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Limitations with cell cultures and experimental animal-based studies have had the scientific and industrial communities searching for new approaches that can provide reliable human models for applications such as drug development, toxicological assessment, and in vitro pre-clinical evaluation. This has resulted in the development of microfluidic-based cultures that may better represent organs and organ systems in vivo than conventional monolayer cell cultures. Although there is considerable interest from industry and regulatory bodies in this technology, several challenges need to be addressed for it to reach its full potential. Among those is a lack of guidelines and standards. Therefore, a multidisciplinary team of stakeholders was formed, with members from the US Food and Drug Administration (FDA), the National Institute of Standards and Technology (NIST), European Union, academia, and industry, to provide a framework for future development of guidelines/standards governing engineering concepts of organ-on-a-chip models. The result of this work is presented here for interested parties, stakeholders, and other standards development organizations (SDOs) to foster further discussion and enhance the impact and benefits of these efforts., QC 20240314

BACKGROUND: Over the past 40 years, the tasks of pharmacists have shifted from logistic services to pharmaceutical care (PhC). Despite the increasing importance of measuring quality of care, there is no general definition of Quality Indicators (QIs) to measure PhC. Recognising this, a working group in a European association of PhC researchers, the Pharmaceutical Care Network Europe (PCNE), was established in 2020. AIM: This research aimed to review existing definitions of QIs and develop a definition of QIs for PhC. METHOD: A two-step procedure was applied. Firstly, a systematic literature review was conducted to identify existing QI definitions that were summarised. Secondly, an expert panel, comprised of 17 international experts from 14 countries, participated in two surveys and a discussion using a modified Delphi technique to develop the definition of QIs for PhC. RESULTS: A total of 182 QI definitions were identified from 174 articles. Of these, 63 QI definitions (35%) cited one of five references as the source. Sixteen aspects that construct QI definitions were derived from the identified definitions. As a result of the Delphi study, the panel reached an agreement on a one-sentence definition of QIs for PhC: "quality indicators for pharmaceutical care are validated measurement tools to monitor structures, processes or outcomes in the context of care provided by pharmacists". CONCLUSION: Building upon existing definition of QIs, an international expert panel developed the PCNE definition of QIs for PhC. This definition is intended for universal use amongst researchers and healthcare providers in PhC.

Tillgång till ett brett sortiment av antibiotika är en nödvändighet för att bedriva en modern sjukvård och bromsa utvecklingen av antibiotikaresistensen. Den europeiska antibiotikamarknaden har sjunkit med ca 40% under perioden 2010-2021. Låga priser på antibiotikaförpackningar har bidragit till att antibiotikaföretagen inte anser det är lönsamt att ha antibiotikaprodukter registrerade i Sverige vilket leder till avregistrering. Läkemedelsverkets målsättning är att säkerställa en fortsatt tillgång till antibiotika och undvika att antibiotikaprodukter som Hälso- och sjukvården behöver avregistreras. Indikatorer som påvisar vilka antibiotikaprodukter som löper risk att avregistreras skulle kunna vara ett verktyg för Läkemedelsverket. Syftet med examensarbetet är att identifiera indikatorer som påvisar vilka antibiotikaprodukter som löper risk att avregistreras. Data har samlats in från olika myndigheters databaser såsom Tandvårds- och läkemedelsförmånsverket, E-hälsomyndigheten samt Läkemedelsverket. Resultatet från analysen påvisar att utträde ur läkemedelsförmånen, takpris, år takpriset sattes, antal Market Authorization Holder/produkt, antal tillverkare av Active Pharmaceutical Ingredient/Market Authorization Holder för en produkt, marknadsföring, försäljningsvärde, samt pris/förpackning påvisar trend för ökad risk av avregistrering. Upprepade restanmälningar, längd av restanmälningar samt försäljningsvolym påvisar inte ökad risk för avregistrering. Lokalisationen av tillverkare av Active Pharmaceutical Ingredient påvisar ökad risk för avregistrering med en viss tveksamhet.

Objectives: To examine life-course models by investigating the roles of childhood and adult socioeconomic position (SEP) in longitudinal changes in a functional aging index. Method: Up to eight waves of testing, covering 25 years, were available from the Swedish Adoption/Twin Study of Aging: N = 654, intake age = 50-82. A two-slope latent growth curve model was applied to the data, and the impact of including childhood and adult SEP as covariates of the intercept (at age 70) and slopes (before and after age 70) was tested. Results: Both childhood and adult SEP contributed to the best-fitting model. Childhood SEP was significantly associated with intercept and Slope 1 (before age 70) of the latent growth curve model (p < .05). Association of adult SEP with Slope 2 (after age 70) trended toward significance (p < .10). There was a significant interaction effect of childhood and adult SEP on the intercept (p < .05). As a result, intercept at age 70 was highest and change after age 70 was fastest for those whose SEP decreased from childhood to adulthood. Conclusions: Both childhood and adult SEP impact change in functional abilities with age, supporting both critical period and social mobility models. The social environment is modifiable by policies at local, national, and international levels, and these policies need to recognize that early social disadvantage can have long-lasting health impacts.

Motor issues are frequently observed accompanying core deficits in autism spectrum disorder (ASD). Impaired motor behavior has also been linked to cognitive and social abnormalities, and problems with predictive ability have been suggested to play an important, possibly shared, part across all these domains. Brain imaging of sensory-motor behavior is a promising method for characterizing the neurobiological foundation for this proposed key trait. The present functional magnetic resonance imaging (fMRI) developmental study, involving children/youth with ASD, typically developing (TD) children/youth, and neurotypical adults, will investigate brain activations during execution and observation of a visually guided, goal-directed sequential (two-step) manual task. Neural processing related to both execution and observation of the task, as well as activation patterns during the preparation stage before execution/observation will be investigated. Main regions of interest include frontoparietal and occipitotemporal cortical areas, the human mirror neuron system (MNS), and the cerebellum.

The regulation of gene expression is a key mechanism that underlies all biological processes, from embryonic development to the onset and progression of various diseases, including cancer. A growing body of evidence places RNA molecules at the center of critical regulatory steps in gene expression. They serve not only as intermediate molecules between DNA and proteins but also act as regulators of processes such as alternative splicing (AS) and translation, among others. This thesis focuses on the role of RNA in gene expression regulation. Specifically, it addresses how intrinsic properties of RNA, RNA chemical modifications, and RNA binding proteins (RBPs) can control gene expression regulatory processes. The first part tackles specific aspects of AS in neurodifferentiation. Paper I shows how RBPs affect AS in mouse embryonic stem cells (ESCs). Within this work, we identified ZFP207, a known transcription factor (TF), as an RBP with a crucial role in modulating the AS of key transcripts for neurodifferentiation. Depletion of ZFP207 in mouse ESCs led to abnormal AS patterns and a differentiated cell phenotype. The second part (Papers II-IV) focuses on the role of RNA modifications in disease. In Paper II, the publicly available literature linking deregulations of RNA modifications and their regulatory proteins with different diseases was curated. The obtained information was integrated into the 2021 update of the MODOMICS database, the most extensive RNA modifications database to date. Papers III and IV exemplify how two different RNA marks contribute to breast cancer. Paper III shows how METTL3, the enzyme responsible for N6-methyladenosine (m6A) deposition on messenger RNA (mRNA), affects tumorigenesis by modulating AS. METTL3-mediated AS regulation can be done either by depositing m6A at the intron-exon junctions of specific transcripts or on transcripts encoding for splicing and transcription factors, such as MYC. Changes in RNA modifications of ribosomal RNA (r

Bioequivalence (BE) studies are conducted to evaluate if drugs with different formulations have the same effect. In this study, whole versus crushed tablets were evaluated with three methods to examine if the formulations were BE. These methods were noncompartmental analysis (NCA), standard model-based (SMB) and individual model averaging (IMA). NCA is a commonly used method to evaluate PK metrics, even though it occasionally produces unreliable results. To improve the unreliability of NCA, a model-based method, SMB can be used to handle smaller sample sizes and carryover effects. SMB can allegedly increase study power, though it might increase type I error. A new model-based method called IMA has been shown in earlier simulation studies to reduce type I error caused by model misspecification. IMA uses mixture modelling to add a calculated probability of study subjects to be divided into different sub-models. The aim of the thesis was to extend the IMA method to the evaluation of real relative bioavailability studies using a model built from Automatic Model Development (AMD). The evaluation of BE using the NCA method was done with the raw data while SMB and IMA used the AMD software to produce the base model. Treatment effect was added on bioavailability and on the rate absorption constant to see if it had any effect. The study showed that the formulations were BE for the SMB method, but not for NCA and IMA. To draw a truthful conclusion, more permutations are needed to increase statistical significance and reduce bias.

Bakgrund: Infertilitet påverkar många par globalt. Orsaken kan vara mångfaldig och inkluderar både kvinnliga och manliga faktorer såsom hormonell obalans, ägglossningsrubbningar, endometrios eller problem med spermiekvaliteten och produktion. För par med infertilitet finns det fler alternativ till behandling där kontrollerad ovariestimulering (COS), in vitro fertilisering (IVF) och intracytoplasmatisk spermieinjektion (ICSI) är några av de. Vilken behandling som rekommenderas beror på grunden till infertiliteten men behandlingarna omfattar oftast någon form av kontrollerad ovariestimulering med gonadotropiner. Gonadotropiner är hormoner som stimulerar äggstockarna hos kvinnor för att ett flertal ägg ska utvecklas. Två vanliga alternativ av gonadotropiner är rekombinant follikelstimulerande hormon (r-FSH) och högrenat humant menopausalt gonadotropin (HP-hMG), där r-FSH är en syntetisk version av det naturligt förekommande FSH-hormonet, medan HP-hMG innehåller en blandning av både FSH och LH som renats ur urin från postmenopausala kvinnor. Syfte: Examensarbetet syftade till att utvärdera effekten och säkerheten av högrenat menotropin jämfört med rekombinant FSH vid IVF-behandling av infertila kvinnor för att bedöma graviditetschanser och identifiera risker. Metod: Metoden involverade sökning och granskning av vetenskapliga studier på Pubmed. Sökordet "follicle stimulating hormone menotropin" genererade 1735 träffar. Efter att ha begränsat sökningen till artiklar publicerade efter 2000, fulltext samt randomiserade kontrollerade studier. Abstracts granskades för att inkludera studier med över 200 infertila deltagare utan diagnoser som polycystisk ovarialsyndrom (PCOS) eller endometrios. Fyra artiklar valdes för analys. Resultat: Samtliga studier som granskades i arbetet visade på att det inte fanns en statistiskt signifikant skillnad i pågående graviditetsfrekvens mellan de två behandlingsformerna oavsett om de administrerats i kombination med en gonadotropinfrisättand

Fever in children, assessment questions, and counseling in community pharmacies: a mystery shopping study Abstract Background The pharmacy staff's lack of knowledge and difficulties in asking key questions lead to incorrect medication recommendations or use. This issue may, for example, apply to fever medicine for children, which this study investigates. Aim The purpose of the study is to evaluate the advisory services of pharmacy personnel regarding fever treatment, by evaluating the use of key patient questions and the accuracy of the advice, with a special focus on fever in children. Method The survey assessed fever treatment advice at 54 pharmacies using mystery shopping, where researchers pose as customers with a scripted scenario to evaluate pharmacy staff conduct, followed by filling out a documentation form after each visit. The results were ultimately analyzed descriptively in Excel. Results The study's results based on the consultations in the 54 pharmacies, age was frequently asked (98%) but detailed questions and full dosing advice were often overlooked. Paracetamol was commonly recommended (78%), (e.g. all pharmacists n=12, pharmacy technicians n=6 gave this advice), and ASA was suggested in 43% of cases if specifically requested, despite not being a first recommendation. However, unclear what educational background 86% of these had. Conclusion This survey shows a gap between guidelines and actual pharmacy practices in managing children's fever, especially in asking specific questions and giving dosing advice. It highlights the critical need for improved, standardized training and resources like checklists and a shared database to ensure consistent and expert advice. There is also a tendency that when the education was unfamiliar less adequate advices were given.

Wihania somnifera L. (Dunal) är en ört med rötter inom den ayurvediska medicinen och dess användning sträcker sig nästan 3000 år tillbaka i tiden. Den kallas även ashwagandha och har fått ökad uppmärksamhet för sina potentiella hälsofördelar. Den anses vara en adaptogen med förmåga att hjälpa kroppen att hantera stress och främja återhämtning, tack vare sina huvudsakliga aktiva föreningar, withanolider. Traditionellt har ashwagandha använts för att lindra stress och främja välbefinnande. Kortisol är ett stresshormon frisatt av binjurarna, som spelar en avgörande roll i kroppens stressrespons. Kronisk exponering för höga nivåer av kortisol kan leda till negativa hälsokonsekvenser. Därför syftade detta litteraturarbete till att undersöka ashwagandhas potentiella effekter på kortisolnivåerna hos människor. Detta undersöktes genom en litteraturanalys av fyra randomiserade, dubbelblinda, placebokontrollerade studier som hittades genom databasen PubMed. Studie 1 visade en signifikant minskning av kortisolnivåerna med 23% hos deltagare som behandlades med ashwagandha jämfört med placebogruppen under en 60-dagarsperiod. Liknande resultat observerades i studie 2, där deltagare som fick ashwagandha visade signifikant minskning av kortisolnivåerna med 22,2% jämfört med placebogruppen under en 8-veckorsperiod. I studie 3 fanns en tendens till minskade kortisolnivåer med 7,8% hos deltagare som behandlades med ashwagandha, även om skillnaden inte var statistiskt signifikant jämfört med placebogruppen. I studie 4 observerades en betydande minskning av morgonkortisolnivåer med 10,9% och eftermiddagskortisolnivåer med 17,1% hos deltagare som fick ashwagandha under 60 dagar. Men när vi jämförde med placebogruppen var endast minskningen av morgonkortisolnivåerna statistiskt signifikant, medan skillnaden i eftermiddagskortisolnivåer inte var signifikant. Sammanfattningsvis tyder resultaten från dessa studier på att ashwagandha kan vara en effektiv metod för att minska kortisolnivåerna, Withania somnifera L. (Dunal), commonly known as ashwagandha, is an herb with roots in ayurvedic medicine dating back almost 3000 years, and it has garnered increased attention for its potential health benefits. It is considered an adaptogen, capable of assisting the body in managing stress and promoting recovery, due to its main active compounds, the withanolides. Traditionally, ashwagandha has been used to alleviate stress and promote well-being. Cortisol, a stress hormone released by the adrenal glands, plays a crucial role in the body's stress response. Chronic exposure to high levels of cortisol can lead to negative health consequences. Therefore, the aim of this literature review was to investigate ashwagandha's potential effects on the cortisol levels in humans. This was examined through the analysis of four randomized, double-blind, placebo-controlled studies found through the PubMed database. Study 1 showed a significant reduction in cortisol levels with 23% in participants treated with ashwagandha, compared to the placebo group over a 60-day period. Similar results were observed in study 2, where participants receiving ashwagandha exhibited decreased cortisol levels with 22,2% compared to the placebo group over an 8-week period. Study 3 indicated a trend towards decreased cortisol levels with 7,8% in participants treated with ashwagandha, although a reduction was detected the difference was not significant compared to the placebo group. In study 4, a significant decrease in morning cortisol levels by 10.9% and afternoon cortisol levels by 17.1% was observed in participants who received ashwagandha for 60 days. However, when compared to the placebo group, only the decrease in morning cortisol levels was statistically significant, while the difference in afternoon cortisol levels was not significant. In conclusion, the results of these studies suggested that ashwagandha may be an effective method for reducing cortisol levels in adult individuals experiencing

Inhalation av torrt pulver är en populär metod för administrering av aerosolläkemedel vid luftvägssjukdomar som Astma och KOL. På den svenskamarknaden finns det flera olika inhalatorer att välja emellan men är alla lika effektiva? I denna litteraturstudie jämförs femton originalartiklar som söktes via Pubmed med syfte att inhämta information om aerosoliserings-förmåga och kapacitet för inhalatorerna Diskus, Easyhaler, Ellipta, Nexthaler, Spiromax och Turbuhaler. Resultaten indikerar att alla inhalatorer har en effektiv aerosolisering, men när det gäller mängden läkemedel som når lungorna, framkommer att Nexthaler och Turbuhaler presterar bäst. Artiklarna visar att inhalatorer som underlättar de-aggregation har en högre andel läkemedel som når lungorna jämfört med de som saknar denna förmåga. En artikel visade också att genom att byta läkemedlet från en inhalators formulering till en annan inhalators formulering förbättrades inhalatorns prestanda. Denna kunskap är av betydelse vid utveckling av inhalatorer och läkemedelsformuleringar för inhalation. Då målet är att öka effektiviteten vid administrering av aerosolläkemedel, med en högre andel läkemedelspartiklar som når lungorna och mindre går förlorad.

PROTAC är en bifunktionell molekyl och används för målinriktad proteindegradation vilket ger upphov till dess specificitet. Detta görs genom att utnyttja cellens egen mekanism för att degradera felveckade eller skadade protein. Mekanismen heter ubikvitin proteasom systemet men förkortas UPS. Syftet med denna studie är att med hjälp av en systematisk litteraturstudie undersöka om PROTAC kan användas som läkemedel mot NSCLC (icke-småcellig lungcancer). För detta inkluderas totalt 9 vetenskapliga artiklar som behandlar PROTAC och jämför dess effekt mot vanliga NSCLC läkemedel. Artiklarna som ligger till grund för denna studie görs både in vitro och in vivo. Slutsatsen i studien visar på att PROTAC har gynnsamma förutsättningar vad gäller ett potentiellt framtida läkemedel. Slutsatsen baseras på resultatet i studien som visar att PROTAC effektivt kan degradera onkoprotein vilket PROTAC dessutom gör utan att påverka vildtypen av målproteinet. Den målinriktade proteindegradationen som PROTAC ger upphov till är effektiv mot degradation av onkoprotein som normalt är mycket svåra att behandla pga. resistensutveckling. Vidare visade resultatet att PROTAC effektivt kunde hämma tumörtillväxt i möss och att detta dessutom skedde effektivare jämfört med möss som behandlades med vanliga läkemedel mot

Regeneration of insulin-producing 13-cells is an alternative avenue to manage diabetes, and it is crucial to unravel this process in vivo during physiological responses to the lack of 13-cells. Here, we aimed to characterize how hepatocytes can contribute to 13-cell regeneration, either directly or indirectly via secreted proteins or metabolites, in a zebrafish model of 13-cell loss. Using lineage tracing, we show that hepatocytes do not directly convert into 13-cells even under extreme 13-cell ablation conditions. A transcriptomic analysis of isolated hepatocytes after 13-cell ablation displayed altered lipid- and glucose-related processes. Based on the transcriptomics, we performed a genetic screen that uncovers a potential role of the molybdenum cofactor (Moco) biosynthetic pathway in 13-cell regeneration and glucose metabolism in zebrafish. Consistently, molybdenum cofactor synthesis 2 (Mocs2) haploinsufficiency in mice indicated dysregulated glucose metabolism and liver function. Together, our study sheds light on the liver-pancreas crosstalk and suggests that the molybdenum cofactor biosynthesis pathway should be further studied in relation to glucose metabolism and diabetes.

This article evaluates the current gaps around the impact of post-manufacturing processes on the product qualities of protein-based biologics, with a focus on user centricity. It includes the evaluation of the regulatory guidance available, describes a collection of scientific literature and case studies to showcase the impact of post-manufacturing stresses on product and dosing solution quality. It also outlines the complexity of clinical handling and the need for communication, and alignment between drug providers, healthcare professionals, users, and patients. Regulatory agencies provide clear expectations for drug manufacturing processes, however, guidance supporting post-product manufacturing handling is less defined and often misaligned. This is problematic as the pharmaceutical products experience numerous stresses and processes which can potentially impact drug quality, safety and efficacy. This article aims to stimulate discussion amongst pharmaceutical developers, health care providers, device manufacturers, and public researchers to improve these processes. Patients and caregivers’ awareness can be achieved by providing relevant educational material on pharmaceutical product handling., This project has received funding from the Innovative MedicinesInitiative 2 Joint Undertaking (JU) under grant agreement N°101007939 (RealHOPE). This Joint Undertaking receives support fromthe European Union’s Horizon 2020 research and innovation programme and EFPIA.

Stimulator of IFN genes (STING) is a promising target for adjuvants utilized in in situ cancer vaccination approaches. However, key barriers remain for clinical translation, including low cellular uptake and accessibility, STING variability necessitating personalized STING agonists, and interferon (IFN)-independent signals that can promote tumor growth. Here, we identify C100, a highly deacetylated chitin-derived polymer (HDCP), as an attractive alternative to conventional STING agonists. C100 promotes potent anti-tumor immune responses, outperforming less deacetylated HDCPs, with therapeutic efficacy dependent on STING and IFN alpha/beta receptor (IFNAR) signaling and CD8+ T cell mediators. Additionally, C100 injection synergizes with systemic checkpoint blockade targeting PD-1. Mechanistically, C100 triggers mitochondrial stress and DNA damage to exclusively activate the IFN arm of the cGAS-STING signaling pathway and elicit sustained IFNAR signaling. Altogether, these results reveal an effective STING- and IFNAR-dependent adjuvant for in situ cancer vaccines with a defined mechanism and distinct properties that overcome common limitations of existing STING therapeutics., We thank Prof. Roger Greenberg for the Tmem173/ B16F10 cells utilized inthis study. We thank Dr. Barry Moran, manager of the Flow Cytometry Facilityat the School of Biochemistry and Immunology, and Comparative MedicineUnit staff at the Trinity Biomedical Sciences Institute for their valuable assistance. E.C.L., R.W.W., J.L.T., and N.M.W. received support from SFI investigator award 12/1A/1421 and Frontiers of the future award 19/FFP/6484.R.W.W. received support from IRC GOIPG/2019/4236. We thank Dr. JohnBrowne and Prof. Stephen Gordon at University College Dublin for providingaccess to NanoString and help with sample preparation and data analysis.

Neurological conditions conquer the world; they are the leading cause of disability and the second leading cause of death worldwide, and they appear all around the world in every age group, gender, nationality, and socioeconomic class. Despite the growing evidence of an immense impact of perturbations in neuroenergetics on overall brain function, only little is known about the underlying mechanisms. Especially human insights are sparse, owing to a shortage of physiologically relevant model systems. With this perspective, we aim to explore the key steps and considerations involved in developing an advanced human in vitro model for studying neuroenergetics. We discuss biological and technological strategies to meet the requirements of a predictive model, aiming at providing a guide and inspiration for future in vitro models of neuroenergetics., QC 20240626

Excessive consumption of sucrose, in the form of sugar-sweetened beverages, has been implicated in the pathogenesis of metabolic dysfunction-associated fatty liver disease (MAFLD) and other related metabolic syndromes. The c-Jun N-terminal kinase (JNK) pathway plays a crucial role in response to dietary stressors, and it was demonstrated that the inhibition of the JNK pathway could potentially be used in the treatment of MAFLD. However, the intricate mechanisms underlying these interventions remain incompletely understood given their multifaceted effects across multiple tissues. In this study, we challenged rats with sucrose-sweetened water and investigated the potential effects of JNK inhibition by employing network analysis based on the transcriptome profiling obtained from hepatic and extrahepatic tissues, including visceral white adipose tissue, skeletal muscle, and brain. Our data demonstrate that JNK inhibition by JNK-IN-5A effectively reduces the circulating triglyceride accumulation and inflammation in rats subjected to sucrose consumption. Coexpression analysis and genome-scale metabolic modelling reveal that sucrose overconsumption primarily induces transcriptional dysfunction related to fatty acid and oxidative metabolism in the liver and adipose tissues, which are largely rectified after JNK inhibition at a clinically relevant dose. Skeletal muscle exhibited minimal transcriptional changes to sucrose overconsumption but underwent substantial metabolic adaptation following the JNK inhibition. Overall, our data provides novel insights into the molecular basis by which JNK inhibition exerts its metabolic effect in the metabolically active tissues. Furthermore, our findings underpin the critical role of extrahepatic metabolism in the development of diet-induced steatosis, offering valuable guidance for future studies focused on JNK-targeting for effective treatment of MAFLD., QC 20240514

Empagliflozin (EMPA) is a potent, competitive and selective sodium glucose cotransporter-2 (SGLT-2) inhibitor that ameliorates blood glucose with the insulin-independent manner. EMPA reduces weight and blood pressure of patients with type 2 diabetes mellitus (T2DM) without developing hypoglycemic risk. To the best of our knowledge, its safety profiling has not been evaluated on human blood cell cultures yet. Again, the embryotoxicity potential by EMPA is still unclear. Therefore, in this investigation we aimed to evaluate the in vitro cytotoxic, genotoxic and embryotoxic damage potential as well as antioxidative/oxidative effects by EMPA in cultured human blood and human pluripotent embryonal carcinoma NT2 cells for the first time. Cell cultures (n = 5) were exposed to different concentrations ranging from 3.25 to 100 mg/L of EMPA for 48 and 72 h. Cell viability was measured by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) release assays. The alterations in antioxidant/oxidant activity were monitored via measuring the total antioxidant capacity (TAC) and total oxidative stress (TOS) levels. For evaluating the genotoxicity of EMPA chromosomal aberration (CA) assay was performed. The present results revealed that EMPA did not induce cytotoxic or genotoxic damage on healthy human blood cells. Moreover, EMPA exerted non-embryotoxic property and supported antioxidative capacity and decreased the oxidative stress in cultured human blood cells. Our results supported the safe and advantageous use of EMPA for the treatment of T2DM., QC 20240513

Statins (3-hydroxy-3-methylglutaryl-CoA reductase, HMGCR, inhibitors) comprise the gold standard for the management of hypercholesterolaemia and prevention of cardiovascular disease (CVDs). However, they are accompanied by potential adverse effects, notably muscle pain and sleep disturbance. These side effects can significantly impact patient adherence to statin therapy and thus increase the risk for CVDs. Despite extensive research, the underlying mechanisms of statin-associated myopathy and sleep disturbance are poorly understood. In Paper I, we conducted a cross-sectional cohort study to investigate the association between statin use and genetic variants for HMGCR with the risk for insomnia and chronotype using UK biobank cohort data. Statin use, insomnia and chronotype were assessed by a self-report touchscreen questionnaire. Statin treatment was associated with an increased risk of insomnia compared to controls, while genetic variants for HMGCR inhibition were associated with a reduced risk for insomnia. No association with late evening chronotype were observed with statin use or genetic variants for HMGCR. In Paper II, we employed Drosophila melanogaster to examine the effect of statins and the role of central inhibition of Hmgcr on sleep behaviour. Flies were treated with fluvastatin for five days and Hmgcr was knocked down in pan neurons and pars intercerebralis (PI), equivalent to the mammalian hypothalamus. Sleep patterns were recorded and analysed. Pan-neuronal- as well as PI inhibition of Hmgcr recapitulates fluvastatin-induced enhanced sleep latency and reduced sleep duration. In Paper III, we deciphered the underlying mechanisms for statin-induced myopathy using D. melanogaster. We found that fluvastatin treatment induced muscular damage, mitochondrial phenotypes, lowered locomotion, reduced climbing activity and was associated with lipotoxicity, impaired muscle differentiation and regeneration, and lowered expression of skeletal muscle chloride cha

Endocrine disruptors (EDs) pose a serious threat to human health and the environment and require a comprehensive evaluation to be identified. The identification of EDs require a substantial amount of data, both in vitro and in vivo, due to the current scientific criteria in the EU. At the same time, the EU strives to reduce animal testing due to concerns regarding animal welfare and sensitivity of animal studies to adequately detect adverse effects relevant for human health. Perfluorooctane sulfonic acid (PFOS) is a persistent organic pollutant that is suspected to be an ED based on academic research, however it is not identified as such from a regulatory perspective. It has previously been shown that PFOS has the potential to cause neurotoxicity as well as affect the thyroid system, and it is known that specific thyroid hormone levels are critical in the development of the brain during. In this work, the aim was to evaluate a mechanism-based approach to identify ED properties of PFOS based on the Adverse Outcome Pathway (AOP) framework and using New Approach Methods (NAMs), by comparing this approach to an ED assessment based on the currently available guidance document. An AOP network (AOPN) was generated for the thyroid modality, and AOPs leading to developmental neurotoxicity (DNT) were identified. A literature search and screening process based on the AOPN, and systematic review methodology, was performed, followed by a rigorous Weight-of-Evidence (WoE) assessment. Evidence was mapped back onto the AOPN used for the literature search, to identify possible endocrine Modes-of-Action (MoAs) for PFOS and data gaps in the two assessments. It could be concluded that PFOS fulfils the criteria for ED classification in the standard ED assessment, but not in the mechanism-based assessment. The need for quantitative information, such as quantitative AOPs, for the mechanism-based approach is discussed. The possibility of a directly neurotoxic alternative MoA was also highl

Understanding the genetic basis of neuro-related proteins is essential for dissecting the molecular basis of human behavioural traits and the disease aetiology of neuropsychiatric disorders. Here the SCALLOP Consortium conducted a genome-wide association meta-analysis of over 12,000 individuals for 184 neuro-related proteins in human plasma. The analysis identified 125 cis-regulatory protein quantitative trait loci (cis-pQTL) and 164 trans-pQTL. The mapped pQTL capture on average 50% of each protein's heritability. At the cis-pQTL, multiple proteins shared a genetic basis with human behavioural traits such as alcohol and food intake, smoking and educational attainment, as well as neurological conditions and psychiatric disorders such as pain, neuroticism and schizophrenia. Integrating with established drug information, the causal inference analysis validated 52 out of 66 matched combinations of protein targets and diseases or side effects with available drugs while suggesting hundreds of repurposing and new therapeutic targets.

The complex formed by Ku70/80 and DNA-PKcs (DNA-PK) promotes the synapsis and the joining of double strand breaks (DSBs) during canonical non-homologous end joining (c-NHEJ). In c-NHEJ during V(D)J recombination, DNA-PK promotes the processing of the ends and the opening of the DNA hairpins by recruiting and/or activating the nuclease Artemis/DCLRE1C/SNM1C. Paradoxically, DNA-PK is also required to prevent the fusions of newly replicated leading-end telomeres. Here, we describe the role for DNA-PK in controlling Apollo/DCLRE1B/SNM1B, the nuclease that resects leading-end telomeres. We show that the telomeric function of Apollo requires DNA-PKcs’s kinase activity and the binding of Apollo to DNA-PK. Furthermore, AlphaFold-Multimer predicts that Apollo’s nuclease domain has extensive additional interactions with DNA-PKcs, and comparison to the cryo-EM structure of Artemis bound to DNA-PK phosphorylated on the ABCDE/Thr2609 cluster suggests that DNA-PK can similarly grant Apollo access to the DNA end. In agreement, the telomeric function of DNA-PK requires the ABCDE/Thr2609 cluster. These data reveal that resection of leading-end telomeres is regulated by DNA-PK through its binding to Apollo and its (auto)phosphorylation-dependent positioning of Apollo at the DNA end, analogous but not identical to DNA-PK dependent regulation of Artemis at hairpins., Funding agencies: Knut and Alice Wallenberg Foundation (F.L. is a Wallenberg Molecular Medicine fellow); Cancerfonden [grant number Can 2018/493 to F.L.]; Vetenskapsrådet [grant number 2018-03215 to F.L]; LiU Cancer (2023 to F.L. with B.W.); National Institutes of Health [grant number AG016642 to T.d.L.]; Carl Tryggers stiftelse för Vetenskaplig Forskning [grant number 20:453 to B.W.]; Vetenskapsrådet [grant number 2020-03352 to B.W.]; Cancer Research UK Career Development Award [grant number C68409/A28129 to M.D.]. Funding for open access charge: Vetenskapsrådet, Cancerfonden and Knut and Alice Wallenberg Foundation.