Prevalence of neural tube defect and its identification during antenatal period: a cross-sectional study in eastern Indian state.

OBJECTIVE: To estimate the prevalence of neural tube defects among all birth outcomes in Odisha during 2016-2022. Additionally, to estimate the identification rate of neural tube defects during Pradhan Mantri Surakshit Matritva Abhiyan sessions. DESIGN: A population-based cross-sectional study with a household survey for neural tube defects using pictorial card as well as a hospital-based study for antenatal ultrasonography data. SETTING: The sample population was selected through multistage random sampling. In the first stage, one district from each zone was selected randomly. In the second stage, using simple random sampling, one community health centre and one urban primary health centre were selected from each district. In the third stage, the population from a block and ward were picked from the selected rural and urban settings, respectively. PARTICIPANTS: All married women in the reproductive age group (18-49 years) residing in these cluster villages in the selected districts were enrolled. RESULTS: The study surveyed 49 215 women and recorded 50 196 birth outcomes, including 49 174 live births, 890 stillbirths and 132 medical terminations of pregnancy. A total of 30 neural tube defect cases were detected. The overall prevalence rate of neural tube defect was 0.59 per 1000 birth outcomes. Spina bifida was the most prevalent neural tube defect with the prevalence of, followed by anencephaly and encephalocele. Despite 26 860 mothers receiving antenatal ultrasonography Pradhan Mantri Surakshit Matritva Abhiyan session, data on neural tube defects and other birth defects detected through these scans is unavailable. CONCLUSION: This study found a low prevalence of neural tube defect in Odisha, which is far lower compared with the older studies from India. There is an urgent need to strengthen the quality of antenatal care services provided under Pradhan Mantri Surakshit Matritva Abhiyan through better training regarding anomaly scans and better data keeping at public healthcare facilities. TRIAL REGISTRATION NUMBER: CTRI/2021/06/034487.

Diagnostic accuracy of flow cytometry in detecting malignant epithelial cells in serous effusions.

INTRODUCTION: This study aims to evaluate diagnostic accuracy of flow cytometry (FCM) in detecting malignant epithelial cells in serous effusions. MATERIALS AND METHODS: Flow cytometric assessment of 96 serous fluids (86 ascitic, 10 pleural) was performed by using epithelial cell adhesion molecule (EpCAM) (in all 96 fluids) and MUC-1 (in a subgroup of 40 fluids) as epithelial markers and CD45 and CD14 as leucocyte markers. The percentage of EpCAM positivity and MUC-1 positivity was calculated in the CD14 and CD45 dual negative population by selective gating. The findings were then correlated with the defined gold standard criteria. RESULTS: The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy for EpCAM was found to be 92.06%, 96.96%, 98.31%, 86.48%, and 93.75%, respectively, while that for MUC-1 was 79.16%, 93.75%, 95%, 71.4%, and 85%, respectively. The sensitivity, specificity, PPV, NPV, and diagnostic accuracy for dual positivity for EpCAM and MUC-1 was found to be 83.3%, 100%, 100%, 80%, and 90% respectively. On combining FCM with cytomorphology the sensitivity, specificity, PPV, NPV, and diagnostic accuracy all increased greatly to 95.3%, 100%, 100%, 91.4%, and 96.8%, respectively. CONCLUSIONS: This study highlights the importance of multicolored flow cytometric analysis in detecting epithelial malignancies in effusions specially in cases belonging to the atypia of undetermined significance and suspicious for malignancy categories and in cases with strong clinical suspicion of malignancy with negative fluid cytology. We recommend the combined use of FCM and cytology for this specific subgroup of patients in routine clinical practice for fast and accurate reporting.

E-cigarettes and myocardial infarction: A systematic review and meta-analysis.

BACKGROUND: With widespread awareness about the harmful effects of traditional smoking, many people are considering using an e-cigarette. However, many studies have shown that e-cigarettes are not entirely harmless, and their use has been implicated in causing major adverse cardiovascular events. METHODS: We adopted the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines to conduct this systematic review. An electronic search was conducted comprehensively through five databases to find the relevant articles. The odds ratio (OR) was used for comparing groups. Meta-analysis was conducted using R statistical software version 3.4.3. A random-effects model was used. RESULTS: A total of 4 studies were included in the analysis incorporating data on 585,306 individuals. Of these, 19,435 were e-cigarettes users, while 1693 used only traditional cigarettes, and 553,095 were non-e-cigarette users. 7.0% of e-cigarettes users suffered an MI (myocardial infarction), while 7.7% and 6.5% of traditional smokers and non-e-cigarettes users suffered an MI. The OR of getting an MI in e-cigarettes (e-cigarettes only or e-cigarettes + traditional smoking) users was 1.33 (95% CI = 1.14-1.56, p-value = 0.01) in comparison to non e-cigarette users (traditional smoking or no smoking). While it is 0.61 (95% CI = 0.40-0.93, p-value 0.02) when compared with traditional smoking. CONCLUSION: Those using e-cigarettes have higher odds of suffering from an MI in comparison to not using e-cigarettes. However, using e-cigarettes is associated with half risk of the risk of MI in comparison to traditional smoking.

Effect of treatment interruptions and outcomes in cancer patients undergoing radiotherapy during the first wave of COVID-19 pandemic in a tertiary care institute.

INTRODUCTION: COVID-19 patients with cancer had poorer outcomes due to immunosuppression during cancer care, poor general condition, and other comorbidities. The study was conducted to present the real-world analysis of the effect of treatment interruptions on the outcomes of patients treated with radiation therapy during the first wave of the COVID-19 pandemic in a tertiary care institute in India. MATERIALS AND METHODS: The study is a retrospective observational cohort study on cancer patients undergoing radiation therapy from March 2020 to January 2021. The study's primary outcome was to analyze the effect of treatment interruptions on the outcomes of patients treated with radiation therapy during the first wave of COVID-19 pandemic. RESULTS: Between March 2020 to January 2021, 218 eligible patients undergoing radiation therapy were found for the study. Among the 218 patients, 25 patients (11.47%) were found positive for COVID-19, while 193 patients (88.53%) were negative for COVID-19. Among COVID-19-positive patients, ten patients had < 3 weeks of treatment interruption, while 15 patients had > 3 weeks of treatment interruptions. After recovering from COVID-19, treatment was resumed and completed for 15 (60.00%) of the COVID-19-positive patients. In comparison, 13 patients (52.00%) were lost to follow-up. Three of the COVID-19-positive patients died. The disease was clinically controlled in 12 (48.00%) of the COVID-19-positive patients, and the patients reported locoregional disease progression in 10 (40.00%). Among the 193 COVID-19-negative patients, 32 patients (16.58%) had treatment interruption. Twelve patients (37.50%) had treatment interruptions for less than 1 week. There was a significant difference in the delay of radiation treatment delivery by 2 weeks (11 fractions) in COVID-19-positive patients compared to only two fractions delay in COVID-19-negative patients. CONCLUSION: COVID-19 impacted the treatment outcomes in both COVID-19-positive and COVID-19-negative cohorts of patients. There was a longer duration of treatment interruptions in the COVID-19-positive patients, leading to fewer patients completing the radiation treatment and thereby increased locoregional disease progression. There was a significant difference in the delay in treatment between the two groups.

Efficiency and kinetics of Assam crude oil degradation by Pseudomonas aeruginosa and Bacillus sp.

We report kinetics of Assam crude oil degradation by Pseudomonas aeruginosa AKS1 and Bacillus sp. AKS2, both isolated from Assam refinery sediments. The isolates exhibited appreciable degrees of hydrophobicity, emulsification index and biosurfactant production. Crude oil degradation efficiency of isolates was assessed in (1) liquid medium amended with 1% v/v crude oil and (2) microcosm sediments (125 mg crude oil/ 10 g sand). In liquid culture, biodegradation rate (k) and half-life (t1/2) values were found to be 0.038 day-1 and 18.09 days for P. aeruginosa AKS1, and 0.020 day-1 and 33.97 days in case of Bacillus sp. AKS2, respectively. In microcosm sediments, the estimated k and t 1/2 values were 0.014 day-1 and 50 days for P. aeruginosa AKS1, and 0.011 day-1 and 61.34 days in case of Bacillus sp. AKS2. The level of nutrient treatment in microcosm sand sediment was 125 µg N and 62.5 µg P/g sediment in case of P. aeruginosa AKS1 and 375 µg N and 37.5 µg P/g sediment in case of Bacillus sp. AKS2. In microcosms without inorganic nutrients, values of k and t1/2 were found to be 0.007 day-1 and 100 days for P. aeruginosa AKS1 and for Bacillus sp. AKS2, the respective values were 0.005 day-1 and 150.68 days. Our data provides important information for predictive hydrocarbon degradation in liquid medium and contaminated sediments.

Role of ivermectin in the prevention of SARS-CoV-2 infection among healthcare workers in India: A matched case-control study.

BACKGROUND: Ivermectin is one among several potential drugs explored for its therapeutic and preventive role in SARS-CoV-2 infection. The study was aimed to explore the association between ivermectin prophylaxis and the development of SARS-CoV-2 infection among healthcare workers. METHODS: A hospital-based matched case-control study was conducted among healthcare workers of AIIMS Bhubaneswar, India, from September to October 2020. Profession, gender, age and date of diagnosis were matched for 186 case-control pairs. Cases and controls were healthcare workers who tested positive and negative, respectively, for COVID-19 by RT-PCR. Exposure was defined as the intake of ivermectin and/or hydroxychloroquine and/or vitamin-C and/or other prophylaxis for COVID-19. Data collection and entry was done in Epicollect5, and analysis was performed using STATA version 13. Conditional logistic regression models were used to describe the associated factors for SARS-CoV-2 infection. RESULTS: Ivermectin prophylaxis was taken by 76 controls and 41 cases. Two-dose ivermectin prophylaxis (AOR 0.27, 95% CI, 0.15-0.51) was associated with a 73% reduction of SARS-CoV-2 infection among healthcare workers for the following month. Those involved in physical activity (AOR 3.06 95% CI, 1.18-7.93) for more than an hour/day were more likely to contract SARS-CoV-2 infection. Type of household, COVID duty, single-dose ivermectin prophylaxis, vitamin-C prophylaxis and hydroxychloroquine prophylaxis were not associated with SARS-CoV-2 infection. CONCLUSION: Two-dose ivermectin prophylaxis at a dose of 300 µg/kg with a gap of 72 hours was associated with a 73% reduction of SARS-CoV-2 infection among healthcare workers for the following month. Chemoprophylaxis has relevance in the containment of pandemic.

Calcifying epithelial odontogenic tumors (Pindborg tumor) of maxilla in pediatric patients.

The calcifying epithelial odontogenic tumor (CEOT) was first described by Pindborg as a distinct entity in 1955. Odontogenic tumors are derived from epithelial, ectomesenchymal, and/or mesenchymal elements that are or have been a part of the tooth-forming apparatus. Of all the odontogenic tumors, CEOT accounts for 1% of the cases. There is no sex predilection, with a 2:1 predilection for the mandible, mostly in the premolar/molar region. The CEOT typically presents clinically as an intraosseous, expansile, and painless mass that exhibits slow growth. It is often locally invasive. Most often, it is associated with an impacted tooth, is asymptomatic, and requires biopsy for diagnosis. Although most of these cases are primarily intraosseous, an extraosseous tumor is also known to occur, first observed by Pindborg in 1966. The lesions were surgically enucleated, and histopathological examination confirmed CEOT. The purpose of this article is to describe one additional case of both variants of CEOT.

Prevalence and determinants of the dangerous selfie among medical and nursing students: a cross-sectional study from eastern India.

BACKGROUND: Globally, there has been an exponential rise in smartphone use and selfie taking among youth. To make selfies exciting, dangerous selfies are often taken that may lead to catastrophic consequences, including death. This study aims to estimate the prevalence of dangerous selfies and to determine the factors associated with dangerous selfies among medical and nursing students in India. METHODS: The study was conducted at the All India Institute of Medical Sciences (AIIMS), Bhubaneswar, India, in April-August 2018. The inclusion criteria were students enrolled in the Bachelor of Medicine and Bachelor of Surgery (MBBS) and nursing courses of AIIMS, Bhubaneswar. Students who did not use smartphones were excluded from the study. The interview schedule and Selfitis Behaviour Scale (SBS) were used to collect information on sociodemographic variables, smartphone use and variables related to selfies and dangerous selfies. Forward stepwise logistic regression was undertaken with the probability of entry and removal as 0.05 and 0.10, respectively. RESULTS: Of 633 eligible participants, 595 were included in the study. The mean (SD) age of the participants was 21.2 (1.6) years. More than half (56.8%) of the participants were female, 384 (64.5%) were medical students and 211 (35.5%) were nursing students. Nearly two-thirds of the participants (70.6%) preferred to take selfie. One hundred thirty three (40.3%) of the participants posted selfies on social media daily. The prevalence of dangerous selfies was 8.74% (95% CI: 6.73-11.28). Eight injury episodes while taking selfies were reported by seven (1.2%) participants. Being male (AOR 4.96, 95% CI 2.53-9.74), posting selfies on social media daily (AOR 3.33, 95% CI 1.71-6.47) and an SBS score > 75 (AOR 4.97, 95% CI 1.43-17.28) were independent predictors of dangerous selfies. CONCLUSION: Nearly one in ten medical and nursing students reported having taken a dangerous selfie, and one in one hundred reported having been injured while attempting to take a selfie. Being male, posting selfies on social media daily and an SBS score > 75 were independent predictors of dangerous selfies. Further research is required to identify the community burden of dangerous selfies and to develop strategies to prevent selfie-related fatalities among youths.

Effect of intravenous iron sucrose on hemoglobin level, when administered in a standard-dose, to anemic pregnant women in rural Northern India.

INTRODUCTION: To combat anemia among pregnant women, in Haryana, Northern India, administration of intravenous iron sucrose (IVIS) was initiated in 2014 as a public-health measure. We assessed the effect of IVIS on hemoglobin (Hb) levels among the pregnant anemic women. METHODS: Pregnant women identified as moderately or severely anemic (Hb <10.0 g/dL) in the second or third trimester during routine antenatal care were prescribed IVIS in a standard-dose of 400 mg given as 100 mg on alternate days. Neither dose calculation, nor allowance for iron-store, was included in this study. We analyzed the data collected between June 2014 and December 2015, at the two primary-health-centers, on Hb level (by HemoCue method) before start of the therapy (baseline), and 4-week or more after the last infusion (endline). RESULTS: A total of 990 women received IVIS. Both baseline and endline Hg-levels were available for 763 (77%), who were included in the analysis. At baseline, the proportion of moderate and severe anemia was 87.94% and 12.06%, respectively. Mean (standard deviation)-Hb increased from 7.85 g/dL (0.80) at baseline, to 9.62 g/dL (1.30) at endline, with a mean increase of 1.76 g/dL (95% confidence interval 1.67, 1.85). The mean increase in Hb-level for pregnant women who had severe and moderate anemia at baseline was 2.54 g/dL and 1.65 g/dL, respectively. Overall, 15.33% of women achieved normal Hb-level by the time of endline measurement. No serious adverse reactions were reported during the observation period. CONCLUSION: Mean increase in Hb-level was 1.76 mg/dL. Severely anemic pregnant women had larger increase in Hb-level when compared with pregnant women with moderate anemia.

Prevalence and correlates of sexual health disorders among adult men in a rural area of North India: An observational study.

BACKGROUND AND OBJECTIVES: Sexual health disorders are an important but less researched public health issue in India. We aimed to estimate the prevalence of sexual health disorders and their associated factors among adult men in a rural community of Haryana, India. MATERIALS AND METHODS: A community-based cross-sectional study was conducted among adult men aged 18-60 years using a multistage stratified random sampling. Information pertaining to sociodemographic characteristics, lifestyle and sexual practices, and self-reported sexual problems were collected. Sexual health disorders were defined based on International Statistical Classification of Diseases-10 classification of mental and behavioral disorders. Step-wise logistic regression was carried out to identify factors independently associated with sexual disorders. RESULTS: At least one sexual health disorder was reported by 81% of the men. The most commonly reported disorder was self-perceived defect in semen (64.4%), followed by loss of libido (21%), masturbation guilt (20.8%), erectile dysfunction (5%), and premature ejaculation (4.6%). Factors significantly associated with sexual health disorders among all men were being never married (odds ratio = 2.04; 95% confidence interval: 1.51, 2.77), smoking (1.57; 1.16, 2.14), cannabis use (4.20; 1.68, 10.48), diabetes (2.40; 1.22, 4.73), and hypertension (3.17; 1.12, 8.92). INTERPRETATION AND CONCLUSIONS: A high burden of sexual health disorders was identified among the rural men. Wider recognition of this issue is needed among the health-care providers and policymakers.

Conformational analysis of a synthetic fish kisspeptin 1 peptide in membrane mimicking environments.

Kisspeptin 1 is a neuropeptide hormone of the RFamide family, which act as an upstream regulator of brain-pituitary-gonad (BPG) axis in most vertebrates including teleosts. In the present study, a 16 amino acid long putative mature bioactive peptide (kiss 1) from preprokisspeptin 1 of golden mahseer, Tor putitora (Hamilton, 1822), was synthesized and characterized using an integrated (experimental and in silico) approach. The far-UV circular dichroism (CD) spectrum of this peptide was evaluated both in aqueous and membrane mimicking solvents (TFE, HFIP and Dioxane). The results indicate that kiss 1 peptide adopted helical, turn and ß conformations in membrane like environments. The near-UV CD spectroscopy was also carried out to examine the tertiary packing around aromatic residues of kiss 1 peptide and the peptide-membrane complex. The kiss 1 peptide exhibited little signal in water, but a prominent negative band was observed at around 275 nm when membrane mimetic solution was added. The observed ordered conformations of kiss 1 peptide in the different solvents indicated its potential biological activity which could enhance the secretion of gonadotropin-releasing hormone (GnRH) at BPG axis. The conformational information generated from the present study reinforces the application prospects of bioactive synthetic peptide analogs of kisspeptin 1 in improving the reproductive performances of important cultivable fish species.

Prevalence of neural tube defects in a rural area of north india from 2001 to 2014: A population-based survey.

BACKGROUND: Neural tube defects (NTDs) are one of the commonest birth defects. There was paucity of community-based data on occurrence of NTDs in India, especially from rural parts of the country. Against this background, the current study was carried out with main objectives to determine the prevalence of NTDs and its specific types (anencephaly, spina bifida and encephalocele) in a rural community setting over the time period 2001 to 2014. METHODS: This was a community-based cross-sectional study carried out in 28 villages of Ballabgarh Tehsil of Faridabad district in north India (population ∼ 96,000). A household survey was undertaken by trained multi-purpose workers who enquired ever-married women about history of conception with outcome as NTD during the study period. The probable case of NTD was determined using a colored pictorial card with photographs of different types of NTDs. These cases were confirmed by doctors. RESULTS: A total of 26,946 live births occurred during the years 2001 to 2014. A total of 140 confirmed cases of NTDs were identified. The live birth prevalence of NTDs was 24.1 per 10,000 live births (95% confidence interval, 18.8-30.6). The birth prevalence of NTDs for the years 2008 to 2014 was 50.8 (95% confidence interval, 39.9-63.8) per 10,000 live and stillbirths. The most common type of NTD was found to be spina bifida followed by anencephaly and encephalocele. CONCLUSION: We found high prevalence of NTDs in rural community settings from north India for years 2001 to 2014.Birth Defects Research 109:203-210, 2017.© 2016 Wiley Periodicals, Inc.

Nasal mask ventilation is better than face mask ventilation in edentulous patients.

BACKGROUND AND AIMS: Face mask ventilation of the edentulous patient is often difficult as ineffective seating of the standard mask to the face prevents attainment of an adequate air seal. The efficacy of nasal ventilation in edentulous patients has been cited in case reports but has never been investigated. MATERIAL AND METHODS: Consecutive edentulous adult patients scheduled for surgery under general anesthesia with endotracheal intubation, during a 17-month period, were prospectively evaluated. After induction of anesthesia and administration of neuromuscular blocker, lungs were ventilated with a standard anatomical face mask of appropriate size, using a volume controlled anesthesia ventilator with tidal volume set at 10 ml/kg. In case of inadequate ventilation, the mask position was adjusted to achieve best-fit. Inspired and expired tidal volumes were measured. Thereafter, the face mask was replaced by a nasal mask and after achieving best-fit, the inspired and expired tidal volumes were recorded. The difference in expired tidal volumes and airway pressures at best-fit with the use of the two masks and number of patients with inadequate ventilation with use of the masks were statistically analyzed. RESULTS: A total of 79 edentulous patients were recruited for the study. The difference in expiratory tidal volumes with the use of the two masks at best-fit was statistically significant (P = 0.0017). Despite the best-fit mask placement, adequacy of ventilation could not be achieved in 24.1% patients during face mask ventilation, and 12.7% patients during nasal mask ventilation and the difference was statistically significant. CONCLUSION: Nasal mask ventilation is more efficient than standard face mask ventilation in edentulous patients.

Toll-like Receptor-4 Polymorphisms and Serum Matrix Metalloproteinase-9 in Newly Diagnosed Patients With Calcified Neurocysticercosis and Seizures.

We evaluated seizure profile, Toll-like receptor (TLR)-4 polymorphisms, and serum matrix metalloproteinases (MMPs) in patients with calcified neurocysticercosis.One-hundred nine patients with calcified neurocysticercosis with newly diagnosed seizures and 109 control subjects were enrolled. TLR-4 Asp299Gly and Thr399Ile polymorphisms and serum MMP-9 levels were evaluated. The patients were followed for 1 year.Asp/Gly (P = 0.012) and Thr/Ile (P = 0.002), Gly (Asp/Gly plus Gly/Gly) (P = 0.008) and Ile (Thr/Ile plus Ile/Ile) (P = 0.003) genotypes were significantly associated with calcified neurocysticercosis compared with controls. Gly/Gly and Ile/Ile genotypes were not significantly associated (P = 0.529 for Gly/Gly, P = 0.798 for Ile/Ile) with either group. The levels of MMP-9 were higher in calcified neurocysticercosis (P =  < 0.001). The levels of MMP-9 were higher in patients with multiple calcified neurocysticercosis compared with single calcified neurocysticercosis (P =  < 0.001).Headache (P = 0.031), status epilepticus (P = 0.029), Todd paralysis (P = 0.039), lesion size >10 mm (P = 0.001), and perilesional edema (P =  < 0.001) were significantly associated with seizure recurrence. Heterozygous form Asp/Gly (P =  < 0.001) and heterozygous form Thr/Ile (P =  < 0.001) were significantly associated with seizure recurrence. The Gly (Asp/Gly plus Gly/Gly) (P =  < 0.001) and Ile (Thr/Ile plus Ile/Ile) (P =  < 0.001) genotypes were also significantly associated with seizure recurrence. Higher serum MMP-9 levels were significantly associated with seizure recurrence (P =  < 0.001).The TLR-4 gene abnormalities may trigger inflammation around calcified neurocysticercosis leading to an increase in perilesional edema and provocation of seizures.

Canine parvovirus NS1 protein exhibits anti-tumor activity in a mouse mammary tumor model.

Many viral proteins have the ability to kill tumor cells specifically without harming the normal cells. These proteins, on ectopic expression, cause lysis or induction of apoptosis in the target tumor cells. Parvovirus NS1 is one of such proteins, which is known to kill high proliferating tumor cells. In the present study, we assessed the apoptosis inducing ability of canine parvovirus type 2 NS1 protein (CPV2.NS1) in vitro in 4T1 cells, and found it to cause significant cell death due to induction of apoptosis through intrinsic or mitochondrial pathway. Further, we also evaluated the oncolytic activity of CPV2.NS1 protein in a mouse mammary tumor model. The results suggested that CPV2.NS1 was able to inhibit the growth of 4T1 induced mouse mammary tumor as indicated by significantly reduced tumor volume, mitotic, AgNOR and PCNA indices. Further, inhibition of tumor growth was found to be because of induction of apoptosis in the tumor cells, which was evident by a significant increase in the number of TUNEL positive cells. Further, CPV2.NS1 was also able to stimulate the immune cells against the tumor antigens as indicated by the increased CD4+ and CD8+ counts in the blood of CVP2.NS1 treated mice. Further optimization of the delivery of NS1 protein and use of an adjuvant may further enhance its anti-tumor activity.

Canine parvovirus NS1 induced apoptosis involves mitochondria, accumulation of reactive oxygen species and activation of caspases.

The non-structural protein (NS1) of parvoviruses plays an important role in viral replication and is thought to be responsible for inducing cell death. However, the detailed mechanism and the pathways involved in canine parvovirus type 2 NS1 (CPV2.NS1) induced apoptosis are not yet known. In the present study, we report that expression of CPV2.NS1 in HeLa cells arrests cells in G1 phase of the cell cycle and the apoptosis is mitochondria mediated as indicated by mitochondrial depolarization, release of cytochrome-c and activation of caspase 9. Treatment of cells with caspase 9 inhibitor Z-LEHD-FMK reduced the induction of apoptosis significantly. We also report that expression of CPV2.NS1 causes accumulation of reactive oxygen species (ROS) and treatment with an antioxidant reduces the ROS levels and the extent of apoptosis. Our results provide an insight into the mechanism of CPV2.NS1 induced apoptosis, which might prove valuable in developing NS1 protein as an oncolytic agent.

Effect of nicotinic acetylcholine receptor alpha 1 (nAChRα1) peptides on rabies virus infection in neuronal cells.

Rabies virus (RABV) is neurotropic and causes acute progressive encephalitis. Herein, we report the interaction of nAChRα1-subunit peptides with RABV and the effect of these peptides on RABV infection in cultured neuronal cells. Peptide sequences derived from torpedo, bovine, human and rats were synthesized and studied for their interactions with RABV using virus capture ELISA and peptide immunofluorescence. The results showed specific binding of the nAChRα1-subunit peptides to the RABV. In the virus adsorption assay, these peptides were found to inhibit the attachment of the RABV to the neuronal cells. The nAChRα1-subunit peptides inhibited the RABV infection and reduced viral gene expression in the cultured neuroblastoma (N2A) cells. Torpedo peptide sequence (T-32) had highest antiviral effect (IC50=14±3.01µM) compared to the other peptides studied. The results of the study indicated that nAChRα1-subunit peptides may act as receptor decoy molecules and inhibit the binding of virus to the native host cell receptors and hence may reduce viral infection.

Characterization of a Highly pH Stable Chi-Class Glutathione S-Transferase from Synechocystis PCC 6803.

Glutathione S-transferases (GSTs) are multifunctional enzymes present in virtually all organisms. Besides having an essential role in cellular detoxification, they also perform various other functions, including responses in stress conditions and signaling. GSTs are highly studied in plants and animals; however, the knowledge regarding GSTs in cyanobacteria seems rudimentary. In this study, we report the characterization of a highly pH stable GST from the model cyanobacterium--Synechocystis PCC 6803. The gene sll0067 was expressed in Escherichia coli (E. coli), and the protein was purified to homogeneity. The expressed protein exists as a homo-dimer, which is composed of about 20 kDa subunit. The results of the steady-state enzyme kinetics displayed protein's glutathione conjugation activity towards its class specific substrate- isothiocyanate, having the maximal activity with phenethyl isothiocyanate. Contrary to the poor catalytic activity and low specificity towards standard GST substrates such as 1-chloro-2,4-dinitrobenzene by bacterial GSTs, PmGST B1-1 from Proteus mirabilis, and E. coli GST, sll0067 has broad substrate degradation capability like most of the mammalian GST. Moreover, we have shown that cyanobacterial GST sll0067 is catalytically efficient compared to the best mammalian enzymes. The structural stability of GST was studied as a function of pH. The fluorescence and CD spectroscopy in combination with size exclusion chromatography showed a highly stable nature of the protein over a broad pH range from 2.0 to 11.0. To the best of our knowledge, this is the first GST with such a wide range of pH related structural stability. Furthermore, the presence of conserved Proline-53, structural motifs such as N-capping box and hydrophobic staple further aid in the stability and proper folding of cyanobacterial GST-sll0067.