bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Peep into The Insulin Signaling Pathway of Palmatine: In Vitro, In Vivo & Ex‐Vivo
Authors: Okechukwu Patrick N. , Sharma Mridula N. , Selvaraja Malarvili N. , Nalweyiso Jacinta N. , Balachandran Abbirami N. , Chan Hor Kuan N. , Gaurav Anand N. 
Published: May 2022

BackgroundInsulin resistance, a characteristic of Diabetes Mellitus (DM), is caused by the inactivation of the insulin signaling pathway: IRS‐ PI3K ‐PKC ‐AKT2, and GLUT4. Conditions such as oxidative stress, glycated proteins, hyperglycemia, endoplasmic stress, fatty acid accumulation are major factors that cause inactivation of the insulin signaling pathway. Palmatine, an alkaloid, has been previously reported to possess antidiabetic, antioxidant, antiglycation properties and inhibition of endoplasmic stress.ObjectiveThe current study evaluated the rejuvenation of insulin signaling pathway by Palmatine in insulin‐resistant L6 skeletal muscle cells, extracted soleus muscle tissue and streptozotocin (STZ)‐induced diabetic rats.MethodDifferentiated L6 skeletal muscle cells and extracted soleus muscle tissue was induced insulin resistance using high insulin and glucose levels media. While STZ was used to induce type 2 DM in Sprague Dawley rats. The cells/tissue/animal was treated with palmatine, while glimepiride and metformin were used as standard drug. The differential gene expression of IRS‐1, PI3K, PKC‐α, AKT2, and GLUT4 was evaluated using qPCR. In silico docking of palmatine, tyrosine phosphatase SHP‐2 (SHP‐2) domains protein and Protein tyrosine phosphatase‐1B (PTP1B) was done using CDOCKER program available in DS V2.0 software.ResultThe results showed that IRS‐1, PI3K, AKT2, and GLUT4 were down‐regulated, while PKC‐α was upregulated in insulin resistant cells, muscle tissue and STZ‐induced diabetic rats. In contrast to the insulin‐resistant cells of the afore mentioned genes were up‐regulated in Palmatine treated cells/tissue/animals, while PKC‐α was down‐regulated. In silico docking of Palmatine showed a good affinity for tyrosine phosphatase SHP‐2 (SHP‐2) domains protein and Protein tyrosine phosphatase‐1B (PTP1B) proteins suggesting the modulation of these proteins.ConclusionPalmatine seem to have rejuvenate the impaired insulin signaling pathway through upregulation of the gene expression of IRS‐1, PI3K, AKT2, and GLUT4 and downregulation of PKC‐α expression. In silico docking shows a reduced modulation of PTP1B and increase affinity to SHP‐2. Reduced expression of PKC‐α and modulation of PTP1B maybe the reason for the observed increased expression of IRS‐1, PI3K, AKT2, and GLUT4.
The Neural basis of Complex Spatial, Social and Acoustic Behaviors – in Freely Behaving and Flying Bats
Authors: Yartsev Michael M. 
Published: May 2022

Research in our lab focused on the neural mechanisms of three main behavioral functions: (i) complex spatial navigation, (2) acoustic communication and (3) group social behaviors. To do so, we utilize an unusual model system, the bat, that offers several advantages.For the study of complex spatial navigation, we utilize bats because they are one of the most exquisite navigators on the planet and the only mammal capable of self‐propelled flight. We developed a human‐free, fully automated experimental setup where we discovered a temporally dissociated code for the representation of space and time in the hippocampus of flying bats. Furthermore, we developed a host of novel technologies, such as wireless calcium imaging in freely flying bats, which enables studying the stability of hippocampal codes in bats for the very first time.For the study of acoustic communication, we utilize bats because they are vocal experts and are one of the only mammals that are believed to acquire their vocalizations through an active learning process. If correct, this would be incredibly important because vocal learning has yet to be rigorously studied in the mammalian brain. Using advanced anatomical mapping methods and in combination with wireless neurophysiological approaches and behavioral monitoring systems we able to study this complex behavior in a detailed manner.Finally, for the study of group social behavior, we build on the fact bats are highly social, long‐lived, mammals that spend their entire life in group social living. We established technologies for multi‐animal wireless neurophysiological approaches that provided us a unique opportunity to study the neural computations underlying group social behavior. Through a series of studies, we uncovered an incredibly rich repertoire of neural representations for group social interactions in bats. These included the representation of social information both within and across the brains of socially interacting individuals as well as a unique representation of individual identity and social relationships between group members during communication.
The Role of Endothelial PARs in Modulating Insulin Signaling
Authors: Rajala Rahul , Griffin Courtney T. 
Published: May 2022

Endothelial cell (EC) dysfunction occurs in diabetes. Thrombin, a circulating serine protease with increased activity in diabetics, signals through the protease‐activated receptors 1 and 4 (PAR1/PAR4). To determine EC‐specific roles of PARs in insulin‐dependent diabetes mellitus (IDDM), we generated Par1fl//fl;Par4fl/fl;iCdh5(PAC)‐CreERT2 (Par1/4ECko) mice, which we induced with tamoxifen to delete Par1/Par4 from ECs and subsequently challenged with streptozotocin (STZ) to induce diabetes.Diabetic Par1/4ECko mice had reduced blood glucose levels (347.6 mg/dl, p<0.001) compared to their diabetic control littermates (538.4 mg/dl). At 12 weeks after STZ treatment, Par1/4ECkomice also weighed significantly more than their diabetic control littermates (~6.5%, p<0.024). Par1/4ECko mice likewise had visible abdominal fat pads, which are normally lost during the progression of diabetes. In addition, serum albumin (Reference Range: 2.5‐4.8 g/dl), which can be an indicator of hyperglycemia‐induced dehydration, was significantly elevated in diabetic control mice (5.2 g/dl, p<0.0007) but attenuated in Par1/4ECko mice (4.46 g/dl). Importantly, diabetic Par1/4ECko mice did not have significantly different (p<0.4518) serum insulin levels than their diabetic control littermates. Therefore, since diabetic Par1/4ECkomice showed better glucose uptake parameters than their control littermates, despite comparably reduced insulin levels, the mutant mice have whole‐body phenotypes that are consistent with heightened insulin sensitivity. Moreover, diabetic Par4ECkomice did not phenocopy the reduced blood glucose levels and weight loss seen Par1/4ECko mutants, indicating that endothelial Par1 deletion likely drives the heightened insulin sensitivity phenotypes.PAR1‐siRNA‐treated primary Human Umbilical Vein Endothelial Cells (HUVECs) showed increased basal insulin receptor (IR) activity (independent of insulin treatment) as measured by the uptake of a fluorescent glucose‐analog. PAR1‐siRNA‐treated HUVECs also showed increased expression of the IR Type A (IR‐A) spliceform, which has high insulin‐independent basal activity. Conversely, treatment of HUVECs with a PAR1 ligand resulted in increased levels of the insulin‐dependent IR Type B (IR‐B)spliceform. These data suggest that endothelial PAR1 can act as a modulator of IR splicing/activity.IR activity in ECs is implicated in insulin transport from the bloodstream to parenchymal tissues. Increased endothelial IR signaling results in increased whole‐body insulin sensitivity. We propose that endothelial PAR1 acts as a modulator of IR splicing/activity and subsequent insulin transcytosis. Therefore, loss of endothelial PAR1 potentially increases whole‐body insulin sensitivity by driving insulin transport to parenchymal tissues and subsequent glucose uptake.
Characterization of Buprenorphine Antagonism at Human Mu Opioid Receptors
Authors: Wedemeyer Michael J. , Berg Kelly A. , Clarke William P. 
Published: May 2022

In 2017 the US government declared a public health emergency to address the national opioid crisis. Despite this focused effort and increased awareness, opioid overdose deaths have since increased by over 50%. Individuals with opioid use disorder have increased risk of opioid related death, and medication assistance can facilitate recovery by reducing opioid cravings and withdrawal effects. One such medication is buprenorphine, an FDA approved drug for the treatment of opioid use disorder sold under the brand Suboxone®, Subutex®, and Zubsolv®. While buprenorphine is clinically described as a partial agonist of the mu opioid receptor (MOR), the literature demonstrates that its pharmacological characteristics are complex. In ongoing studies we are rigorously characterizing the pharmacological actions of buprenorphine at the human MOR.A genetically encoded biosensor was used to monitor cAMP levels in real time in HEK293 cells expressing a low density of the human MOR. In these cells, buprenorphine acted as an antagonist, shifting the concentration response curve of the MOR agonist, DAMGO, to the right (reduced potency) in a concentration‐dependent manner, similar to the action of the MOR antagonist naloxone. Unlike the effect of naloxone, buprenorphine also decreased the maximal response of DAMGO, an effect that saturated with increasing concentrations of buprenorphine. Similar antagonism by buprenorphine was observed when fentanyl was used as the MOR agonist. The effect of buprenorphine to decrease the potency of DAMGO was reduced by washout of buprenorphine. In contrast, the reduction in the maximal response to DAMGO was resistant to washout of buprenorphine. Both effects of buprenorphine (on DAMGO potency and efficacy) were prevented by pre‐treatment with naloxone, followed by rigorous washout. This suggested that buprenorphine’s effects were mediated by a naloxone‐sensitive binding site, presumably the orthosteric site. Given the slow rate of dissociation of buprenorphine binding to MOR in several published studies and the rapid time‐course involved in the cAMP response, we considered the possibility that the observed effects of buprenorphine were due to hemi‐equilibrium conditions. Computations conducted using hemi‐equilibrium equations support the hypothesis that the antagonist effects of buprenorphine in this system are due to hemi‐equilibrium. These kinetics were further supported in high‐expressing hMOR CHO cells, as buprenorphine agonism was resistant to reversal by naloxone or washing. As a frontline treatment for opioid use disorder, an understanding of buprenorphine’s pseudo‐irreversible binding may elucidate buprenorphine’s unique properties and facilitate future drug discovery.
Assessment of Metabolic Response During High‐Intensity Interval Exercise and Resting Vascular and Mitochondrial Function in Recreational CrossFit Participants
Authors: Pearson Regis C. , Olenick Alyssa A. , Jenkins Nathan T. 
Published: May 2022

BackgroundHigh‐intensity functional training (HIFT) can play a major role in physiological adaptations to prevent cardiometabolic disease. A majority of HIFT interventions incorporate a CrossFit (CF) training methodology, in which the goal is to increase work capacity over time while using mono‐structural, body weight, and weightlifting movements.MethodsWe measured aerobic capacity, metabolic response during high‐intensity interval exercise (HIIE), resting mitochondrial oxidative capacity, and resting large vessel function and microvascular reactivity in adults who have participated in a CF exercise for greater than one year vs. a sedentary group completing <2 h·wk‐1 of structure exercise for at least one year (SED).Results21 healthy, non‐smoking, participants were recruited (CF n = 13 vs. SED n = 8). CF participants had 40.4% greater absolute and 33.0% relative VO2 peak (P < 0.001) and lower body composition (CF = 18.6 (3.8) vs. SED = 30.3 (8.4); P < 0.001). CF participants had lower ratings of perceived exertion (P = 0.022) with higher respiratory responses (P < 0.05) and a lower heart rate response during HIIE (P = 0.024). CF participants had higher fat and carbohydrate oxidation when expressed as absolute and body weight relative values (P < 0.013), but not when expressed relative to lean mass (P > 0.200), indicating that skeletal muscle mass is a primary driver in substrate oxidation. CF participants had greater mitochondrial oxidative capacity (CF n = 12; P = 0.014). There were no differences in large artery function, but CF participants had greater baseline arterial diameter (CF n = 12; P = 0.004) and faster reperfusion following arterial occlusion (CF n = 12; P < 0.05), suggesting some influence of exercise habits on microvascular function.ConclusionOverall, these data support the effectiveness of HIFT programs, like CF exercise, to improve fitness and weight status and metabolic, mitochondrial, and vascular function.
Polyunsaturated fatty acids may decrease cancer risk in rural midwestern post‐menopausal women on vitamin D and calcium supplementation
Authors: Jackson Mariah , Angell Kathleen , Smith Lynette , Lappe Joan , Armas Laura , Ehlers Diane , D'Angelo Christopher , Hanson Corrine 
Published: May 2022

ObjectivesDetermine the extent to which fatty acid intakes at baseline are independently associated with cancer development by final study visit in a cohort of rural midwestern post‐menopausal women. Our hypothesis was higher polyunsaturated fatty acid (PUFA) intake at baseline will be protective in cancer development.MethodsThis study was a secondary analysis of participants enrolled in a randomized controlled trial evaluating the effect of a four‐year vitamin D and calcium supplementation intervention (2000 IU/d vitamin D3 and 1500 mg/d calcium) on cancer development in rural, midwestern post‐menopausal women from June 2009‐August 2015. The primary outcome was any‐type cancer assessed at 6‐month intervals over four years. Intakes of five fatty acid categories (saturated fat, monounsaturated fatty acids (MUFAs), PUFA, omega (n)‐3 fatty acids, n‐6 fatty acids) and seven individual fatty acids (linoleic acid, linolenic acid, stearidonic acid, arachidonic acid, eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA)) were evaluated via a Block Food Frequency Questionnaire at baseline. Multivariable logistic regression models, adjusted for body mass index (BMI), age, hormone replacement therapy, physical activity score, smoking status and treatment, were utilized to determine if consumption levels of fatty acids at baseline were associated with development of cancer. Interaction with treatment group was examined.ResultsThere were 2,109 participants available for analysis; 109 developed cancer. The majority of participants (99.5%) were White, with a mean age of 65.1 years and mean BMI of 29.9 kg/m2. Participants who developed cancer were significantly older (Cancer cases: 67.9 years, No cancer: 65 years; p<0.001) and had lower BMIs (Cancer cases: 28.4 kg/m2, No Cancer: 30.0 kg/m2; p= 0.015). There were no differences in demographics between treatment and placebo groups. In adjusted logistic regression models, PUFAs were associated with an increased risk of cancer development in the placebo group [OR 1.04; 95% CI (1.00, 1.08); p=0.048]. In the vitamin D and calcium treatment group, PUFA [OR 0.95; 95% CI (0.90, 1.00); p=0.042], linoleic acid [OR 0.93; 95% CI (0.88, 1.00); p=0.035], DHA [OR 0.98; 95% CI (0.97, 1.00); p=0.045] and n‐6 fatty acids [OR 0.93; 95% CI (0.88, 1.00); p=0.036] were found to decrease the odds of cancer development.ConclusionsWhen combined with vitamin D and calcium supplementation, PUFAs, including DHA, linoleic acid, and grouped n‐6 fatty acids, may be protective against cancer development in rural midwestern post‐menopausal women. In the absence of vitamin D and calcium supplementation, PUFA intake may increase odds of developing cancer. Further research should investigate fatty acid intake in the context of healthy diet patterns, including adequate vitamin D and calcium, for cancer prevention.
Reserve‐adaptive potential of sperm plasma in stallions of different ages
Authors: Atroshchenko Mikhail M. , Engalycheva Maria G. , Shitikova Anna M. 
Published: May 2022

During the aerobic metabolism of sperm, various reactive oxygen species are produced. Sperm cells are extremely vulnerable to oxidative stress (OS), which causes loss of membrane integrity, resulting in increased cell permeability, enzyme inactivation, structural DNA damage and cellular apoptosis. The aim of this study was to assess the reserve‐adaptive potential (RAP) of sperm plasma in stallions of various ages. RAP is the difference between the level of spontaneous and induced oxidative protein modification (OMP), reflecting the degree of OS. Spontaneous OMP reflects the accumulation of OS products in vivo. To determine the level of spontaneous OMP, a sperctrophotometric analysis of 2,4‐dinitrophenylhydrazones (DNPH) formed during the interaction of carbonyl derivatives of sperm plasma proteins with 2,4‐dinitrophenylhydrazine was carried out (spectrophotometer “SF‐2000”, Russia). R.L. Levine, modified by E.E. Dubinina. Measurements of DNPH derivatives were carried out at 14 wavelengths: in the range 260‐280nm for the registration of aldehyde dinitrophenylhydrazones of a neutral nature (ADNPHn), of a basic nature (ADNPHb) – 258‐264 and 428‐520nm, for the determination of ketone‐dinitrophenylhydrazones of a neutral nature (CDNPHn) 363‐370nm, basic character (CDNFGb) – 430‐434 and 524‐535nm. After that, the total amount of carbonyl derivatives was found. Induced OMP was determined in a similar way, but after adding iron (II) sulfate and hydrogen peroxide solutions to the reaction mixture. This provoked the formation of new carbonyl groups. The more RAP, the higher the ability of the biological material to resist the OS.Two groups of stallions were formed: the first group consisted of 15 stallions aged 13 to 20 years (average age 15.2±1.8 years), the second group consisted of 15 stallions aged 3 to 6 years (average age 4.0±0.5 years). The spontaneous OMP of sperm plasma in aged stallions was 495.6 Eod/ g protein, the induced one – 573.4 Eod / g protein. In the group of young stallions, spontaneous OMP – 347.2 Eod / g protein, induced – 485.3 Eod / g protein. The OMP level of sperm plasma in older stallions is initially higher than in young stallions (p = 0.04), which indicates the accumulation of carbonyl derivatives in vivo as a result of the prevalence of OS over the antioxidant capabilities of the organism. But when additional oxidizing components are added to the reaction mixture, the OMP of aged stallions does not increase as significantly as in young ones. RAP of sperm plasma of old stallions is 13.56%, of young stallions – 28.75%. The ability to withstand OS, the reserve of antioxidant capabilities of sperm plasma in young stallions is 2.1 times higher than in older stallions.
Influence of pole‐like kinase 1 on controlling contractility of the urinary tract and gene activated patterns of human bladder smooth muscle cells
Authors: Wang Xiaolong 
Published: May 2022

BackgroundPole like kinase (PLK)‐mediated promotion of proliferation and smooth muscle contraction in the lower urinary tract has been recently suggested and may offer putative targets and novel compounds for treatments of lower urinary tract symptoms. However, PLK function for contractility of smooth muscle and proliferation of bladder smooth muscle cells are unknown and the specificity of PLK inhibitors has been questioned.MethodsHere, human bladder smooth muscle cells (hBSMCs) inhibited with five PLK inhibitors (TAK960, MLN0905, Wortmannin, GW843682X, Centrinone‐B) and hBSMCs cultured in smooth muscle medium were compared. hBSMCs characteristics according to criteria set by the International Society of Cellular Therapy were confirmed by flow cytometry. Proliferation and viability were assessed by CCK‐8 assay. Contractions of human ureterovesical junction smooth muscle from donation after circulatory death renal transplantation were studied in an organ bath, where they were induced by the α1‐adrenoceptor agonists noradrenaline, phenylephrine, and electric field stimulation. Transcriptome sequencing of hBSMCs were obtained using NEBNext®UltraTM RNA Library Prep Kit for Illumina®. Gene set enrichment analysis was performed using the Kyoto Encyclopedia of Genes and Genomes and gene ontology terms.ResultsViability and proliferation of hBSMCs were reduced in a concentration‐ and time‐dependent manner. TAK960 inhibits adrenergic contractions of human ureterovesical junction smooth muscle, and proliferation of hBSMCs. No significant differences in other PLK inhibitors combined with control were found. Compared to hBSMCs in control medium, 323 upregulated genes and 160 downregulated genes were significantly altered by TAK960 24hours 500nM, respectively (FDR=0.05, FC=1.5). Genes most prominently and significantly upregulated by PLK inhibiting condition were IFI30, GBP4, VTN, SCG2, OAS2, RRAD, TNFSF10, KRT18, SFN, meanwhile, downregulated was CCL11 (log2FC ≥1.5 or ≤ ‐1.5). The most significantly relative signaling pathway included cytokine‐cytokine receptor interaction, systemic lupus erythematosus and transcriptional misregulation cancer (enrichment factor ≥0.05). Accession to cite for these sequences read archive data is PRJNA714246.ConclusionOur findings point to a certain role of PLK‐1 in of proliferation in hBSMCs and contractility in ureterovesical junction smooth muscle, which may be of relevance in lower urinary tract symptom. PLK‐1 inhibitors block these effects and show shared and divergent effects. PLK‐1 regulation of bladder smooth muscle growth and contraction may include genomic and nongenomic mechanisms.