bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sex Differences in GABAergic Neurotransmission to Gastric‐Projecting DMV Neurons
Authors: Jiang Yanyan , D'Angelo Michael Paul , Anselmi Laura , Travagli R. Alberto 
Published: November 2023

Functional gastrointestinal disorders, including delayed gastric emptying and decreased gastric motility, are more prevalent in women than men. It is suggested that the abundance of circulating estrogen in women may account for this difference. Gastric motility is finely tuned by the inputs arising from the dorsal motor nucleus of the vagus (DMV), which is itself controlled by a tonic, inhibitory GABAergic input. Estrogen has been shown to increase GABA expression in various CNS area, however the effect of estrogen to modulate GABAergic inputs onto DMV neurons, has not been investigated.The aims of the present study were to test the hypotheses that: i) in female rats the GABAergic tone controlling gastric motility is higher than in male rats; ii) estrogen levels in female rats influence the gastric motility response; and, iii) the response to exogenous application of estrogen in the DMV varies between male and female rats.In vivo anesthetized preparations of Sprague‐Dawley rats were used to compare gastric tone and motility in age‐matched males and virgin females; female rats were further subdivided according to the high estrogen (pro‐estrus and estrus) or low‐estrogen (meta‐ and di‐estrus) period of their cycle. To record gastric tone and motility, miniature strain gauges were sutured onto both the anterior corpus and antrum portions of the stomach. Following exposure of the dorsal brainstem, microinjections of the GABA‐A selective antagonist bicuculline methiodide (BIC, 0.1–5 pmoles/60nL) increased gastric tone and motility in a dose‐dependent manner; the increase was significantly higher in high estrogen (BIC 0.1pmoles increased gastric tone by 192±51 mg and motility by 158±53%; N=5) than in low estrogen females (100±29 mg and 175±112%; N=4) or males (56±17 mg and 111±19%; N=6). Conversely, microinjections of 17b‐estradiol (10pmoles/60nl) decreased gastric motility and tone by −141±26 mg and 145±33% (N=9) in males and by −111±54 mg and 153±40% (N=4) and −205±64 mg and 84±15% (N=5) in low and high‐estrogen females, respectively. The tone and motility effects of 17b‐estradiol were antagonized by pretreatment with the GPER receptor antagonist ICI182,720 (3nmol).The data suggests that i) in female rats the GABAergic tone controlling gastric motility is higher than in male rats; ii) estrogen levels in female rats influence the gastric motility response; and, iii) the response to exogenous application of estrogen in the DMV varies between male and female rats. Taken together, the data supports the hypothesis that estrogen mediates a greater inhibitory input to the DMV through increased expression GABAergic inputs.Support or Funding InformationSupported by ANMS and by AHA grants
NHE8 is required for goblet cell function
Authors: Xu Hua , Li Jing , Ghishan Fayez 
Published: November 2023

We reported previously that NHE8 plays an important role in mucosal protection. We found that mice lacking NHE8 are susceptible to DSS treatment and that loss of NHE8 expression in mice reduced muc2 expression and the number of PAS‐positive goblet cells. To increase our understanding of the impact and mechanism of these cellular changes, we studied cell proliferation using BrdU labeling. Our data showed that the rate of intestinal cell proliferation was enhanced in NHE8KO mice compared with their wild‐type littermates. We also analyzed additional genomic alterations by microarray and real‐time PCR. At the gene expression level, the intestinal expression of E2F2, a transcriptional factor involved in cell cycle regulation, Lgr5, an adult stem cell marker, and Wnt3, a secreted signaling protein, were elevated in NHE8KO. Therefore, the reduced mucosal protection in NHE8KO mice is most likely due to the disruption of normal goblet cell function.Support or Funding InformationThis study is supported by NIH grant R01DK073638.
Impact of Dietary Fat on Murine Gut Microbiota and Fatty Liver
Authors: Shang Yue , Derakhshani Hooman , Sarna Lindsei K , Siow Yaw L , Khafipour Ehsan , Karmin O 
Published: November 2023

Overconsumption of fat‐rich diets is a potential risk for obesity and metabolic disease such as non‐alcoholic fatty liver disease (NAFLD). Gut microbiota is an important factor that plays a role in regulating energy homeostasis and body weight control. An obesity‐associated gut microbiota can be induced by a high‐fat diet (HFD). Dietary intervention is one of the approaches for obesity management. The aim of our study was to investigate the impact of dietary fat intervention on the composition of gut microbiota. Male C57BL/6J mice were fed a control (10% kcal fat) or a HFD (60% kcal fat) for 7 weeks. A third group of mice was fed a HFD for 5 weeks followed by a control diet for 2 weeks (HFD + Control). DNA was extracted from cecal and colonic mucosa samples for amplification of V4 region of bacterial 16S rRNA genes and subjected to Illumina sequencing. Bioinformatics analyses were performed using QIIME. Statistical difference was determined by one‐way ANOVA and Permutational MANOVA at P < 0.05. At the end of 7 week trial, body weight gain of the control or the HFD+Control group was significantly lower than that of the HFD. Significant differences were observed on richness, abundance, diversity and functional properties of the gut microbiota in cecal and colonic mucosa between the control and the HFD group. The HFD+Control diet partially shifted microbiota profile to the control and reduced hepatic lipid accumulation. In conclusion, our results suggested that 5‐week HFD induced obesity‐associated microbiota. Withdrawal of HFD could prevent excess body weight gain and attenuated fatty liver while only partially restored the composition, diversity and profile of gut microbiota.Support or Funding InformationThe study was supported, in part, by NSERC, CIHR and St. Boniface Hospital Research Centre.
A Systematic Examination of the Association between Parental and Child Obesity: Systematic Review and Meta‐Analysis
Authors: Wang Youfa , Min Jungwon , Khuri Jacob , Li Miao 
Published: November 2023

ObjectivesChildhood obesity has become a global epidemic. Many factors contribute to childhood obesity. Parents and family influence children's energy‐balanced related behaviors and thus likely affect their weight status. However, the reported intergenerational association in obesity varies and no systematic examination has been reported. This study systematically examined the association between parental and child obesity/overweight and its variation by demographic characteristics and countries.MethodsA systematic literature search was conducted using PubMed for relevant studies published worldwide between Jan 1, 2000 and July 31, 2015. Strength of overall parent‐child (P‐C) obesity (including overweight, body mass index) associations was evaluated and compared across demographic factors and country economic development level including by meta‐analysis of meta‐regression models with random effects.ResultsIn total, 32 studies from 21 countries met inclusion criteria, and 27 of them reported odds ratios (ORs) as the key measure of P‐C obesity association and were included in meta‐analysis. About 2/3 of the 32 studies reported moderate or strong P‐C association in obesity. Meta‐analysis showed overall P‐C weight status association was significant (pooled OR=2.22 (2.09–2.36), ie, children with overweight/obese parents were twice more likely to be overweight/obese than those with normal weight parents), and the association varied by type of P‐C pairs, child age, parent/child weight status, and country economic development level. The association with both parents examined was more likely to be strong (63.0%) than those with either father (24.5%) or mother (30.2%) examined separately. The proportion of studies reporting strong P‐C association in high‐ and middle‐income countries were 40.2% and 16.1%, respectively. Meta‐analysis shows a stronger association (ie, higher ORs) was more likely to be reported for older children (β (SE)= 0.02 (0.01)), parental obesity (0.26 (0.10)) and child obesity (0.28 (0.12)) than if younger children and parental and child overweight were examined.ConclusionsAvailable research from multiple countries and with various study design show association between parent‐child obesity, but the association strength varied according to child age, types of parent‐child pair, their weight statuses and country‐level economic development. Families and parents are a key target for obesity prevention.Support or Funding InformationFunding: NIH U54 center grant HD070725.
Berberine Ameliorates Hepatic Steatosis and Suppresses Liver and Adipose Tissue Inflammation in Obesity Mice Independent of AMPK
Authors: Woo Shih‐Lung , Guo Ting , Guo Xin , Li Honggui , Zheng Juan , Botchlett Rachel , Cai Yuli , Li Xiaodong , Li Qifu , Xiao Xiaoqiu , Huo Yuqing , Wu Chaodong 
Published: November 2023

Much evidence has demonstrated the beneficial effects of berberine (BBR) on obesity‐associated non‐alcoholic fatty liver disease (NAFLD). However, the mechanisms underlying BBR actions remain to be explored. In the present study, we provided evidence to support an AMP‐activated protein kinase (AMPK)‐independent mechanism through which BBR suppresses obesity‐associated inflammation and improves hepatic steatosis. To induce obesity and obesity‐associated NAFLD, C57BL/6J mice were fed a high‐fat diet (HFD) for 12 weeks. During the last 4 weeks of the feeding regimen, the mice also were treated with BBR or vehicle control. Compared with the control, treatment with BBR decreased inflammation in both the liver and adipose tissue as indicated by reduction of the phosphorylation state of JNK1 and the mRNA levels of proinflammatory cytokines. Treatment of HFD‐fed mice with BBR also decreased hepatic steatosis, as well as the expression of lipogenesis‐related enzymes such as acetyl‐CoA carboxylase and fatty acid synthase. Interestingly, treatment of HFD‐fed mice with BBR did not significantly alter the phosphorylation state of AMPK in both the liver and adipose tissue. Consistently, BBR treatment significantly decreased the phosphorylation state of JNK1 in both hepatoma H4IIE cells and mouse primary hepatocytes in dose‐dependent and time‐dependent manners. BBR treatment also caused a decrease in palmitate‐induced fat deposition in primary mouse hepatocytes. However, BBR treatment did not alter the phosphorylation state of AMPK in H4IIE cells. Although increasing AMPK phosphorylation in primary mouse hepatocytes, BBR treatment suppressed the inflammatory responses in a manner irrelevant to AMPK. Taken together, these results suggest that BBR improves aspects of NAFLD mainly by suppressing inflammation in an AMPK‐independent manner.Support or Funding InformationThis study was support in part by NIH grants.
Deficiency of Vascular TLR4 Protects Against Lipopolysaccharide‐Induced Endothelial Dysfunction and NADPH oxidase‐derived Superoxide
Authors: Didion Sean 
Published: November 2023

The goal of this study was to test the hypothesis that homozygous toll‐like receptor 4 (TLR4) deficiency protects against lipopolysaccharide (LPS)‐induced endothelial dysfunction. Carotid arteries from wild‐type (TLR4+/+) and TLR4−/− mice were incubated in DMEM with either vehicle (saline) or LPS (0.05–0.50 μg/ml) for 22 hrs. LPS produced concentration‐dependent impairment (P<0.05) of endothelium‐dependent relaxation to acetylcholine in wild‐type mice. Both basal and NADPH‐stimulated superoxide levels were selectively increased in arteries from wild‐type mice treated with LPS. Tempol, a superoxide scavenger, improved endothelial responses and lowered superoxide levels in vessels from wild‐type mice treated with LPS. In contrast, LPS had no effect (P>0.05) on responses to acetylcholine or on basal or NADPH‐stimulated superoxide levels in TLR4−/− mice. The effects of LPS were selective for endothelium as responses to nitroprusside were unaltered (P>0.05) in either LPS‐treated wild‐type or TLR4−/− mice. Taken together, these genetic and pharmacological findings demonstrate that TLR4 expressed within the vascular wall contributes to LPS‐induced impairment of endothelial function, which is mediated in large part by increases in NADPH oxidase‐derived superoxide.Support or Funding InformationSupported by NIH HL‐089884 and HL‐107632.
Molecular Targets for Nitric Oxide Deficient Priapism
Authors: La Favor Justin D , Fu Zongming , Bivalacqua Trinity J , Van Eyk Jennifer E , Burnett Arthur L 
Published: November 2023

Priapism is a disorder in which prolonged penile erection persists uncontrollably, potentially leading to tissue damage. Priapism commonly afflicts patient populations with severe nitric oxide (NO) deficiency. As NO is a primary mediator of erection, the molecular mechanisms involved in the pathophysiology of NO‐deficient priapism are not well understood. The objective of this study was to identify novel molecular targets and signaling pathways in a mouse model of NO deficiency. Double NO synthase (NOS) knockout mutant mice (dNOS−/−), in which endothelial NOS and the α‐splice variant of neuronal NOS are deficient, were used as a model of NO‐deficiency. Intracavernosal pressure (ICP) was measured in response to low level electrical stimulation of the cavernous nerve in male dNOS−/− and age‐matched wild type (WT) mice (n=7/group). Priapic activity was demonstrated in the dNOS−/− mice, as the duration of erection during 4 minutes immediately following electrical stimulation was significantly elevated (64 ± 23 sec vs. 2.6 ± 2.6 sec; P = 0.02) relative to WT. Penile tissue was harvested, and samples processed and analyzed by reversed‐phase liquid chromatography tandem mass spectrometry. Proteins corresponding to differentially (P < 0.05) detected spectral counts were imported into Ingenuity Pathway Analysis (IPA) software for analysis of divergent protein networks and signaling pathways. 1518 total proteins were identified, 53 of which were significantly downregulated and 107 of which were significantly upregulated in dNOS−/− vs. WT. IPA revealed 11 protein networks, 98 transcription factors, and 81 signaling pathways predicted to be significantly dysregulated. Increased myosin light chain phosphatase protein content, protein kinase A signaling, and decreased adenosine catabolism in dNOS−/− penis all represent potential compensatory mechanisms to NO‐deficiency that may lead to priapism. In conclusion, these results identify novel candidate proteins that may play a role in priapism secondary to NO‐deficiency.Support or Funding InformationSupported by a mini grant from the National Kidney Foundation of Maryland and National Institutes of Health grant 1F32DK100082.
Deficiency in the Anti‐Aging Gene Klotho Promotes Aortic Valve Fibrosis through AMPKα‐mediated Activation of RUNX2
Authors: Chen Jianglei , Lin Yi , Sun Zhongjie 
Published: November 2023

BackgroundFibrotic aortic valve disease (FAVD) is an important cause of aortic stenosis, yet currently there is no effective treatment for FAVD due to its unknown etiology. The purpose of this study is to investigate whether deficiency in the anti‐aging Klotho gene (KL) promotes high‐fat‐diet‐induced FAVD and to explore the underlying molecular mechanism.Methods & ResultsHeterozygous Klotho‐deficient (KL+/−) mice and WT littermates were fed with a high‐fat diet (HFD) or normal diet for 13 weeks, followed by treatment with the AMPKa activator (AICAR) for an additional 2 weeks. A HFD caused a greater increase in collagen levels in the aortic valves of KL+/− mice than of WT mice, indicating that Klotho deficiency promotes HFD‐induced aortic valve fibrosis (AVF). AMPKa activity (pAMPKα) was decreased while protein expression of collagen I and RUNX2 was increased in the aortic valves of KL+/− mice fed with a HFD. Treatment with AICAR markedly attenuated HFD‐induced AVF in KL+/− mice. AICAR not only abolished the downregulation of pAMPKα but also eliminated the upregulation of collagen I and RUNX2 in the aortic valves of KL+/− mice fed with HFD. In cultured porcine aortic valve interstitial cells, Klotho‐deficient serum plus cholesterol increased RUNX2 and collagen I protein expression, which were attenuated by activation of AMPKa by AICAR. Interestingly, silencing of RUNX2 abolished the stimulatory effect of Klotho deficiency on cholesterol‐induced upregulation of matrix proteins, including collagen I and osteocalcin.ConclusionKlotho gene deficiency promotes HFD‐induced fibrosis in aortic valves, likely through the AMPKa–RUNX2 pathway.Support or Funding Informationsupported by NIH R01 HL118558, DK093403, and HL105302.
Lethal H5N1 Virus Invades Medullary Respiratory‐related Neurons Responsible for Respiratory Chemoreflexes and Rhythm/Pattern Generation
Authors: Zhuang Jianguo , Gao Xiuping , Ye Chunyan , Xu Fadi 
Published: November 2023

Highly pathogenic H5N1 avian influenza virus is neurotropic and its infection is lethal mainly due to the respiratory failure. Studies have shown that lethal H5N1 HK483 (A/HK/483/97) virus invades the central nervous system, blunts ventilatory responses to hypoxia and hypercapnia, and induces apneas, leading to death at 6–8 days postinfection (dpi) in mice (Gao et al, Journal of virology 1999, Zhuang et al., ATS 2013, EB 2015). However, the relevant mechanisms remain unclear. We asked if HK483 H5N1 virus: 1) invaded neurons and/or astrocytes in the medullary respiratory‐related nuclei; 2) infected those respiratory neurons expressing neurokinin 1 receptor (NK1R), tyrosine hydroxylase (TH, a marker of catecholaminergic neuron), or tryptophan hydroxylase (TPH, a marker of serotonergic neuron); and 3) altered the these neural expressions of NK1R, TH, or TPH. Female Balb/c mice were intranasally inoculated with vehicle (Ctrl) or HK483 virus (100 PFU) and euthanized at 6–7 dpi. After fixation, the medullary sections were immunohistochemically processed to detect the nucleoprotein of H5N1 influenza A (NP) in neurons and/or astrocytes marked by glial fibrillary acidic protein (GFAP) and its co‐labeling with immunoreactivity (IR) of NK1R, TH, or TPH. We found that: 1) HK483 virus mainly invaded the respiratory‐related nuclei including the nucleus of solitary tract (NTS), ambiguus nucleus (Amb), pre‐Botzinger complex (PBC), Botzinger complex (BC), retrotrapzoid nucleus (RTN) and their neighboring areas with the raphe less infected and the invasion in astrocytes was limited; 2) approximately 20–50% of NK1R neurons (the NTS, PBC, and RTN), 10–30% of TH‐IR neurons (C1/2, A1 areas, the NTS, and locus ceruleus), and < 10% TPH (the raphe) were co‐labeled with NP; and 3) significantly reduced the density of NK1R‐IR in the corresponding nuclei with less effect on reduction of the raphe neural population expressing TPH and no effect on TH‐IR. We conclude that HK483 is capable of invading the medullary respiratory nuclei, particularly those neurons expressing NK1R or TPH to depress NK1R or TPH expression, which likely contributes to depression of the ventilatory responses to hypoxia and hypercapnia and introduction of apneas/respiratory failure, leading to the death.Support or Funding InformationSupported by HL119683 and HL107462
Pulmonary Vascular Pressures and Ventilatory Control during Exercise in Heart Failure with Preserved or Reduced Ejection Fraction
Authors: Van Iterson Erik H. , Snyder Eric M. , Olson Thomas P. 
Published: November 2023

IntroductionA disproportionate increase in ventilation relative to carbon dioxide output (VE/VCO2) slope is a strong prognostic indicator in heart failure with reduced (HFrEF) or preserved ejection fraction (HFpEF) patients. Similarly, secondary pulmonary hypertension also poses a profound mortality risk in these patients. Exaggerated ventilation and pulmonary pressures appear to parallel one another during exercise in HF. However, an underlying pathophysiologic mechanism relating these two outcomes remains incompletely understood. Thus, the aim of this study was to determine associations between anatomical and physiologic metrics of ventilatory control and pulmonary pressures during exercise in HFrEF and HFpEF.Methods32 HFpEF and 33 HFrEF (EF: 62±2 vs 22±1%; male: 17 and 31; age: 70±2 and 55±2 yrs; BMI: 34±1 and 28±1 kg/m2; weight: 98±4 and 86±3 (all P<0.05), respectively, participated in cardiopulmonary exercise testing during right heart catheterization. Measures of pulmonary pressures included: systolic (PASP); diastolic (PADP); mean (mPAP); wedge (PCWP); pulse (PP); transpulmonary gradient (TPG). Partial pressure of arterial oxygen (PaO2) and carbon dioxide (PaCO2) were measured via radial arterial catheterization. Ventilation was measured continuously using a breath by breath metabolic measurement system. We calculated dead space to tidal volume [Vd/Vt=(VE×PaCO2)‐(863×VCO2)/(VE×PaCO2)] and Alveolar‐arterial oxygen gradient (A‐a)=PAO2‐PaO2; VE/VCO2 slope=rest to peak exercise.ResultsAt rest, pulmonary pressures did not differ between groups (P>0.05); whereas Vd/Vt was lower in HFpEF vs HFrEF (0.42±0.02 vs 0.48±0.01, respectively, P=0.03). At peak exercise, HFpEF had higher PCWP (34.8±1.2 vs 30.2±1.5 mm Hg, P=0.02) and PADP (35.4±1.3 vs 29.8±1.6 mm Hg, P=0.02), but lower Vd/Vt (0.32±0.02 vs 0.38±0.01, P=0.03) and VE/VCO2 slope (35.3±1.6 vs 43.5±2.0, P<0.01) vs HFrEF. At peak in HFpEF, Vd/Vt was related to PASP (r=0.40, P=0.03), PP (r=0.44, P=0.01), and TPG (r=0.53, P<0.01), and A‐a was related to PASP (r=0.43, P=0.01), PP (r=0.44, P=0.01), and TPG (r=0.46, P<0.01). At peak in HFrEF, Vd/Vt was related to PADP (r=0.41, P=0.02), and A‐a was related to PASP (r=0.43, P<0.01), PADP (r=0.42, P=0.01), and mPAP (r=0.44, P<0.01). There were no significant relationships between measures of pulmonary pressures and PaCO2. Remaining correlations between pulmonary pressures and anatomical or physiologic metrics of ventilatory control were negligible in both groups (P>0.05).ConclusionThese data suggest pulmonary pressures play a role in dead space ventilation during exercise in HF. Moreover, as suggested by the A‐a gradient, augmented pulmonary pressures may contribute to an exacerbation of ventilation/perfusion mismatch during exercise in both HFrEF and HFpEF patients.Support or Funding InformationNone