Nazish Bostan

Livestock is the backbone of the economy in an agricultural country like Pakistan, with cattle serving as a milk and protein source. In 2021/22, Pakistan was hit by the first major outbreak of lumpy skin disease (LSD) in cattle, in all four provinces. LSD is characterized by the development of skin nodules, leading to severe illness, decreased milk production, and mortality, causing huge economic losses. This study aimed to analyze and classify the lumpy skin disease virus (LSDV) strains involved in the outbreak in the Punjab province at the molecular and phylogenetic levels to develop effective control strategies. A combination of different real-time PCRs was used for the identification and differentiation between vaccine, wild-type, and recombinant LSDV strains. This was mented with the sequence determination and phylogenetic analysis of ten genomic loci from two selected isolates from the 2021/22 Pakistan outbreak. The combined data showed that these isolates belonged to LSDV clade 1.2 and were clearly different from the vaccine clade 1.1 (Neethling-like), as well as from the recombinant clade 2 strains. In addition, using a fit-for-purpose gel-based PCR, the isolates from the outbreak were also shown to be different from KSGP0240-based vaccines.

Water filtration plants are a main source of drinking water for local populations in almost all big cities of Pakistan but these filtration plants are ineffective in eliminating contaminants from water. This study was conducted to evaluate water quality with specific focus on physicochemical parameters and bacterial contamination including occurrence of multidrug-resistant bacteria in drinking water collected from filtration plants (N = 64) of the twin cities of Pakistan. Physicochemical parameters were analyzed following American Public Health Association standard guidelines and total coliform count was determined using membrane filtration method. Gram-negative bacteria were isolated and characterized in terms of species and antibiotic susceptibility. Elevated levels of total dissolved solids, nitrates, total coliforms, lead and copper were found in several samples. Multivariate correlation coefficient revealed significant positive correlation between nitrate levels and total coliform count in bacteriologically contaminated samples. Antibiotic resistance patterns revealed multidrug-resistant and extensively drug-resistant bacteria with high resistance against third generation cephalosporins. Our results highlight the urgent need for improved water filtration processes and regular monitoring to provide safe drinking water to the local community.

Genes of folate-mediated 1 carbon metabolism are found to be highly upregulated in tumor cells and promote cancer cell proliferation. The current study aimed to determine the expression of the MTHFD2 gene in bladder and breast cancers. Furthermore, the determination of potential ligand-based inhibitors against MTHFD2 was performed in comparison with those of chemotherapeutic drugs and natural plant-based compounds. Semiquantitative expression analysis along with structure-based virtual ligand library screening was done to find plausible inhibitors. MTHFD2 expression was significantly increased with tumor stage progression both in low- and high-grade bladder cancer and especially in triple-negative breast cancer. Virtual ligand-based library screening against the three-dimensional MTHFD2 protein structure led to the identification of plausible inhibitors like MCULE-8109969891-0 and MCULE-9715677418-0-1 that displayed lower binding free energy as compared to that of already documented LY345899. Similar scaffold commercial drugs leucal (LEU), epirubicin (EPI), and lometrexol also displayed strong binding to the active site of MTHFD2. EPI and LEU in combinatorial therapy were also tested in vitro on MDA-MB-231 cells. The high doses of LEU in combination with EPI showed a significant reduction in cell viability at 2 and 3 μM concentrations. The interaction of breast cancer serum with high expression of MTHFD2 also showed an increase in binding of epirubicin in the presence of leucovorin. The decrease in the absorbance spectra of epirubicin at 37 and 53 °C displayed the stability induced by LEU on the interaction of EPI with the MTHFD2 binding pocket. Leucovorin tends to stabilize the interaction as the binding affinity is high even at 53 °C. Thus, MTHFD2 might be used as a cancer biomarker since its expression level changes drastically with tumor progression. Further experimental studies are required to establish the potential mode of inhibition of the novel small ligands. Future in vivo trials may validate the effectiveness of the combinatorial therapy.

Background: Pakistan is 1 of the 2 countries endemic for wild poliovirus type 1 (WPV1). Active clinical and environmental wastewater surveillance along with laboratory investigation is an integral and primary component of polio eradication strategies. The current study is mainly focused on virological data to understand the current epidemiology of WPV1 in Pakistan during 2019–2022. Methods: In total, 141 037 stool specimens from patients reported with acute flaccid paralysis and 3171 wastewater samples were tested for detection of poliovirus using cell culture and polymerase chain reaction (PCR). Phylogenetic analysis of WPV1 was performed using MEGA and Nextstrain. Results: Poliovirus isolates were classified into 15 distinct genetic clusters with multiple transmission lineages. Spatiotemporal trends indicated a significant decline in the incidence of poliomyelitis, reported in 58 districts in 2019 and just 3 in 2022. The historical reservoirs in Peshawar, Quetta, and Karachi successfully eliminated the indigenous transmission chains of wild poliovirus that had been active there for years Conclusions: Our findings reinforce the evolving epidemiology of poliovirus in Pakistan, which is now confined to south-KP. All historically known reservoirs in Peshawar, Karachi, and Quetta block are now free of poliovirus. Intensified clinical and environmental surveillance should be maintained to eliminate the very few remaining transmission lineages and certify poliovirus eradication by 2026.

Data published in 2020 by the International Agency for Research on Cancer (IARC) of the World Health Organization show that breast cancer (BC) has become the most common cancer globally, affecting more than 2 million women each year. The complex tumor microenvironment, drug resistance, metastasis, and poor prognosis constitute the primary challenges in the current diagnosis and treatment of BC. Magnetic iron oxide nanoparticles (MIONPs) have emerged as a promising nanoplatform for diagnostic tumor imaging as well as therapeutic drug-targeted delivery due to their unique physicochemical properties. The extensive surface engineering has given rise to multifunctionalized MIONPs. In this review, the latest advancements in surface modification strategies of MIONPs over the past five years are summarized and categorized as constrast agents and drug delivery platforms. Additionally, the remaining challenges and future prospects of MIONPs-based targeted delivery are discussed.

Salmonella enterica includes enteric pathogens of zoonotic potential, possessing one of the largest pools of temperate phages in their genomes. One such class of phages previously called Podoviridae (Now genera placed directly under class Caudoviricetes temporarily) is the largest group of temperate phages that lysogenize clinically and economically important Salmonella enterica serovars. These phages are capable of generalized transduction and are well known for carrying foreign DNA (mobile genetic elements and resistance genes) to new bacterial species that play a major role in the evolution, pathogenicity, and host adaptability of Salmonella serovars. Here we report a novel species of podo viruses; Salmonella phage BIS08P22 (BIS08) that infects Salmonella enterica serovar Typhimurium strain SE-BS17 (Acc. NO: MZ503545). Phage BIS08 was viable only at biological pH and temperature (pH7 and 37 °C) and features 41,574 -base pair (bp) linear ds DNA genome with 47% GC content. It encodes 73 putative Open Reading Frames (ORFs), has a mosaic arrangement, and shares only 16 core genes with its closest homologs. BIS08 genome possesses only 52 % homology (genome-wide Nucleotide homology by VIRIDIC analysis) with members of the genus Lederbergvirus which is not sufficient to place it in the same genus. This phage has a unique terminase enzyme (DNA packaging motor) with no nucleotide and protein homology with any known member of the Lederbergvirus genus. We carried out a detailed phylogenetic and genome analysis of this novel phage and proposed its placement in a new genus. Our study also suggests a revision in the classification of the genus Lederbergvirus. We also performed in silico structural analysis of the BIS08 unique terminase enzyme comparing it to its close homologs.

Tamoxifen is the drug of choice as hormonal therapy forhormonereceptor-positive breast cancers and can reduce the risk of breastcancer recurrence. However, oral tamoxifen has a low bioavailabilitydue to liver and intestinal metabolic passes. To overcome this problemand utilize the potential of this drug to its maximum, inorganic nanoparticlecarriers have been exploited and tested to increase its bioavailability.Biocompatibility and unique magnetic properties make iron oxide nanoparticlesan excellent choice as a drug delivery system. In this study, we developedand tested a "green synthesis" approach to synthesizeiron nanoparticles from green tea extract and coated them with agarfor longer stability (AG-INPs). Later, these hybrid nanoparticleswere conjugated with tamoxifen (TMX). By using this approach, we synthesizedstable agar-coated tamoxifen-conjugated iron nanoparticles (TMX-AG-INPs)and characterized them with Fourier-transform infrared (FTIR) spectroscopy.The average particle size of AG-INPs was 26.8 nm, while the averageparticle size of tamoxifen-loaded iron nanoparticles, TMX-AG-INPs,was 32.1 nm, as measured by transmission and scanning electron microscopy.The entrapment efficiency of TMX-AG-INPs obtained by the drug releaseprofile was 88%, with a drug loading capacity of 43.5%. TMX-AG-INPswere significantly (p < 0.001) efficient in killingbreast cancer cells when tested in vitro on the establishedbreast cancer cell line MCF-7 by cell viability assay, indicatingtheir potential to control cell proliferation.