bluuebunny/crossref_metadata_2025_split huggingface dataset:
Impaired mechanotransduction of small cerebral arteries after traumatic brain injury. Role of arachidonic acid pathway dysfunction
Authors: Koller Akos , Szenasi Annamaria , Amrein Krisztina , Czeiter Endre , Szarka Nikolett , Toth Peter 
Published: May 2022

BackgroundHealthy functioning of the brain requires a precise autoregulation of cerebral blood flow, which in part is achieved by the pressure and flow sensitive vasomotor mechanisms. Traumatic brain injury (TBI) frequently occurs worldwide, resulting in brain diseases with high morbidity and mortality. Here, we hypothesized that TBI impairs the autoregulatory mechanisms, namely the pressure‐ and flow‐induced constrictions of isolated rat middle cerebral arteries (MCAs).MethodsTBI was induced in anaesthetized rats by weight drop model, and then MCAs were isolated and transferred into a pressure‐flow chamber. Changes in internal diameter in response to hemodynamic forces and vasoactive drugs were measured by a video‐microscopy.ResultsIn MCAs from intact rats, increases in pressure‐, flow‐ and pressure + flow elicited constrictions (max.: ‐26 ± 1.7; ‐26 ± 1.9; and ‐52 ± 2.8 µm, p < 0.05). Flow‐induced constrictions were significantly reduced by HET0016, an inhibitor of cytochrome P450 4A (CYP450 4A) or in the presence of thromboxane‐prostanoid (TP receptor) antagonist SQ 29,548. Both responses were significantly reduced after TBI. Arachidonic acid, (AA, 10−7 M), and CYP450 4A metabolite 20‐hydroxyeicosatetraenoic acid (20‐HETE) elicited constrictions of intact MCA (‐26 ± 2.3% and ‐31 ± 3.6%), which were significantly reduced after TBI (to ‐11 ± 1.3% and ‐16 ±2.5%). The TP receptor agonist U46619 (10−7 M) elicited substantial constrictions of MCA from intact rats (‐21 ± 3.3%), which were also significantly reduced, after TBI (to ‐16 ± 2.4%).ConclusionsBoth pressure and flow‐induced constrictions of MCA were impaired by traumatic brain injury, likely due to the reduced ability of cytochrome P450 4A to convert arachidonic acid to constrictor prostaglandins (cPGs) and the mitigated sensitivity of thromboxane‐prostanoid receptors to cPGs. We propose that the impairment of autoregulatory vasomotor mechanisms contribute to the TBI‐induced human brain diseases, such as headache, disruption of blood brain barrier, brain edema, Alzheimer‐type diseases and vascular dementia.
Estrogen Sulfotransferase is Required for the Female Mice Sensitivity to T Cell‐Mediated Hepatitis in an Estrogen‐Independent Manner
Authors: Wang Jingyuan , Guan Jibin , Zhang Ziteng , Xie Wen 
Published: May 2022

IntroductionAutoimmune hepatitis (AIH) is a severe T cell‐mediated chronic liver disease with a higher incidence in females. However, the pathological mechanism of AIH remains elusive and there is no effective therapy except for nonspecific immunosuppression. Estrogen sulfotransferase (EST or SULT1E1) is a phase II enzyme best known for its function in sulfonating and deactivating estrogens. Previous work from our lab showed that EST is inducible and functional in several other inflammation diseases. However, the possible role and mechanism of EST in T‐cell mediated hepatitis remain unknown. The goal of this study is to investigate the role of EST in the female preference of T‐cell mediated hepatitis.MethodsConcanavalin A (ConA) was used to induce T cell‐mediated hepatitis in female C57BL/6J mice. For Est loss of function models, we used the whole‐body or liver‐specific Est knockout mice, or treating WT mice with the Est inhibitor Triclosan. We also used Est whole‐body knockout mice with Est specifically reconstituted in the hepatocytes. The severity of liver injury was evaluated through the serum levels of Alanine transaminase (ALT), Aspartate transaminase (AST) and the necrosis in liver histology. Inflammatory responses were characterized by the levels of cytokines and infiltrated immune cells in the liver by real‐time PCR, ELISA, and flow cytometry. To test the role of estrogens in the regulation, we depleted estrogens by ovariectomy in female mice prior to the ConA challenging.ResultsEst was highly induced in the liver of ConA‐treated mice. Knockout or pharmacological inhibition of Est significantly protected the female mice from ConA‐induced hepatitis regardless of ovariectomy. Moreover, ESTKO mice exhibited a more robust inflammatory response with a larger spleen‐to‐body‐weight ratio and a higher level of pro‐inflammatory cytokines production. ESTKO mice had a higher frequency of CD4+ T cells, including a higher level of Treg population, and less cytotoxic CD8+ T cells. The protective phenotype of ESTKO mice was abolished when the expression of EST was reconstituted in the hepatocytes.ConclusionsHepatic EST is required for female sensitivity to Con A‐induced T‐cell mediated hepatitis in an estrogen‐independent manner. Pharmacological inhibition of EST might be a potential strategy for the treatment of AIH.
Antisense oligonucleotides (ASO) to increase progranulin production in frontotemporal dementia
Authors: Ebenezer Sarah , Ebenezer Joel , Nguyen Andrew 
Published: May 2022

Frontotemporal Dementia (FTD) is a debilitating disease that causes the individual to slowly lose their personalities, emotions, and control over social behaviors as disease progresses. FTD is caused due to significant neuronal loss in the frontal and temporal lobes of the brain. Though there is no treatment or cure yet for FTD, research has identified that the protein progranulin may be responsible for this neuronal loss, as a lack of progranulin has been linked to the degradation of neurons in FTD patients. Therefore, an effective treatment for FTD must aim to increase the production of progranulin in the brain. Previous research papers (Jiao J et al, 2010*) found that MicroRNA 29 (miR‐29) was responsible for regulating the translation of hPGRN (human progranulin) RNA through binding to the 3’UTR and preventing translation, suggesting that preventing miR‐29 from binding to the 3’UTR of hPGRN RNA should increase the overall production of progranulin. Antisense oligonucleotides (ASO) achieve this through binding to the hPGRN RNA and disrupting the binding site of miR‐29 and therefore preventing miR‐29 from binding. In our study, neuroblastoma cells were transfected with luciferase DNA as a surrogate measure of progranulin translation and then treated with increasing concentrations of M5 ASO and evaluated for luciferase expression. The results indicated that there was an increase in luciferase activity with the increasing concentrations of ASO, specifically concentrations of 10μM and 20μM which had a significant increase in progranulin production. Further tests were done in an attempt to confirm that the ASOs are working through binding to the 3’UTR and preventing miR‐29 from binding, however these tests came back inconclusive. These findings provide a beneficial insight into further development of these ASOs as possible treatments and therapies for FTD.*Jiao J, Herl LD, Farese RV, Gao FB. MicroRNA‐29b regulates the expression level of human progranulin, a secreted glycoprotein implicated in frontotemporal dementia. PLoS One. 2010 May 10;5(5):e10551
Anti‐obesity effect of Premature Citrus Extract in high‐fat diet induced obesity mice
Authors: Natraj Premkumar , Rajan Priyanka , Ranaweera Sachithra S. , Dayarathne Lakshi A. , Han Chang‐Hoon 
Published: May 2022

BackgroundObesity is a worldwide health threat. Premature citrus extract (PCE) are known to contain high concentrations of flavonoids, polyphenols, and organic acids with health functions. The current study aimed to investigate the anti‐obesity effect of PCE on a high‐fat diet (HFD)‐induced obesity mice.MethodMice were fed on chow or HFD with or without Orlistat (ORL), Hesperidin (HES), and PCE treatment for 10 weeks. Triglyceride (TG) content and total cholesterol (TC), and low‐density lipoprotein cholesterol (LDL‐c), and high‐density lipoprotein cholesterol (HDL‐c) levels were determined by TG, HDL and LDL Assay Kit (BioAssay System). The phosphorylation levels of AMP‐activated protein kinase (AMPK) and acetyl Co‐A carboxylase (ACC); the expression of sterol regulatory element binding protein‐2 (SREBP‐2), 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase (HMGCR), Acyl‐coenzyme A: cholesterol acyltransferases (ACAT‐1) and mitochondrial uncoupling proteins (UCP‐3) were observed via western blot analysis.ResultsPCE and HES administered group significantly reduced the weights of body, liver, and white adipose tissue compared to the HFD group. Supplementation with PCE and HES significantly decreased the levels of TG, TC, and (LDL‐c), and increased (HDL‐c) levels in serum compared to the HFD group. Furthermore, treatment with PCE and HES significantly decreased the levels of TG and TC in the liver whereas increased the levels of TG and TC in feces compared to the HFD group. Western blot analysis revealed that PCE and HES increased the phosphorylation levels of both AMPK and ACC and significantly decreased the expression levels of SREBP‐2, HMGCR, and ACAT‐1 in the liver compared to the HFD group. In addition, both PCE and HES increased the expression level of UCP‐3 in skeletal muscle tissue.ConclusionOverall results suggest that PCE and HES exerts an anti‐obesity effect via activation of AMPK and associated signaling cascades related to lipid metabolism in HFD‐induced obesity mice
Moringa oleifera extract protects against rhabdomyolysis‐induced acute kidney injury
Authors: Falayi Olufunke , Etim Utibe , Ogunpolu Blessing , Oyagbemi Ademola , Omobowale Temidayo , Adedapo Adeolu 
Published: May 2022

Acute kidney injury (AKI) is a life‐threatening abrupt disruption in kidney function. The pathophysiological mechanisms of AKI are poorly understood. Also, there are currently no effective drugs to treat AKI. Phytochemicals from Moringa oleifera plants exhibit antihypertensive, cholesterol‐lowering, diuretic, and hepatoprotective properties. This study aims to determine if Moringa oleifera extract can ameliorate rhabdomyolysis‐induced AKI and its possible mechanisms of action.Experimental rats were randomly divided into five groups. Rhabdomyolysis was induced by intramuscular glycerol injection. Group A received distilled water only. Group B received distilled water, followed by 50% glycerol on the 8th day. Group C and D received 50 and 100 mg/kg methanol stem extract of Moringa oleifera (orally) for seven days and glycerol on the 8th day. Group E received 100 mg/kg of methanol stem extract of Moringa oleifera alone for seven days and normal saline on the 8th day. Necropsy was performed 24 h after glycerol treatment. Blood and kidneys samples were collected for biochemical, histopathological, and immunohistochemical analyses.Significant decreases in superoxide dismutase, glutathione peroxidase, glutathione S‐transferase, glutathione, protein thiols, and non‐protein thiols were observed in animals treated with only glycerol when compared with the control groups. Glycerol treatment also caused a significant increase in markers of AKI and inflammation, including creatinine, nitric oxide, urea, hydrogen peroxide, malondialdehyde, protein carbonyl, and advanced oxidative protein products. Immunohistochemical staining of renal NF‐ƙβ and KIM‐1 was also elevated in glycerol‐treated rats. Treatment with methanol stem bark extract of Moringa oleifera ameliorated glycerol‐induced oxidative stress, kidney damage, and inflammation.The methanol stem extract of Moringa oleifera alleviated rhabdomyolysis‐induced AKI in rats through its antioxidant and anti‐inflammatory properties.
Increased Glycolysis and ATP Production Are Not Prerequisites for Cardiomyocytes Cell Cycle Induction
Authors: Tang Ling , Zhu Wuqiang 
Published: May 2022

BackgroundIt has been reported that metabolic switch from fatty acid oxidation to glycolysis is associated with cardiomyocyte cell cycle induction. However, it is unknown if increased glycolysis and ATP production are indispensable for cardiomyocyte cell cycle induction. Here, we aim to address this question.MethodsCell cycle was induced by overexpression of CCND2 gene or treatment of FGF1 (100 ng/mL) and CHIR99021 (5 µM) on human induced pluripotent stem cells‐derived cardiomyocytes (hiPSC‐CMs). Contraction inhibition was induced by treatment of verapamil (0.5 μM). Cell cycle was evaluated by immunostaining using antibodies against cell cycle markers. Real‐time ATP production and mitochondrial oxidative phosphorylation were determined by Seahorse assay. Metabolic profile was assessed by liquid chromatography–mass spectrometry.ResultsTreatment of FGF1 and CHIR99021 induces cell cycle of hiPSC‐CMs, reduced carbohydrate metabolism and real‐time ATP production from both mitochondria and cytoplasm. Data from RNA Sequencing showed this treatment increased expression of CCND2 and CDK4 and decreased expression of sarcomeric contractile related genes. Overexpression of CCND2 in hiPSC‐CMs also induces cell cycle, increased aerobic glycolysis (Warburg effect) and no change of real‐time ATP production. Inhibition of contraction via treatment of verapamil resulted in reduced overall metabolism and real‐time ATP production.ConclusionOur data suggest that cardiomyocyte cell cycle may be induced without increased glycolysis or real‐time ATP production.
Inhibition of Escherichia coliATP synthase by dietary pomegranate phenolics
Authors: Lakhani Muhaib , Ahmad Zulfiqar 
Published: May 2022

BackgroundA wide range of natural and synthetic compounds bind and inhibit ATP synthase. Dietary polyphenols found in fruits, vegetables, and spices such as tea, olives, grapes, cantaloupes, garlic, and pomegranate possess antimicrobial activities. Dietary pomegranate polyphenols (DPPs) have been shown to exhibit anticancer, antioxidant, and antimicrobial properties. In the current study we are exploring the connection between antimicrobial properties of DPPs and the inhibition of microbial ATP synthase. ATP synthase is the main source of ATP in almost all organisms from bacteria to man. Selective inhibition of bacterial ATP synthase could help in combating microbial infections.MethodsWild type E. coli pBWU13.4 is grown on minimal media to late log phase. Cells are harvested, French Pressed and centrifuged to isolate and purify membrane bound F1FO ATP synthase. Our null control is E. coli pUC118 with deleted ATPase gene. DPPs and its analogs induced inhibition of wild‐type and mutant membrane‐bound ATP synthase is performed at room temperature. Wild‐type, mutant, and null cell growth assays on limiting glucose are evaluated at OD595 using AccuSkan Plate reader over a period of 24 hours.ResultsVariable oxidative phosphorylation and ATPase activity pattern was observed between wild type phytochemical binding site mutant strains. Western blot showed purity and integrity of wild type and mutant enzymes. DPPs and its analogs caused variable inhibition of ATP synthase. The wild‐type enzyme was fully inhibited while mutant enzyme showed variable resistance in presence of DPPs. Variation in the growth pattern was in agreement with the inhibitory profiles of isolated enzymes.ConclusionsThe inhibitory profiles of isolated E. coli membrane‐bound ATP synthase shows that DPPs are potent inhibitors of ATP synthase. Inhibition of E. coli cell growth was also related to DPPs caused ATP synthase inhibition suggesting that ATP synthase is a valuable molecular drug target to combat microbial infection.
Microvascular anatomy and physiology laboratory class using the Australian cane toad
Authors: Murphy Timothy V. , Perry Michael A. , Ulman Lesley G. , Marden Nicole A. , Daniel Balu , Hanna Ghada , Housley Gary D. 
Published: May 2022

The microvasculature is a key part of the cardiovascular system and teaching microvascular structure and function is a key part of any cardiovascular science course. The Australian cane toad Bufo marinus / Rhinella marina is an economical and widely available species within Australia which can be used to demonstrate basic principles of blood flow and microvascular anatomy and physiology in an in vivo setting.In the laboratory class, cane toads of either gender (approx. 120 g) are anesthetized by immersion in 0.3 % ethyl 3‐aminobenzoate methanesulfonate (MS‐222) for 15 min and for the duration of the class (about 2 hr) swabs soaked in MS‐222 are placed on the skin. Level of anaesthesia is regularly assessed by the corneal reflex. At the end of the class toads are euthanized by cardiac removal (UNSW ACEC approval #19/67B). The surgical procedure involves a small incision in the upper abdomen, through which a lung, the bladder and the mesentery may be exteriorized and the microvasculature examined under a microscope equipped with a camera for video streaming to high definition monitors.The class is designed to demonstrate principles delivered in lectures and allow students to integrate knowledge of these principles into microvascular homeostasis and adaptation to interventions. Microvascular features which can be clearly demonstrated include microvascular anatomy and structural adaptations in different tissues, including the lung (pulmonary microcirculation), urinary bladder and mesentery. Observable physiological phenomena include blood flow velocity and its’ effect blood cell behaviour, axial streaming, plasma skimming, leukocyte rolling and adhesion, vasoconstriction and the response of the mesenteric microvasculature to haemorrhage.Student learning is assessed through pre‐ and post‐class exercises completed on‐line. The pre‐class exercise includes demonstrative videos and a brief quiz. This is intended to ensure students have read and understood the laboratory class notes, along with some basic vascular science, prior to the class, avoiding time‐wasting. The post‐class exercise includes video material recorded during the class accompanied by a quiz aimed at testing the students’ understanding of the observations made; for example, the physiological advantage of unique microvascular anatomy in certain tissues, or the mechanism underlying capillary swelling and hemodilution during haemorrhage. The videos can be edited into a package and used to deliver the class in an on‐line format if required. Thus, the cane toad preparation allows key aspects of microvascular anatomy, physiology, pharmacology and rheology to be demonstrated and taught in a clear and compelling fashion.
The long noncoding RNA uc.230/CUG‐binding protein 1 axis sustains intestinal epithelial homeostasis and response to tissue injury
Authors: Yu Tingxi , Kalakonda Sudhakar , Liu Xiangzheng , Han Naomi , Chung Hee K. , Xiao Lan , Jaladanki Rao N. , He Tong‐Chuan , Raufman Jean‐Pierre , Wang Jian‐Ying 
Published: May 2022

BACKGROUND & AIMSThe mammalian intestinal epithelium is colonized by complex microbiota and exposed to a wide variety of luminal noxious substances. Intestinal epithelial integrity is commonly disrupted in patients with critical disorders, but the exact underlying mechanisms are unclear. Long noncoding RNAs transcribed from ultraconserved regions (T‐UCRs) control different cell functions and are involved in gut pathologies. Here, we investigated the role of T‐UCRs in intestinal epithelial homeostasis and identified T‐UCR uc.230 as a major regulator of epithelial renewal, apoptosis, and barrier function.METHODSIntestinal mucosal tissues were collected from mice following dextran sodium sulfate (DSS) treatment or fasting, and from patients with ulcerative colitis and Crohn’s disease. We isolated primary enterocytes from the small intestine of mice and developed intestinal organoids. Levels of uc.230 were silenced in intestinal epithelial cells (IECs) and organoids by transfection with small interfering RNAs or elevated using a plasmid vector that overexpressed uc.230. Association of uc.230 with miR‐503 was determined by biotinylated RNA pulldown assays.RESULTSCompared with controls, intestinal mucosal tissues from mice with DSS‐induced colitis or fasted for 48 h and from patients with ulcerative colitis had increased levels of uc.230. The uc.230 content in Crohn’s disease tissues was the same as in control tissue. Silencing uc.230 inhibited the growth of IECs and intestinal organoids and resulted in epithelial barrier dysfunction. Silencing uc.230 also increased IEC vulnerability to apoptotic cell death, whereas increasing uc.230 levels protected IECs against apoptosis. Mechanistic studies revealed that uc.230 increased CUG‐binding protein 1 (CUGBP1) by acting as a natural decoy RNA for miR‐503, which interacts with Cugbp1 mRNA and represses its translation.CONCLUSIONSOur results indicate that uc.230 sustains intestinal mucosal homeostasis by promoting epithelial renewal and barrier function and protects IECs against apoptosis at least in part by serving as a natural sponge for miR‐503, thereby preserving CUGBP1 expression.
An organism‐wide atlas of tissue crosstalk in physical activity
Authors: Wei Wei 
Published: May 2022

Physical activity is a powerful physiologic stimulus that evokes tissue crosstalk and provides benefits for multiple organ systems. However, our understanding of how exercise remodels endocrine communication pathways remains largely incomplete due to a lack of tools. We have previously developed a chemical biology technology to label the collection of secreted proteins (e.g., secretome) from blood plasma in a cell type‐selective manner in mice1. Here we apply our approach to generating a 10‐organ, 21‐cell type plasma proteome atlas of secreted proteins in response to treadmill running in mice. 3% of detected protein‐cell type pairs exhibit selective bidirectional changes in response to exercise. Our secretome atlas validates known exercise‐regulated cell type–protein pairs, identifies new cellular sources for exercise‐regulated plasma proteins and discovers undescribed protein hormones that recapitulate the potent anti‐obesity effect of physical activity. We further show that the change in protein secretion from multiple cell types is induced in part by the elevation of extracellular lactate. On the cell type level, surprisingly, PDGFRa+ cells exhibit the most dynamic exercise‐regulated secretome, exceeding that of other classically exercise‐responsive cell types including myocytes, adipocytes, and hepatocytes. Further functional analysis reveals dramatic downregulation of several complement proteins from PDGFRa+ cells and may explain the anti‐fibrotic effect of physical activity across multiple organs. Taken together, these data provide a molecular and cell type‐specific map and uncover fundamental regulation of the dynamic changes to secreted polypeptides that mediate tissue crosstalk in physical activity.1. Wei, W*., Riley, NM* et al. Cell type‐selective secretome profiling in vivo. Nat Chem Biol (2021).