A biochemical and histological evaluation of in vivo exposure of bisphenol P for multi-organ toxicity and pathology in rats.

Bisphenol P (BPP) is a structural analog of bisphenol A (BPA) and is increasingly used as a substitute of BPA in commercial and household applications. In recent years, BPP has been frequently detected in terrestrial and aquatic ecosystems. Very little epidemiological and experimental information are available on the toxicity potential of BPP in human and animal systems, which is very concerning in view of its increasing use. The current study evaluated the biochemical and histopathological effects of BPP in rats. The seven experimental groups (n = 5 rats/group) included BPA5 (5 mg), BPA50 (50 mg), BPA100 (100 mg), BPP5 (5 mg), BPP50 (50 mg), and BPP100 (100 mg) while the remaining one group served as untreated control. At the end of treatment, the organs (liver, kidney, heart, and lung) of rats were harvested for oxidative stress and histopathological analyses. A significant (p < .05) decrease was observed in the weight of the liver, lungs, and kidneys in the BPP100 group similar to the BPA100 group compared with the control group. Further, a significant (p < .05) decrease was also observed for concentrations of antioxidant enzymes (catalase, peroxidase, superoxide dismutase, and glutathione peroxidase) in the liver, lungs, kidneys, and heart at the highest two doses of BPP similar to the respective BPA groups compared with the control group. The two highest doses of BPP induced histopathological changes in the liver such as nuclei distortion, excessive necrosis of hepatocytes, nuclei shrinkage and pyknosis of cells with disrupted cell structure (BPP100), and cellular congestion and degeneration of hepatocytes (BPP50) similar to the two respective doses of BPA. The BPP treated groups also showed varying histopathological changes in kidney tissue, heart tissue, and lung tissue similar to BPA treated rats. In conclusion, the present study indicated that BPP has the potential to induce oxidative stress and alter the histomorphological architecture of different organs and is as deleterious as BPA.

Evaluation of phytochemicals from Thymus serpyllum as potential drug candidates to manage oxidative stress in transition dairy cows.

Dairy cows undergo immense stress and experience autoimmune reactions during the transition period, majorly due to the generation of ROS in the body. So, pharmacological approaches are needed to manage oxidative stress in the transition cows. Recently, the use of phytochemicals as feed additives in cows' nutrition has gained interest in managing various disease conditions. In the current study, we have evaluated the potential effects of phytochemicals derived from methanolic extract of Thymus serpyllum against oxidative stress and autoimmunity via inhibition of bovine nuclear factor kappa B (NF-κB). The free radical scavenging activity of Thymus serpyllum seed and leaf extracts was 71.8 and 75.6%, respectively at 100 µg/mL concentration. Similarly, both extracts displayed radicals reducing power and inhibition of lipid-peroxidation maximally at 100 µg/mL. A total of 52 bioactive compounds were identified when the plant extract was characterized by the GC-MS analysis, and five (Thymol, Luteolin 7-o-glucuronide, Rosmarinic acid, Apigenin 6,8-di-c-glucoside, Kaempferol) had binding free energy values of -11.6433, -10.002, -8.2615, -7.1714, -6.4870, respectively, in complexes with bovine NF-κB. Through computational analysis, the screened compounds showed good pharmacokinetic parameters, including non-toxicity, non-carcinogenic, high gastrointestinal absorption and thus can serve as potential drug candidates. MD simulation studies predicted the stability of complexes and the complex of Kaempferol was most stable based on RSMD value and MM/GBSA binding energy. The biochemical assays and computational studies indicated that Thymus serpyllum could be used as a promising feed additive in dairy cows to manage oxidative stress during the transition period.Communicated by Ramaswamy H. Sarma.

Metabolic alterations unravel the maternofetal immune responses with disease severity in pregnant women infected with SARS-CoV-2.

Pregnancy being an immune compromised state, coronavirus disease of 2019 (COVID-19) disease poses high risk of premature delivery and threat to fetus. Plasma metabolome regulates immune cellular responses, therefore we aimed to analyze the change in plasma secretome, metabolome, and immune cells with disease severity in COVID-19 positive pregnant females and their cord blood. COVID-19 reverse transcriptase-polymerase chain reaction positive pregnant females (n = 112) with asymptomatic (Asy) (n = 82), mild (n = 21), or moderate (n = 9) disease, healthy pregnant (n = 18), COVID-19 positive nonpregnant females (n = 7) were included. Eighty-two cord blood from COVID-19 positive and seven healthy cord blood were also analyzed. Mother's peripheral blood and cord blood were analyzed for untargeted metabolome profiling and cytokines by using high-resolution mass spectrometry and cytokine bead array. Immune scan was performed only in mothers' blood by flow cytometry. In Asy severe acute respiratory syndrome coronavirus 2 infection, the amino acid metabolic pathways such as glycine, serine, l-lactate, and threonine metabolism were upregulated with downregulation of riboflavin and tyrosine metabolism. However, with mild-to-moderate disease, the pyruvate and nicotinamide adenine dinucleotide (NAD+ ) metabolism were mostly altered. Cord blood mimicked the mother's metabolomic profiles by showing altered valine, leucine, isoleucine, glycine, serine, threonine in Asy and NAD+ , riboflavin metabolism in mild and moderate. Additionally, with disease severity tumor necrosis factor-α, interferon (IFN)-α, IFN-γ, interleukin (IL)-6 cytokine storm, IL-9 was raised in both mothers and neonates. Pyruvate, NAD metabolism and increase in IL-9 and IFN-γ had an impact on nonclassical monocytes, exhausted T and B cells. Our results demonstrated that immune-metabolic interplay in mother and fetus is influenced with increase in IL-9 and IFN-γ regulated pyruvate, lactate tricarboxylic acid, and riboflavin metabolism with context to disease severity.

One Health Assessment of Bacillus anthracis Incidence and Detection in Anthrax-Endemic Areas of Pakistan.

Anthrax, a severe zoonotic disease, is infrequently reported in anthrax-endemic regions of Pakistan. Despite clinical reports indicating its presence, particularly cutaneous anthrax, there is insufficient laboratory evidence regarding disease occurrence and environmental persistence. The present study aimed to confirm Bacillus anthracis presence, accountable for animal mortality and human infection, while exploring environmental transmission factors. Between March 2019 and July 2021, a total of 19 outbreaks were documented. Of these, 11 affected sheep/goats in Zhob district and 8 affected cattle/sheep in Bajour Agency. Clinical signs suggestive of Bacillus anthracis outbreak were observed in 11 animals. Blood and swab samples were collected for confirmation. The study followed a One Health approach, analyzing animal, environmental (soil/plant), and human samples. Of the 19 outbreaks, 11 were confirmed positive for anthrax based on growth characteristics, colony morphology, and PCR. Soil and plant root samples from the outbreak areas were collected and analyzed microscopically and molecularly. Cutaneous anthrax was observed in six humans, and swab samples were taken from the lesions. Human serum samples (n = 156) were tested for IgG antibodies against PA toxin and quantitative analysis of anthrax toxin receptor 1 (ANTXR1). Bacillus anthracis was detected in 65 out of 570 (11.40%) soil samples and 19 out of 190 (10%) plant root samples from the outbreak areas. Four out of six human samples from cutaneous anthrax lesions tested positive for Bacillus anthracis. Human anthrax seroprevalence was found to be 11% and 9% in two districts, with the highest rates among butchers and meat consumers. The highest ANTXR1 levels were observed in butchers, followed by meat consumers, farm employees, meat vendors, veterinarians, and farm owners. These findings highlight the persistence of anthrax in the region and emphasize the potential public health risks.

Identification of phytochemicals as putative ligands for the targeted modulation of peroxisome proliferator-activated receptor α (PPARα) in animals.

The PPAR family of transcription factors are ligand-activated and regulate diverse functions including metabolic, neurological, and inflammatory diseases, neurodegenerative disorders, fertility or reproduction in the body. Specifically, PPARα is known to play a role in reducing the levels of circulating triglycerides and regulating energy homeostasis in livestock animals. This study aimed to identify phytochemicals that could serve as ligands for modulation of the bovine nuclear peroxisome proliferator-activated receptor alpha (PPARα) using molecular docking studies. Therefore, we investigated 1000 flavonoids belonging to different groups for their ability to bind to PPARα using molecular docking. Out of 1000, 6 top lead compounds with maximum binding affinity, evaluated through molecular docking, were further analysed for physicochemical properties and drug-likeness attributes. The results revealed that two flavonoids, Quercetin-3-o-rhamnoside and (-)- epicatechingallate, which are known fatty acid synthase inhibitors, demonstrated high docking scores with PPARα (-8.66 kcal/mol and -8.49 kcal/mol, respectively) and low RMSD values with PPARα (1.61 kcal/mol and 1.28 kcal/mol, respectively) as compared to PPARα agonist (synthetic), fenofibrate (-6.24 kcal/mol and 2.19 kcal/mol) and thus analyzed further for prediction of stability of docked complexes through MD simulations. MD simulation studies predicted the stability of complexes and the complex of Quercetin-3-o-rhamnoside and (-)- epicatechingallate were found to be stable at 100 ns based on RSMD value and RMSF residue index. Through computational analysis, the screened compounds showed good pharmacokinetic parameters, including non-toxicity, non-carcinogenic, high gastrointestinal absorption and thus can serve as potential drug candidates. Finally, the findings suggest that these phytochemicals have the potential to act as potent PPARα pharmacological agonists to prevent disease mechanisms and their related complications, providing insights into the role of phytochemicals as feed additives in animals for modulating PPARα functions.Communicated by Ramaswamy H. Sarma.

In-vitro Augmentation of Mesenchymal Stem Cells by Using Adult Bovine Serum.

BACKGROUND: Umbilical cord mesenchymal stem cells (UC-MSCs) are increasingly being utilized for immune-related disease therapies due to their low immunogenicity. However, the primary culture of UC-MSCs requires the supplementation of serum in the growth medium, which has posed a challenge due to ethical issues related to the collection method of the fetal bovine serum (FBS) that is routinely used in cell culture. AIM: In order to address this, the purpose of this research was to assess the effectiveness of adult bovine serum (ABS) as a different and more affordable source of serum for the in-vitro cultivation of UC-MSCs. UC-MSCs were isolated from the umbilical cord of Wharton's jelly of cow immediately after birth, by digestion with Collagenase type I. METHOD: ABS was collected from fresh bovine sources and heat-inactivated. The morphology of UC-MSCs was observed under an inverted microscope, and growth patterns, proliferative index, and doubling time were calculated every two days to compare the efficacy of ABS with FBS. Immunocytochemistry for specific markers was also conducted on the MSCs. RESULT: The results showed a notable difference in morphology, growth rate, population doubling, and proliferative index between ABS and FBS. CONCLUSION: Intriguingly, ABS proved to be an effective supplement in the growth medium for expanding UC-MSCs in vitro, providing a viable alternative to FBS.

Antifungal Activity of Juglans-regia-Mediated Silver Nanoparticles (AgNPs) against Aspergillus-ochraceus-Induced Toxicity in In Vitro and In Vivo Settings.

Aflatoxins produced by some species of Aspergillus are considered secondary toxic fungal by-products in feeds and food. Over the past few decades, many experts have focused on preventing the production of aflatoxins by Aspergillus ochraceus and also reducing its toxicity. Applications of various nanomaterials in preventing the production of these toxic aflatoxins have received a lot of attention recently. The purpose of this study was to ascertain the protective impact of Juglans-regia-mediated silver nanoparticles (AgNPs) against Aspergillus-ochraceus-induced toxicity by exhibiting strong antifungal activity in in vitro (wheat seeds) and in vivo (Albino rats) settings. For the synthesis of AgNPs, the leaf extract of J. regia enriched with high phenolic (72.68 ± 2.13 mg GAE/g DW) and flavonoid (18.89 ± 0.31 mg QE/g DW) contents was used. Synthesized AgNPs were characterized by various techniques, including TEM, EDX, FT-IR, and XRD, which revealed that the particles were spherical in shape with no agglomeration and fine particle size in the range of 16-20 nm. In vitro antifungal activity of AgNPs was tested on wheat grains by inhibiting the production of toxic aflatoxins by A. ochraceus. According to the results obtained from High-Performance Liquid Chromatography (HPLC) and Thin-Layer Chromatography (TLC) analyses, there was a correlation between the concentration of AgNPs and a decrease in the production of aflatoxin G1, B1, and G2. For in vivo antifungal activity, Albino rats were administrated with different doses of AgNPs in five groups. The results indicated that the feed concentration of 50 µg/kg feed of AgNPs was more effective in improving the disturbed levels of different functional parameters of the liver (alanine transaminase (ALT): 54.0 ± 3.79 U/L and aspartate transaminase (AST): 206 ± 8.69 U/L) and kidney (creatinine 0.49 ± 0.020 U/L and BUN 35.7 ± 1.45 U/L), as well as the lipid profile (LDL 22.3 ± 1.45 U/L and HDL 26.3 ± 2.33 U/L). Furthermore, the histopathological analysis of various organs also revealed that the production of aflatoxins was successfully inhibited by AgNPs. It was concluded that the harmful effects of aflatoxins produced by A. ochraceus can be successfully neutralized by using J. regia-mediated AgNPs.

Spatial distribution and computational modeling for mapping of tuberculosis in Pakistan.

BACKGROUND: Tuberculosis (TB) like many other infectious diseases has a strong relationship with climatic parameters. METHODS: The present study has been carried out on the newly diagnosed sputum smear-positive pulmonary TB cases reported to National TB Control Program across Pakistan from 2007 to 2020. In this study, spatial and temporal distribution of the disease was observed through detailed district wise mapping and clustered regions were also identified. Potential risk factors associated with this disease depending upon population and climatic variables, i.e. temperature and precipitation were also identified. RESULTS: Nationwide, the incidence rate of TB was observed to be rising from 7.03% to 11.91% in the years 2007-2018, which then started to decline. However, a declining trend was observed after 2018-2020. The most populous provinces, Punjab and Sindh, have reported maximum number of cases and showed a temporal association as the climatic temperature of these two provinces is higher with comparison to other provinces. Machine learning algorithms Maxent, Support Vector Machine (SVM), Environmental Distance (ED) and Climate Space Model (CSM) predict high risk of the disease with14.02%, 24.75%, 34.81% and 43.89% area, respectively. CONCLUSION: SVM has a higher significant probability of prediction in the diseased area with a 1.86 partial receiver-operating characteristics (ROC) value as compared with other models.

Air Pollution and Cerebrovascular Disorders with Special Reference to Asia: An Overview.

Background: Among the primary environmental issues affecting global health, air pollution is considered the leading cause of concern. Globally, around 800,000 deaths were attributed to air pollution according to WHO. Evidence suggests that there has been a strong association of air pollution with stroke. Approximately, 25% of stroke mortality was due to air pollution according to a study in 2013. Objective: The aim of this review was to analyze the association between stroke, intracerebral hemorrhage, and subarachnoid hemorrhage and air pollution and its burden globally with a special focus on South Asia along with its association with the COVID-19 pandemic. Results: There is growing research data linking air pollution to cardiovascular disorders including stroke. Short-term and long-term air pollution exposures have been shown to increase stroke incidence in epidemiological data. Air pollution, both gaseous and particle, show a strong and tight temporal relationship with stroke hospitalizations and death. The link between ICH and SAH to air pollution is less strong and less well studied as compared to ischemic stroke. Stroke and air pollution both are highly prevalent in South Asia. It is possible that the high prevalence of stroke in south Asia may be linked to the high frequency of air pollution in addition to other conventional risk factors. Decreased stroke admissions and mortality and reduced cardiovascular mortality reported during coronavirus disease 2019 (COVID) lockdown may be attributable to decreased levels of air pollution. Conclusion: Even though air pollution poses a significant threat to human health, a great number of countries still fail to achieve internationally agreed air quality standards. Air pollution should be recognized among the most significant controllable risk factors for cardiovascular and cerebrovascular disease prevention and treatment.

Genomic analysis of arginine vasopressin gene in riverine buffalo reveals its potential association with silent estrus behavior.

BACKGROUND: Poor estrus expression behavior causes suboptimal reproductive efficiency through poor conception rate. Various signaling pathways are involved in estrus expression but arginine vasopressin (AVP) gene with oxytocin predominantly regulates estrus behavior. This study aimed to perform genomic characterization and evolutionary dynamics of AVP gene through association testing of the novel polymorphic loci and comparative genomic analysis to explore the potential effect of AVP gene on estrus behavior of Nili-Ravi buffaloes. METHODS AND RESULTS: 198 Nili-Ravi buffaloes were screened for the quest of novel polymorphism in the AVP gene. In exon-1, five polymorphic sites were detected including deletion of two (c.47delA and c.57delA) nucleotides that caused drastic variation in subsequent amino acid sequence due to frame shift including functional short peptide of nine residues. The 3-D structure revealed a loss of transmembrane loop between 16 and 31 residues in Nili-Ravi buffalo AVP protein sequence, suggesting that missing loop apparently reduced the gene functionality in Nili-Ravi buffalo by inhibiting cellular reactions and muting the animal estrus cyclicity. Three polymorphisms detected in AVP gene were significantly associated with silent estrus (P < 0.05). The comparative genomic analysis revealed that AVP gene is present on chromosome 14 having one conserved motif (Neurohypophysial) in buffalo. CONCLUSIONS: This study suggested the potential use of polymorphic sites as promising genetic markers for selection of buffaloes with pronounced estrus expression.

Cytotoxic and genotoxic evaluation of bisphenol S on onion root tips by Allium cepa and comet tests.

Bisphenol S (BPS) is an analog of bisphenol A, which is used as substitute of BPA in many products like airport luggage tags, baby bottles, plastics, and epoxy resins etc. Bisphenol S can cause toxic effects in different organisms, i.e., mice, rat, zebrafish, and C.elegans, etc. Bisphenol S is also known as "endocrine disruptor" due to its ability to mimic the endocrine receptors. So, the aim of this study was to evaluate the cytotoxic and genotoxic effects of bisphenol S on meristematic cells present in onion root tips through Allium cepa (A.cepa) and comet tests. Root growth inhibition was evaluated by root growth inhibition assay. Mitotic index (MI) and chromosomal aberrations (CAs) were assessed by A.cepa assay. DNA damage was evaluated by comet assay. Root growth of A.cepa was inhibited due to bisphenol S. LC50 value calculated by root growth inhibition assay for bisphenol S was (2.6±0.63, 50 µg/ml). Mitotic index was reduced, and chromosomal aberrations were observed, i.e., stickiness, polyploidy, and disturbed ana-telophase in anaphase and telophase stages of mitosis. In case of comet assay, DNA damage was increased in statistically significant manner (p ≤ 0.05). It was concluded that bisphenol S constitutes cytotoxic and genotoxic effects on A. cepa root meristematic cells. Moreover, it is suggested to explore more toxicity studies of bisphenol S at molecular level.

Nutrigenomic Interventions to Address Metabolic Stress and Related Disorders in Transition Cows.

For dairy cattle, the period involving a shift from late pregnancy to early lactation termed transition or periparturient is an excruciating phase. Health-related disorders are likely to happen in this time frame. Timely postpartum and metabolic adjustments to this new physical state demands correct management strategies to fulfill the cow's needs for a successful transition to this phase. Among the management strategies, one of the most researched methods for managing transition-related stress is nutritional supplementation. Dietary components directly or indirectly affect the expression of various genes that are believed to be involved in various stress-related responses during this phase. Nutrigenomics, an interdisciplinary approach that combines nutritional science with omics technologies, opens new avenues for studying the genome's complicated interactions with food. This revolutionary technique emphasizes the importance of food-gene interactions on various physiological and metabolic mechanisms. In animal sciences, nutrigenomics aims to promote the welfare of livestock animals and enhance their commercially important qualities through nutritional interventions. To this end, an increasing volume of research shows that nutritional supplementation can be effectively used to manage the metabolic stress dairy cows undergo during the transition period. These nutritional supplements, including polyunsaturated fatty acids, vitamins, dietary amino acids, and phytochemicals, have been shown to modulate energy homeostasis through different pathways, leading to addressing metabolic issues in transition cows.

Genomic and Computational Analysis of Novel SNPs in TNP1 Gene Promoter Region of Bos indicus Breeding Bulls.

Transition nuclear proteins (TNPs), the principal proteins identified in the condensing spermatids chromatin, have been found to play a key role in histone displacement and chromatin condensation during mammalian spermatogenesis. One such gene belonging to the TNP family called TNP1 gene is abundantly expressed in the regulation of spermatogenesis, and its sequence is remarkably well conserved among mammals. Genomic analysis, by sequencing and computational approach, was used to identify the novel polymorphisms and to evaluate the molecular regulation of TNP1 gene expression in Sahiwal cattle breeding bulls. DNA samples were sequenced to identify novel single nucleotide polymorphisms (SNPs) in the TNP1 gene. Modern computational tools were used to predict putative transcription factor binding in the TNP1 promoter and CpG islands in the TNP1 promoter region. In the TNP1 gene, four SNPs, three TATA boxes, and one CAAT box were identified. One CAAT box was discovered at 89 bp upstream of start site ATG. The computational analyses indicated that the polymorphisms inside the promoter sequence results in an added HNF-1 transcription factor binding site. In contrast, the other variations may remove the naturally occurring SRF transcription factor binding site. The CpG islands in the TNP1 promoter region were predicted to be absent by the MethPrimer program before and after SNP site mutations. These findings pave the way for more research into the TNP1 gene's promoter activity and the links between these SNPs and reproductive attributes in the Sahiwal breeding bulls.

Investigating the role of carbohydrate-binding module 34 in cyclomaltodextrinase from Geobacillus thermopakistaniensis: structural and functional analyses.

Carbohydrate-binding modules (CBMs) are noncatalytic regions found in several enzymes of glycoside hydrolase family 13 and are proposed to orient substrates to the catalytic site. In this study, a substantial information on the conserved aromatic residues in CBM34 regions of characterized bacterial cyclolmaltodextrinases (CDases) has been presented. Molecular modeling of CDase from Geobacillus thermopakistaniensis (CDase Gt ) revealed a change in the active site geometry due to CBM34 truncation. The binding energies of full-length (CDase Gt ) and CBM34 truncated (CDase Gt -ΔN) models showed opposite trends. The least preferred substrate molecule by the full-length model was the most preferred by the CBM34 truncated one. These exciting in silico findings were experimentally verified by recombinant production and characterization of the full-length and the CBM34 truncated proteins. Both the enzymes showed similar optimum pH and temperature. However, substrate specificity was in the reverse order. These experimental verifications matched the homology modeling and docking predictions. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03089-9.

Role of Peroxisome Proliferator-Activated Receptors (PPARs) in Energy Homeostasis of Dairy Animals: Exploiting Their Modulation through Nutrigenomic Interventions.

Peroxisome proliferator-activated receptors (PPARs) are the nuclear receptors that could mediate the nutrient-dependent transcriptional activation and regulate metabolic networks through energy homeostasis. However, these receptors cannot work properly under metabolic stress. PPARs and their subtypes can be modulated by nutrigenomic interventions, particularly under stress conditions to restore cellular homeostasis. Many nutrients such as polyunsaturated fatty acids, vitamins, dietary amino acids and phytochemicals have shown their ability for potential activation or inhibition of PPARs. Thus, through different mechanisms, all these nutrients can modulate PPARs and are ultimately helpful to prevent various metabolic disorders, particularly in transition dairy cows. This review aims to provide insights into the crucial role of PPARs in energy metabolism and their potential modulation through nutrigenomic interventions to improve energy homeostasis in dairy animals.

Comparison Of Operative Complications Of Direct Trocar Access Versus Veress Needle Insertion Technique For Initial Peritoneal Entry In Patients Undergoing Gynecological Laparoscopic Surgery.

BACKGROUND: Laparoscopy is considered to be a good diagnostic and surgical way to replace open surgeries in all surgical fields. As it is a least invasive method, with advantages for patients and all associated with medical care. There is a significant increase in the use of laparoscopic surgery in gynecology. This study was done to compare the frequency of operative complications of direct trocar access versus Veress needle insertion technique for initial peritoneal entry in patients undergoing gynecological laparoscopic surgery. METHODS: This Randomized Controlled Trial was conducted at the Department of Obstetrics & Gynecology, Lahore General Hospital, Lahore from 26-12-2019 to 25-6-2020. Four hundred consecutive patients were enrolled and divided randomly into two groups by blocked randomization. Direct trocar insertion (DTI) technique was used for initial peritoneal access in group A and Veress needle (VNI) was inserted first for peritoneal access in group B. RESULTS: The mean age of the patients in DTI group was 35.76±8.38 years whereas that in VNI group was 35.85±8.38 years. In DTI group there were 8(4%) patients with extra-peritoneal insufflation and in VNI group there were 19(9.5%) patients with extra-peritoneal insufflation. There were 8(4%) patients in DTI group with Omental injury and VNI group there were 13(6.5%) patients with Omental injury. In DTI group there were 7(3.5%) patients with port-site infection compared to 15(7.5%) in VNI group. CONCLUSIONS: According to findings the rate of complications observed with VNI, DTI can be a preferred procedure for gynecological surgeries.

Exploring the Potential of Interferon Gamma Gene as Major Immune Responder for Bovine Tuberculosis in River Buffalo.

Bovine tuberculosis (bTB) is a widespread zoonotic infection targeting the livestock sector, especially in developing countries, and posing a risk to humans and animal populations. Its recent prevalence in river buffaloes has been estimated as higher as 33.7%. In emergent countries like Pakistan, there is likeliness of human-livestock interfaces extensively and lacking of effective preventive measures that illustrate the risk of spreading the infection at a remarkable rate. The river buffalo (Bubalus bubalis) is an upkeep host of Mycobacterium bovis and is responsible for disease transmission among buffaloes and other livestock species. In this study, potential molecular biomarkers in the Interferon-gamma gene (IFNg) were identified after genomic screening of river buffaloes. Unique genomic loci in river buffalo proved the novelty of the genomic structure of this phenomenal animal but also highlighted its significance in natural immunity against the Mycobacterium. A total of eight single nucleotide polymorphisms were identified in the coding region of IFNg. The SNPs in the exonic region were all transitions, i.e., the conversion of purines to purines. These SNPs were analyzed for Hardy Weinberg Equilibrium, chi2 test, gene diversity, and protein structural conformation. Pathway analysis in tuberculosis revealed that IFNg inhibits the antigen-presenting cells (APC) through JAK and STAT pathways. Network analysis of IFNg proteins in both species showed strong associations among the immunity-related proteins (interleukins, tissue necrosis factors) and receptors of interferons. The identified polymorphic sites might be novel-potentiated markers for the selection of animals with superior immune response against bTB and can be exploited as promising genomic sites for breeding the resistant animal herds to combat Mycobacterium infection in a long run.

Genomic Identification, Evolution and Sequence Analysis of the Heat-Shock Protein Gene Family in Buffalo.

Heat-shock proteins (HSP) are conserved chaperones crucial for protein degradation, maturation, and refolding. These adenosine triphosphate dependent chaperones were classified based on their molecular mass that ranges between 10-100 kDA, including; HSP10, HSP40, HSP70, HSP90, HSPB1, HSPD, and HSPH1 family. HSPs are essential for cellular responses and imperative for protein homeostasis and survival under stress conditions. This study performed a computational analysis of the HSP protein family to better understand these proteins at the molecular level. Physiochemical properties, multiple sequence alignment, and phylogenetic analysis were performed for 64 HSP genes in the Bubalus bubalis genome. Four genes were identified as belonging to the HSP90 family, 10 to HSP70, 39 to HSP40, 8 to HSPB, one for each HSPD, HSPH1, and HSP10, respectively. The aliphatic index was higher for HSP90 and HSP70 as compared to the HSP40 family, indicating their greater thermostability. Grand Average of hydropathicity Index values indicated the hydrophilic nature of HSP90, HSP70, and HSP40. Multiple sequence alignment indicated the presence of highly conserved consensus sequences that are plausibly significant for the preservation of structural integrity of proteins. In addition, this study has expanded our current knowledge concerning the genetic diversity and phylogenetic relatedness of HSPs of buffalo with other mammalian species. The phylogenetic tree revealed that buffalo is more closely related to Capra hircus and distantly associated with Danio rerio. Our findings provide an understanding of HSPs in buffalo at the molecular level for the first time. This study highlights functionally important HSPs and indicates the need for further investigations to better understand the role and mechanism of HSPs.

Polymorphism association and expression analysis of alpha-lactalbumin (LALBA) gene during lactation in Nili Ravi buffalo.

Alpha-lactalbumin has been reported as a highly polymorphic gene that potentially alters the gene expression and is associated with milk composition in dairy breeds. Current study was conducted in two phases. In the first phase, polymorphisms identification in alpha-lactalbumin (LALBA) gene and its association with milk composition was performed. To identify the genetic polymorphism, Nili Ravi buffaloes at their second lactation were selected from Government livestock farm (Buffalo Research Institute, Pattoki). Genomic DNA was extracted from blood samples. After PCR amplification, products were sequenced, and data was analyzed. Results showed that the identified polymorphisms at chromosomal position 34310940 were found associated with major whey protein. In the second phase of study, milk samples were collected from five healthy mastitis-free Nili Ravi buffaloes in their second lactation for expression analysis of alpha-lactalbumin gene at their transition (day 15), mid (day 90), and late (day 250) lactation. Gene expression was observed highest in transition phase with a gradual decrease of expression in mid and late phase of lactation. Further studies are needed to explore the regulation of milk production genes and their translational efficiency during the course of lactation.

Molecular phylogenetics of Black Cobra (Naja naja) in Pakistan

Background: Snakes are found on every continent in the world except Antarctica, and on smaller land masses. Being ecologically important, they also cause a large number of bites, leading to millions of deaths. Mitochondrial and nuclear gene sequences are being used to identify, characterize, and infer genetic biodiversity among different snake species. Furthermore, phylogenetics helps in inferring the relationships and evolutionary histories among these species. Black cobra is one of the four most venomous snakes in Pakistan. Four mitochondrial (ND4, Cytochrome b, 12S rRNA, and 16S rRNA) and four nuclear (C-mos, RAG-1, BDNF, and NT3) genes were used to trace diversity and infer the phylogenetic relationship of black cobra in Pakistan. Results: Almost similar phylogenies were obtained through maximum likelihood and Bayesian inference, showing two species of cobra in Pakistan, namely, black cobra (Naja naja) and brown cobra (Naja oxiana). All Naja species were divided into three clades: black cobra (N. naja) and brown cobra (N. oxiana) cladding with different species of Naja; N. naja (Pakistan) cladding with N. naja from Nepal; and N. oxiana showed close relationship with Naja kaouthia from Thailand and Naja siamensis from Thailand. Conclusion: It was confirmed genetically that there are two cobra species in Pakistan, i.e., black and brown cobras. This study will help in not only genetic conservation but also developing anti-venom against snake species.