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Background Citrullinated proteins formed by peptidyl arginine deiminases (PADIs) deimination of arginine residues in proteins are of particular interest in arthritis pathogenesis. Polymorphisms on the PADI-4 gene lead to the malfunctioning of PADIs leading to the onset of arthritis. Objective The present study was conducted to determine the polymorphisms on the PADI-4 gene and their association with rheumatoid arthritis (RA) as well as Osteoarthritis (OA). Methodology To achieve the above-mentioned objective a case-control study was conducted. Blood samples were collected from RA, OA, and control subjects. DNA was extracted from each blood sample by modified organic method and was quantified as well as qualified by DNA gel electrophoresis and Nanodrop. Patients were tested for rs874881, rs11203366, rs11203367, rs2240336, rs2240337, rs2240339, rs1748033 and rs2240340 polymorphic sites by amplifying targeted regions through PCR with site-specific primers. Genotyping was performed by Restriction Fragment Length Polymorphism and direct sequencing method. Mutations were identified by analyzing sequences on BioEdit software. Allelic, genetic, and multiple site analysis were performed by SHEsis and PLINK software. Change in the amino acid sequence was identified by MEGA 6.0 software. Results Polymorphisms were identified on all targeted polymorphic sites except rs2240337 in both RA and OA individuals. In addition, two novel mutations were also identified in exon 4 identified i-e SCV000804840: c.218T > C and SCV000807675: c.241G > T. All the SNPs except rs11203366 were found to be significantly associated with RA at an allelic level whereas all SNP’s have been significant risk factors in the onset of OA. At genotypic level rs874881, rs11203366, rs2240339, SCV000804840 and SCV000807675 were significantly associated to RA development whereas rs874881, rs11203366, rs11203367, rs2240339, SCV000804840 and SCV000807675 were genetic risk factors in OA onset. Haplotype analysis indicated that GACCACGCC and GACCACGCT were highly significant in disease development. Polymorphisms identified altered the functioning of PADIs by altering their amino acid sequence. Conclusion In conclusion, it was found that PADI-4 gene polymorphism was not only involved in the onset of RA but was also found to be a significant risk factor in OA onset.

Introduction: In Pakistan, for the patients of Hepatitis C virus (HCV), Direct-Acting Antiviral (DAA) therapy for 12 weeks and 24 weeks had been reported to be highly efficacious for genotype 3. We currently carried out an observational study to predict the rate of efficacy of sofosbuvir and ribavirin in hepatitis C patients to establish concrete or authentic data on this combination of DAA for long-term treatment. Materials and Method: Among 2000 subjects who attended tertiary care unit in Lahore-Pakistan from November 2018 to February 2019, 1990 satisfied the criteria set for the present investigation i.e. SVR12 after being treated with sofosbuvir and ribavirin combination. Results: It was noted that genotype 3a were more common among all the subjects under observation with 50.65 % (1008/1990) in females and 49.35% (982/1990) in males. Overall efficacy analysis was found to be 95.4% (1900/1990) in patients while the moderate response was noted in elderly subjects including both genders (61-90 years). DAA responders (male: female percentages) shown the following stats; 66.63 (42/66):36.36 (24/66) in 11-20 years, 56.6(240/424):43.39 (184/424) in 21-30 years and 44.73(272/608):55.27(336/608) in 31-40 years. Conclusion: Collectively, this combinational drug therapy was observed to be successful among the Pakistani population. However, more comprehensive follow-up studies are needed on a larger pool of population nationwide to check only this combinational therapy (sofosbuvir and ribavirin) would be beneficial or not? Or next-generation DAA regimes would be the choice for the Pakistani population.

Objective: To assess the clinical profile and outcome of patients admitted with the diagnosis of tuberculous meningitis (TBM) at neurology unit of a tertiary care teaching hospital of Pakistan. Study Design: Cross-sectional study. Place and Duration of Study: Neurology department, Pak Emirates Military Hospital Rawalpindi, from Dec 2018 to Jun 2019. Methodology: This study was conducted on 69 patients of tuberculous meningitis admitted. Demographic profile included age, gender, and the symptoms with which the patient presented. Neurological complications were also documented among the target population. Outcomes included recovery, shifting to intensive care unit and death. Results: A total of 69 patients admitted in neurology ward diagnosed and managed as tuberculous meningitis in the study duration time. Male to female ratio was 1.4:1. Mean age of patients diagnosed with tuberculous meningitis in our study was 34.23 ± 3.915 years. Most of the patients presented with the fever followed by headache. Out of 69 patients, 41 (59.4%) recovered, 22 (31.9%) were shifted to the intensive care unit and 6 (8.7%) died. Seizures were the commonest neurological complication faced by the patients followed by cranial nerve palsies. Conclusion: Fever and headache were common symptoms with which patients of tuberculous meningitis presented in our study population. Most of the patients recovered in the ward but a high percentage required intensive care unit admission as well. Seizures and cranial nerve palsies were commonly experienced complication by patients included in our study.

The trend of consuming food high in calories, fat, and sugar with little nutritional value and reduced physical exercise has resulted in an alarming ratio of overweight and obese subjects worldwide. Low-grade chronic inflammation is the key feature of obesity that causes an increase in pro-inflammatory cytokines and decrease in anti-inflammatory cytokines in circulation. The current study was aimed to investigate the effect of high-fat diet on the architecture of spleen, liver, and skeletal muscle and changes in the expression of hepatic cytokines. Two groups of experimental rats were established, against control that were given different percentage of fats in their diet. After a period of sixteen weeks, rats were dissected and their organs were excised out and processed accordingly. Spleen sections of experimental groups, revealed increased recruitment of lymphocytes, sinusoidal dilatations, necrotic lymphocytes, increased ratio of white-to-red pulp, and hemosiderin and iron deposits in red pulp indicating immune system activation. Hepatic sections showed enlarged sinusoidal spaces, disruptive hepatocytes, necrosis and dilation of portal veins. Sections of skeletal muscle showed degenerating fibers, increased fat accumulation, and recruitment of macrophages. Elevated expression of IFN-γ and decreased expression of IFN-α and IFN-β cytokines verified the adverse effect of high-fat diet on immune system as well. Fats tend to accumulate in organs due to increased intake of fat-rich diet disturbing their normal function and histology. In addition, gene expression analysis of cytokines confirmed the effect of high-fat diet as an inflammatory agent.

Objective: To determine the diagnostic accuracy of USG in detecting hepatocellular carcinoma. Methodology: A total of 70 patients with liver cirrhosis with mass and age 18-60 years of either gender were included. Nodular lesion

In this study, Zn(II) centered complexes with aldimine derivatives were synthesized using green solvent, polyethylene glycol (PEG-400) and amorphous complexes were characterized by FT-IR, multinuclear (1H and 13C NMR), elemental and thermal analysis. Thermogravimetric analysis indicated the extended thermal stability of the synthesized complexes. All the Zn(II) complexes show very significant photoexcitation in the range of 318 – 384 nm and photoemission in the range of 502 – 562 nm. Among all the complexes, Zn(II) complex (3Zn) showed minimum band gap value, 2.35 eV. These amorphous complexes have been reported for their wide applications in biomedical sciences. The synthesized aldimine ligands and Zn(II) complexes were investigated for anti-inflammatory activity and these complexes showed more anti-inflammatory potential than the corresponding aldimine ligands. The solubilization of zinc complexes in sodium dodecyl sulphate was also investigated to reveal the interaction of metal complexes by using UV-Visible spectroscopy and electrical conductivity measurements.

SIRT6, a nuclear protein, has been implicated in a number of essential cellular processes, such as the DNA damage response, metabolic homeostasis, inflammation, tumorigenesis and aging. However, the role of Sirt6 in the regulation of spermatogenesis is yet unknown. In the present study, we successfully generated Sirt6-/- mice on a C57BL6/ICR mixed background and found that some Sirt6-/- mice survived beyond eight weeks. We further revealed that spermatogenesis in Sirt6-/- mice was arrested at the elongated spermatid stage. Sirt6-/- male mice were completely infertile and had an increased number of apoptotic spermatids. To our surprise, deacetylation activities of SIRT6 on H3K9ac, H3K18ac and H3K56c were not required for spermatogenesis. Therefore, our findings establish a novel link between Sirt6 and male fertility, suggesting an essential role of Sirt6 in spermatogenesis.

Objective: To highlight the prevalence and epidemiology of New Delhi metallo-ß-lactamase-1.producers in pus samples. Methods: The cross-sectional study was conducted from April to August 2018at the Biotechnology Laboratory, Balochistan University of Information Technology Engineering and Management Sciences, Quetta, Hi-tech Laboratory, Centre for Advance Studies in Vaccinology and Biotechnology, University of Balochistan, Quetta, and Microbiology Laboratory, Bolan Medical Complex Hospital, Quetta, Pakistan. Biochemical and molecular approaches were used for the identification of the isolates and Modified Hodge Test for the phenotypic detection of class-A carbapenemase activity. Minimum Inhibitory Concentration was performed using E-test and broth microdilution method. Molecular basis of carbapenemase activity was ascertained by the recognition of blaNDM-1 gene in the isolates. Results: Of the 300 pus samples taken from surgical/burn units,6(2%) blaNDM-1 harbouring isolates were found; 3(50%) each being Klebsiellapneumoniae andPseudomonas aeruginosa. Klebsiella. pneumoniaeisolates were extensivelydrug-resistant. The Pseudomonas aeruginosaisolates displayed resistance against 21 antibiotics of tetracyclines, quinolones, ?-lactams, aminoglycosides, monobactams, sulphonamides, macrolides, cephalosporins, phosphonic acid and polypeptide groups, suggesting pan-drug resistance. Conclusion: The resistance pattern of the bacterial isolates poses a significant clinical threat in the region. Key Words: blaNDM-1, Carbapenemase, XDR, PDR, Quetta.

Severe acute pancreatitis (SAP) has a complex course and a worse prognosis. Immune response imbalance is an important cause of severe pancreatitis or even death in patients. Immunomodulation therapy can regulate the imbalance of inflammatory response, alleviate SAP-related organ damage and improve the prognosis of patients. There are some problems in early immune regulation measures, such as single target and simple way. In recent years, new treatment methods, such as regulating the maturation and apoptosis of immune cells, the application of mesenchymal stem cells (MSCs) and multifactor combination therapy, have provided new ideas and hope for the future treatment of SAP. This article reviews the development of SAP immunoregulation and its recent progress.

Objectives: This study aims at measuring compliance to iron supplements because iron deficiency anemia (IDA) is worldwide problem among pregnant women, so Iron supplements are advised usually in once daily regimen but compliance to iron supplements therapy is a major problem due to possible side effects and long-term duration of therapy. Study Design: Retrospective questionnaire-based study. Setting: Mola Bakhsh Hospitals, Sargodha, Pakistan, Period: From February to December, 2018. Material & Methods: This study was design to check compliance to iron supplementation during pregnancy and various factors related to directly and indirectly noncompliance to iron supplementation. A total of 267 pregnant females were recruited in study. Data was collected through a questionnaire translated in local language. On this basis sample population was divided into three groups compliant, partially compliant and non-compliant major factors associated with the prevalence of anemia and non-compliance to iron supplements were education, side effects, financial issues, forgetfulness, personal beliefs, disliking of pharmaceutical preparation etc. Results: After thorough analysis it was found that iron deficiency anemia is prevalent in about 64% of females in the third trimester. Improvement in Hb level in third trimester compared to 1st trimester was taken as parameter for measurement of compliance. Conclusion: side effects due these iron supplements and certain human habits act as hurdle to fight anemia during pregnancy and to eradicate these hurdles certain steps like education of women about anemia, its causes, health implication and imparting nutritional education with special emphasis to improve dietary intake of proteins and iron.

Severe acute pancreatitis (SAP) is an inflammatory disorder that progresses with local and systemic difficulties accompanied by a relatively high mortality rate. In recent years, maresin 1 (MaR1) has been shown to be a macrophage mediator with effective proresolving and anti-inflammatory properties that prevents the occurrence of various inflammatory conditions. The purpose of this study was to investigate the role of MaR1 in SAP and related lung injury. Experimental SAP was induced in mice with a combination of cerulean and lipopolysaccharide. MaR1 was administered 30 min before the primary injection of cerulean. Biochemical markers and histological injury scores were used to evaluate the severity of acute pancreatitis. To determine the degree of inflammation, serum cytokines and myeloperoxidase activity in pancreas and lung tissues were measured. Western blot analysis detected the activation of NF-κB. After MaR1 pretreatment, the activities of amylase, lipase, TNF-α, IL-1β, and IL-6 were decreased in serum, and the myeloperoxidase activity both in pancreas and in lung tissues significantly decreased, whereas the activity of anti-inflammatory cytokine IL-10 in serum was increased. MaR1-pretreated mice reduced the activation of pancreatic NF-κB and decreased the severity of pancreatic and lung-related injuries. These results confirm that MaR1 alleviated inflammation of the pancreas and lung by inhibiting the activity of NF-κB in experimentally induced acute pancreatitis and exerted anti-inflammatory effects. These findings suggest that MaR1 could be a new and useful drug in the treatment of SAP. NEW & NOTEWORTHY These results provided us evidence to confirm that maresin 1 (MaR1) can alleviate inflammation of the pancreas and lung by inhibiting the activity of NF-κB in experimental induced acute pancreatitis and exerts certain anti-inflammatory effects. These findings suggest that MaR1 could be a new and useful drug in the treatment of severe acute pancreatitis.

Tributyltin(IV) derivative (1) was synthesized by reacting the ligand (A(1)) {2-[(2-methylpropanoyl)amino]propanoic acid} with tributyltin(IV)chloride in presence of potassium hydroxide. The compou ...

Objectives: To determine the mode of penetrating injuries to abdomen and to determine the effect of these injuries on outcome and to formulate recommendations for management of patients sustaining penetrating abdominal trauma. Study Design: Descriptive, cross sectional. Setting: Surgical Department Sandeman Provincial hospital, Quetta. Period: 1 year 2012-2013. Methodology: 147 consecutive cases of abdominal trauma presenting to emergency were studied for pattern of injury and management outcome. Data was recorded and analyzed using SPSS v10. Frequency tables were generated for various variables. Results: The commonest mode of injury was stabbing occurring in 76 cases (51.7%) followed by gunshot injuries in 60 cases (40.8%), 11 patients (7.4%) sustained blast pellet injuries. Patients were either managed conservatively or underwent laparotomy depending on mechanism of injury and clinical presentation. Stab and blast pallet wounds which were superficial were managed by local wound exploration those with peritoneal breach, hemodynamic instability and visceral evisceration were managed by Laprotomy. All gunshot injuries underwent mandatory Laprotomy. Gut was most commonly injured viscus followed by liver and kidney. Types of procedure performed were primary repair, bowel resection, protective ileostomy, splenectomy, nephrectomy, 2 patients were managed by damage control i.e. liver packing. Overall mortality was 5%. Conclusion: Mandatory laprotomy for all gunshots, and stabs and pellets that penetrate the peritoneal cavity proves to be safe and a prudent policy.

Sexual reproduction is the most common form of reproduction among eukaryotes, which is characterized by a series of massive cellular or tissue renovations. Recent studies have revealed novel functions of autophagy during sexual reproductive processes, ranging from yeast to mammals. In mammals, autophagy is indispensable for spermatogenesis and oogenesis, and it participates in early embryonic development and maternal-fetus crosstalk to ensure the development of embryos or fetuses. Thus, autophagy provides the molecular basis for resource allocation among parents and their offspring, providing an important way to benefit the next generation. Abbreviations: ATG: autophagy-related; Becn1: beclin 1, autophagy related; CMA: chaperone-mediated autophagy; epg: ectopic PGL granules; ES: ectoplasmic specialization; EVTs: extravillous trophoblasts; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; PCD: programmed cell death; PTB: preterm birth; STB: syncytiotrophoblast

The present study investigated the (i) socio-demographic predictors of psychological distress, (ii) socio-demographic predictors of satisfaction from online classes, and (iii) the relationship between psychological distress and satisfaction from online classes among university students of Pakistan during the COVID-19 pandemic. An online questionnaire-based survey was conducted. A total of 2220 respondents that was enrolled at the University of the Punjab (PU), University of Management and Technology (UMT), and the University of Central Punjab (UCP) were involved in the current study. Data were collected at a 64% response rate and analyzed with SPSS IBM Version 21.0. Results revealed that approximately 41% of the students were facing severe psychological distress while about 65% were found unsatisfied with online classes. Besides, a linear negative relationship between the independent variable, i.e. psychological distress and the dependent variable, i.e. satisfaction from online classes was found. Therefore, to minimize the level of psychological distress and increase students’ satisfaction with online classes it is highly recommended to take precautionary measures by the relevant stakeholders., COVID-19, Pandemic, Panic, Stress management, Psychological impact, Students.

Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) gene plays a vital role in the activation of T-cells as a down regulator. CTLA-4 gene polymorphisms have implicated a potential risk factor for autoimmune disorders like arthritis. Therefore the current study was designed to determine the association of CTLA-4 gene polymorphism in the onset of rheumatoid and osteoarthritis in Pakistani individuals. Genotyping was performed on 300 RA, 316 OA, and 412 control subjects by direct sequencing method as well as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. It was observed that allelic and genotypic frequency of rs5742909, rs231775, rs4553808, rs733618, and rs3087243 were significantly varied among patients and controls and considered as a significant risk factor in the onset of RA as well as OA. However, no mutation was identified on the rs11571317 polymorphic site. Haplotype CAGTCA and CAG TCG act as a protectant against disease onset whereas CAACCG was significant in disease onset. Mutation on rs231775 polymorphic site lead to the change of threonine into alanine It was concluded that CTLA-4 gene polymorphism is a significant risk factor in the onset of RA as well as OA. Large scale survey is required for the screening of the genetic markers for pre-diagnosis of the disease.SUMMARY STATEMENTThe study summarized that CTLA-4 gene polymorphism plays a key role in the arthritis onset in Pakistani population.

Background Glioblastoma is the most common primary brain tumor and remains uniformly fatal, highlighting the dire need for developing effective therapeutics. Significant intra- and inter-tumor heterogeneity and inadequate delivery of therapeutics across blood–brain barrier continue to be significant impediments towards developing therapies which can significantly enhance survival. We hypothesize that microRNAs have the potential to serve as effective therapeutics for glioblastoma as they modulate the activity of multiple signaling pathways, and hence can counteract heterogeneity if successfully delivered. Methods Using a computational approach, we identified microRNA-34a as a microRNA that maximally reduces the activation status of the three core signaling networks (the receptor tyrosine kinase, p53 and Rb networks) that have been found to be deregulated in most glioblastoma tumors. Glioblastoma cultures were transfected with microRNA-34a or control microRNA to assess biological function and therapeutic potential in vitro. Nanocells were derived from genetically modified bacteria and loaded with microRNA-34a for intravenous administration to orthotopic patient-derived glioblastoma xenografts in mice. Results Overexpression of microRNA-34a strongly reduced the activation status of the three core signaling networks. microRNA-34a transfection also inhibited the survival of multiple established glioblastoma cell lines, as well as primary patient-derived xenograft cultures representing the proneural, mesenchymal and classical subtypes. Transfection of microRNA-34a enhanced temozolomide (TMZ) response in in vitro cultures of glioblastoma cells with primary TMZ sensitivity, primary TMZ resistance and acquired TMZ resistance. Mechanistically, microRNA-34a downregulated multiple therapeutic resistance genes which are associated with worse survival in glioblastoma patients and are enriched in specific tumor spatial compartments. Importantly, intravenous administration of nanocells carrying miR-34a and targeted to epidermal growth factor receptor (EGFR) strongly enhanced TMZ sensitivity in an orthotopic patient-derived xenograft mouse model of glioblastoma. Conclusions Targeted bacterially-derived nanocells are an effective vehicle for the delivery of microRNA-34a to glioblastoma tumors. microRNA-34a inhibits survival and strongly sensitizes a wide range of glioblastoma cell cultures to TMZ, suggesting that combination therapy of TMZ with microRNA-34a loaded nanocells may serve as a novel therapeutic approach for the treatment of glioblastoma tumors.

Background: Extremophiles are a hot topic in the field of biotechnology for their immense potential and applications in multiple industries. Objectives: The present review aims to sum up the potential applications of alkalophilic protease-producing bacteria and their optimized growth requirement. The isolation, characterization, and optimization of various isolates (especially of genus Bacillus) from different harsh niches, including soil samples from deserts and soil having decaying matters, wastewaters from industries, soda lakes, and alkaline springs have been reported in this review. Methodology: All the relevant papers published from 2013-2020 were looked over numerous sources like Google Scholar, Medline, PubMed, Research Gate, Science Direct, Scopus and Web of Science. Results: Most of the microbial life found in extreme alkaline habitats are found to form a variety of enzymes and an array of other substances of biotechnological interests. These enzymes, especially proteases, are exploited in industries globally because of their ability to withstand rigorous industrial reactions and conditions. Conclusion: Though a number of alkalophilic protease-producing bacteria have been isolated, still a large number of these micro-organisms are unidentified. The current demand for biotechnological products from them appeals to the need for isolation of unidentified bacteria.

Vaccines develop the immune response against disease and are the intricate biological entities containing one or more than one antigen beside an adjuvant In order to ensure that manufactured vaccines are safe potent and of consistent quality four primary controls of bio safety namely PPE facility design SOPs and leadership are required at every step of manufacture and quality control testing until the administration of vaccine to the target population The safety and efficacy of testing methods are based upon the data validation of several years With time the advancements in vaccine production techniques has brought about advancements in testing techniques as well These improvements have enabled scientists to detect highly sensitive potential hazards which were a dream few years back An increasing assemblage of physico chemical methods permits much better characterization of the product In addition to advance tests vaccine regulations necessitate quite a few numbers of strategies to ensure the safety and potency of the vaccine serum These measures include cell banking seed lot system supplier audits master seed compliance with good manufacturing practices independent lot release from National Regulatory Authority and increased pre and post marketing surveillance to look for possible adverse effects following immunization AEFI Together all these methods ensure the quality of vaccine for the purpose it is manufactured against

The process of paper production requires a huge quantity of water and energy and in turn contributes a number of effluents in the form of phenolics, toxic organic compounds and heavy metals in wastewater (leachate). The present investigation was aimed to assess the toxic effects of leachate on liver micro-architecture and heavy metal elements of the liver. Eighteen (18) healthy male Wistar rats (240 ± 10g) were selected and acclimatized prior to experimental treatment. These rats were randomly divided into three groups viz, Control group (received 4ml/ kg normal saline), Group 1 (4ml/ kg leachate) and Group 2 (4ml/ kg 1:10 diluted leachate). All the animals were dissected and liver tissues were collected and processed accordingly after 24 h of leachate treatment. High level of cadmium and chromium were found in Group 1 as compared to the control group upon liver metal contents analysis found out by flame atomic absorption spectrometer. A clear disruption of micro-architecture of the liver, congested sinusoids, damaged central vein, and perturbed morphology was observed in Group-1 as revealed by H & E staining. Moreover, loss of polarity, congestion, and disruption of hepatocytes and pronounced vacuolization in the cytoplasm was observed in Group 2 compared to control sections. On the basis of above findings, it can be concluded that paper industry leachate is highly toxic and its intraperitoneal injection results in hepatotoxicity that not only affects the hepatic micro-architecture but also results in perturbed liver metal contents. Therefore, proper treatment of such wastewater is required before its disposal.

This study investigates the effect of barley (Hordeum vulgare), black plum (Syzygium cumini) and Chinese tree leaves (Pistacia chinesis) aqueous extracts on serum glucose, cholesterol, urea, uric acid level and body weight of normal and alloxan-induced diabetic male rabbits. Rabbits were kept in fasting condition for 12 h, then diabetes was induced using alloxan monohydrate (150 mg/Kg) . From the 7th day of diabetes induction, blood serum was collected for glucose analysis. Rabbits having fasting glucose level ≥ 250 mg/dL were considered as diabetic. Rabbits were treated with 1% barley, black plum and Chinese tree aqueous extract to check their effects on the serum glucose, serum cholesterol, serum urea, uric acid and body weight of the animals. Samples were collected after every 72 h for biochemical analysis. It was noted that 1% barley, black plum and Chinese tree aqueous extract reduced 43.92, 39.05 and 32.47% glucose level. Results suggest that oral administration of aqueous extract of barley, black plum and Chinese tree reduced glucose, cholesterol, urea, and uric acid level. Therefore, these medicinal plants should be supplemented as herbal drugs in the treatment of various complex diseases and specially to control diabetes. Key words: Chinese tree leaves, antidiabetic agents, barley, black plum, rabbits, diabetes mellitus.

Assessment of medical images and diagnostic decision making of lumbar associated diseases by clinicians is invariably subjective, time consuming and challenging task. Presently, clinicians make use of either manual or semi-automated computer-aided tools to make relevant measurements for adding vote of confidence to their grading and evaluation. Lacking reliability and offering substantive dissimilarity once performed by different clinicians, these methods complicate the evaluation process. In an effort to support the decision making process of clinicians, in this paper we present a lumbar assessment framework with autonomous extraction of spinal measurements. Furthermore, an effort is made to address the challenges faced by clinicians while assessing disorders including spondylolisthesis and assessment of lumbar lordosis (LL) by proposing novel disease classification methodologies. For spondylolisthesis classification, we achieved an accuracy of 89% by using angular deviation metric whereas, 93% accuracy for determining adequacy/inadequacy in LL assessment through computation of area within enclosed lumbar curve region. Our framework involves semantic segmentation of vertebral bodies (VBs) using ResNet-UNet where we achieved DSC of 0.97 and IoU of 0.86. Subsequently, we achieved a statistically significant correlation coefficient R and encouraging mean absolute error (MAE) with clinicians’ grading for measurements involving lumbar lordotic angle (LLA), lumbosacral angle (LSA), VB dimensions and lumbar height. In addition to this, we have publicly released the dataset with all the clinicians markings at https://data.mendeley.com/datasets/k3b363f3vz/2 .

Cisplatin is a DNA-damaging chemotherapeutic agent used for treating cancer. Based on cDNA dataset analysis, we investigated how cisplatin modified gene expression and observed cisplatin-induced dysregulation and system-level variations relating to insulin resistance and type 2 diabetes mellitus (T2DM). T2DM is a multifactorial disease affecting 462 million people in the world, and drug-induced T2DM is a serious issue. To understand this etiology, we designed an integrative, system-level study to identify associations between cisplatin-induced differentially expressed genes (DEGs) and T2DM. From a list of differential expressed genes, cisplatin downregulated the cyclin-dependent kinase inhibitor 1 (CDKN1A), tumor necrosis factor (FAS), and sestrin-1 (SESN1) genes responsible for modifying signaling pathways, including the p53, JAK-STAT, FOXO, MAPK, mTOR, P13-AKT, Toll-like receptor (TLR), adipocytokine, and insulin signaling pathways. These enriched pathways were expressively associated with the disease. We observed significant gene signatures, including SMAD3, IRS, PDK1, PRKAA1, AKT, SOS, RAS, GRB2, MEK1/2, and ERK, interacting with source genes. This study revealed the value of system genetics for identifying the cisplatin-induced genetic variants responsible for the progression of T2DM. Also, by cross-validating gene expression data for T2DM islets, we found that downregulating IRS and PRK families is critical in insulin and T2DM signaling pathways. Cisplatin, by inhibiting CDKN1A, FAS, and SESN1, promotes IRS and PRK activity in a similar way to rosiglitazone (a popular drug used for T2DM treatment). Our integrative, network-based approach can help in understanding the drug-induced pathophysiological mechanisms of diabetes.

Therapeutic resistance stemming from inter and intra-tumoral heterogeneity is a significant impediment towards development of effective therapeutics for glioblastoma. We hypothesized that microRNAs can potentially counteract resistance emanating from such heterogeneity as they simultaneously modulate the expression of multiple proteins. We identified microRNA-34a as a unique microRNA which modulates multiple oncoproteins in GBM using two different in silico approaches. We investigated the therapeutic effects of microRNA-34a in three primary patient-derived xenografts (PDX) representing classical (GBM6), proneural (GBM118) and mesenchymal (GBM118) subtypes; four established cell lines (T98G, U251, A172, LN229) and two cell lines with acquired resistance to temozolomide (A172-TR, LN229-TR) in vitro. Glioblastoma cell cultures showed variable responses to temozolomide but microRNA-34a inhibited proliferation in all cell cultures. Furthermore, microRNA-34a also sensitized all tested cell lines to temozolomide (combination index < 0.8, p=.03) and radiation treatment (dose enhancement factor 1.7–2.2, p=0.02). Mechanistically, microRNA 34a down-regulates at least six distinct therapeutic resistance proteins. Importantly, these resistance proteins are expressed in distinct spatial niches and are prognostic for patient survival based on our analysis of the cancer genome atlas (TCGA) data. For in vivo delivery of microRNA-34a, we utilized nanocells which are derived from genetically modified bacteria, loaded with microRNA-34a and tagged with a bispecific antibody targeting EGFR. Nanocells were injected intravenously while temozolomide was administered by oral gavage in an orthotopic PDX model. We confirmed delivery of microRNA-34a to tumor by observing down-regulation of cMet and phosphorylated Akt in treated mice. Importantly, microRNA-34a nanocells resulted in significant reduction in tumor growth (p=0.021), increased survival (p

During sexual reproduction, two haploid gametes fuse to form the zygote, and the acrosome is essential to this fusion process (fertilization) in animals. The acrosome is a special kind of organelle with a cap-like structure that covers the anterior portion of the head of the spermatozoon. The acrosome is derived from the Golgi apparatus and contains digestive enzymes. With the progress of our understanding of acrosome biogenesis, a number of models have been proposed to address the origin of the acrosome. The acrosome has been regarded as a lysosome-related organelle, and it has been proposed to have originated from the lysosome or the autolysosome. Our review will provide a brief historical overview and highlight recent findings on acrosome biogenesis in mammals.

Objective: Extensive drug resistant Gram-negative bacilli, harboring New Delhi metallo-β-lactamase-1 (blaNDM-1) having the ability to hydrolyze β-lactams, have become a vital global clinical threat. The present study was, therefore, designed to investigate the prevalence and epidemiology of NDM-1 producers in Quetta, Pakistan. Methods: This study was carried out in Microbiology Laboratory, Bolan Medical Complex Hospital Quetta, Biotechnology laboratory, BUITEMS Quetta and Hi-tech laboratory, CASVAB, University of Balochistan, Quetta, from March to June 2018, during the hot season. Biochemical and molecular approaches were applied for the identification of bacterial isolates. Minimum Inhibitory Concentrations (MICs) were determined using E-test method. Carbapenemase activity was ascertained by Modified Hodge Test (MHT) and the presence of blaNDM-1 gene was recognized by Polymerase Chain Reaction (PCR). Results: We isolated five blaNDM-1 harboring isolates of three different species namely Morganella morganii (n=2) Enterobacter cloacae (n=2) and Citrobacter freundii (n=1), from 300 pus samples. These isolates were found extensive drug resistant (XDR). Strikingly, two isolates of M. morganii were displaying resistance against 23 antibiotics of sulphonamides, aminoglycosides, polypeptide, monobactams, tetracyclines, quinolones, macrolides, cephalosporins, phosphonic acid and β-lactams groups, suggesting Pan Drug Resistance (PDR). Conclusion: This is the first report on emergence of PDR strain of M. morganii producing NDM1 in the province of Balochistan, Pakistan. The presence of blaNDM-1 in different bacterial species and their extensive rather pan drug resistance pattern poses a momentous clinical threat. doi: https://doi.org/10.12669/pjms.35.4.372 How to cite this:Din M, Babar KM, Ahmed S, Aleem A, Shah D, Ghilzai D, et al. Prevalence of extensive drug resistance in bacterial isolates harboring blaNDM-1 in Quetta Pakistan. Pak J Med Sci. 2019;35(4):1155-1160. doi: https://doi.org/10.12669/pjms.35.4.372 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Acute pancreatitis (AP) is an acute gastrointestinal disorder that is the most common and requiring emergency hospitalization. Its incidence is increasing worldwide, thus increasing the burden of medical services. Approximately 20% of the patients develop moderate to severe necrotizing pancreatitis associated with pancreatic or peri‐pancreatic tissue necrosis and multiple organ failure. There are many reports about the anti‐inflammatory effect of mesenchymal stem cells (MSCs) on pancreatitis and the repair of tissue damage. MSCs cells come from a wide range of sources, autologous MSCs come from bone marrow and allogeneic MSCs such as umbilical cord blood MSCs, placenta‐derived MSCs, etc. The wide source is not only an advantage of MSCs but also a disadvantage of MSCs. Because of different cell sources and different methods of collection and preparation, it is impossible to establish a unified standard method for evaluation of efficacy. The biggest advantage of iMSCs is that it can be prepared by a standardized process, and can be prepared on a large scale, which makes it easier to commercialize. This paper reviews the present status of diagnosis and progress of MSCs therapy for AP.

Extracellular vesicles (EVs) are a heterogeneous group of membrane-bounded vesicles that are believed to be produced and secreted by presumably all cell types under physiological and pathological conditions, including tumors. EVs are very important vehicles in intercellular communications for both shorter and longer distances and are able to deliver a wide range of cargos including proteins, lipids, and various species of nucleic acids effectively. EVs have been emerging as a novel biotherapeutic platform to efficiently deliver therapeutic cargos to treat a broad range of diseases including cancer. This vast potential of drug delivery lies in their abilities to carry a variety of cargos and their ease in crossing the biological membranes. Similarly, their presence in a variety of body fluids makes them a potential biomarker for early diagnosis, prognostication, and surveillance of cancer. Here, we discuss the relatively least and understudied aspects of EV biology and tried to highlight the obstacles and limitations in their clinical applications and also described most of the new warfronts to beat cancer at multiple stages. However, much more challenges still remain to evaluate EV-based therapeutics, and we are very much hopeful that the current work prompts further discovery.