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Vascular endothelial cell (EC) dysfunction plays a key role in diabetic complications. This study discovers significant upregulation of Quaking-7 (QKI-7) in iPS cell-derived ECs when exposed to hyperglycemia, and in human iPS-ECs from diabetic patients. QKI-7 is also highly expressed in human coronary arterial ECs from diabetic donors, and on blood vessels from diabetic critical limb ischemia patients undergoing a lower-limb amputation. QKI-7 expression is tightly controlled by RNA splicing factors CUG-BP and hnRNPM through direct binding. QKI-7 upregulation is correlated with disrupted cell barrier, compromised angiogenesis and enhanced monocyte adhesion. RNA immunoprecipitation (RIP) and mRNA-decay assays reveal that QKI-7 binds and promotes mRNA degradation of downstream targets CD144, Neuroligin 1 (NLGN1), and TNF-α-stimulated gene/protein 6 (TSG-6). When hindlimb ischemia is induced in diabetic mice and QKI-7 is knocked-down in vivo in ECs, reperfusion and blood flow recovery are markedly promoted. Manipulation of QKI-7 represents a promising strategy for the treatment of diabetic vascular complications. Vascular endothelial cell (EC) dysfunction contributes to the occurrence of diabetic complications. Here the authors report that in diabetic conditions, upregulation of the RNA binding protein QKI-7 in ECs due to the imbalance of RNA splicing factors CUG-BP and hnRNPM contributes to EC dysfunction, and that in vivo QKI-7 silencing improves blood flow recovery in diabetic mice with limb ischemia.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.An amendment to this paper has been published and can be accessed via a link at the top of the paper.

The growing prevalence of diabetes highlights the urgent need to study diabetic cardiovascular complications, specifically diabetic cardiomyopathy, which is a diabetes-induced myocardial dysfunction independent of hypertension or coronary artery disease. This review examines the role of mitochondrial dysfunction in promoting diabetic cardiac dysfunction and highlights metabolic mechanisms such as hyperglycaemia-induced oxidative stress. Chronic hyperglycaemia and insulin resistance can activate harmful pathways, including advanced glycation end-products (AGEs), protein kinase C (PKC) and hexosamine signalling, uncontrolled reactive oxygen species (ROS) production and mishandling of Ca2+ transient. These processes lead to cardiomyocyte apoptosis, fibrosis and contractile dysfunction. Moreover, endoplasmic reticulum (ER) stress and dysregulated RNA-binding proteins (RBPs) and extracellular vesicles (EVs) contribute to tissue damage, which drives cardiac function towards heart failure (HF). Advanced patient-derived induced pluripotent stem cell (iPSC) cardiac organoids (iPS-COs) are transformative tools for modelling diabetic cardiomyopathy and capturing human disease’s genetic, epigenetic and metabolic hallmarks. iPS-COs may facilitate the precise examination of molecular pathways and therapeutic interventions. Future research directions encourage the integration of advanced models with mechanistic techniques to promote novel therapeutic strategies.

IntroductionDiabetes and associated cardiovascular diseases (CVDs) are a class of disorders affecting the heart or blood vessels. Despite progress in clinical research and therapy, CVDs still represent the leading cause of mortality and morbidity worldwide. The hallmarks of cardiac diseases include heart dysfunction and cardiomyocyte death, inflammation, fibrosis, scar tissue, hyperplasia, hypertrophy, and abnormal ventricular remodelling. The loss of cardiomyocytes is an irreversible process that leads to fibrosis and scar formation, which, in turn, induce heart failure with progressive and dramatic consequences. Both genetic and environmental factors pathologically contribute to the development of CVDs, but the precise causes that trigger cardiac diseases and their progression are still largely unknown. The lack of reliable human model systems for such diseases has hampered the unravelling of the underlying molecular mechanisms and cellular processes involved in heart diseases at their initial stage and during their progression. In this study we use induced pluripotent stem cells (iPSCs) from diabetic and non-diabetic donors to recapitulate an iPSC-driven cardiac model with the aim of underlining the potential of stem-cell biology-based approaches in the elucidation of the pathophysiology of cardiac disease.MethodsCardiomyocytes were generated from iPS cells from both diabetic (DiPSC-CMs) and non-diabetic donors (NDiPS-CMs) within a thirteen day differentiation protocol. Cardiac commitment was assessed by Flow cytometry, PCR analysis, and Immunofluorescence staining. Morphological features of the sarcomere arrangement and mitochondria size were assessed using TEM microscopy. Cardiac function was measured by assessing calcium flux using Flexstation. Beating qualities were assessed by Nikon 6D Life imaging.ResultsUpon differentiation both DiPSC-CMs and NDiPSC-CMs presented strong cardiac commitment as evident by the significant expression of cardiac markers evaluated by PCR and flow cytometry. iPS-CM from both donors presented no significant differences in the expression of cardiac markers brachyury and cardiac troponin as assessed by flow cytometry (p > 0.7078) and sarcomere proteins a-Actinin, myosin light chain MLCA2 assessed by immunofluorescence staining. Cardiomyocytes derived from diabetic donors (DiPS-CMs) showed differences in the uptake of calcium when these compared to the nondiabetic counterparts (NDiPS-CMs). Calcium flux was measure by capturing fluorescence intensity of Fura-2 calcium dye by Flexstation (p > 0.0156). TEM imaging and assessment of mitochondria between DiPSC-CMs and NDiPS-CMs showed differences in mitochondrial morphological features such as aspect ratio, perimeter and length (p > 0.05) between our two groups indicating a morphological change that may underly be related to an underlying mitochondrial disorder associated to a metabolic dysfunction.Abstract BS62 Figure 1ConclusionsDiPS-CMs and NDiPS-CMs showed no differences in cardiac marker expression related to cardiomyocyte phenotype and cardiac commitment however, when iPS-CMs are subjected to functional assessments, there have been differences observed in calcium uptake and mitochondrial phenotypes. These observations might potentially be the reflection of an underlying dysfunctional metabolic fingerprint that further needs to be investigated and evaluated in detail. In vitro generation of functional CMs hold a great potential for personalised medicine as these cells recapitulate the structure and function of human cardiac tissue and are amenable to model and identify new biomarkers of diabetic cardiomyopathy, affecting hundreds of millions of patients world-wide.Conflict of Interestno conflict declared so far

The relationship between MMR vaccine and ASD to this day has been a big problem in the public eye. The presumed causality of ASD from MMR vaccine has led to a big portion of parents worldwide to not vaccinate their children with the MMR vaccine. If there was a correlation between the MMR vaccine and ASD, then there should have been a rise in ASD cases the more children get vaccinated. To fully analyze the connection between ASD and the MMR vaccine, both case studies and qualitative studies were accessed. Qualitative studies provided information regarding the repercussions of under-vaccination over the years, while case studies were used to study any potential causal effects of the MMR vaccine and ASD. Many studies and journals were accessed from PubMed and Medline Plus. Through all the accessed studies there has been no significant statistical correlation between MMR vaccine and autism. Health care professionals need to play a big role in educating parents about vaccines and the unproven claims of disease that the general public might think they cause. Hypothesis: If there was a correlation between the MMR vaccine and ASD, then there should have been a rise in ASD cases the more children get vaccinated.

IntroductionImpaired function of blood vessels can lead to cardiovascular diseases (CVDs), which is a major cause of death worldwide. The presence of both endothelial cells (ECs) and mural cells is central to the proper function of blood vessels in health and pathological changes in diseases including diabetes. Although iPSCs-derived vascular organoids (VOs) provide an appealing source for in vitro vascular disease modelling and drug testing, whether these organoids can recapitulate human vascular disease is yet to be determined.MethodsBlood mononuclear cells from six donors with diabetes (DB) and three with non-diabetes (ND) were subjected to reprogramming into iPSCs, and subsequent differentiation to VOs. DB-VOs and ND-VOs were compared using immunohistochemistry, angiogenic array, ROS production assay, acetylated-LDL uptake assay, transmission electron microscopy, western blotting, flow cytometry and single-cell RNA sequencing. Additionally, the regenerative potential of DB-VOs vs ND-VOs was compared by assessment of blood recovery via Laser Doppler imaging in mice ischaemic hindlimb model, and organoid cells’ integration into host vasculature was tracked by Bruker fluorescent imaging.ResultsWe showed diabetic derived-iPSCs-VOs represent impaired vascular function including enhanced ROS level, with higher mitochondrial content and activity, increased pro-inflammatory cytokines, and less regenerative potential in vivo. Using single-cell RNA sequencing, we identified all specialized types of vascular cells (artery, capillary, vein, lymphatic and tip cells, as well as pericytes and vSMCs) within vascular organoids, while demonstrating the dichotomy landscape of ECs and mural cells. Furthermore, we revealed basal heterogeneity within vascular organoids and demonstrated differences between diabetic and non-diabetic VOs. Of note, a subpopulation of ECs significantly enriched for ROS and oxidative phosphorylation hallmarks in DB-VOs, representing early signs of aberrant angiogenesis in diabetes. For the first time, we report that GAP43 (Neuromodulin) is expressed in ECs, and GAP43+ ECs are distinctly increased in DB-VOs. Therefore, GAP43 is possibly a biomarker for the onset and progression of diabetic-related blood vessel dysfunction.Abstract BS16 Figure 1ConclusionsWe comprehensively characterized DB-VOs versus ND-VOs at a single-cell resolution, demonstrated cellular components of blood vessel organoids for the first time, and represented molecular and functional differences between their vascular cell types and clusters.Conflict of InterestNo

Diabetic endotheliopathy is the main cause for impaired angiogenesis and reduced neovascularization that lead to microvascular injury and vascular complications. The pathogenic basis for vascular complications arising from diabetes is complex. Elucidation of key underlying mechanisms will help the development of novel therapies and the discovery of potential biomarkers. The ability to generate functional endothelial cells (ECs) from induced pluripotent stem cells (iPSCs) from small amounts of blood is a novel and powerful tool for cell-based therapies. Human iPSC-derived ECs (iPS-ECs) have a broad range of clinical applications including cell-based therapy, disease modelling and drug screening; they can be used in mechanistic studies towards the development of novel therapies and in the discovery of new biomarkers to be applied in regenerative medicine and treatment of diabetic vasculopathy. Here we utilize transcriptomic and proteomic technologies to assess patient-specific iPS-ECs from diabetic (DiPS-ECs) and non-diabetic (NiPS-ECS) donors 1,2,3,4 in order to investigate the mechanisms driving endotheliopathy in diabetes. Our in vitro and in vivo models recapitulate the effects of hyperglycaemia on the vasculature in the clinical setting. RNA-seq data showed that genes and proteins involved in angiogenesis and EC function were significantly downregulated in DiPS-ECs in comparison to NiPS-ECS (n=3, p<0.05). Specific epsins regulating VEGF-mediated angiogenesis were downregulated in DiPS-ECs, leading to increased signalling VEGF pathway activation6. Moreover factors involved in E-cadherin signalling, endothelial-to-mesenchymal transition and fibrosis were increased in DiPS-ECs. We detected abnormal capillary permeability and barrier integrity in DiPS-ECs using xCELLigence®. DiPS-ECs had significantly reduced barrier integrity and barrier recovery (n=3, p<0.001, ±SEM) and also displayed impaired tube formation in vitro (n=3, ±SEM, p<0.05). DiPS-ECs displayed impaired function demonstrated by decreased blood flow recovery (BFR) compared to NiPS ECs (n=3) when injected to the hindlimb of mice following femoral artery ligation. Finally, our proteomic and transcriptomic analysis confirmed imbalances in several angiogenic genes including endothelial specific Roundabout protein 4 (ROBO4) that is highly involved in pathways related to angiogenesis, barrier stability and endothelial health 7. Expression of ROBO4 was found to be impaired in DiPS-ECs and transcriptomic analysis along with in vitro and in vivo studies revealed its importance in vascular development and angiogenesis. Our data support the impaired angiogenic functionality of DiPS-ECs cells in vitro and in vivo and show that DiPS-ECs carry an imprint of the diabetic milieu which is reflected in their dysfunction. To the best of our knowledge, we have identified a novel disease-specific signature in diabetic iPS-ECs, therefore our human iPS-EC model may serve as a valuable tool to study biological pathways and identify new treatments for diabetes-induced endotheliopathy.Conflict of InterestNone

Introduction The palmaris longus muscle (PLM), located in the forearm's anterior compartment, plays an essential role in wrist flexion. Its tendon is often used for grafting because of its accessibility and minimal function. However, its anatomy varies, with congenital absence being the most common variation. This study aims to detect the incidence of the palmaris longus tendon (PLT) and compare physical examination methods with standard ultrasonography, as previous studies have only utilized physical examinations. Methods In this study, 61 participants were examined bilaterally (122 wrists) using three different physical examination methods: the Schaeffer, Thompson, and Mishra I tests. The ultrasonography test was conducted by a single observer using a GE LOGIQ e ultrasound system (GE HealthCare Technologies, Inc., Chicago, IL, USA), equipped with a linear transducer with a frequency range of 12-15 Hz, attached transversely to the anterior distal forearm. The tendon was visualized anterior to the median nerve, medial to the flexor carpi radialis, and superficial to the flexor retinaculum. To assess the difference between physical examination and ultrasound, statistical analyses were conducted on subgroups using a t-test. Additionally, PLT incidence was evaluated according to gender, ethnicity, and hand dominance. Results On ultrasound examination, the PLT was detected in 72.13% of wrists bilaterally (n = 88), 5.74% unilaterally (n = 7), and was absent in 22.13% of the 122 wrists examined (n = 27). Physical examination methods detected an average of 56.28% bilaterally (n = 68.66), 6.56% unilaterally (n = 8), and 37.16% as absent (n = 45.33). Total detection by ultrasound was 77.87% (n = 95), while the physical examination average was 62.84% (n = 76.66). Overall, there is a significant difference between the physical examination methods and ultrasonography in detecting PLT, with ultrasonography demonstrating greater accuracy. It should also be noted that the prevalence of PLT is not affected by gender. Conclusion Ultrasonography is crucial in clinical settings to confirm the presence of the PLT, even when a physical examination is inconclusive. Physical and ultrasound approaches can, therefore, be combined to avoid producing incorrect negative results when locating the PLT for tendon grafting.

Focused assessment with sonography for trauma (FAST) ultrasound (US) is a valuable medical examination used in trauma settings, particularly for rapid responses to events such as natural disasters. Although the efficacy and benefits of FAST in patient care have been extensively studied, there is limited research on training medical students in FAST. Previous studies have found that medical students can proficiently perform a FAST US after two days of training. However, these studies exclusively included first-year medical students without considering variations in their medical knowledge. Particularly, the advantage of medical students having US experience before undergoing FAST training has not been previously examined.INTRODUCTIONFocused assessment with sonography for trauma (FAST) ultrasound (US) is a valuable medical examination used in trauma settings, particularly for rapid responses to events such as natural disasters. Although the efficacy and benefits of FAST in patient care have been extensively studied, there is limited research on training medical students in FAST. Previous studies have found that medical students can proficiently perform a FAST US after two days of training. However, these studies exclusively included first-year medical students without considering variations in their medical knowledge. Particularly, the advantage of medical students having US experience before undergoing FAST training has not been previously examined.Assess the performance and knowledge acquisition of medical students with and without prior US experience after completing a FAST training course.OBJECTIVESAssess the performance and knowledge acquisition of medical students with and without prior US experience after completing a FAST training course.The study included a total of 71 students, consisting of 33 males and 38 females, who were between the ages of 18 and 31, with an average age of 24.6 and a standard deviation of 2.4. The inclusion criteria targeted first- and second-year medical school students who participated on a volunteer basis. Students were divided into two groups: group A, consisting of those without prior US experience, and group B, made up of those who had previous US experience. All students completed a pre-training survey to share their comfort and confidence in US use and knowledge. A baseline FAST exam was conducted to establish initial performance. A comprehensive three-hour training session was then provided. Post-training, students performed another FAST exam to assess improvement, followed by a post-training survey to evaluate comfort and confidence.METHODSThe study included a total of 71 students, consisting of 33 males and 38 females, who were between the ages of 18 and 31, with an average age of 24.6 and a standard deviation of 2.4. The inclusion criteria targeted first- and second-year medical school students who participated on a volunteer basis. Students were divided into two groups: group A, consisting of those without prior US experience, and group B, made up of those who had previous US experience. All students completed a pre-training survey to share their comfort and confidence in US use and knowledge. A baseline FAST exam was conducted to establish initial performance. A comprehensive three-hour training session was then provided. Post-training, students performed another FAST exam to assess improvement, followed by a post-training survey to evaluate comfort and confidence.Medical students who had prior experience in the US (group B) performed significantly better (p<0.01) in both the pre- and post-training FAST exams when compared to students without previous US experience. Specifically, in locating the liver, right kidney, hepatorenal recess, and left kidney, as well as detecting fluid accumulation when in a supine position. Additionally, medical students with prior US experience (group B) exhibited higher baseline confidence (p<0.005-p<0.01) in their ability to perform a FAST exam, as indicated by the results of the pre-testing survey.RESULTSMedical students who had prior experience in the US (group B) performed significantly better (p<0.01) in both the pre- and post-training FAST exams when compared to students without previous US experience. Specifically, in locating the liver, right kidney, hepatorenal recess, and left kidney, as well as detecting fluid accumulation when in a supine position. Additionally, medical students with prior US experience (group B) exhibited higher baseline confidence (p<0.005-p<0.01) in their ability to perform a FAST exam, as indicated by the results of the pre-testing survey.Previous experience with US significantly boosted confidence and knowledge gains following FAST training. This emphasizes the value of including US training in medical school programs after earlier exposure, offering evident benefits. The study reveals the unexplored benefit of having prior US experience for medical students undergoing FAST training, thus addressing a previously unexplored area in current research. The conclusions stress the necessity of integrating US training into medical school curricula after initial exposure. This understanding can direct medical educators in refining the education process, enabling students to be better equipped for real-world medical situations involving FAST.CONCLUSIONPrevious experience with US significantly boosted confidence and knowledge gains following FAST training. This emphasizes the value of including US training in medical school programs after earlier exposure, offering evident benefits. The study reveals the unexplored benefit of having prior US experience for medical students undergoing FAST training, thus addressing a previously unexplored area in current research. The conclusions stress the necessity of integrating US training into medical school curricula after initial exposure. This understanding can direct medical educators in refining the education process, enabling students to be better equipped for real-world medical situations involving FAST.

The presence of both endothelial cells (ECs) and mural cells are central to the proper function of blood vessels in health and pathological changes in diseases including diabetes. Although iPSCs-derived vascular organoids (VOs) provide an appealing in vitro disease model and platform for drug screening, whether these organoids recapitulate human disease remains debatable. Here, we show human diabetic (DB)-VOs represent impaired vascular function including enhanced ROS activity, with higher mitochondrial content and activity, increased pro-inflammatory cytokines, and less regenerative potential in vivo. Using single-cell RNA sequencing, we identify all specialized types of vascular cells (artery, capillary, vein, lymphatic and tip cells, as well as pericytes and vSMCs) within vascular organoids, while demonstrating the dichotomy landscape of ECs and mural cells. Furthermore, we reveal basal heterogeneity within vascular organoids and demonstrate differences between diabetic and non-diabetic VOs. Of note, a subpopulation of ECs significantly enrich for ROS and oxidative phosphorylation hallmarks in DB-VOs, may represent early signs of aberrant angiogenesis in diabetes. This study helps to identify key biomarkers for diabetic disease progression and find signalling molecules amenable to drug intervention.