Validation of an LC-MS/MS method for quantitation of fostemsavir in plasma.

BACKGROUND: A novel, sensitive and specific LC-MS/MS technique was developed and validated for the quantification of fostemsavir in human plasma and its pharmacokinetic application in rabbits. METHODS: Chromatographic separation of the fostemsavir and fosamprenavir (internal standard) were achieved on Zorbax C18 (50 mm × 2 mm × 5 µm) column with 0.80 mL/min flow rate and coupled with API6000 triple quadrupole MS in multi reaction monitoring mode by applying mass transitions m/z 584.16/105.03 for fostemsavir and m/z 586.19/57.07 for the internal standard. RESULTS: The calibration curve exhibited linearity in concentration range of 58.5-2340.0 ng/mL for fostemsavir. The LLOQ was 58.5 ng/mL. The validated LC-MS/MS process was effectively applied for the analysis of plasma in healthy rabbits for determinations of Fostemsavir. From the pharmacokinetic data, the mean of Cmax and Tmax were 198.19 ± 5.85 ng/mL and 2.42 ± 0.13, respectively. Plasma concentration reduced with t1/2 of 7.02 ± 0.14. AUC0→Last value obtained was 2374.87 ± 29.75 ng. h/ml, respectively. CONCLUSION: In summary, the developed method has been successfully validated and pharmacokinetic parameters were demonstrated after oral administration of Fostemsavir to healthy rabbits.

Cardioprotective Effect of Tangeretin by Inhibiting PTEN/AKT/mTOR Axis in Experimental Sepsis-Induced Myocardial Dysfunction.

Sepsis aggregates undesirable immune response causing depression of ventricular myocardium and diastolic dysfunction. This present study examined the effect of a plant-derived flavone tangeretin (TG) on autophagy and reduction in myocardial dysfunction. The sepsis was induced by cecum ligation and puncture (CLP) in male Sprague-Dawley rats. Abnormal changes were seen in the heart after the sepsis induction. These abnormalities were analyzed based on the cardiac markers, namely Cardiac myosin light chain-1 (cMLC1) and Cardiac troponin I (cTnl), echocardiography, and plasma parameters, like Lactate dehydrogenase (LDH) and Creatinine kinase (CK). Microanatomy of the heart was studied using hematoxylin and eosin stained histopathological samples of cardiac tissue. Western blot technique was used to detect the nature and extent of protein with the amount of a specific RNA (gene expression) in the cardiac homogenate. Oxidative damage was analyzed using redox marker, reduced glutathione. This study successfully showed that TG attenuated sepsis-induced myocardial dysfunction by inhibiting myocardial autophagy via silencing the Phosphatase and tensin homolog (PTEN) expression and acting on the AKT/mTOR pathway. The present findings supported that TG is a novel cardioprotective therapeutic target for sepsis induced myocardial dysfunction.

Apigenin-Loaded Solid Lipid Nanoparticle Attenuates Diabetic Nephropathy Induced by Streptozotocin Nicotinamide Through Nrf2/HO-1/NF-kB Signalling Pathway.

BACKGROUND: Apigenin is known to have a broad-spectrum efficacy in oxidative stress and conditions due to inflammation, although weak absorption, fast metabolic rate and a fast elimination (systemic) limit the pharmacological efficacy of this drug. Hence, we propose the usage of highly bioavailable Apigenin-solid lipid nanoparticles (SLNPs) to recognize such limitations. The defensive function of Apigenin-SLNPs on renal damage induced by streptozotocin (STZ) in animals was studied. MATERIALS AND METHODS: We initially injected the rats with 35 mg kg-1 streptozocin intraperitoneally, and after 7 days, the rats were then injected 150 mg kg-1 of metformin intragastrically followed by a once-daily intragastric dose of Apigenin-SLNP (25 or 50 mg kg-1) for a continuous period of 30 days. We then measured the level of insulin and blood glucose, superoxide dismutase, catalase and malondialdehyde in the tissues of the kidney. We also observed messenger-RNA expression of Interleukin-1ß, Interleukin-6 and Tumor Necrosis Factor-alpha in renal tissue through RT-PCR technique. Moreover, H&E staining and Western blotting observed the histopathological variations and protein expression of nuclear factor erythroid 2-related factor 2/heme oxygenase/Nuclear Factor-κB signaling pathway, respectively. RESULTS: An enhancement in the expressing of nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 and a suppression in the expression of Nuclear Factor-κB occurred due to Apigenin-SLNPs treatment, which was a result of the protective mechanism of Apigenin-SLNPs which is because of not only its anti-inflammatory function (by inhibition of release of inflammatory factors) but also their anti-oxidant activity (through reduction of lipid peroxidation production). CONCLUSION: We found that a protective effect on diabetic nephropathy was shown due to Apigenin-SLNPs, in rats induced with streptozocin maybe through the pathway of nuclear factor erythroid 2-related factor 2/heme oxygenase-1/Nuclear Factor-κB.

Evaluation of self-reported medication adherence and its associated factors among epilepsy patients in Hospital Kuala Lumpur.

INTRODUCTION: Reports on medication adherence and its associated factors in patients with epilepsy in South East Asian countries are lacking. The primary purpose of this study was to assess the degree of medication adherence and its relationship with patient's satisfaction, psychosocial factors, quality of life and mental health in a sample of Malaysian epilepsy patients. METHODOLOGY: It is a cross-sectional study and was carried out in the outpatient Neurology Department of Hospital Kuala Lumpur, Malaysia (n=272). Data was collected by administering the structured questionnaire. RESULTS AND DISCUSSION: Results showed that 49.3% of the epilepsy patients were non-adherent to their prescribed regimen. Univariate analysis showed significant associations between medication adherence and the following factors: race, seizure frequency, overall patient satisfaction, medication taste and smell, medication cost and physical appearance, medication effectiveness, complexity of medication regimen, patient barrier, patient understanding, patient role functioning, patient positivity, vitality and general interest. Multiple regression analysis indicated that factors that are influencing medication adherence are seizure frequency (P = 0.048), overall patient satisfaction (P = 0.043) and patient understanding about their illness (P = 0.001). The model chosen for testing the relationship between medication adherence and its associated factors give an R2 value of 25.2% with an adjusted R2 of 21.4%. The F value was also significant (P = 0.000). Based on the research findings, the researchers recommends that clinicians need to play a vital role in educating the patients on their disease conditions. By educating the patients on nature of epilepsy, different modalities of treatment and benefits of adherence to treatment will help in the better adherence and management.

Effectiveness of flipped classroom with Poll Everywhere as a teaching-learning method for pharmacy students.

OBJECTIVES: Flipped classroom (FC) is a pedagogical model to engage students in learning process by replacing the didactic lectures. Using technology, lectures are moved out of the classroom and delivered online as means to provide interaction and collaboration. Poll Everywhere is an audience response system (ARS) which can be used in an FC to make the activities more interesting, engaging, and interactive. This study aims to study the perception of undergraduate pharmacy students on FC activity using Poll Everywhere ARS and to study the effectiveness of FC activity as a teaching-learning tool for delivering complementary medicine module in the undergraduate pharmacy program. MATERIALS AND METHODS: In this nonrandomized trial on interrupted time series study, flipped class was conducted on group of 112 students of bachelor of pharmacy semester V. The topic selected was popular herbal remedies of the complementary medicine module. Flipped class was conducted with audio and video presentation in the form of a quiz using ten one-best-answer type of multiple-choice questions covering the learning objectives. Audience response was captured using web-based interaction with Poll Everywhere. Feedback was obtained from participants at the end of FC activity and debriefing was done. RESULTS: Randomly selected 112 complete responses were included in the final analysis. There were 47 (42%) male and 65 (58%) female respondents. The overall Cronbach's alpha of feedback questionnaire was 0.912. The central tendencies and dispersions of items in the questionnaire indicated the effectiveness of FC. The low or middle achievers of quiz session (pretest) during the FC activity were three times (95% confidence interval [CI] = 1.1-8.9) at the risk of providing neutral or negative feedback than high achievers (P = 0.040). Those who gave neutral or negative feedback on FC activity were 3.9 times (95% CI = 1.3-11.8) at the risk of becoming low or middle achievers during the end of semester examination (P = 0.013). The multivariate analysis of "Agree" or "Disagree" and "Agree" or "Strongly Agree" was statistically significant. CONCLUSION: This study provides insight on how the pharmacy students learn and develop their cognitive functions. The results revealed that the FC activity with Poll Everywhere is an effective teaching-learning method.

Protective effect of pioglitazone, a PPARγ agonist against acetaminophen-induced hepatotoxicity in rats.

Acetaminophen has a reasonable safety profile when consumed in therapeutic doses. However, it could induce hepatotoxicity and even acute liver failure when taken at an overdose. Pioglitazone, PPARγ ligand, is clinically tested and used in treatment of diabetes. PPARγ is a key nuclear hormone receptor of lipid metabolisms and regulates several gene transcriptions associated with differentiation, growth arrest, and apoptosis. The aim of our study was to evaluate the hepatoprotective activity of pioglitazone on acetaminophen-induced hepatotoxicity and to understand the relationship between the PPARγ and acetaminophen-induced hepato injury. For the experiment, Sprague-Dawley rats (160-180 g) were used and divided into four groups. Groups I and II were normal and experimental controls, respectively. Groups III and IV received the pioglitazone 20 mg/kg for 10 days. Hepatotoxicity was induced in Groups II and III on the eighth day with acetaminophen (i.p. 350 mg/kg body weight). The hepatoprotective effect was evaluated by performing an assay of the total protein, total bilirubin, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and α-fetoprotein as well as glutathione peroxidase, lipid peroxidation, catalase, superoxide dismutase, and glutathione transferase and liver histopathology. The assay results were presented as mean and standard error of mean for each group. The study group was compared with the control group by one-way ANOVA test. A p value of <0.05 was considered significant. Pioglitazone significantly reduced the elevated level of above serum marker enzymes and also inhibits the free radical formation by scavenging hydroxyl ions. It also restored the level of LPO and significantly elevated the levels of endogenous antioxidant enzymes in acetaminophen-challenged hepatotoxicity. Liver histopathological examination showed that pioglitazone administration antagonized acetaminophen -induced liver pathological damage. Various biochemical estimations of different hepatic markers and antioxidant enzymes and histopathological studies of liver tissues glimpse a support to its significant hepatoprotective activity on acetaminophen -induced hepatotoxicity.