Journal Staff

Adenovirus encodes for the pVII protein, which interacts and modulates virus DNA structure in the infected cells. The pVII protein is synthesized as the precursor protein and undergoes proteolytic processing by viral proteinase Avp, leading to release of a propeptide sequence and accumulation of the mature VII protein. Here we elucidate the molecular functions of the propeptide sequence present in the precursor pVII protein. The results show that the propeptide is the destabilizing element targeting the precursor pVII protein for proteasomal degradation. Our data further indicate that the propeptide sequence and the lysine residues K26 and K27 regulate the precursor pVII protein stability in a co-dependent manner. We also provide evidence that the Cullin-3 E3 ubiquitin ligase complex alters the precursor pVII protein stability by association with the propeptide sequence. In addition, we show that inactivation of the Cullin-3 protein activity reduces adenovirus E1A gene expression during early phase of virus infection. Collectively, our results indicate a novel function of the adenovirus propeptide sequence and involvement of Cullin-3 in adenovirus gene expression

Implementing Test Automation with Selenium WebDriver

Many software programs, such as applications for designing, modeling, simulating, and analyzing systems, are now commonly available as web-based applications. The testing of such sophisticated web applications is highly challenging and can be extremely tedious and error-prone if done manually. Recently automation tools have become increasingly used for testing web-based applications, as they minimize human involvement and repetitive work. For this problem report project, we have built and implemented an automation testing framework for web applications. The project specifically uses a tool called Selenium WebDriver, which has been used to develop the testing framework. By using this framework, testers may quickly and effectively write their test cases. The benefits of Selenium WebDriver include that it does not require in-depth research and training by testers, and due to the framework\u27s ability to take screenshots, it provides a useful way for developers to study their code. The framework relies on the Chrome web browser, along with Java running in Eclipse, to provide a user-friendly interface for constructing and running test suites. To validate the testing framework, we performed a case study involving NanoHub (nanoHUB.org), which is a well-known platform that provides valuable resources for those involved in nanotechnology research and education. NanoHub serves as an open-access repository for a wide range of tools, simulations, and information related to nanoscale science and engineering, and it is designed particularly to model and simulate electronic systems and nanoscale phenomena. Testing a website such as NanoHub.org typically encompasses a blend of functional testing, usability testing, and performance testing. Based on the results of this testing, several observations are made about the testing framework in general, and its application to NanoHub in particular. The comprehensive testing approach documented in this report is aimed at ensuring the platform functions as intended, provides a user-friendly experience, and delivers optimal performance. This testing is particularly crucial when dealing with tools and simulations related to electronic systems

A sensitive bioanalytical method development and validation of cabozantinib in human plasma by LC-ESI-MS/MS

A simple, sensitive and specific liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of cabozantinib (CZ) in human plasma using cabozantinib-d4 (CZD4) as an internal standard (IS). Chromatographic separation was performed on Xbridge C18, 50 x 4.6 mm, 5 mm column with an isocratic mobile phase composed of 10mM Ammonium formate and Methanol in the ratio of (20:80 v/v), at a flow-rate of 0.7 mL/min. CZ and CZD4 were detected with proton adducts at m/z 502.2 → 391.1 and 506.3 → 391.2 in multiple reaction monitoring (MRM) positive mode respectively. Liquid-Liquid extraction method was used to extract the drug and IS. The method was validated over a linear concentration range of 5.0-5000.0 pg/mL with correlation coefficient (r2 ) ≥ 0.9994. This method demonstrated intra and inter-day precision within 1.95 to 2.37 and 2.93 to 9.3 % and Accuracy within 101.4 to 102.4 and 99.5 to 104.8 %. Cabozantinib was found to be stable throughout freeze-thawing cycles, bench top and postoperative stability studies

PHARMACOKINETIC INVESTIGATION OF REMOGLIFLOZIN IN RAT PLASMA SAMPLES BY HIGH-THROUGHPUT HPLC-MS-MS

Objective: Remogliflozin (REMO), a selective inhibitor of the renal sodium-dependent glucose transporter 2 channel, which could increase urine glucose excretion and lower plasma glucose in humans. To establish a simple, sensitive and completely validated HPLC-MS-MS approach for the analysis of Remogliflozin in rat plasma samples. Methods: The method was developed after simple step protein precipitation by acetonitrile and Empagliflozin (EMPA) was used as internal standard. Separation was done on an CORTECS C18, 90 Å, 2.7 µm, 4.6 mm X 150 mm with an isocratic mobile phase consisting of 0.1% Formic acid: acetonitrile (20:80%, v/v) and pumped at a flow stream of 0.8 ml/min at ambient temperature. Results: The approach developed showed fine calibration curve in the quantity range of 5-1000 pg/ml with correlation coefficient (r2) of ≥ 0.9997 and the intra-run accuracy and precision was 99.91 to 109.07% and 0.17 to 1.34, inter-run accuracy and precision was 99.8 to 101.54 and 0.17 to 1.66 according to FDA guidelines. Conclusion: The newly designed and validated approach was simple, fast and applied effectively for single-dose oral pharmacokinetic investigation in Wistar male rats for the quantification of REMO in biological matrix

Trial of ORG 10172 in acute stroke treatment classification and associated risk factors of ischemic stroke: a prospective study from a tertiary care center in South India

Background: Stroke is the leading cause of acquired disability worldwide. Better understanding of risk factors helps to reduce the community burden of stroke. Aim of the study was to determine pattern of ischemic stroke subtype and associated risk factors.Methods: Of 220 patients with first ever stroke, 168 patients of ischemic stroke were enrolled prospectively from January 2016 to May 2016 at ESIC Superspeciality hospital, Hyderabad. Patients were categorized in accordance with the Trial of Org 10172 in Acute Stroke treatment (TOAST) criteria.Results: Of 168 patients, 110 (65.4%) were males; male to female ratio was 1.8:1. The mean age was 53.912.3 years. Small vessel occlusion (SVO) occurred in younger age group as compared to other stroke subtypes. Frequency of large-artery atherosclerosis (LAA) (46.4%) was highest (Extracranial: 41.1%, intracranial: 36.9%), followed by SVO (32.4%), cardio embolism (15.4%), undetermined (4.7%) and other determined causes (1.1%). Risk factors included dyslipidemia (79%), hypertension (67.8%), smoking (58.3%), alcohol use (54.7%), diabetes (40.4%) and physical inactivity (27.3%). Common association observed was smoking (P - 0.004) and alcohol (P - 0.003) with LAA and SVO, and dilated cardiomyopathy with cardio embolism.Conclusions: LAA was observed frequently than SVO. Frequency of SVO was lower than Asian but higher than western, while cardio embolism was lower than Western and similar to Asian stroke studies. Dyslipidemia was the predominant risk factor than hypertension reported from Asian and Western stroke registry. This unique pattern can be attributed to differences in demographic and risk factor profiles. The study confirms the need for vigorous primary and secondary prevention measures targeting modifiable risk factors of stroke

Blood lipid levels, statin therapy and risk of intracerebral hemorrhage versus ischemic vascular events: a prospective case control study from tertiary care center of south India

Background: Intracerebral hemorrhage (ICH) is characterized by high mortality and morbidity. A little is known about the association between blood lipids, statin use and risk of ICH. Objective of the study was to investigate the relation between blood lipid levels and risk of hemorrhagic stroke.Methods: Prospectively compared the lipid levels of primary ICH patients (case) with ischemic group (control) patients, i.e. age and sex matched individuals admitted from January 2014 to January 2015 and outcome analyzed.Results: Of the 678 acute stroke patients, 78 (11.5%) had ICH who was enrolled. Mean age was 53±14.4. ICH was frequent in older age (57.6%) with male gender predominance (73%). Most frequent location of bleed was in thalamus (30.7%). Low density lipoprotein (LDL), triglyceride (TG) and very low density lipoprotein (VLDL) cholesterol were significantly low in ICH patients compared to controls. There was no significant difference in the high density lipoprotein (HDL) levels in both groups. Mean total cholesterol was significantly low in a subset of study group that included male gender, younger onset stroke (<50 years) and with prior history of hypertension. Subgroup analysis in ICH group showed significantly low mean total cholesterol, LDL and TG cholesterol in statin group compared to non-statin group.Conclusions: Lower blood lipid levels are associated with an increased risk of ICH. The reduction of blood lipids due to statin therapy might increase the risk of ICH, especially in hypertensive individuals and those with alcohol use; hence there should be a cautious use of statins

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE DETERMINATION OF BAMIFYLLINE HYDROCHLORIDE IN TABLET DOSAGE FORM

Objective: The objective of the current study was to develop and validate a novel RP-HPLC method for determination of bamifylline hydrochloride in pharmaceutical dosage form.Methods: Chromatographic separation was conducted on Agilent technologies-1260 series with the G1311C quaternary pump, eclipse XDB C18 column (4.6 mm i.d. X 250 mm, 5 µm particle sizes) and equipped with photodiode array detector G1315D. Mobile phase consisted of methanol and acetonitrile were mixed in the ratio of 90:10 v/v, was used at a flow rate of 1 ml/min and detection wavelength was set at 263 nm.Results: The retention time for bamifylline hydrochloride was found to be 2.913 min. The calibration was linear (r2= 0.9996) in the concentration range of 2-10 µg/ml. The limit of detection and the limit of quantitation were found to be 0.4825 μg/ml and 1.4621 µg/ml respectively. Recovery of bamifylline hydrochloride in tablet formulation was observed in the range of 99.6-99.8 %. Percentage assay of bamifylline hydrochloride (Bamifix) was found to be 99.4 % w/w.Conclusion: Thus the novel proposed method for bamifylline hydrochloride was found to be feasible for the estimation of bamifylline hydrochloride in bulk as well as a pharmaceutical dosage form.Â

DESIGN, SYNTHESIS, IN VITRO ANTIOXIDANT AND IN VIVO ANTI-INFLAMMATORY ACTIVITIES OF NOVEL OXADIAZOLE DERIVATIVES

Objective: In the present study, a series of novel 1,3,4-oxadiazole derivatives (3a-3q) were designed, synthesized and evaluated for antioxidant and anti-inflammatory activities.Methods: The title compounds were designed and docked onto the COX-2 enzyme (3LN1) protein using SYBYLX 2.1. 2-substituted-5-(5-nitrobenzofuran-2-yl)-1,3,4-oxadiazole derivatives (3a-3p) were synthesized from acid catalyzed dehydrative cyclization of 5-nitrobenzofuran-2-carbohydrazide (2) with various heteroaryl/aryl/aliphatic carboxylic acid derivatives. And 5-(5-nitrobenzofuran-2-yl)-1,3,4-oxadiazole-2-thiol (3q) was synthesized on reacting the hydrazide derivative 2 with carbon disulfide. The synthesized compounds were evaluated for in vitro antioxidant property by DPPH radical scavenging assay method and in vivo anti-inflammatory activity by carrageenan induced paw edema method.Results: The synthesized 1,3,4-oxadiazole derivatives (3a-3q) were characterized on the basis of LCMS, 1HNMR [13]CNMR, IR and elemental analysis. The title compounds 3a-3q exhibited significant antioxidant efficacy ranging from 34 to 86%and the results of anti-inflammatory evaluation revealed that compounds 3c, 3e and 3d exhibited substantial anti-inflammatory activity of 72, 68 and 65%, respectively, at a dose of 50 mg kg-1.Conclusion: A significant correlation was observed between the in silico study and the anti-inflammatory results. The anti-inflammatory results highlight the synthesized compounds 3c, 3e and 3d could be considered as possible hit as therapeutic agents.Â

3D-QSAR AND MOLECULAR DOCKING STUDIES ON 1, 2, 4 TRIAZOLES AS MetAP2 INHIBITORS

Objective: Angiogenesis inhibitors are a novel class of promising therapeutic agents for treating cancer and other human diseases. Biological transformations and pathways that play a role in angiogenesis are, therefore, particularly attractive targets as potential methods for inhibiting solid tumors. MetAP2 is of particular interest because the enzyme plays a key role inÂ&nbsp;angiogenesis, the growth of new blood vessels, which is necessary for the progression of diseases including solid tumorÂ&nbsp;cancersÂ&nbsp;and rheumatoid arthritis. In this paper we report the quantitative structure activity relationship and docking studies of 1, 2, 4 triazole derivatives for designing novel MetAP2 inhibitors. Methods: Tripos Sybyl X 2.1 program was used to conduct docking based CoMFA, CoMSIA and Topomer CoMFA QSAR modeling for a dataset of 77 triazoles. Results: The CoMFA, CoMSIA and Topomer CoMFA models demonstrated good statistical results with cross-validated coefficient (q2) of 0.703, 0.704, 0.746 and correlation coefficient (r2) of 0.894, 0.889, 0.886 respectively and these models have been externally validated. Conclusion: Based on the statistical results obtained from the above model, the CoMFA, CoMSIA and Topomer CoMFA model can be utilized to design new molecules having 1, 2, 4 triazoles as common core with significant MetAP2 inhibitory activity

Prevalence, pattern, risk factors and outcome of stroke in women: a clinical study of 100 cases from a tertiary care center in South India

Background: Stroke is the leading cause of acquired disability and the third leading cause of death in women worldwide. There had been relatively few studies of stroke in women. Objective of the study was to study the prevalence, patterns, risk factors and outcome of stroke in women. A cross sectional study with case control comparison and prospective follow up at one month at ESIC Super speciality hospital, Hyderabad in South India.Methods: Total 100 stroke patients were identified over a period of 3 months and data collected on the basis of clinical proforma developed for the purpose.Results: Of 100 stroke patients, 31 were females. Ischemic stroke was seen in 25 (80.64%) females. Age was an important non-modifiable risk factor for stroke. Stroke was predominant among older women 23 (74.19%).  Mean age of stroke in females was 57 years. Menopause 27 (87.09%) was the predominant risk factor followed by hypertension in 25 (80.6%), dyslipidemia in 19 (70.3%) physical inactivity in 17 (54.8%) and diabetes in 12 (38.7%) females respectively. Majority of females 27 (87.09%) were uneducated as compared to males 27 (39.13%). The overall mortality in females was 3 (9.6%) as compared to males 3 (4.3%). 25 (89.2%) of females were ambulatory when compared to males 62 (93.9%). Conclusion: Stroke was common in older women and ischemic stroke was the predominate type of stroke.  Physical   inactivity was the significant risk factor in women when compared to men. Women are more likely to be disabled after stroke than men.