In silico annotation of a hypothetical protein from Listeria monocytogenes EGD-e unfolds a toxin protein of the type II secretion system

The gram-positive bacterium Listeria monocytogenes is an important foodborne intracellular pathogen that is widespread in the environment. The functions of hypothetical proteins (HP) from various pathogenic bacteria have been successfully annotated using a variety of bioinformatics strategies. In this study, a HP Imo0888 (NP_464414.1) from the Listeria monocytogenes EGD-e strain was annotated using several bioinformatics tools. Various techniques, including CELLO, PSORTb, and SOSUIGramN, identified the candidate protein as cytoplasmic. Domain and motif analysis revealed that the target protein is a PemK/MazFlike toxin protein of the type II toxin-antitoxin system (TAS) which was consistent with BLASTp analysis. Through secondary structure analysis, we found the random coil to be the most frequent. The Alpha Fold 2 Protein Structure Prediction Database was used to determine the three-dimensional (3D) structure of the HP using the template structure of a type II TAS PemK/MazF family toxin protein (DB ID_AFDB: A0A4B9HQB9) with 99.1% sequence identity. Various quality evaluation tools, such as PROCHECK, ERRAT, Verify 3D, and QMEAN were used to validate the 3D structure. Following the YASARA energy minimization method, the target protein's 3D structure became more stable. The active site of the developed 3D structure was determined by the CASTp server. Most pathogens that harbor TAS create a crucial risk to human health. Our aim to annotate the HP Imo088 found in Listeria could offer a chance to understand bacterial pathogenicity and identify a number of potential targets for drug development.

Association of CAPN10 gene (rs3842570) polymorphism with the type 2 diabetes mellitus among the population of Noakhali region in Bangladesh: a case-control study

Type 2 diabetes mellitus (T2DM) is a multifactorial, polygenic, and metabolically complicated disease. A large number of genes are responsible for the biogenesis of T2DM and calpain10 (CAPN10) is one of them. The association of numerous CAPN10 genetic polymorphisms in the development of T2DM has been widely studied in different populations and noticed inconclusive results. The present study is an attempt to evaluate the plausible association of CAPN10 polymorphism SNP-19 (rs3842570) with T2DM and T2DM-related anthropometric and metabolic traits in the Noakhali region of Bangladesh. This case-control study included 202 T2DM patients and 75 healthy individuals from different places in Noakhali. A significant association (p 0.05). Additionally, SNP-19 genetic variants showed potential associations with the anthropometric and metabolic traits of T2DM patients in terms of body mass index, systolic blood pressure, diastolic blood pressure, total cholesterol, and triglycerides. Our approach identifies the 2R/3R genotype of SNP-19 as a significant risk factor for biogenesis of T2DM in the Noakhali population. Furthermore, a large-scale study could be instrumental to correlate this finding in overall Bangladeshi population.

An in-silico approach to design potential siRNAs against the ORF57 of Kaposi’s sarcoma-associated herpesvirus

Kaposi's sarcoma-associated herpesvirus (KSHV) is one of the few human oncogenic viruses, which causes a variety of malignancies, including Kaposi's sarcoma, multicentric Castleman disease, and primary effusion lymphoma, particularly in human immunodeficiency virus patients. The currently available treatment options cannot always prevent the invasion and dissemination of this virus. In recent times, siRNA-based therapeutics are gaining prominence over conventional medications as siRNA can be designed to target almost any gene of interest. The ORF57 is a crucial regulatory protein for lytic gene expression of KSHV. Disruption of this gene translation will inevitably inhibit the replication of the virus in the host cell. Therefore, the ORF57 of KSHV could be a potential target for designing siRNA-based therapeutics. Considering both sequence preferences and target site accessibility, several online tools (i-SCORE Designer, Sfold web server) had been utilized to predict the siRNA guide strand against the ORF57. Subsequently, off-target filtration (BLAST), conservancy test (fuzznuc), and thermodynamics analysis (RNAcofold, RNAalifold, and RNA Structure web server) were also performed to select the most suitable siRNA sequences. Finally, two siRNAs were identified that passed all of the filtration phases and fulfilled the thermodynamic criteria. We hope that the siRNAs predicted in this study would be helpful for the development of new effective therapeutics against KSHV.

Prevalence and Associated Factors of Hypertension Subtypes Among the Adult Population in Nepal: Evidence from Demographic and Health Survey Data

OBJECTIVES: This study aims to determine the prevalence, and associated factors of undiagnosed hypertension [Systolic Diastolic Hypertension (SDH), Isolated Systolic Hypertension (ISH) and Isolated Diastolic Hypertension (IDH)] in the Nepalese adult population.METHODS: Nepal Demographic and Health Survey 2016 data from adults (≥ 18 years) was used in this study. The final weighted sample size was 13,393. Blood pressure (BP) was measured 3 times and the average of the second and third measurement was reported. SDH (systolic BP (SBP) ≥ 140 mmHg and diastolic BP (DBP) ≥ 90 mmHg), ISH (SBP ≥ 140 mmHg and DBP < 90 mmHg), and IDH (SBP < 140 mmHg and DBP ≥ 90 mmHg) were measured. Multilevel logistic regression analyses were conducted to find the association between the independent variables and the covariates.RESULTS: The prevalence of SDH, IDH and ISH were 8.1%, 7.5%, and 3.3% respectively. The odds of having SDH and ISH increased with old age. However, the odds of having IDH decreased with increasing age. Females has lower odds of having SDH and IDH compared with male participants. Individuals that had been married, resided in Province 4 (p < 0.05) or 5 (p < 0.01) were statistically significantly associated with having IDH. Being overweight or obese was statistically significantly associated with all 3 HTN subtypes (p < 0.001).CONCLUSION: The necessary steps should be taken so that public health promotion programs in Nepal may prevent and control undiagnosed hypertension.

The usefulness of osteocalcin measurements in Malaysian patients with rheumatoid arthritis

Objective: Rheumatoid arthritis (RA) is a chronic inflammatory condition that can be associated with abnormal bone turnover and hence osteoporosis. Osteocalcin (OC) levels are increased in conditions with high bone turnover, including high RA disease activity. Thus, OC levels could possibly be used as a marker to assess bone health and disease activity in RA patients. As there have been no previous studies looking at serum OC levels in Malaysian RA patients, this study was performed to examine possible correlations between OC, bone mineral density (BMD) and disease activity in this population.Methods: A cross-sectional study of 75 female RA patients and 29 healthy controls was performed. Serum OC was measured using a Quantikine® ELISA kit. Dual-energy x-ray absorptiometry (DXA) was used to assess BMD.Results: Serum OC levels were not significantly different between RA patients (median 14.44 ng/mL, interquartile range [IQR 12.99]) compared to healthy controls (median 11.04 ng/mL IQR 12.29) (p=0.198). Serum OC increased with age (Spearman’s rho r=0.230, p=0.047). There was no significant correlation between serum OC and body mass index (BMI), menopause status, BMD, DAS28, swollen or tender joint counts. Overall, there were 11 (14.7%) patients with osteoporosis and 27 (36.0%) with osteopenia. Menopause status was significantly associated with BMD at all sites (lumbar spine p=0.002, femoral neck p=0.004, total hip p=0.002). Conclusions: Serum OC were similar in RA patients compared to healthy controls. In RA patients, serum OC did not correlate with RA disease activity or BMD. Menopause status remains an important influence on BMD. Thus, measuring serum OC levels in Malaysian RA patients was not useful in identifying those at risk of low BMD.

Metastatic ganglioneuroma: A misnomer.

Neuroblastoma, ganglioneuroblastoma and ganglioneuroma atumors arising from the neural crest cells. Ganglioneuroma is considered as the most mature amongst the three and usually has no metastatic potential. Spontaneous maturation of neuroblastoma into ganglioneuroma is, however, quite well-known. Here, we present a case of an 8-year-old girl child with evidence of metastasis of ganglioneuroma into a lymph node.

Use of the WOMAC questionnaire in Mumbai and the challenges of translation and cross cultural adaptation.

Patient-reported outcome measures (PROMs) are disease specific questionnaires that are being increasingly used in clinical practice and research. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), is a widely used PROM in patients with hip or knee osteoarthritis. A validated WOMAC was used by us, and significant challenges were faced in administering it as several questions did not have a cultural connect. Functionally equivalent items in the Indian context had then to be used to complete the score.

Hepatoprotective and antioxidant activity of methanolic extract of flowers of Nerium oleander against CCl4-induced liver injury in rats

OBJECTIVE@#To investigate the antioxidant and hepatoprotective activity of methanolic flower extract of Nerium oleander against CCl(4)-induced hepatotoxicity in rats.@*METHODS@#In vitro antioxidant activity of methanolic extract of flowers of Nerium oleander (MENO-F) was evaluated by various assays, including reducing power, lipid peroxidation, DPPH, ABTS, superoxide anion, hydroxyl radicals and metal chelation. The hepatoprotective and in vivo antioxidant activity of MENO-F were evaluated against CCl(4)-induced hepatic damage in rats. The MENO-F at dose of 100, 200 and 400 mg/kg were administered orally once daily for seven days. Serum enzymatic levels of serum glutamate oxaloacetate transaminase (AST), serum glutamate pyruvate transaminase (ALT), serum alkaline phosphatase (ALP) and total bilirubin were estimated along with estimation of superoxide dismutase (SOD) and malondialdehyde (MDA) levels in liver tissues. Further histopathological examination of the liver sections was carried out to support the induction of hepatotoxicity and hepatoprotective efficacy.@*RESULTS@#The extract showed potent activities on reducing power, lipid peroxide, DPPH, ABTS, superoxide anion, hydroxyl radical and metal chelation. The substantially elevated serum enzymatic levels of AST, ALT, ALP and total bilirubin were found to be restored towards normalization significantly by the MENO-F in a dose dependent manner with maximum hepatoprotection at 400 mg/kg dose level. The histopathological observations supported the biochemical evidences of hepatoprotection. Elevated level of SOD and decreased level of MDA further strengthen the hepatoprotective observations.@*CONCLUSIONS@#The results of the present study strongly reveal that MENO-F has potent antioxidant activity and hepatoprotective activity against CCl(4)-induced hepatic damage in experimental animals.

Van der Knaap syndrome, a case from West Bengal, India

Leucodystrophies are a group of white matter diseases caused by an abnormality in the formation or maintenance of one or more components of the myelin sheath. The devastating course of the disease coupled with the lack of any defi nitive treatment render them potentially fatal and incurable. Van der Knaap Syndrome, also known as megaloencephalic leucoencephalopathy with subcortical cyst, is a relatively new and rare entity. Although reported sporadically from Europe, Asia and the Agarwal community in India, there is practically no case report from the eastern part of India. We present a case of van der Knaap syndrome in an adult female with some atypical features.

Clinico-etiologic correlates of onychomycosis in Sikkim.

AIMS AND OBJECTIVES: The etiological spectrum of any superficial mycosis is largely dependent on the flora in the immediate environment of the individual. It is influenced by the geographic, climatic and occupational factors. The study is basically to formulate baseline data for the species prevalence of various dermatophytes in patients with onychomycosis attending the Central Referral Hospital, Gangtok, Sikkim. MATERIALS AND METHODS: Thirty-four clinically suspected cases of onychomycosis were subjected to mycological studies. RESULTS: Thirty-two (94.12%) cases were positive for fungal elements by direct microscopy and 28 (82.35%) by culture. Young adults in the age group of 21-30 years were mainly affected. The male:female ratio was 1.125:1. Dermatophytes were isolated in 18 cases (64.29%). Trichophyton tonsurans (44.44%) was the most common isolate followed by T. mentagrophytes (22.22%), T. rubrum (11.11%), T. verrucosum (11.11%) and Microsporum audouinii (11.11%). Apart from dermatophytes, Aspergillus niger (21.43%) and Penicillium marneffei (14.28%) were also isolated. CONCLUSION: Dermatophytes, mainly T. tonsurans, as well as moulds other than dermatophytes were isolated from onychomycosis patients in Gangtok, Sikkim.

Are Indians and females less tolerant to pain? An observational study using a laboratory pain model

In Malaysia, it is a common belief among health care workers that females and Indians have lower pain threshold. This experience, although based on anecdotal experience in the healthcare setting, does not allow differentiation between pain tolerance, and pain expression. To determine whether there is a difference in the tolerance to pain between the three main ethnic groups, namely the Malays, Chinese and Indians as well as between males and females. This was a prospective study, using a laboratory pain model (ischaemic pain tolerance) to determine the pain tolerance of 152 IMU medical students. The mean age of the students was 21.8 years (range 18-29 years). All of them were unmarried. The median of ischaemic pain tolerance for Malays, Chinese and Indians were 639s, 695s and 613s respectively (p = 0.779). However, statistically significant difference in ischaemic pain tolerance for males and females Indian students were observed. Possible ethnic difference in pain tolerance in casual observation is not verified by this laboratory pain model. Difference in pain tolerance between genders is shown only for Indians.

Common laboratory tests for rheumatologicaldisorders: how do they help the diagnosis?

No screening test is ideal for detecting rheumatic diseases; diagnosis depends on appropriate history and thorough physical examination. Sometimes, laboratory investigations may be useful in confirming or ruling out rheumatic disease after a clinical diagnosis is considered. Once a rheumatic disease has been diagnosed, certain laboratory tests can help in assessing prognosis or determining the extent of the disease. Laboratory tests may also help the physician monitor certain rheumatic diseases, guide treatment or assess potential drug toxicity.

Effect of estradiol on expression of cytoskeletal proteins during spermatogenesis in testis of sexually mature rats.

Earlier studies had shown that long term treatment with estradiol arrests spermatogenesis in adult male rats, at a dose of 0.1 mg/kg/day. The present study was therefore undertaken to ascertain the causes underlying the reduction in sperm counts by administering estradiol for a short term, at the dose of 0.1 mg/kg/day. Estradiol valerate was injected at a dose of 0.1 mg/kg/day, for a period of 10 days to one group of adult male rats, which were administered saline for 12 days prior to estradiol injection, and sacrificed after 22 days. The control group was administered saline for 22 days. The sera were analyzed for testosterone and FSH levels. One testis of each male was immersion fixed for histology, and for immunohistochemistry of two testicular cytoskeletal proteins, vimentin and vinculin. The contralateral testes were used for analysis of vimentin and vinculin gene expression by reverse transcriptase polymerase chain reaction (RTPCR) and western blotting. Another group exposed to estradiol for 10 days was injected with bromodeoxyuridine (BrdU), at a dose of 100 mg/kg/day, to ascertain the effect on germ cell proliferation, and sacrificed 12 days later, while estradiol treatment was continued till sacrifice. BrdU, at a dose of 100 mg/kg/day was injected i.p. to a group of control rats treated with saline for 10 days, and sacrificed 12 days later. The testes from both groups were immersion fixed for immunohistochemical detection of BrdU. Histology of estradiol treated testis showed predominance of tubules with round spermatids with accumulation of lipid droplets in Sertoli cell cytoplasm and decreased cell height, whereas controls showed elongating spermatids. BrdU immunolocalization in the testis, irrespective of treatment, indicated its incorporation in deoxyribonucleic acid (DNA) suggesting that estradiol sustained germ cell proliferation. Both vimentin and vinculin could be immunolocalized to the testis. The testicular levels of vimentin and vinculin, quantified after western blotting, were unaffected. The testicular expression of vimentin and vinculin seen by RTPCR was also unaffected. The study suggested that estradiol induced reduction in sperm counts was not due to adverse effects on proliferation. The observed predominance of seminiferous tubules showing round spermatids, accumulation of lipid droplets as compared to controls suggested that reduction in elongated spermatids occurred through reduced spermiation and phagocytosis. The study also suggested that reduction in Sertoli cell height after short-term estradiol treatment was not due to reduced expression of vimentin and vinculin, which could be maintained by estradiol. However, reduction in Sertoli cell height could have been due to suppression of FSH and testosterone, implicated in the polymerization of vimentin and organization of vinculin, two cytoskeletal proteins involved in inter-Sertoli or Sertoli-germ cell junctions. The study suggested that disorganization of Sertoli cell cytoskeleton and reduction in the volume of Sertoli cells could be an important factor for reduced efficiency of spermatogenesis after exposure to estrogenic molecules.

Differential depression of spinal synaptic transmission in vitro by different hypoxic insults.

The effects of hypoxia (O2-free), aglycemia (glucose-free), ischemia (O2- and glucose-free) and chemical anoxia (by 3-nitropropionic acid; 3-NPA) were evaluated on the synaptic transmission in vitro. Stimulation of a dorsal root in hemisected spinal cord from neonatal rat, evoked monosynaptic (MSR) and polysynaptic reflexes (PSR) in the segmental ventral root. In all the hypoxic conditions, the reflexes were depressed in a time-dependent manner. Hypoxia took longer time (> 240 min) to abolish the reflexes where as, aglycemia and ischemia abolished them within 35 min. Recovery after wash was complete in hypoxia, 60-70% in aglycemia and 20-25% in ischemia. The time required for 50% depression of reflexes (T-50) was also in the same order (100, 23 and 13 min). The elimination of O2 in hypoxic or ischemic solution by N2 bubbling abolished the reflexes within 16 min. The T-50 values in both the conditions were between 5-8 min. Superfusion of 3-NPA (an irreversible inhibitor of succinate dehydrogenase) depressed the reflexes. The abolition time and T-50 values were shorter with the increasing concentrations of 3-NPA. The present results reveal that the energy production in hypoxic condition with normal glucose level can sustain the synaptic activity for a longer time while the glucose deficiency even in normoxic conditions drastically impair the synaptic activity. Further, aglycemia depressed the reflexes almost in a similar time as seen with ischemia.

Changing scenario of transfusion-related viral infections.

OBJECTIVE: To study the changing incidence of blood transfusion-related viral infections consequent to compulsory screening of blood and greater awareness of the problem, over the last five years. METHODS: This is a cross-sectional study carried out at Medical College, Calcutta. Three groups each consisting of 100 subjects were selected for this study. Group A comprised multiple transfused patients who have also received transfusion before 1995. Group B comprised patients who had received transfusions only since 1995. Group C comprised of control patients who have never been transfused. The incidence of HBsAg +ve, anti-HCV +ve and HIV +ve cases were calculated and expressed as percentages and compared using the chi square test. RESULTS: The incidences of HBsAg +ve and anti-HCV +ve cases in the three groups were 20% and 16% in Group A, 7% and 6% in Group B and 4% and 2% in Group C. The difference between Group A and Group B were statistically significant. CONCLUSION: The incidence of HBsAg and anti-HCV positive cases among the multi-transfused has decreased over the last five years.

Post-marketing surveillance of arteether in malaria.

OBJECTIVE: To undertake post-marketing surveillance (PMS) of arteether (E-Mal) with the aim of obtaining feedback from clinicians regarding its safety, tolerability, efficacy and adverse event profile in patients of P. falciparum malaria. METHOD: Post-marketing surveillance proforma to collect data from clinicians using arteether (E-Mal) was provided to institutions/nursing homes and hospitals where Arteether (E-Mal) was available for use in treatment of P. falciparum malaria. These clinicians were informed about the need and relevance of providing this feedback regarding their reexperience on E-Mal therapy on predesigned proforma. Duly filled proformas were received by Central Drug Research Institute, Lucknow for data analysis, documentation and conclusions regarding E-Mal therapy. RESULT: A total of 300 reports were received for analysis from states of Bihar, Gujarat, Madhya Pradesh, Maharashtra, Rajasthan and Uttar Pradesh. The results show that 294 cases (98%) were cured, five cases improved and one patient did not show any change in the clinical status. The side effects (headache, nausea, vomiting and giddiness) reported in the proforma of 14 cases were mild in nature and no causal relationship with arteether could be ascertained. CONCLUSION: An indepth analysis of these 300 reports confirmed the safety, highlighted excellent tolerability and further proved the efficacy of three-day schedule of arteether (IMI) for treatment of malaria. Arteether should not be used in P. Vivax malaria (E-Mal) except when smear is positive for both (P. falciparum and P. vivax). In such a situation risk-benefit should be carefully evaluated before advocating the use of E-Mal therapy. The post-marketing surveillance is continuing and it is hoped that with more feedback from the clinicians from various parts of the country PMS data on this novel antimalarial drug (E-Mal) would further be documented.

Mitochondrial myopathy presenting as ataxia with dilated cardiomyopathy.

Mitochondrial disorders are multisystem diseases with very heterogeneous clinical manifestations. Common cardiac features include cardiomyopathy and conduction defects. We report a five-year-old boy who presented with signs of congestive cardiac failure and was diagnosed to have dilated cardiomyopathy. Six months later, he developed progressively worsening ataxia, hypotonia, other cerebellar signs, hearing loss, severe sensory peripheral neuropathy and lactic acidosis. Electronmicroscopy of skeletal muscle biopsy was consistent with mitochondrial myopathy.