and the docking analysis was carried out between vaccine model and immune cell receptors. The analysis showed the vaccine construct as immunogenic, non-allergen and non-toxic. Determination of physiochemical parameters, prediction of immunodominant epitopes and results of molecular docking depicted the potential of OmpC to be used as vaccine candidate. The OmpC gene was then expressed in tobacco plants via transformation. Gateway® cloning strategy was used to clone OmpC gene. Integration of transgene within tobacco plants was confirmed through conventional PCR. qRT-PCR was done and copy number was calculated to be 2. The expressed OmpC protein level was calculated to be 0.42% of total soluble protein and its expression was confirmed via western blotting. Immunization of mice with the TSP and purified OmpC protein resulted in generation of strong humoral immune response as verified by ELISA.

Scorpion Venom Peptides Caused Mitochondrial-Driven Apoptosis in Cervical, Prostate, and Brain Cancers

Humaira Hassan1, Munazza Raza Mirza1, Almas Jabeen1, Junaid Ahmed Kori1, Rabia Saultan1, M. Iqbal Choudhary1

Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences (ICCBS), University of Karachi, Karachi, 75270, Pakistan.

Emails: hassanhumaira8@gmail.com, munzihyder@yahoo.com

In this study, we explored the therapeutically potential of venom of yellow scorpions (Buthus sindicus) from Sindh, Pakistan, uncovering its remarkable anti-cancer and anti-inflammatory properties. Our investigation revealed that crude venom effectively inhibits the oxidative burst of human phagocytes, with up to 28.3% inhibition at a concentration of 300 μg/mL. When tested against human prostate (PC3), cervical (HeLa), and neuroblastoma (U87-MG) cancer cell lines, as well as normal human fibroblasts (BJ), the venom exhibited significant cytotoxic effects against cancerous cell lines, particularly against PC3 cells showing IC50 values as 170.8 ± 0.1, 171.7 ± 0.04, 237.4 ± 0.06 μg/mL, respectively. whereas no significant cytotoxicity was found against BJ normal cell line at 100 μg/mL. In order to obtain the effective peptide from crude venom RP-UPLC fractionation was performed, and later we assessed their anticancer potential in-vitro. Notably, Fraction I emerged as a standout, dramatically reducing cell viability in all cancer cell lines (P < 0.001) while non- cytotoxic to normal cells at the effective concentration. Flow-cytometry explored the apoptotic pathways triggered by crude venom and Fraction I in the highly sensitive PC3 cell line. Key assays performed included ROS production, the Bax/Bcl-2 ratio, and p53 gene expression. The findings demonstrated that both the crude venom and Fraction I induce mitochondrial-mediated apoptosis, leading to effective cancer cell death. Mass spectrometry analysis also resulted in the identification of over 35 distinct proteins in the yellow scorpion venom, including potassium channel toxins, proteases, and other functional proteins. Fractions 1 contained potassium channel toxins which potentially were involved in apoptosis and cell proliferation. This study is the first to report on the anti-cancer and anti- inflammatory potential of Pakistani yellow scorpion venom and its relatively purified form, highlighting its promising therapeutic applications and underscoring the need for continued purification and validation research.

Association of Skin Type with Genetic polymorphism in patients with Acne Vulgaris

Quratul Ain1,2, Sidra Younis1, Qamar Zaman1

Quaid I Azam University Islamabad
Shifa Tameer-e-Millat University Islamabad

Email: Quratulain.qau@gmail.com

Acne vulgaris is a chronic, multifactorial inflammatory skin disorder affecting the pilosebaceous follicles, predominantly during adolescence. It is characterized by both non-inflammatory and inflammatory lesions. The primary pathophysiological factors contributing to acne development include abnormal hyper ductal keratinization, elevated sebum secretion, proliferation of Propionibacterium acnes (P. acnes), and an intense inflammatory response.Among the five skin types, oily skin is most susceptible to acne development, followed by combination, sensitive, normal, and dry skin. Interleukin-6 (IL-6), a multifunctional cytokine involved in inflammation and various immune responses, is thought to play a role in acne pathogenesis. Several single nucleotide polymorphisms (SNPs) have been identified in the human IL-6 gene, with four located in its promoter region. The G/C promoter polymorphism at position -174 has been implicated in various inflammatory disorders, including psoriasis vulgaris.This study aimed to investigate the IL-6-174G/C polymorphism in patients with different skin types. The study included 65 patients and 70 healthy controls. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques were employed to analyze the IL-6 gene (-174G/C) promoter polymorphism, using N/aIII for restriction digestion.Results showed no statistically significant differences in genotypic and allelic frequencies between acne patients and healthy controls (P = 0.30 and P = 1.00, respectively). Additionally, no association was found between genotypic and allelic frequencies of acne patients with different skin types (P = 0.38). Similarly, no correlation was observed between IL-6 (-174) genotypes and acne severity (P = 0.38).In conclusion, this preliminary study suggests that the IL-6 (-174G/C) promoter polymorphism is not associated with skin type or acne development in the Pakistani population. Further studies are needed to elucidate the relationship between skin types and the -174G/C IL-6 promoter polymorphism in acne pathogenesis.

In Silico Analysis of 6 Novel Hydrazone Derivatives to Predict their Therapeutic Potential

Hakmeen Khawaja, Sadia Tariq, Fizza Ahmed Tariq, Khadija Shafqat, Mohammad Tahir Waheed*

Department of Biochemistry, Quaid-i-Azam University, Islamabad

tahirwaheed@q au.edu.pk

With the increased technological advancements, Computer-Aided Drug Design (CADD) is gaining attraction because of numerous advantages. With the help of in silico based drug design, it becomes easier to develop cost-effective, efficient, and potential therapeutic drugs. In this study, six synthetic hydrazones (E)-N'-(2-propoxybenzylidene) benzohydrazide (HZ1), (E)-N'-(4- butoxybenzylidene) benzohydrazide (HZ3), (E)-N'-(4-butoxybenzylidene)-4- methoxybenzohydrazide (HZ4), (Z)-N'-(4-methoxybenzylidene)-2-((E)-(2- hydroxynaphthalen-1-yl) diazenyl) benzohydrazide (HZ13), (Z)-N'-(3,4,5- trimethoxybenzylidene)-2-((E)-(2-hydroxynaphthalen 1-yl) diazenyl) benzohydrazide (HZ14), and (E)-2-((E)-(2-hydroxynaphthalen-1-yl) diazenyl)-N'- (furan-2 ylmethylene) benzohydrazide (HZ17) were studied computationally for their pharmacological potential in coagulation, depression, and tuberculosis pathways. Four enzymes, COX 1, MAO-A, KasA, and Pks-13 were used as targets for docking. Molview, AutoDock Vina, Discovery Studio BIOVIA, OSIRIS Property Explorer, and PyMOL were used for the computational investigation of these compounds. HZ1 proved best for coagulation and depression pathways. While HZ3 and HZ4 showed best results in tuberculosis pathway. However, HZ13, HZ14, and HZ17 need certain modifications to make these compounds potentially effective.

Identification of Novel Biomarkers and Main Signaling Pathway in Pakistani Fibroadenoma Patients

Email: mohsinbiostudent18@gmail.com

Background: Fibroadenoma is the most rare benign breast neoplasm in women of childbearing age. The exact molecular pathways of pathology are not well understood yet because of the rare occurrence of this tumor type. This work will seek to investigate the molecular changes in fibroadenoma compared with normal breast tissues to identify specific biomarkers and signal pathways associated with this disease.

Methods: Total RNA and protein will be prepared from these fresh samples of Pakistani Fibroadenoma patients. The expression of all genes of cell proliferation, apoptosis, and extracellular matrix remodeling will be measured using the qRT-PCR technique. Evaluation of such factors as ER, PR, HER2, p53, and Bcl-2 expression levels will be done using Western blot analysis for quantification. The novel mutations in differentially expressed genes will be found using Sanger sequencing and confirmed using ARMS PCR.

Expected Results: Information based on prior investigation revealed that qRT- PCR is expected to indicate differential expressions of several genes linked to proliferative and anti-apoptotic markers compared to the normal tissue. We anticipate that Western blot analysis will reveal the over/under-expression of the proteins described. Such data should pinpoint new mutations in the splice isoforms that are characteristic of fibroadenoma using the Sanger Sequencing technique.

Conclusion: This work will also help in the molecular analysis of fibroadenoma and discover new biomarkers that may lead to the disease. These results are expected to provide novel information concerning the development of fibroadenoma and its possible treatment.

HPV and Cancer in Pakistan: Investigating Renal and Prostate Cancer Associations

Maham Ayman 1, Ayesha Irfan 1†, Khalid Farooq 2, Nuzhat Sultana 3, Sobia Asghar 1, Aneela Javed 1, Saira Justin 1, *

Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST), H-12 Sector, Islamabad, Pakistan. 44000.
Urology and Renal Transplantation, Foundation University Medical College/Fauji Foundation Hospital, Rawalpindi,
Department of Pathology, Northwest School of Medicine, Peshawar,

† Co-first Author

Email: justinsaira@hotmail.com

Abstract

Human papillomavirus (HPV)-attributable cancers are more pronounced in low- and middle-income countries (LMICs), where limited access to screening and vaccination programs poses a significant challenge. The etiological association of HPV infections with anogenital and urologic cancers is under- researched in Pakistan. Therefore, this study aims to investigate the link between HPV and renal and prostate cancers. After obtaining informed consent, 50 biopsy samples from each cancer type, confirmed by H&E and IHC staining, were collected along with histopathological and clinical data. DNA was extracted and processed for HPV detection using GP5+/6+ consensus primers followed by HPV genotyping using E6-E7 specific primers for HPV16 and HPV18, via conventional PCR and Sanger sequencing. For comparative structural analysis of HPV-L1 protein, HPV-positive samples were amplified with MY09/11 primers and sequenced using the Sanger method. Protein sequences were translated using MEGA 11 software followed by homology modeling via trROSETTA, and 3D structure alignment via MOE. For renal cancer, 30% (15) of the cases tested positive for HPV, of which 93% of the cases exhibited HPV16&18 co-infection. Contrarily, all the prostate cancer samples tested negative for HPV. Additionally, no significant structural variations were seen in the HPV-L1 proteins of the HPV-positive samples. This is the first study to explore the link of HPV with renal and prostate cancer in Pakistan. To gain a deeper understanding, more comprehensive research is needed. Given the HPV burden in Pakistan, globally available HPV vaccines are an effective preventive option to reduce cancer and socioeconomic impact.

Keywords: HPV; HPV16; HPV18; Prostate cancer; Renal cell carcinoma (RCC); Sexually transmitted diseases (STD)

Prevalence Of Genetic Disorders In Pakistan: A Systematic Review

Muhammad Ammar1, Sahar Safdar1*, and Syeda Marriam Bakhtiar1

Genetic and Molecular Epidemiology Research Group, Department of Bioinformatics and Biosciences,
Capital University of Science and Technology (CUST), Islamabad, Pakistan

E-mail: saharsafdarss26@gmail.com

Pakistan is characterized by significant genetic diversity due to its complex population structure where the prevalence of genetic disorders poses a substantial public health challenge. A genetic disorder is defined as a health condition resulting from abnormalities in the genome, originating from mutations in genes, chromosomal changes, or multifactorial influences that encompass both genetic and epigenetic factors. The incidence of non- communicable diseases is on the rise in Pakistan, contributing to approximately 58% of the total mortality rate. With a population exceeding 245 million and a growth rate of 1.89%, Pakistan is ethnically diverse and the world's 5th most populous country in 2024, comprising various ethnogeographic groups include Punjabis, Pashtuns, Sindhis, Saraikis, Muhajirs, Balochs, Hindkowans, Brahuis, Meos, Kohistanis and many other minorities, each contributing to the intricate genetic landscape of the nation. This diversity is further compounded by cultural practices by significant numbers of, particularly the highest practice of consanguineous marriages globally which is more than 63%, which significantly elevates the risk of genetic disorders. This study aimed to analyze 104 genetic disorders, predicting their prevalence through advanced artificial intelligence methodologies and literature mining their prevalence through conventional practices. A comprehensive systematic literature review was subsequently conducted, mining data from 130 articles and selecting 24 that provided relevant prevalence statistics for 65 highly prevalent genetic disorders in Pakistan. Later applied multiple tools on extracted data to gain statistical and graphical insights. Our findings reveal a discrepancy between AI estimated prevalence and those derived from conventional literature mining, primarily attributed to the scarcity and limitation of published data about the Pakistani population. The AI-based predictions were found to lack authenticity, underscoring the challenges associated with relying solely on computational methods in the absence of robust empirical data. The literature shows that over 90% of the identified genetic disorders in Pakistan are directly or indirectly associated with consanguineous marriages, highlighting a significant concern given that Pakistan has one of the highest rates of such unions globally. This association underscores the urgent need for targeted public health strategies to address genetic disorders, particularly in communities with elevated rates of consanguinity. Challenges of the study include a small sample size and also the not reported or incomplete nature of the epidemiological genetic data, necessitating further research to comprehensively understand the prevalence and impact of genetic disorders in Pakistan. This research not only elucidates the current state of genetic disorders in Pakistan but also paves the way for future investigations into unpublished genetic diseases related epidemiological data. It serves as a critical call to action for policymakers to prioritize genetic health within their agendas and develop strategies aimed at mitigating the burden of genetic disorders on the population. By enhancing awareness comprehensively regarding the causes, accurate diagnosis, prevalence, available treatments, and counseling for genetic disorders will enable stakeholders to more effectively address the public health challenges posed by these conditions, ultimately improving health outcomes for affected Pakistani individuals and communities.

Exploring Gene Expression Profiles in HER2 Positive Breast Cancer using GEO2R and RT PCR

Email: rubabayesha21@gmail.com

Background: Breast Cancer is the abnormal growth and multiplication of cells in the breast tissues due to various mutations in the pathways of different genes. Breast cancer is the second most common cancer in women. HER2 breast cancer is a primary global health concern caused by excessive gene amplification, a member of ERBB family. The given study investigates gene expression patterns linked with HER2-positive breast cancer to improve our knowledge and find potential biomarkers for targeted therapies.

Method: Differential gene expression was conducted on the dataset taken from the Gene Expression Omnibus (GEO) database using the GEO2R program. The primary breast tissue samples were taken from the GEO database and were subsequently divided into two subgroups, i.e., HER2 positive and Negative breast cancer, depending on their HER2 status. The hub genes filtered from this bioinformatics analysis will be validated for their up- regulation or down-regulation expression by RT PCR.

Results: The hub genes TP53, PTEN, ESR1, MHY7, ACTN2, are specific for HER2- positive breast cancer pathogenesis. These were identified using bioinformatics tools such as GEO2R, STRING and protein-protein interaction networks. These genes are differentially expressed and play important roles in the progression of HER2-positive breast cancer. We will perform RT PCR of targeted hub genes to check their differential expression.

Conclusion: The current study comprehensively examines gene expression and mutation status in HER2-positive breast cancer patients, offering guided targeted therapy options and insights for future research.

Novel therapeutic to Combat Tuberculosis: A combination of CAF01 conjugated multi-subunit Vaccine and BCG

Saila Batool

University of Manchester

Email: batoolsaila.00@gmail.com

Till present times, tuberculosis (TB) remains as one of the significant diseases in low middle income incomes of the South Asia including Pakistan. WHO recognises TB as a global epidemic, one with a global prevalence skewed by age. The Bacillus Calmette-Guerin (BCG) vaccine usually administered in the first decade of life remains as the standard of care treatment for preventing TB. A live attenuated vaccine, BCG was derived from Mycobacterium Bovis (the main causative agent of bovine TB) through multiple serial passages. However, whilst a cornerstone in its’ time, the prophylactic use of the BCG vaccine is increasingly unfit for purpose. Multiple studies have proven that likely through increased serial passages over time, BCG has deviated away from the structure of Mycobacterium Tuberculosis (MTB), the causative agent of TB. Moreover, it immunogenicity wanes down to ineJective level by the 4- 5th decade of life, thereby leaving the middle-aged and elderly unprotected. A proposed design of a multi-subunit vaccine made up of well-defined antigens could potentially replace the current outdated BCG vaccine being used. Recent studies have proposed and trialed subunit designs that have used a maximum of 8. Herein we propose a vaccine design incorporating 13 subunits with greater specificity to MTB to induce a stronger and long-lived antiMTB immune response. Compared to primarily cell based, or live attenuated vaccinations, subunit has innumerate benefits. The most critical benefit, of course is the markedly reduced incidence of allergic or autoimmune reactions, which elderly persons are inherently predisposed to, due to processes such as inflammageing and immunosenescence.

Computational Exploration of Berberis Lycium Royle : A Hidden Treasure Trove for Antiviral Development

Mamuna Mukhtar1, Haris Ahmed Khan1,2, Tope Abraham Ibisanmi3, Ayodele Ifeoluwa Faleti4, Najam us Sahar Sadaf Zaidi1

National University of Sciences and Technology (NUST)

Email: mamunakhan26@gmail.com

Viral infections and associated illnesses account for approximately 3.5 million global fatalities and public health problems. Medicinal plants, with their wide therapeutic range and minimal side effects, have gained limelight particularly in response to growing concerns about drug resistance and sluggish development of antiviral drugs. This study computationally assessed eleven chemical compounds from Berberis lycium along with two antiviral drugs to inhibit SARS CoV 2 (Covid-19) RNA-dependent RNA polymerase (RdRP), Influenza virus RdRP, and two crucial dengue virus enzymes (NS2B/NS3 protease and NS5 polymerase). Berberine and Oxyberberberine passed all pharmacokinetics analysis filters including Lipinski's rule, BBB permeant, and cytochrome suppression and demonstrated druglikeness, bioavailability, and a non-toxic profile. Docking of phytochemicals from Berberis lycium returned promising results with selected viral proteins i.e., DENV NS2BNS3 (Punjabine-10.9 kcal/mol), DENV NS5 (Punjabine -10.4 kcal/mol), covid 19 RdRP (Oxyacanthine -9.5 kcal/mol), and Influenza RdRP (Punjabine -10.4 kcal/mol). The optimal pharmacokinetics of berberine, exhibited good binding energies with NS2BNS3 (-8.0 kcal/mol), NS5 (-8.3 kcal/mol), covid-RdRP (-7.7 kcal/mol) and Influenza RdRP (-8.3 kcal/mol) while MD simulation of a 50ns time scale by GROMACS software package provided insights into the flexibility and stability of the complexes. A hidden treasure trove for antiviral research, berberine, berbamine, berbamunine, oxyberberine, oxyacanthine, baluchistanamine, and sindamine have showed encouraging findings as possible lead compounds. Pharmacological analyses provide credence for the proposed study; nevertheless, as the antiviral mechanisms of action of these phytochemicals are not well understood, additional research and clinical trials are required to demonstrate both their efficacy and toxicity through in vitro and in vivo studies.

Non-synonymous Polymorphic Variant of AP-2REP Impacting Protein Structural and Functional Stability

Ghulam Murtaza Kamal

National University of Sciences and Technology

Email: mmurtaza4498@gmail.com

Liver cancer is the third most common cause of death globally, accounting for 7.8% of deaths by cancer. Early-stage liver cancer is frequently asymptomatic and aggressive, hindering the timely and effective diagnostic and prognostic approach. Diagnosis at an advanced stage frequently makes liver cancer less curable as it exhibits drug resistance. One of the most common genetic predispositions, single nucleotide polymorphism (SNP) has been linked to the cancer pathogenesis. However, pathogenic impact of non-synonymous SNP variants of AP-2REP has not been evaluated yet in liver cancer development. AP-2REP belongs to the KLF family, acts as a transcription factor, and multiple targeted genes of AP-2REP are involve in cell cycle regulation, cell proliferation, cell growth and apoptosis. Hence, it is postulated that alterations in AP-2REP structure might impair the AP-2REP functions and interaction and could exacerbate the key cellular processes. This dysfunction may contribute to the development and progression of cancer. The aim of this study is to investigate pathogenicity, structural and functional impairments induced by non-synonymous SNP variants of AP-2REP and to establish a relation to liver cancer susceptibility, development, and progression. Various in silico tools were employed to investigate SNP variants’ effect on AP-2REP protein stability, structural and functional alterations. The most pathogenic non- synonymous SNP variant rs1257857115 (R397W) genotype was identified in 100 HCC patients and 100 healthy controls by employing tetra ARMS PCR. The SNPs in AP-2REP affect the protein’s stability, structure, and function.There was a positive association between AP-2REP variant rs1257857115 genotype (TT) in HCC patients with higher susceptibility risk, in contrast, the protective role of wildtype genotype (CC) was found in healthy controls. Hence, it is concluded that AP-2REP variant rs1257857115 is significantly associated with HCC and can surge cancer development and could be used as a diagnostic biomarker in HCC.

Exploring the Complex Interplay between Transcription Factor Cooperation and the Milk Traits of Murciano-Granadina Goats Muhammad Imran Khan1,2,3 , Hendrik Bertram1 , Armin Otto Schmitt2,4 , Faisal Ramzan5 , and Mehmet Gültas2,4 1.

Faculty of Agriculture, South Westphalia University of Applied Sciences, Lübecker Ring

2, 59494 Soest, Germany 2. Breeding Informatics Group, Department of Animal Sciences, Georg-August University, Margarethe von WrangellWeg 7, 37075 Göttingen, Germany 3. Department of Livestock Production and Management, PMAS-Arid Agriculture University, Rawalpindi 4. Center for Integrated Breeding Research (CiBreed), Carl-Sprengel-Weg 1, Georg-August University, 37075 Göttingen, Germany 5. Institute of Animal and Dairy Sciences, Faculty of Animal Husbandry, University of Agriculture, Faisalabad, Pakistan Email: khan.imran@fh-swf.de

The Murciano-Granadina goat (MUG), a prominent dairy breed, stands out for its adaptability, resilience, and distinguished milk qualities, including high protein and fat levels combined with low somatic cell counts. These traits are shaped by complex biological processes, essential for phenotypic variation. Therefore, understanding the regulatory mechanisms of milk production and lactation in this breed is vital. This research delves into the genetic framework governing seven milk traits in MUG through a two-phase analysis assessing genotype-phenotype associations. Initially, a Random Forest algorithm identified the relative importance of each SNP in determining the traits of interest. The second step applies an information theory-based approach, Mutual Information-Based Detection of Epistatic SNP Pairs, to explore the complex genetic architecture of quantitative milk traits, focusing on epistatic interactions that might be overlooked in the first step. This methodology helped to identify nearly unique sets of candidate genes for each trait. In contrast, by analyzing the promoter regions of these genes, we have revealed common regulatory mechanisms among the milk traits under study. These traits include dry matter percentage, length of lactation/milk production days, fat percentage, protein percentage, lactose percentage, milk yield at 210 days, and somatic cell count. These insights are pivotal for grasping the molecular mechanisms of gene regulation, emphasizing the critical role of transcription factors (TFs) and their preferential partner choice in the development of these traits. Notably, TFs such as DBP, HAND1E47, HOXA4, PPARA, and THAP1 were consistently identified for all traits, highlighting their important roles in immunity within the mammary gland and milk production during lactation.

Computational Analysis of BDNF Gene Variants and Their Role in Neurodegenerative Disorders

Nazia Hadi

Department of Biotechnology, Abdul Wali Khan University Mardan KPK Pakistan

Email: naziahadi123@gmail.com

Brain-derived neurotrophic Factor (BDNF) is an essential growth factor in the neurotrophin family and is crucial for developing the nervous system. It supports the generation of new neurons and helps maintain the nervous system's overall health. Alterations in BDNF levels in the blood and central nervous system are linked to the onset of disorders such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, ischemic stroke, etc. This study aims to identify potentially harmful single nucleotide polymorphisms (SNPs) and assess their effects on protein function and stability. We employed various bioinformatics tools and methodologies to pinpoint the most damaging nsSNPs that could significantly affect the structural integrity of the BDNF protein, including SNP & GO, PolyPhen 2, and PANTHER, SIFT, PHD-SNP, Predict-SNP, and SNAP2. Moreover, IMutant, MUPro and ConSurf respectively. The 3-D protein structures were predicted using I-TASSER. Gene-gene interactions by gene- MANIA and protein-protein interactions were explored through the STRING database. Our study discovered ten harmful missense single nucleotide polymorphisms (SNPs) (rs866172975, rs771341699, rs1590217373, rs751698045, rs758638310, rs1330439007, rs77787410, rs1852795747, rs1590215885 and rs780128716) in the BDNF gene coding region. These nsSNPs drastically reduced BDNF protein stability and modified its structural arrangements, causing potential susceptibility towards various neurological ailments. Gene-gene interaction analysis has underscored the importance of BDNF across multiple pathways and its associations with other genes. These findings suggest the potential impact of harmful nsSNPs on BDNF-related disorders by opening new avenues to understand further the role of BDNF gene mutation in neurological disease development.

Relationship of Anxiety, Depression, Screen Time and Sleep Quality among adolescent: Cross sectional study

1Nur Zakiah Mohd Saat, 1Nur Atikah Mustaim, 2Mahadir Ahmad, 3Siti Aishah Hanawi, 2Nor Farah Muhammad Fauzi

1Programme Biomedical Science, Centre of Community Health Studies(ReaCH), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Aziz, Kuala Lumpur 50300, Malaysia; 2 Centre of Community Health Studies (ReaCH),, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Kuala Lumpur 50300, Malaysia; 3SOFTAM, Faculty of Information Science and Technology, Universiti Kebangsaan Malaysia, Bangi 43600, Malaysia

Nur Zakiah Mohd Saat

Email: nurza@ukm.edu.my

In the current digital age, people's use of electronic devices has significantly increased screen time, which may have an impact on different aspects of their lives. Two important areas of concern are sleep quality and overall quality of life in terms of anxiety and depression, which have only been partially studied to screen time. The purpose of this study is to examine the relationships between screen time, sleep quality, and quality of life. This is a cross sectional stud among 353 secondary school students in the Klang Valley using a questionnaire the Hopkins Symptoms Checklist-25 (HSCL-25), SCREEN-Q to evaluate screen time and Pittsburgh Sleep Quality Index for Malaysian populations (PSQI-M). The results revealed that there was significant difference of depression and anxiety level between male and female adolescent (p<0.05). Meanwhile gender was a significant moderating variable for relationship between depression level and duration of phone. Furthermore, age group was a significant moderating factor between anxiety level and depression level with sleep quality. (p<0.05 Understanding the effects of excessive screen time on sleep and well-being may have a substantial impact on public health policies and interventions. Enacting policies that promote better screen habits and sleep hygiene could improve people's overall quality of life and well-being in the digital age. However, more longitudinal research is needed to confirm the causality of these relationships and investigate potential intervention strategies.

Protective Antioxidant Role of Dodoneae viscosa Against Oxidative Stress in Liver Damage

Anam Urooj* , Muhammad Rashid Khan * , Shahzad Khan ** , Maryam Mushtaq *

, Misbah Farooq * , Saira Bibi * , Kashif Awan **

*Department of Biochemistry, Quaid-i-Azam University (QAU), Islamabad **Department of Pathobiology, University of Poonch (UPR), Rawalakot

Tenacity of our study is to investigate the antioxidant property of Dodonaea viscosa flower against CCl4.Dried flower of Dodonaea viscosa. To study the antioxidant potential of methanol and all its derivatives, different in vivo and in vitro antioxidant assays were performed. One being the Ethyl acetate, which was the active fraction in all assays, has expressed best results. Protective role of crude methanolic extract of Dodonaea viscosa flower was determined against oxidative stress as liver and DNA damage in rats was initiated by carbon tetrachloride. Treatment with plant extract (DVME) against oxidative damage induced by CCl4 was given on each alternate day for one month. Concentration of glutathione (GSH) and enzymes (CAT, SOD, POD) activity in liver and brain were initiated in CCl4 group in comparison to plant treated group. While for peroxidation (TBARS), lower values were noted in extract group and CCl4 group has higher values. Similarly, H2O2 and nitrite levels were significantly.

Enhancing Immunogenicity and Stability of Vaccine Against Newcastle Disease Virus; an emerging infectious disease in Pakistan

Zainab Kamran1, Maaz Waseem1, Muqadas Mirza2, Saifullah Khan1, Muhammad Asghar1,3

Atta-Ur-Rahman School of Applied Biosciences (ASAB), National University of Science and Technology (NUST),
Department of Biological Sciences (DBS), International Islamic University,
Biology Department, Lund University,

Newcastle Disease Virus (NDV) is a major threat to poultry industry causing millions of deaths of chickens worldwide. The poultry industry in Pakistan also faces huge danger as NDV causes recurrent epidemics due to virus rapid genomic evolution. This study focuses on development of multiepitope vaccine against NDV strains in Pakistan. For vaccine we had domain-based approach which gave us promising B and T cells epitopes. The 3D structure of the vaccine was constructed, refined, and validated. To improve immunogenicity, these epitopes were merged into a vaccine design together with the appropriate adjuvants and linkers. High potential for immunogenicity, non-allergenicity, and antigenicity was demonstrated by the vaccination. Molecular docking confirmed strong binding to chicken Toll-like receptor-4, and in silico immunological simulations showed a robust immune response. Computational analyses suggest the vaccine is physically stable and highly immunogenic, making it a promising candidate against NDV strains from Pakistan. The final vaccine construct was designed by linking selected epitopes with adjuvants and linkers to enhance immunogenicity. This study highlights the efficacy of vaccine design, offering a solution for NDV outbreaks in Pakistan and similar regions in the time of emerging infectious diseases.

Alleviation of Dexamethasone and BaCl 2 -induced Skeletal Muscle Injury by Mitochondrial Transplantation.

Fazal Wahid

Department of Biochemistry, QAU, Islamabad

Aim: Evaluating the impact of allogeneic mitochondrial transfer as a therapeutic strategy in skeletal muscle injury in mice.

Objectives: The development of skeletal muscle atrophy model using dexamethasone. The establishment of skeletal muscle myotoxic injury model using BaCl2. MT in injured skeletal muscles to assess its effect on dexamethasone and BaCl2-induced muscle injuries.

Materials and Methods: Liver Mitochondria Isolation, OCR Assay, MTT Assay, Healthy Mitochondria, Mitochondrial Transplantation.

Conclusion: Dexamethasone and Bacl2 can be used as skeletal muscle atrophy and myotoxic injury mice models.

Mitigating Depression through Natural Supplementation: A Comparative Study of Behavioural Patterns in Rodent Model Muhammad Nouman Zahid Magray, Naila Naz

National University of Sciences and Technology, Islamabad Pakistan

Chronic daily life stress is one of the leading causes of depression, which is a prevalent psychological disorder affecting millions of people globally. To treat depression, various known and commercially available antidepressants, such as SSRIs, SNRIs, and TCAs, exist. However, all of these have unpleasant side effects. Apart from these conventional antidepressants, various vitamins are associated with depression management such as folates. Folate is a key component of brain health involved in various pathways, particularly in neurotransmitter synthesis, such as serotonin and dopamine. The aim of this study is to explore the role of folate as a prevention strategy for depression. Components such as L-Methyl folate, Vitamin B2, and Vitamin D3 were used in combination on Sprague Dawley rats’ model of chronic unpredictable mild stress. Both male and female rats, were divided into three groups; the control group was given neither stress nor vitamins, the second group was subjected to stress and saline injections, and the third group was subjected to stress and subcutaneous vitamins injections daily for 21 days. Anhedonic behaviour, locomotor activity, and despairlike behaviour were examined using the sucrose preference test, open field test, and forced swimming test. The data revealed that the combination of L-Methyl folate with vitamins significantly prevented anhedonia, as evidenced by a sucrose preference of 80% (p < 0.0001). Moreover, it effectively preserved locomotor abilities, with rats covering a total distance of 80 meters in the open field test, in contrast to the 50% observed in depressed rats (p < 0.03). Additionally, the combination therapy prevented despair-like behaviour, as indicated by a reduction in immobility percentage to 50% compared to the 70% observed in depressed rats (p < 0.01). These results suggest that folate in combination with vitamin B2 and D3 can serve as a potential preventative candidate for stress-related mental health disorders.

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