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Introduction: Neuroglioma, a classification encompassing tumors arising from glial cells, exhibits variable aggressiveness and depends on tumor grade and stage. Unraveling the EGFR gene alterations, including amplifications (unaltered), deletions, and missense mutations (altered), is emerging in glioma. However, the precise understanding of emerging EGFR mutations and their role in neuroglioma remains limited. This study aims to identify specific EGFR mutations prevalent in neuroglioma patients and investigate their potential as therapeutic targets using FDAapproved drugs for repurposing approach. Methods: Neuroglioma patient's data were analyzed to identify the various mutations and survival rates. High throughput virtual screening (HTVS) of FDA-approved (1615) drugs using molecular docking and simulation was executed to determine the potential hits. Results: Neuroglioma patient samples (n=4251) analysis reveals 19% EGFR alterations with most missense mutations at V774M in exon 19. The Kaplan-Meier plots show that the overall survival rate was higher in the unaltered group than in the altered group. Docking studies resulted the best hits based on each target's higher docking score, minimum free energy (MMGBSA), minimum kd, ki, and IC50 values. MD simulations and their trajectories show that compounds ZINC000011679756 target unaltered EGFR and ZINC000003978005 targets altered EGFR, whereas ZINC000012503187 (Conivaptan, Benzazepine) and ZINC000068153186 (Dabrafenib, aminopyrimidine) target both the EGFRs. The shortlisted compounds demonstrate favorable residual interactions with their respective targets, forming highly stable complexes. Moreover, these shortlisted compounds have drug-like properties as assessed by ADMET profiling. Conclusion: Therefore, compounds (ZINC000012503187 and ZINC000068153186) can effectively target both the unaltered/altered EGFRs as multi-target therapeutic repurposing drugs towards neuroglioma. [ABSTRACT FROM AUTHOR]

The 2019 coronavirus pandemic prompted extensive research into the molecular composition of the virus. The SARS-CoV-2 virus contains small noncoding RNAs (sNCR, miRNAs) targeting crucial host genes. This study identified 24 potential precursor and mature Viral-miRNAs (V-miRNAs) in the SARS-CoV-2 genome with favorable thermodynamic energies associated with 1883 human genes, 334 of which are involved in the COVID-19 infection. Out of 24, 14 predicted mature SARS-CoV-2 V-miRNAs target 28 host genes of the 334 genes involved in COVID-19 based on their robust molecular hybridization (thermodynamic stability, canonical base pairing, minimal noncanonical base pairs, loops, and bulges). Eight of the 14 mature V-miRNAs showed sequence conservancy with other organisms based on predicted 3D structures. Gene ontology enrichment analysis revealed these 28 host genes are involved in 42 biological processes, 4 cellular components, and 10 molecular processes. Further, the 14 predicted mature V-miRNAs were screened against FDA-approved (COVID-19 drug dataset) to identify the top best-hit compounds, which include Bemcentinib and Zavegepant. These two drugs show strong binding affinities (Δ G = − 9.2 to −12.4 kcal/mol) with 13 and 12 predicted mature V-miRNAs, respectively. Furthermore, molecular docking of V-miRs-host AGO protein interactions suggested strong binding affinities and extensive hydrogen bonding interactions between V-miRNAs and the host AGO protein (involved in miRNA biogenesis and mRNA regulation). These findings suggest a potential regulatory role of Bemcentinib and Zavegepant through strong binding interaction with various V-miRNAs and in modulating host gene expression through their interactions with the AGO protein. This study identified the potential 14 mature Viral miRNAs (V-miRNAs) in the SARS-CoV-2 genome which targets 28 host genes involved in COVID-19 infection. These V-miRNAs demonstrated robust molecular hybridization with their target host genes, and eight of V-miRNAs were found to be conserved in other organisms. The mature V-miRNAs has potential binding role in Argonaute protein (AGO), involved in the regulation of host genes expression. FDA drugs, Bemcentinib and Zavegepant, exhibited strong binding affinities with 12 V-miRNAs showing antagonistic behavior with the AGO-miRNA role thereby reducing the COVID-19 infection. [ABSTRACT FROM AUTHOR]

Background: Smoking is one of the leading causes of millions of deaths worldwide. During cigarette smoking, most affected and highly exposed cells are the alveolar epithelium and generated oxidative stress in these cells leads to death and damage. Several studies suggested that oxidative stress causes membrane remodeling via Phospholipase A2s but in the case of cigarette smokers, mechanistically study is not yet fully defined. In view of present perspective, we evaluated the involvement of cytosolic phospholipase A2 (cPLA2) IVA as therapeutic target in cigarette smoke induced pathologies in transformed type I and type II alveolar epithelial cells. Methods: Transformed type I (WI26) and type II (A549) alveolar epithelial cells were used for the present study. Cigarette smoke condensate (CSC) was prepared from most commonly used cigarette (Gold Flake with filter) by the Indian population. CSC-induced molecular changes were evaluated through cell viability using MTT assay, reactive oxygen species (ROS) measurement using 2,7 dichlorodihydrofluorescin diacetate (DCFH-DA), cell membrane integrity using fluorescein diacetate (FDA) and ethidium bromide (EtBr) staining, super oxide dismutase (SOD) levels, cPLA2 activity and molecular involvement of specific cPLA2s at selected 24 h time period. Results: CSC-induced response on both type of epithelial cells shown significantly reduction in cell viability, declined membrane integrity, with differential escalation of ROS levels in the range of 1.5-15 folds and pointedly increased cPLA2 activity (p < 0.05). Likewise, we observed distinction antioxidant potential in these two types of lineages as type I cells had considerably higher SOD levels when compared to type II cells (p < 0.05). Further molecular expression of all cPLA2s increased significantly in a dose dependent manner, specifically cytosolic phospholipase A2 IVA with maximum manifestation of 3.8 folds. Interestingly, CSC-induced ROS levels and cPLA2s expression were relatively higher in A549 cells as compared to WI26 cells. Conclusions: The present study indicates that among all cPLA2s, specific cPLA2 IVA are the main enzymes involved in cigarette smoke induced anomalies in type I and type II lung epithelial cells and targeting them holds tremendous possibilities in cigarette smoke induced lung pathologies. [ABSTRACT FROM AUTHOR]

Background and Aims 14 C-urea breath test (14 C- UBT) is considered as 'gold standard' for detection of active gastric H. pylori infection. However, till date no comparative study using encapsulated and non-encapsulated 14 C- UBT protocols has been conducted in same subjects in identical conditions. We monitored gastric fate of capsule containing 14 C-urea with real time display and compared sensitivities of these protocols at different time points of breath collection. Methods Non-encapsulated 14 C- UBT was performed using 74 k Bq of 14 C-urea in 100 dyspeptic patients by collecting breath samples at 10, 15 and 20 minutes. Thereafter, within 2 days a gelatin capsule containing 14 C-urea along with 6.0 MBq of 99m Tc-diethylene triamine penta-acetic acid was administered to each patient for real time display of capsule movement and its fate in gastrointestinal tract by gamma camera. Simultaneously, breath samples were collected for 14 CO2 measurement during image acquisition. Results Employing non-encapsulated 14 C- UBT, 74 out of 100 dyspeptic patients were found to be H. pylori positive. Discordant 14 C- UBT results were obtained in 4/74 (5.4%) cases using these two protocols. By employing encapsulated and nonencapsulated 14 C- UBT protocols, sensitivities of 14 C- UBT were found to be 90.5 versus 98.6% at 10 and 91.8 versus 97.2% at 15 minutes respectively; while these were 94.6 versus 100, 90.7 versus 98.6 and 83.7 versus 93.2% considering any one, two or all three positive values respectively. Conclusions Incomplete/non-resolution of 14 C-urea capsule in stomach during the phase of breath collections appears to decrease sensitivity of encapsulated 14 C- UBT as compared to nonencapsulated protocol for detection of H. pylori infection. [ABSTRACT FROM AUTHOR]

Background and Aim: Previous studies have shown that while performing the 14C-urea breath test (14C-UBT) for the detection of Helicobacter pylori (H. pylori), there is possibility of false-positive results due to the other urease producing bacteria present in oropharynx, if breath samples are obtained within 30 min after administration of non-capsulated 14C-urea. Therefore, we have exclusively evaluated the kinetics of 14carbon dioxide (14CO2) excretion by oral commensal flora to theoretically propose optimum breath collection timings for 14C-UBT. Methods: Multiple breath samples up to 15 min were collected in 0.25 mmol benzethonium hydroxide from 25 healthy volunteers after they withheld 37 kBq (1 μCi) of 14C-urea in their mouths for 15 s and then expectorated the tracer. The test was repeated on the same subjects without and with mouth cleansing protocols. Breath 14CO2 content was measured by the Liquid Scintillation Counter (1409; Wallac, Turku, Finland) and results were expressed as 14CO2 excretion per mmol breath CO2 (% administered dose). Results: Peak breath radioactivity at 1 min in the former protocol was 3.53 times higher than the latter which declined subsequently with a half time of 1 min and 2.5 min, and reached baseline levels by 15 and 10 min, respectively. The peak radioactivity (100%) at 1 min declined by 94% and 97.8% in the former and later protocols, respectively, at 15 min. Although magnitude of the peak varied in different subjects, the shape of curve remained almost similar in all cases. Conclusions: Without mouth cleansing, oral micro flora excreted more 14CO2 up to 15 min after administration of non-capsulated 14C-urea. Therefore, it is proposed that two breath samples may be obtained either at 15 and 20 min without or at 10 and 15 min with mouth cleansing protocols for reliable analysis of 14C-UBT data for H. pylori detection. [ABSTRACT FROM AUTHOR]

14C-urea breath test (UBT) is considered to be an accurate diagnostic test for the detection of active Helicobacter pylori infection. Various test meals are used in 14C-UBT to slow down gastric emptying, and to enhance the gastric distribution, in order to increase the time and area of contact between microorganisms and the tracer substrate. The aim of the present paper was to evaluate the effect of gastric environment on the performance of 14C-UBT using an alkaline and an acidic liquid test meal having gastric emptying retardant effect. The comparison of 14C-UBT was done with liquid test meals (200 mL water) comprising (i) plain drinking water (PDW); (ii) 1.3 g or 3.0 g citric acid (CA); and (iii) 3.0 g trisodium citrate (TSC). Eighteen patients (37 ± 12 years, range 18–57 years) with complaints of dyspepsia participated in the study. The status of H. pylori was confirmed by histology and rapid urease test. A total of 93 kBq of 14C-urea (0.5 mL) in a gelatin capsule was orally administered along with liquid test meals to the overnight fasting subjects. Breath samples were collected and radioactivity measured. Results were expressed as 14CO2/mmol exhaled CO2 as percentage of administered radioactive urea. Higher acidic gastric environment (pH approx. 2.0) with CA was found to increase the exhaled 14CO2 level in a dose-dependent manner as compared to PDW and TSC meal ( P < 0.05) at all time points. With TSC test meal, the expired 14CO2 level decreased in the lower acidic gastric environment (pH approx. 5.3). The peaks of exhaled 14CO2 with TSC test meal were observed at the same time points as that with PDW and CA test meals. The 14C-UBT with TSC was found to be positive in 77% of patients (10/13). Better interaction between the microbial urease and 14C-urea, caused by a test meal that retards gastric emptying and that changes gastric pH, plays an important role in hydrolysis of the administered 14C-urea by H. pylori urease. © 2005 Blackwell Publishing Asia Pty Ltd [ABSTRACT FROM AUTHOR]

A letter to the editor is presented in response to the article "Increased Superoxide Formation Induced by Irradiation Preconditioning Triggers Kidney Resistance to Ischemia-Reperfusion Injury in Mice," by Jinu Kim et al. in a 2009 issue.

Aims: To find out the role of secretory phospholipase A2 (sPLA 2 ) isozymes as potential targets in tobacco condensate-induced colon damage., Background: The effects of cigarette smoke condensate (CSC) and the molecular mechanisms involved in the regulation of phospholipase A2 (PLA 2 ) and its isozymes in colon cells, which are still unclear and emerging, are studied., Objectives: The study aimed to check the effect of CSC on cell viability and reactive oxygen species (ROS) and superoxide. Also, the effect of CSC on gene expression of different secretory phospholipase A2 (sPLA 2 ) was evaluated. Moreover, the impact of inhibition of sPLA 2 on various cell properties i.e. cell viability, cell proliferation, membrane damage and free radicals' generation is also studied., Methods: CSC-induced changes were evaluated in cell viability by MTT assay, followed by the evaluation of membrane modulation by flow cytometry, free radical generation by fluorescent dyes, PLA 2 isoforms gene expression patterns and their suppression by small interfering RNA (siRNA) studied in HCT-15 male and HT-29 female colon cells., Results: Our results demonstrate that HCT-15 and HT-29 cells treated with CSC significantly reduced the cell viability by 50% within 48 h and significantly enhanced the total reactive oxygen species (ROS) by 2 to 10-fold, and mitochondrial ROS (mtROS) and superoxide radicals (SOR) by 2-fold each. Treatment with CSC significantly unregulated secretory phospholipase A2 (sPLA 2 ) IID group and down-regulated IB and cytosolic phospholipase (cPLA 2 ) IVA groups in HCT-15 cells without affecting them in HT-29 cells. Silencing the sPLA 2 IID group results in an increase in cell viability and a decrease in ROS. Silencing the PLA 2 IVA gene in the HCT-15 cells showed a reduced expression which had no impact on the CSC-induced cell proliferation, membrane damage and free radicals (ROS, mtROS, and SOR) generation., Conclusion: Therefore, identifying cell-specific sPLA 2 isozymes seems to play a key role in controlling the ROSinduced damage by CSC and helps develop specific therapeutic strategies., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background and Aims: (14) C-urea breath test ((14) C-UBT) is considered as "gold standard" for detection of active gastric H. pylori infection. However, till date no comparative study using encapsulated and non-encapsulated (14) C-UBT protocols has been conducted in same subjects in identical conditions. We monitored gastric fate of capsule containing (14) C-urea with real time display and compared sensitivities of these protocols at different time points of breath collection., Methods: Non-encapsulated (14) C-UBT was performed using 74 kBq of (14) C-urea in 100 dyspeptic patients by collecting breath samples at 10, 15 and 20 minutes. Thereafter, within 2 days a gelatin capsule containing (14) C-urea along with 6.0 MBq of (99m) Tc-diethylene triamine penta-acetic acid was administered to each patient for real time display of capsule movement and its fate in gastrointestinal tract by gamma camera. Simultaneously, breath samples were collected for (14) CO2 measurement during image acquisition., Results: Employing non-encapsulated (14) C-UBT, 74 out of 100 dyspeptic patients were found to be H. pylori positive. Discordant (14) C-UBT results were obtained in 4/74 (5.4%) cases using these two protocols. By employing encapsulated and nonencapsulated (14) C-UBT protocols, sensitivities of (14) C-UBT were found to be 90.5 versus 98.6% at 10 and 91.8 versus 97.2% at 15 minutes respectively; while these were 94.6 versus 100, 90.7 versus 98.6 and 83.7 versus 93.2% considering any one, two or all three positive values respectively., Conclusions: Incomplete/non-resolution of (14) C-urea capsule in stomach during the phase of breath collections appears to decrease sensitivity of encapsulated (14) C-UBT as compared to nonencapsulated protocol for detection of H. pylori infection., (© 2013 John Wiley & Sons Ltd.)

Background: Inflammatory bowel disease (IBD) is an idiopathic, chronic inflammatory condition, which affects the gastrointestinal tract and has no curative treatment. The present study aimed to investigate the effect of different doses of pioglitazone alone and in combination with sulfasalazine in TNBS (trinitrobenzenesulfonic acid)-induced inflammatory bowel disease (IBD) in rats., Methods: A total of 36 animals were included in the study. Animals were divided into five groups (n = 6): group I--vehicle (ethanol), group II--TNBS + ethanol, group IIIA--TNBS + pioglitazone (15 mg/kg), group IIIB--TNBS + pioglitazone (30 mg/kg), group IV--TNBS + sulfasalazine (360 mg/kg), group V--TNBS + sulfasalazine (360 mg/kg) + pioglitazone (least effective dose found in group III). Group III was divided into two subgroups, namely IIIA and IIIB, on the basis of different doses of pioglitazone used. After completion of two weeks of treatment, rats were sacrificed under ether anesthesia by cervical dislocation for assessment of intestinal inflammation, histological analysis, myeloperoxidase assay, malondialdehyde assay and TNF-α estimation., Results: All the drug-treated groups showed both gross morphological and microscopic score either 1 or 2. None of them showed score of > 2 on both gross and microscopic morphological examination. Both MDA levels and MPO activity were significantly reduced in the drug-treated groups, with maximum reduction seen in the combination group. TNF-α was reduced in pioglitazone group. It was highly reduced in sulfasalazine group (group V) as compared to TNBS group thereby indicating that pioglitazone is protective in TNBS-induced inflammatory bowel disease., Conclusion: The present study showed reduction in lipid peroxidation, malondialdehyde levels and TNF-α levels in pioglitazone-treated group and hence, there was significant improvement in gross and microscopic features, too. However, combination of pioglitazone and sulfasalazine has shown greater efficacy.

Background: 14C-urea breath test (14C-UBT) is employed as a 'gold standard' technique for the detection of active gastric Helicobacter pylori infection and is recommended as the best option for "test-and-treat" strategy in primary health care centers., Aim: To compare the performance of capsulated and non-capsulated 14C-UBT protocols for the detection of H. pylori infection in patients., Methods: Fifty eight H. pylori infected patients underwent routine upper GI endoscopy and biopsies were processed for rapid urease test (RUT) and histopathology examination. Capsulated 14C-UBT was done in a novel way by using 74 kBq of 14C-urea along with 6.0 MBq of 99mTc-diethylene triamine penta-acetic acid (99mTc-DTPA) to simultaneously monitor the movement and the fate of ingested capsule after delineating the stomach contour by using 20.0 MBq of 99mTechnetium pertechnetate (99mTcO4-) under dual head gamma camera. Non-capsulated 14C-UBT was performed within 2 days of the previous test and the results of these protocols were compared., Results: In 3 out of 58 H. pylori positive cases (5.17%), 14C-UBT results were found to be negative by using the capsulated method. Interestingly, on monitoring the real time images of the capsule in these cases it was found that misdiagnosis of H. pylori infection occurred mainly due to either rapid transit of the 14C-urea containing capsule from the upper gastric tract or its incomplete resolution in the stomach during the phase of breath collection., Conclusion: Use of non-capsulated '4C-UBT protocol appears to be a superior option than the conventional capsule based technique for the detection of H. pylori infection.

Objectives: The aim of the study was to evaluate the correlation between colitis and susceptibility to seizures., Methods: Colitis was induced in Wistar rats by a single intracolonic administration of trinitrobenzene sulfonic acid (TNBS; 20 mg in 35% ethanol). The control group were given intracolonic vehicle. One group of rats with colitis were treated with thalidomide (150 mg/kg p.o.) daily for 14 days. The other colitis group received vehicle only. On day 15, seizure susceptibility was tested by administration of pentylenetetrazole (40 mg/kg i.p.). Colonic tissue was collected for estimation of morphological score, and malondialdehyde, superoxide dismutase, catalase and glutathione peroxidase. Tumour necrosis factor (TNF)-alpha levels were measured in serum and brain samples., Key Findings: The colitis group showed a significant increase in seizure score and reduction in onset time compared with the control group. Thalidomide was protective against seizures, resulting in decreased seizure score and significantly delaying the onset of seizures. Thalidomide also provided significant protection against TNBS-induced colonic damage in terms of morphological and histological score and levels of lipid peroxidation, superoxide dismutase, catalase and glutathione peroxidase in colonic tissue. The level of TNF-alpha in serum was also reduced significantly whereas brain TNF-alpha level was reduced but not significantly., Conclusions: TNBS-induced colitis increased seizure susceptibility to a subconvulsive dose of pentylenetetrazole; the immunomodulator thalidomide was protective.

Increased levels or overexpression of ornithine decarboxylase (ODC), a rate-limiting enzyme in polyamine biosynthesis pathway, is characteristic of tumor cells. Similarly, prostaglandins (PGs) appear to be important in the pathogenesis of cancer because these affect mitogenesis, cellular adhesion, immune surveillance and apoptosis. Cancers form much more PGs than the original tissue from which they have arisen. This study has revealed that pretreatment of mice with resveratrol at a dose of 2.5 mg/kg body weight for two weeks blocked the N-nitrosodiethylamine(NDEA)-induced cytosolic ODC levels in the liver and lungs. The blockage was pronounced in hepatic tissue compared to pulmonary tissue. Resveratrol feeding caused a significant reduction in microsomal cyclooxygenase (COX) activities in the liver and lungs, while the dosage of NDEA (200 mg/kg body weight) induced COX activity 24 h after its administration. In any case, resveratrol pretreatment turned out to effectively block the induction of COX activity in the lungs by NDEA.

Antiradical activity of nimesulide, a commonly used COX-2 specific inhibitor, was estimated in vitro by 1, 1 diphenyl-2-picrylhydrazyl (DPPH) assay, nitroblue tetrazolium reduction assay and lipid peroxidation assay, respectively. The biochemistry of antioxidant functions of nimesulide was also investigated under control and inflammatory conditions, caused by intratracheal instillation of lipopolysaccharide (LPS). Pro-inflammatory conditions generally end up in oxidative insults, which have been suggested to be the cause of multiple organ failure in inflammation. A primary defense, constituted of antioxidant enzymes, against this oxidative damage has evolved in the body. In this study, male Wistar rats were orally administered with nimesulide (9 mg/kg/twice daily for 1 week), followed by intratracheal instillation with 2 microg of LPS and after 18 hr, antioxidant defense system and lipid peroxidation were measured in liver, lungs and kidneys. Nimesulide pretreatment was found to protect the tissue from enhanced levels of lipid peroxidation, and also stimulated the levels of glutathione-S-transferase (GST) in liver and glutathione reductase in kidneys. Surprisingly, nimesulide oral feeding also significantly suppressed superoxide dismutase (SOD) activity in all the three organs. Although, in our study, nimesulide proved to be an inducer of GST (a marker for chemoprevention) and a scavenger of superoxide anions at higher concentrations (> 250 microM), but the relevance of suppression of SOD enzyme activity, which may contribute to the drug's toxic effects cannot be ignored. The work suggests that further long term studies are needed to confirm nimesulide as a safe drug.