Isolation and Characterization of an Unknown Process-Related Impurity in Furosemide and Validation of a New HPLC Method.

Furosemide is a widely used loop diuretic in the treatment of congestive heart failure and edema. During the preparation of furosemide, a new process-related impurity G in the levels ranging from 0.08% to 0.13% was detected in pilot batches by a new high performance liquid chromatography (HPLC) method. The new impurity was isolated and characterized by comprehensive analysis of FT-IR, Q-TOF/LC-MS, 1D-NMR (1H, 13C, and DEPT), and 2D-NMR (1H-1H-COSY, HSQC, and HMBC) spectroscopy data. The possible formation pathway of impurity G was also discussed in detail. Moreover, a novel HPLC method was developed and validated for the determination of impurity G and the other six known impurities registered in the European Pharmacopoeia as per ICH guidelines. The HPLC method was validated with respect to system suitability, linearity, the limit of quantitation, the limit of detection, precision, accuracy, and robustness. The characterization of impurity G and the validation of its quantitative HPLC method were reported for the first time in this paper. Finally, the toxicological properties of impurity G were predicted by the in silico webserver ProTox-II.

Effect of Co-Treatment of Olanzapine with SEP-363856 in Mice Models of Schizophrenia.

Olanzapine is a commonly used drug in the treatment of schizophrenia, but its clinical application has been restricted by metabolic-related side effects. In order to mitigate the weight gain side effects caused by olanzapine, other drugs with different targets were selected for combined use and evaluated in animal models of schizophrenia. SEP-363856 is a novel psychotropic agent which is under phase III clinical trials for schizophrenia treatment. The aim of the research was to evaluate whether co-administration of olanzapine and SEP-363856 exerts synergistic anti-schizophrenic effects in the apomorphine (APO)-induced climbing test, the MK-801-induced hyperactivity test, and the Morris water maze test, and therefore reduces the weight gain side effects induced by olanzapine. Through isobolographic analysis, the results showed a synergistic interaction in the climbing test; the experimental ED30 (3 mg/kg) was significantly smaller (p < 0.05) than the theoretical ED30 (5 mg/kg). Additionally, such potentiating effects appeared additive in the MK-801 challenge experiment. Co-treatment with an effective dose of olanzapine and a low dose of SEP-363856 reversed MK-801-induced cognitive impairment symptoms in mice. Moreover, combination treatment with olanzapine and SEP-363856 controls sustained weight gain in mice with chronic exposure to olanzapine. These results support further clinical trials to test the effectiveness of co-treatment of olanzapine and SEP-363856 for controlling symptoms and weight gain in patients with schizophrenia during antipsychotic treatments.

An evaluation of cascading mentorship as advocacy training in undergraduate medical education.

BACKGROUND: Physicians are in a position of great influence to advocate for health equity. As such, it is important for physicians-in-training to develop the knowledge and skills necessary to fulfil this role. Although various undergraduate medical programs have implemented health advocacy training, they often lack experiential learning and physician involvement. These aspects are foundational to the Advocacy Mentorship Initiative (AMI) which utilizes cascading mentorship as a novel approach to advocacy training. Medical students develop advocacy competency as peer mentors to youth raised in at-risk environments, while also being mentored themselves by physician residents. We aim to determine whether there are specific advantages to utilizing cascading mentorship to facilitate the attainment of advocacy competencies in undergraduate medical education. METHODS: Medical students participating in AMI between 2017 to 2020 completed pre- and post-exposure questionnaires. Questionnaires assessed confidence in advocacy-related skills and knowledge of youth advocacy concepts, as well as learning goals, skills gained, benefits of AMI and resident mentors, and impact on future career. Sign tests were utilized to analyze quantitative results, and content analysis was used for open-ended responses. A triangulation protocol was also utilized. RESULTS: Fifty mentors participated, 24 (48%) of which completed both pre- and post-exposure questionnaires. Participants gained confidence in advocacy-related skills (p < 0.05) such as working with vulnerable populations and advocating for medical and non-medical needs. They also reported significant improvements (p < 0.01) in their understanding of social determinants of health and concepts related to children's health and development. Content analysis showed that participants built meaningful relationships with mentees in which they learned about social determinants of health, youth advocacy, and developed various advocacy-related skills. Participants greatly valued mentorship by residents, identifying benefits such as support and advice regarding relations with at-risk youth, and career mentorship. AMI impacted participants' career trajectories in terms of interest in working with youth, psychiatry, and advocacy. CONCLUSIONS: AMI offers a unique method of advocacy training through cascading mentorship that engages medical students both as mentors to at-risk youth and mentees to resident physicians. Through cascading mentorship, medical students advance in their advocacy-related skills and understanding of social determinants of health.

GYS1 or PPP1R3C deficiency rescues murine adult polyglucosan body disease.

OBJECTIVE: Adult polyglucosan body disease (APBD) is an adult-onset neurological variant of glycogen storage disease type IV. APBD is caused by recessive mutations in the glycogen branching enzyme gene, and the consequent accumulation of poorly branched glycogen aggregates called polyglucosan bodies in the nervous system. There are presently no treatments for APBD. Here, we test whether downregulation of glycogen synthesis is therapeutic in a mouse model of the disease. METHODS: We characterized the effects of knocking out two pro-glycogenic proteins in an APBD mouse model. APBD mice were crossed with mice deficient in glycogen synthase (GYS1), or mice deficient in protein phosphatase 1 regulatory subunit 3C (PPP1R3C), a protein involved in the activation of GYS1. Phenotypic and histological parameters were analyzed and glycogen was quantified. RESULTS: APBD mice deficient in GYS1 or PPP1R3C demonstrated improvements in life span, morphology, and behavioral assays of neuromuscular function. Histological analysis revealed a reduction in polyglucosan body accumulation and of astro- and micro-gliosis in the brains of GYS1- and PPP1R3C-deficient APBD mice. Brain glycogen quantification confirmed the reduction in abnormal glycogen accumulation. Analysis of skeletal muscle, heart, and liver found that GYS1 deficiency reduced polyglucosan body accumulation in all three tissues and PPP1R3C knockout reduced skeletal muscle polyglucosan bodies. INTERPRETATION: GYS1 and PPP1R3C are effective therapeutic targets in the APBD mouse model. These findings represent a critical step toward the development of a treatment for APBD and potentially other glycogen storage disease type IV patients.

The Impact of Mobile Apps on Alcohol Use Disorder: A Systematic Review Protocol.

BACKGROUND: Alcohol use disorder (AUD) is among the most prevalent mental disorders worldwide and is associated with a diverse range of physical and psychological comorbidities. Despite various types of treatment, there are many barriers to accessing treatment (ie, stigma, cost, accessibility of service, etc). Mobile apps have the potential to overcome these barriers and provide support to those who need it. OBJECTIVE: The purpose of this systematic review is to assess the effectiveness of mobile apps in reducing alcohol consumption for individuals with AUD and understand the psychological outcomes of using the apps (ie, client empowerment, self-efficacy, etc). METHODS: The search strategy was applied to 7 health sciences and interdisciplinary databases. Two reviewers will independently assess all titles and abstracts for relevance and then full texts of relevant articles for eligibility. To be included, the article must be a quantitative evaluation of clinical outcomes using the intervention and the intervention must be a consumer-facing app focused on supporting individuals with AUD. Two reviewers will independently extract data from all eligible articles using a standardized extraction worksheet and will independently assess the study quality. A meta-analysis will be conducted if appropriate. Depending on outcomes reported, pooled risk ratios or standardized mean differences will be calculated and reported in the review. RESULTS: The search strategy yielded 699 unique citations. Of those, 63 (9.0%, 63/699) articles were assessed as relevant for full-text review. The full-text reviews are currently underway and the final review is projected to be completed in the summer of 2017. CONCLUSIONS: There is potential for mobile apps to support individuals with AUD to reduce their alcohol consumption. This review will be the first to assess the effectiveness of AUD mobile apps and client experiences using the apps.

SGK1 (glucose transport), dishevelled2 (wnt signaling), LC3/p62 (autophagy) and p53 (apoptosis) proteins are unaltered in Lafora disease.

Glycogen forms through the concerted actions of glycogen synthase (GS) which elongates glycogen strands, and glycogen branching enzyme (GBE). Lafora disease (LD) is a fatal neurodegenerative epilepsy that results from neuronal accumulation of hyperphosphorylated glycogen with excessively long strands (called polyglucosans). There is no GBE deficiency in LD. Instead, the disease is caused by loss-of-function mutations in the EPM2A or EPM2B genes, encoding, respectively, a phosphatase, laforin, and an E3 ubiquiting ligase, malin. A number of experimentally derived hypotheses have been published to explain LD, including: The SGK1 hypothesis - Phosphorylated SGK1 (pSGK1) raises cellular glucose uptake and levels, which would activate GS. Based on observing increased pSGK1 in LD mice it was proposed that raised pSGK1 leads to polyglucosan generation through GS hyperactivation. The Dishevelled2 hypothesis - Downregulating malin in cell culture was reported to increase levels of dishevelled2, which through the wnt/glycogen synthase kinase-3 pathway would likewise overactivate GS. The Autophagic defect hypothesis - Polyglucosans may be natural byproducts of normal glycogen metabolism. LD mice were reported to be autophagy-defective. LD would arise from failed autophagy leading to failed polyglucosan clearance. Finally, the p53 hypothesis - laforin and malin were reported to downregulate p53, their absence leading to increased p53, which would activate apoptosis, leading to the neurodegeneration of LD. In the present work we repeat key experiments that underlie these four hypotheses. We are unable to confirm increased pSGK1, dishevelled2, or p53 in LD mice, nor the reported autophagic defects. Our work does not support the above hypotheses in understanding this unique and severe form of epilepsy.