bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of the Momordica charantia (MC) in the alterations caused by administration of a high fat diet (HL) as a model of non‐alcoholic fatty liver disease (NAFLD)
Authors: Santos Maria Emilia Soares Martins , Lemos Bruno Silva Oliveira , Teixeira Keyt Braz , Silva Lilian Nayra , Souza Geovana Alves , Pinto Flávia Carmo Horta , dos Santos Lima Luciana Alves Ribeiro , Santos Maria Emilia Soares Martins
Published: June 2021

The aim of this work was to study the effects of MC on metabolism from NAFLD animals. Mice received a control (C) or a HL diet/8 weeks. One HL group was administered with MC extract/5days (HLT5) after the diet period. Other HL group received MC tea (HLTT) instead of water at the same period of diet. Blood was collected for analysis of glucose (GLU), cholesterol (CTO), high density lipoprotein (HDL), triacylglycerol (TAG), alanine aminotransferase (ALT) and very low density lipoprotein (VLDL). Liver was removed for histology. HL showed an increase in body weight (35.0 ±0.8 vs 48.3 ±1.0), liver weight (C 1.7 ±0.07 vs HL 2.6 ±0.01), GLU (562.1±13.4 vs 701.8 ±20.1), ALT (57.3 ±6.4 vs 105.1 ±6.1), CTO (145.8 ±9.9 vs 195.2±14.9), HDL (107.7± 0.6 vs 171.9 ±8.7), VLDL (21.5 ±1.54 vs 34.8 ± 6.8) and TAG (75.0 ±6.6 vs 171.5±34.4) in comparison to C. MC extract administration decreased: GLU (701.8 ±20.1 vs 306.7 ±31.0), CTO (195.2 ±14.9 vs 103.6 ±4.6), VLDL (34.8 ± 6.8 vs 8.6 ± 0.5), HDL (171.9 ±8.7 vs 97.1 ±7.8), TAG (171.5 ±34.4 vs 43.4 ± 2.6), liver weight (2.6 ±0,01 vs 1.7 ±0.13). HLTT animals decreased: GLI (661.8±26.2 vs. 528.5±57.9), VLDL (37.5±4.142 vs. 9.7±1.1) and TAG (174.07±16.71 vs. 48.9±5.64) in comparison to HL. Significant improvement was observed histologically in liver from animals with NAFLD when they received MC. MC seems to be efficient in treatment/preventing hyperglycemia, hyperlipidemia and NAFLD.

Financial support: CNPq
Effect of prolonged, mild cold exposure on metabolic regulation in insulin resistant overweight and obese men
Authors: Chondronikola Maria , Hurren Nicholas M. , Porter Graig , Annamalai Palam , Volpi Elena , Borsheim Elisabet , Sidossis Labros
Published: June 2021

The purpose of this study was to investigate the effect of prolonged, mild cold exposure (CE) on metabolic regulation in middle‐aged and older, overweight and obese males. Eight men (51.9 ± 4.4 years old, BMI 29.7 ± 1.4 kg/m2) completed two trials: a) 8 h CE (17 – 19°C) above the shivering threshold and b) 8 h in thermoneutral (TN) conditions (26 – 28 °C). In the final two hours of each trial, hyperinsulinemic euglycemic clamps were performed to determine insulin sensitivity. Insulin sensitivity was higher after CE compared with TN conditions (1.5 ± 0.3 *10−3 dL−1.kg−1.min−1 vs 1.0 ± 0.2*10−3 dL−1.kg−1.min−1, p=0.02). Mild CE decreased skin (31.1 ± 1.1 °C vs 34.5 ± 0.3 °C, p < 0.001), but not core temperatures (36.5 ± 0.3 °C vs 36.8 ± 0.6 °C) compared with TN conditions. Next day fasting plasma triglycerides and VLDL‐cholesterol were lower after the CE trial compared with TN (164 ± 17 mg.dl−1 vs 197 ± 22 mg.dl−1, p = 0.02 and 33±4 mg.dl−1 vs 39.3 ± 4.4 mg.dl−1, p = 0.01, respectively). Prolonged, mild CE improves the metabolic profile of insulin resistant obese men. Future studies will determine if these findings are mediated via activation of brown adipose tissue, as it has recently been suggested that CE activates brown adipose tissue in many humans. Research Support: CTSA UL1TR000071, Pepper Center Pilot Grant, Sealy Center on Aging, Shriners Hospitals for Children Grant 84090, Onassis Foundation
Diosgenin administration improve hyperglycemia through the activation of muscular glucose metabolism
Authors: Sato Koji , Fujita Satoshi , Iemitsu Motoyuki
Published: June 2021

The purpose of this study was to investigate whether administration of diosgenin, a steroid structurally similar to DHEA (dehydroepiandrosterone) and contained highly in dioscorea, improved hyperglycemia in T1DM model rats through the enhancement of muscular GLUT4 signaling. The study was conducted in conformance with the FASEB Statement of Principles for the use of animals in research and education. After one week of STZ injection, fasting glucose level was taken from the tail vein and was monitored for 150min after diosgenin or discorea was injected (3mg/kg). The other day, muscle was taken after 150min of diosgenin or discorea injections. Serum DHEA level was significantly increased after 120min of diosgenin or discorea injections, with concomitant and significant decrease in blood glucose level. Moreover, GLUT4 translocation, as well as phosphorylation of Akt and PKC ζ/λ improved significantly with diosgenin or dioscorea administration. However, these effects of diosgenin and dioscorea were blocked by a dehydrotestosterone DHT inhibitor which inhibited binding to androgen receptor. These results suggest that diosgenin‐induced increase of DHEA level may contribute to the improvement of hyperglycemia through muscular GLUT4 signaling pathway activation in type1 diabetes rats. This work was supported by grants‐in‐aid for Scientific Research of Japan (21300254, 22650166)
The appearance of the gastric submucosal microvessels and the degree of gastric mucosal atrophy may predict prognosis in gastric cancer patients
Authors: Tomchuk Olesya
Published: June 2021

This study examined the morphological changes of the gastric mucosa around the stomach cancer in patients to correlate them with tumor progression. Twenty‐three patients (average age 61.6±2.68 years) with histologically confirmed gastric cancer were selected for study. Tissue samples, taken at least 3–5cm away from the edge of the tumor, were processed for histopathological examination. Sections were stained with Mayer's hematoxylineosin, and CD34 antibody. We found that the tumor size positively correlated with the severity of atrophic changes in the gastric mucosa (r=0.670, p=0.03) and negatively correlated with the completion of the eradication therapy prior to surgery (r=−0.540, p=0.022). For instance, the tumors were significantly larger in patients with the higher degree of gastric mucosal atrophy (5.53±1cm vs. 3.5±1.9cm, respectfully; p=0.016) and were significantly smaller in the patients with the history of eradication therapy (3.4±2.45cm vs. 5.2±1.74cm, respectfully; p=0.065). The presence of metastases in regional lymph nodes positively correlated with the presence of dilated capillaries in the gastric submucosa (r=0.863, p=0.001; in 85.7% of cases) and severe gastric mucosal atrophy (r=0.560, p=0.031; in 71.4% of cases). Therefore, we propose that the marked dilatation of submucosal microvessels and the high degree of gastric mucosal atrophy may be indicative of gastric cancer progression.
Effect of leptin on HCT‐116 colon cancer cell growth
Authors: Tanner Stephen , Osborne David L
Published: June 2021

Obesity is a growing health problem of particular importance because it predisposes individuals to a number of diseases. One major disease linked with obesity is colon cancer. Obesity leads to increased concentrations of leptin and leptin may be one of the factors that leads to growth and progression of colon cancer. This study sought to analyze the effect of leptin on the proliferative activity of HCT‐116 colon carcinoma cells. Positive control cells were grown in RPMI 1640 media supplemented with 10 % FCS. Negative control cells were grown under similar conditions but using RPMI1640 + 2% FCS. All experimental cells were grown under similar conditions but using RPMI1640 + 2% FCS supplemented with various concentrations of leptin ranging from 50–400 ng/mL. Cells were counted on days 1, 3, and 5 following initiation of culture with 50,000 cells per well in each well of a 6 well plate. The results showed that while a trend of increased growth with increased leptin concentration could be inferred, statistical analysis indicated that there was no difference between RPMI 1640 +2% FCS media controls and groups treated with leptin. Results indicate that variability between cell types and origin may lead to differing responses to leptin in colon cancer cells. Our results indicate that HCT116 human colon cancer cells may not express leptin receptors or do not respond to leptin and therefore, do not serve as a good model for further studies.
Genome‐wide combinatorial transcriptional regulatory dynamics during early onset of liver regeneration and chronic alcohol intake
Authors: PATRA BISWANATH , Kuttippurathu Lakshmi , COOK DAN , HOEK JAN B , Vadigepalli Rajanikanth
Published: June 2021

The objective of this study is to identify genome‐scale transcriptional regulatory network dynamics affected by chronic alcohol intake during the early phase of liver regeneration. Rats were fed liquid diet containing 36% of total calories derived from ethanol for 5 weeks, and controls were pair‐fed an isocaloric liquid diet. A 70% partial hepatectomy (PHx) was performed. Genome‐wide binding targets for NF‐kB, C/EBP‐beta, and STAT3 were detected either by Roche NimbleGen ChIP‐chip or ABI SOLiD sequencing approach. Our findings indicate significant differences in the binding profiles of all the three regulators in chronic ethanol compared to isocaloric controls, at baseline as well as in response to PHx. Pathway analysis revealed that potential target genes specific to the chronic ethanol group participate in key processes such as mitochondria processing, DNA methylation, cell death, proteolysis, histone modification, lipid biosynthetic process, and regulation of cell cycle etc. Our results revealed combinatorial binding dynamics of multiple TFs from ChIP‐seq and DNA binding motif. Our results provide a first system‐wide view of the regulatory network dynamics in liver regeneration as it is affected by alcohol, and implicate combinatorial regulatory modules as participating in the adverse liver regeneration response. Research Support: NIH R01 AA018873, K05 AA017261, and T32 AA007463.
Roles of RSK1 in LPA5R‐induced NHE3 regulation
Authors: Yoo Byong Kwon , Yun C. Chris
Published: June 2021

Na+/H+ exchanger 3 (NHE3) mediates bulk of Na+ and fluid absorption in the small intestine and colon. We have previously shown that lysophosphatidic acid (LPA) receptor LPA5R regulates NHE3 by two distinct signaling pathways involving Pyk2 and Erk1/2. The goal of this study is to identify downstream effectors of Pyk2 and Erk1/2. In Caco‐2bbe cells stably expressing NHE3 and LPA5R, Caco‐2bbe/NHE3V/LPA5R, LPA treatment increased phosphorylation level of PDK1. Knockdown of Pyk2 blocked LPA‐induced phosphorylation of PDK1, suggesting that PDK1 is dependent on Pyk2. Knockdown of PDK1 subsequently abrogated LPA‐induced NHE3 activation demonstrating a role of PDK1 in NHE3 regulation. RSK1 is a known target of Erk1/2. We found that LPA activates RSK1 in Caco‐2bbe cells. To determine whether RSK1 is a downstream target of PDK1 or Erk1/2, we used two distinct antibodies to RSK1 that recognize phosphorylation at S221 by PDK1 or at S380 by Erk1/2. Phosphorylation at S221 and S380 was attenuated by PDK1 knockdown and PD98059, respectively, suggesting that both PDK1 and Erk1/2 are involved in LPA‐induced RSK1 activation. In addition, LPA induced colocalization of RSK1 and NHE3 at the plasma membrane, and knockdown of RSK1 diminished LPA‐induced NHE3 activation. In summary, RSK1 is the converging target of Pyk2 and Erk1/2 and is involved in NHE3 activation by LPA5R.
Farnesoid X receptor activation inhibits colonic epithelial CFTR expression ‐ a new target for antidiarrheal therapy?
Authors: Mroz Magdalena Slawa , Keating Niamh , Ward Jospeh BJ , Amu Sylvie , Sarker Rafiquel , Fallon Padraic Gerard , Donowitz Mark , Keely Stephen J
Published: June 2021

Cl− secretion, a main driving force for intestinal fluid secretion, can become dysregulated in conditions of disease, leading to diarrhea. While diarrheal diseases constitute a huge global health and economic burden, available medications are unsatisfactory. We previously showed that, at physiological concentrations, deoxycholic acid (DCA), inhibits intestinal secretory function. Here, we investigated the nuclear bile acid receptor, farnesoid X receptor (FXR), as a potential target for development of antidiarrheal drugs.

GW4064 was used as a specific FXR agonist. Cl− secretion was measured in Ussing chambers as increase in short‐circuit current and protein expression was determined by western blotting and qPCR.

Treatment of T84 colonic epithelial cells with either GW4064 (5μM) or DCA (50μM) for 6–24 hrs induced nuclear translocation of FXR and inhibited agonist‐induced Cl− secretory responses. Furthermore, in vivo administration of GW4064 (50mg/kg) also attenuated colonic secretion and reduced stool water in a mouse model of diarrheal disease. At the molecular level, GW4064 inhibited CFTR Cl− channel currents and reduced mRNA, protein, and membrane expression of CFTR. In contrast, GW4064 did not alter the activity of ENaC or NHE3.

These data reveal a new role for FXR in regulating epithelial CFTR expression and secretory function and suggest that FXR agonists may be useful in treating diarrheal diseases.
NANC neuron mediated relaxation of murine gastric fundus smooth muscle: Phosphorylation of MYPT1 on T696 (ROK site) and S695 (PKG site) are mutually exclusive
Authors: Pfitzer Gabriele , Hunger Mona S. , Puetz Sandra , Metzler Doris , Lubomirov Lubomir T , Somlyo Avril V.
Published: June 2021

Myosin phosphatase is the target of activating and relaxing pathways which modulate its activity by phosphorylation of its regulatory subunit, MYPT1, at several sites, and by telokin. Here, we investigated whether phosphorylation of MYPT1‐S695 is involved in relaxation of murine gastric fundus by NANC neurons known to act through cAMP/cGMP. Endothelin‐1 (ET‐1) preconstricted SM strips were relaxed by electrical field stimulation (EFS, 10 Hz, 37°C, blocked adrenergic and cholinergic responses) and shock frozen for western blot analysis with phosphospecific antibodies at desired time points. EFS induced relaxation (half time ~8s) was preceded by dephosphorylation of MLC‐20 (S19) and of MYPT1 (pMYPT1) at T696/T853 indicating de‐inhibition of MLCP. Dephosphorylation of T696 preceded the EFS‐induced increase in pMYPT1‐S695. During force recovery (half time ~150s) pMYPT1‐S695 was dephosphorylated within 30s whereas re‐phosphorylation of T696/T853 paralleled force recovery. Relaxation was associated with phosphorylation of telokin and ablation of telokin reduced relaxation by ~25% (at 2 Hz).To conclude, our results indicate that phosphorylation of S695 is not involved in the initiation of PKA/PKG mediated relaxation but may prevent re‐phosphorylation of T696. Our results are consistent with the notion that phosphorylation of T696/S695 is mutually exclusive. Funding: German Research Foundation.
Pharmacologically‐Induced Alterations in Gastrointestinal Motility Affects Regional Colonic Microbial Assemblage
Authors: Touw Ketrija , Wang Yunwei , Huang Edmond , Leone Vanessa , Chang Eugene
Published: June 2021

Recent studies have implicated a role of gut microbes in the pathogenesis of irritable bowel syndrome (IBS). However, it is largely unknown how changes in motility affect the gut microbiome. In this study we sought to determine how delayed gastrointestinal (GI) motility affects the colonic microbial population. Adult C57Bl/6 mice treated with 0.1% Loperamide in drinking water for 7 days to induce constipation. Loperamide treated mice had two fold increase in whole gut transit time and two fold decrease in stool output, indicating constipation. Bacterial DNA from stool and cecum samples were extracted and analyzed by T‐RFLP, HiSeq and 16S rRNA clone‐library screening. Taxon assignments based on 16S rRNA profiles showed greater abundance of Bacteroidetes in stool than Firmicutes in cecal samples of constipated mice versus controls. In contrast, Loperamide increased Firmicutes and decreased Bacteroidetes in the cecum. The most significant changes in bacterial species were Prevotella, Parabacteroides and Lactobacillus. Overall, these results suggest that delayed GI transit significantly alters the enteric microbiome in a regional and taxon specific manner. These changes are likely to be associated with alterations in functional profiles of gut microbiota that can impact physiological host response, and, in some instances, cause motility disturbances that become symptomatic and contribute to IBS.
Human prorenin induces Ang II‐independent pressor response in neuron‐specific human (pro)renin receptor transgenic mice
Authors: Peng Hua , Li Wenchen , Sigmund Curt D , Feng Yumei
Published: June 2021

We previously showed that (pro)renin receptor (PRR) mediated Ang II‐independent signals in neuronal cells. To elucidate the significance of these signaling pathways in vivo, we developed a novel neuron‐specific human PRR transgenic mouse model (syn‐hPRR). Despite high hPRR expression level selectively in brain of syn‐hPRR mice, baseline blood pressure (BP, mmHg) and heart rate (HR, bpm) were similar in syn‐hPRR and NT mice measured by telemetry (n=8/group). However, the cardiac (ÄHR: −68.4±1.3 VS. −45.3±1.4) and vasomotor (ÄMAP: −41.1±2.1 VS. −28.9±0.4) sympathetic tones were significantly higher in syn‐hPRR compared with NT mice. Interestingly, ICV infusion (0.3μl/min) of human prorenin (300ng) significantly increased BP in syn‐hPRR mice (ÄMAP: 22.5±4.6, P<0.05), but not in NT mice (ÄMAP: 1.6±0.8). Although human prorenin‐induced pressor response in syn‐hPRR mice could not be blocked by ICV infusion of losartan (30μg/μl) or captopril (10μg/μl), the pressor response was abolished by ICV infusion of either reactive oxygen species (ROS) scavenger tempol (100μM, ÄMAP: 4.5±0.5 P<0.05) or NADPH oxidase inhibitor diphenyleneiodonium (10μM, ÄMAP: 4.7±1.0 P<0.05). These data suggest that human prorenin via binding to hPRR mediates Ang II‐independent pressor response in syn‐hPRR mice through increase in brain ROS levels, suggesting a novel role of Ang II‐independent pathways of human PRR in hypertension.
Loss of cilia increases angiotensin II mediated apical angiotensin‐1‐receptor localization in collecting duct cells
Authors: Saigusa Takamitsu , Dang Yujing , Morinelli Thomas A , Bell P. Darwin
Published: June 2021

Hypertension (HTN) is frequently present early on in Polycystic Kidney Disease (PKD) before a significant cystic burden has occurred. However, the mechanism for PKD‐induced HTN has not been fully elucidated. The renin‐angiotensin‐system (RAS) is one factor contributing to HTN in PKD, although there are no consistent changes in plasma renin/aldosterone levels in animals or humans with PKD. We hypothesized that intra‐renal‐RAS rather than the systemic RAS may be important in PKD. As an initial approach, we used immortalized collecting duct (CD) cell line derived from Orpk mice with a hypomorphic Ift88 gene resulting in stunted/absent cilia: cilia (−), and Ift88 rescued cells: cilia (+) with normal cilia. There was increased abundance of the angiotensin‐1‐receptor (AT1R) and in (pro) renin in cilia (−) compared to cilia (+) cells. Using biotinylation of apical membranes, 30 min treatment with Ang II (1nM) increased apical AT1R expression in cilia (−) cells but not in cilia (+) cells. Using a GFP‐AT1R construct transfected into these CD cells, we found that with Ang II administration cilia (−) but not cilia (+) cells had predominant luminal membrane localization of GFP‐AT1R. Finally, with addition of Ang II, phospho ERK was increased in cilia (−) compared to cilia (+). Thus, loss of cilia leads to increased abundance of AT1R and the targeting of AT1R to the apical membrane which may lead to increased RAS activity in the CD in PKD.
Phosphoproteomic Analysis of Signaling Responses to Angiotensin II in Proximal Tubules of NHE3‐deficient Mice
Authors: Li Xiao C , Miguel‐Qin Elisa , Shull Gary E , Zhuo Jia L
Published: June 2021

Mice with the Na+/H+ exchanger 3 deficiency (Nhe3−/−) develop hypotension, suggesting an important role of NHE3 in the proximal tubule of the kidney. Using control (Nhe3+/+) and Nhe3−/− mice, we determined blood pressure (SBP) and signaling responses to angiotensin II (ANG II) for 2 weeks (40 ng/min, i.p., via osmotic minipump). Na+‐K+‐ATPase α1, Na+/HCO3−, phosphorylated PKCα (p‐PKCα), glycogen synthase kinase 3α/β (p‐GSK3α/β), MAP kinases ERK1/2 (p‐ERK1/2), CREB (p‐ CREB), and aquaporin 1 (AQP‐1) proteins in the proximal tubule of the kidney were measured by western blot analysis. Under basal conditions, Nhe3−/− mice had lower SBP (p<0.01) and 24 h urinary water, Na+ and K+ excretion (p<0.01). Plasma ANG II and aldosterone were significantly elevated (p<0.01), but kidney ANG II was unaltered in Nhe3−/− mice. Na+‐K+‐ATPase α1, Na+/HCO3−, p‐PKCα, p‐GSK3α/β, p‐ERK1/2, and AQP‐1 proteins were significantly increased in Nhe3−/− mice compared with Nhe3+/+ mice (p<0.01). ANG II increased SBP and 24 h urinary Na+ and K+ excretion to greater extents in Nhe3+/+ than in Nhe3−/− mice. Although ANG II increased Na+‐K+‐ATPase α1, Na+/HCO3−, p‐ PKCα, p‐ERK1/2, and AQP‐1 signaling responses in Nhe3+/+ mice, it had no effects on these signaling proteins in Nhe3−/− mice. We concluded that global deletion of the NHE3 gene induces significant compensatory transporter and signaling responses in the proximal tubule of the kidney.
Angiotensin II receptor signaling modulates vascular smooth muscle sensitivity to nitric oxide in an adiposity‐specific manner in healthy adults
Authors: Christou Demetra D. , Hwang Moon‐Hyon , Walker Ashley E. , Zigler Melanie , Seals Douglas R.
Published: June 2021

Vascular function (VF) is a predictor of cardiovascular disease (CVD) risk. Angiotensin II (Ang II) contributes to impaired VF in patients with CVD, but its influence on healthy adults is unknown. We tested the hypothesis that Ang II modulates VF in adults free of CVD and that this influence is related to adiposity. Acute Ang II receptor blockade (ARB; 16 mg candesartan) was administered in a randomized, double‐blind, placebo‐controlled, cross‐over study in 27 healthy men varying in adiposity (body mass index; 18 to 38 kg/m2). We measured total body fat (dual energy x‐ray absorptiometry) and abdominal fat (computed tomography) at baseline and vascular endothelial function (brachial artery flow‐mediated dilation; FMD) and vascular smooth muscle sensitivity to nitric oxide (NO) (brachial artery dilation to 0.4 mg sublingual nitroglycerin; NID) after ARB and placebo. In the overall group, ARB had no effect on FMD (6.2±0.5 vs. 5.8±0.6%, P=0.4; ARB vs. placebo) and NID (16.5±0.8 vs. 16.8±0.9%, P=0.6). Among individuals, the change in NID with ARB was inversely related to total and abdominal body fat measures (r=−0.43 to −0.49, P=0.01 to 0.04). In subgroups, ARB increased NID in normal/overweight men (< 30 kg/m2), whereas, ARB decreased NID in obese (≥ 30 kg/m2) men (1.80±0.61 vs. −2.18±0.87%, P=0.001). In conclusion, Ang II modulates vascular smooth muscle sensitivity to NO in healthy men in an adiposity‐specific manner.
Dexamethasone Differentially Modulates the Human Angiotensinogen Gene (hAGT) Expression and Blood Pressure in Transgenic Mice Containing ‐6A and ‐6G Haplotype of the hAGT Gene
Authors: Pandey Varunkumar G. , Jain Sudhir , Kumar Ashok
Published: June 2021

Human angiotensinogen gene (hAGT) has ‐6A/G polymorphism and ‐6A allele is associated with increased blood pressure. We have found that hAGT gene has three additional SNPs. Variants ‐ 1670A, ‐1562C, and ‐1561T almost always occur with ‐6A haplotype (Hap‐6A); and variants ‐1670G, ‐1562G, and ‐1561G almost always occur with ‐6G haplotype (Hap‐6G). We have shown that these polymorphisms affect the binding of glucocorticoid receptor to the promoter. Reporter construct containing 1.8Kb of the hAGT gene promoter with Hap‐6A has 4 fold increased glucocorticoid induced promoter activity as compared to Hap‐6G. Therefore, we have generated transgenic (TG) mice containing either Hap‐6A or Hap‐6G of the hAGT gene. These TG mice were treated with dexamethasone (DEX) (2.5μg/mL in drinking water) for 1 week. Our Q‐RTPCR results show that mice with Hap‐6A has 2 and 2.06 fold DEX induced mRNA expression of hAGT in liver and kidney respectively whereas no significant changes were observed in Hap‐6G. Q‐RTPCR results were confirmed by western blots showing 1.80, 1.94 and 1.40 fold induction of hAGT protein in liver, kidney and plasma respectively in TG mice with Hap‐6A. DEX treatment also increased the SBP of TG mice containing Hap‐6A by 12 mmHg as compared to 5 mmHg in the mice with Hap‐6G. We subjected these TG mice to variable doses of DEX and observed a dose dependent modulation of SBP. DEX dependent increase in the SBP was obliterated by losartan.

Research Support: NIH grants HL081752 and HL66296 to AK
ACE2 activation ameliorates endothelial function in the right ventricles of pulmonary hypertensive rats
Authors: Sapp Glenn Herod , Abbatematteo Joseph , Cole Colleen , Shenoy Vinayak , Katovich Michael , Raizada Mohan , Muller‐Delp Judy
Published: June 2021

Discovery of angiotensin‐converting enzyme 2 (ACE2), which converts angiotensin II to angiotensin‐(1–7) has led to investigation of ACE2 activators as therapeutic agents for improved lung and heart function in pulmonary hypertension (PH). The purpose of this study was to investigate the effects of experimental PH on endothelium‐dependent vasodilation of right ventricular resistance arteries (RVRA) and the possibility that the ACE2 activator, diminazene aceturate (DIZE), ameliorates endothelial function of RVRA of rats with experimental PH. Rats were divided into four groups: control (no treatment), monocrotaline‐treated (MCT) to induce PH, MCT and DIZE, and DIZE alone. After 4 weeks of daily treatments, relative pulmonary pressure was assessed. Endothelium‐dependent and ‐independent vasodilation were also assessed with acetylcholine and the nitric oxide donor, Dea‐NONO‐ate, respectively. We observed that endothelium‐dependent vasodilation was impaired in RVRA while responses to Dea‐NONO‐ate were not altered in RVRA from MCT‐treated rats. DIZE treatment showed a restored responsiveness to acetylcholine in RVRA from MCT‐treated rats. This suggests that the impairment of the endothelial function in resistance vasculature due to PH may be restored by activation of endogenous ACE2.

NIH grants‐ HL 56921 and HL 102033
Angiotensin II (ANG II) receptors modulate nitric oxide (NO) effects on renal sodium (Na+) handling in conscious lambs
Authors: Vinturache Angela Elena , Smith Francine Gabriel
Published: June 2021

ANG II and NO are considered of primary importance in maintaining Na+ homeostasis in adult mammals. Evidence suggests that NO effects on renal handling of Na+ are modulated by ANG II, yet the roles of AT1Rs and AT2Rs are not well understood. In previous studies we reported a robust natriuresis associated with the removal of endogenous NO in six week old lambs. The present experiments were designed to evaluate whether there is an interaction between AT1Rs and AT2Rs in modulating the effects of NO on tubular Na+ handling in the immature kidney. In conscious, chronically instrumented six week old lambs (N=11), renal function was measured before and after selective pharmacological inhibition of AT1Rs with ZD 7155, of AT2Rs with PD 123319, and both receptors with ZD 7155 and PD 123319, before and after removal of endogenous NO with the L‐arginine analogue, L‐NAME. The increase in fractional excretion of Na+ and Na+ clearance with L‐NAME observed previously also occurred after pretreatment with ZD 7155; this response was enhanced by combined treatment with PD 123319. Fractional Na+ reabsorption fell after ZD 7155 and L‐NAME, and decreased further after addition of PD 123319. These findings suggest that AT2Rs may potentiate the effects of AT1R on NO‐mediated Na+excretion, providing the first evidence of a cross‐talk between ATRs in modulating NO effects on Na+ homeostasis under physiologic conditions in the immature animal.
Comparative Microscopic Study of Human and Rat Lungs after Exposure to Welding Fume
Authors: Antonini James Michael , Roberts Jenny R , Meighan Terence G , Schwegler‐Berry Diane , Mercer Robert R
Published: June 2021

Welding generates complex aerosols of hazardous metal fumes. Animal models and the ability to control the welding fume (WF) exposure have helped in developing an understanding of how WF affects health. There are no studies that have performed a side‐by‐side comparison of the pulmonary responses from an animal toxicology study with the lung responses associated with WF exposure by a welder. Post‐mortem lung tissue was donated from a long‐time welder with a history of extensive WF exposure. To simulate long‐term WF exposure in an animal model, Sprague‐ Dawley rats were treated for 28 weeks by intratracheal instillation with 2 mg of WF. Lung tissues from the welder and treated rats were examined by enhanced‐darkfield light microscopy and electron microscopy. Pathological analysis of lung tissue from the welder demonstrated inflammatory cell influx and pulmonary alveolar proteinosis, likely due to long‐term exposure to potentially toxic metals. The lung toxicity profile for the treated rats was similar. Aggregates of deposited welding particles were easily visualized, persisted in macrophages, and were composed of Fe, Cr, and Ni. In summary, long‐term exposure to specific WF can lead to serious chronic lung disease characterized by significant particle persistence as demonstrated in both a human case study and rat model. Not only were lung responses the same, but the WF composition in situ was similar.
Complementary roles of KCa3.1 channels and β1‐integrin in the regulation of alveolar epithelial repair
Authors: GIRAULT alban , BASTIEN Rachel , TRINH Nguyen Thu Ngan , PRIVE Anik , MAILLE Emillie , BROCHIERO Emmanuelle
Published: June 2021

A common feature of acute lung injury is extensive damage and remodeling of alveolar epithelium. It has been shown that epithelial regeneration and secondary lung edema resorption are crucial to patient recovery. We previously reported that KvLQT1 and KATP K+ channels regulated alveolar epithelial repair processes, but the role of another candidate, the KCa3.1, has never been evaluated.

We first demonstrated that the wound‐healing rates were higher in primary rat alveolar cell monolayers cultured on a fibronectin‐collagen (FC) matrix, than on non‐coated supports. KCa3.1 inhibition, with TRAM‐34, or activation, with 1‐EBIO, respectively down‐ and up‐regulated the alveolar wound‐healing rates on FC matrix, but were without effect in absence of FC coating. Accordingly, single cell migration rates were also dependent, at least in part, of FC matrix coating and KCa3.1 activity. A possible relationship between KCa3.1 and migratory cell proteins such as the integrin, activated by fibronectin, was then assessed. Co‐immunoprecipitation experiments thus revealed a physical interaction between KCa3.1 and the β‐1 integrin subunit. Altogether, our data showed for the first time a complementary role of KCa3.1 channels, extracellular matrix and β‐1 integrin, in the regulation of alveolar repair after injury.

Project funded by CIHR, RHN of FRQS, and FESP of Université de Montréal.
Impact of P.aeruginosa bacterial filtrate on CFTR channels and bronchial epithelial repair.
Authors: Trinh Nguyen Thu Ngan , Bilodeau Claudia , Maillé Émilie , Brochiero Emmanuelle
Published: June 2021

Cystic fibrosis (CF) pathology, due to mutations in the CFTR, is characterized by bacterial colonisation and chronic inflammatory response leading to a progressive damage of the airways. It then becomes crucial to better understand the impact of these deleterious phenomena on the repair processes in attempt to restore the CF airways integrity.

First, we confirmed, as in our previous studies, that the repair rates of airway epithelial CFBE‐ΔF508 (CF) monolayers were slower than that of CFBE‐wt (non‐CF) monolayers, which was, at least in part, dependent on CFTR function. We now observed that P. aeruginosa bacterial filtrate (PAF) inhibited wound closure of both non‐CF and CF CFBE monolayers. We also observed that wt‐ and ΔF508‐CFTR protein expressions, as well as Cl− currents through CFTR channels in CFBE‐wt monolayers were decreased after PAF exposure. Finally, CFTR corrector VRT‐325, initially developed to promote CFTR folding and function, was shown to be able to also improve the repair capacity of CF cell monolayers. However, its beneficial effect was abolished in the presence of PAF.

This study showed that exoproducts from P. aeruginosa alter CFTR expression/function and also exhibit a deleterious effect on airway repair capacity, in basal condition or after CFTR correction. Project supported by Cystic Fibrosis Canada, the Canadian Lung Association, the CORAMH and Fondation Go.
R‐Equol‐induced non‐genomic estrogenic effects and interactions with estradiol signaling
Authors: Saraf Manish Kumar , Jeng Yow‐Jiun , Watson Cheryl S
Published: June 2021

We previously reported that phytoestrogens, including daidzein, are potent inducers of nongenomic responses acting via membrane receptors for estrogens (mERs). R‐Equol (R‐Eq), a metabolite of daidzein, preferentially binds mERα. Therefore, we examined R‐Eq‐ induced nongenomic effects in mERα‐enriched GH3/B6/F10 pituitary tumor cells at the cellular signaling and function levels. R‐Eq (15 nM, dietary level), compared to 1 nM estradiol (E2, a female physiologic level), induced mitogen‐activated protein kinase (MAPK) activities (pERK, pJNK and p38) with variant initial peaks in the 2.5–15 min range and subsequent oscillations, as we have observed for other estrogens previously. In contrast to E2, R‐Eq did not produce a dose‐dependent increase in cell proliferation. R‐equol, like E2, augmented intracellular Ca++ levels and prolactin release from pituitary tumor cells. R‐Eq and E2 both alone and combined significantly activated Gαi by GTP‐charging. R‐Eq (10−16 to 10−7M) combined with 1 nM E2 significantly suppressed pERK, pJNK and p38 activities and cell proliferation, especially at concentrations of 10−10to 10−7M. We conclude that R‐Eq, like E2, is a rapid activator of nongenomic signals, but with slight differences in timing patterns and efficacies. However, when R‐Eq is combined with E2, it sometimes interferes with E2‐induced nongenomic estrogenic effects.
The effects of (−)‐epigallocatechin‐3‐gallate (EGCG) and phosphatidylinositol 3‐kinase (PI(3)K) inhibition on Burkitt lymphoma cells
Authors: Domina Aaron M. , Van Doorn Carl‐Christian
Published: June 2021

Burkitt lymphoma, an aggressive non‐Hodgkins B cell lymphoma, is comprised of three major subtypes: sporadic, Epstein‐Barr virus‐associated, and HIV‐associated. A recent study has shown that an overactive phosphatidylinositol 3‐kinase (PI(3)K) pathway occurs in many Burkitt lymphomas and inhibition of this pathway leads to apoptosis (Schmizt R. et. al., 2012). It has also been shown that the green tea compound, (−)‐epigallocatechin‐3‐gallate (EGCG) induces apoptosis in Burkitt lymphoma cells (Nakazato T. et. al., 2005 and Noda C. et. al., 2007). Therefore, the goal of this current study is to determine the relationship between EGCG treatment and the PI(3)K pathway. To this end, the effects of EGCG and the PI(3)K inhibitor, LY294002, on three sporadic Burkitt lymphoma cell lines, BL41–3, DG75, and Ramos were evaluated using an MTT assay and Western blot analysis. EGCG and LY294002 each inhibited Burkitt lymphoma cell growth in a dose‐dependent manner, although the degree of inhibition for each dose and the effect of combination treatment varied between the three cell lines, with the Ramos cell line showing the greatest decrease in viable cells. These results provide novel insight into the use of EGCG for the treatment of sporadic Burkitt lymphomas. This research was funded by the Husson University School of Pharmacy.
The Inhibitory Effects of Clitoria ternatea Extract on Protein Glycation, Amadori Product and Amyloid Cross β‐structure Formations.
Authors: Yibchok‐anun Sirintorn , Chayaratanasin Poramin , Adisakwattana Sirichai
Published: June 2021

Various pharmacological activities of Clitoria ternatea (CT) have been reported in literatures such as anti‐oxidant and anti‐diabetic properties. The accumulation of advanced glycation end products (AGEs) in body tissue leads to several degenerations, atherosclerosis and diabetic complications. The objective of this study was to determine the inhibitory effect of CT extract on AGEs formation In Vitro using bovine serum albumin (BSA)‐fructose assay. The aqueous extract of CT was incubated with BSA and fructose at 37 □ for 14 days. The Amadori product (fructosamine) and amyloid cross β structure were determined by nitroblue‐tetrazolium and thioflavin T assay. The results showed that the fluorescent intensity of BSA incubated with fructose increased by 3.6 folds compared to BSA alone, indicating an evidently formation of AGEs. The CT extract (0.25–1.0 mg/ml) significantly inhibited formation of AGEs in a concentration‐dependent manner by approximately 24.11 ± 0.3% −55.97 ± 2.93 %. In addition, CT markedly reduced levels of fructosamine and amyloid cross β‐structure formation. Our results suggest that CT extract may exert anti‐glycation effect that could be offered as a compound for further study as a new natural product for prevention of AGE‐mediated diabetic complications.
ANTIHYPERALGESIC EFFECT OF LATEX PROTEINS FROM CALOTROPIS PROCERA: INVOLVEMENT OF INFLAMMATORY MEDIATORS
Authors: Luz Patricia Bastos , Pineiro Rachel Sindeaux Paiva , Freitas Lyara Barbosa Nogueira , Bitencourt Flavio Silveira , Aragão Karoline Saboia , Carmo Luana Davi , Couto Tatiana Souza , Ramos Marcio Viana , de Alencar Nylane Maria Nunes
Published: June 2021

Background and aims

Calotropis procera is a tropical plant found worldwide and has been used in the traditional medicinal for the treatment of various diseases. This study aimed to evaluate the effects of protein fraction from Calotropis procera latex (LP) in the mechanical hypernociception (increase in the sensitivity of nociception).

Methods

Swiss male mice were used. b) Mechanical hypernociception (MH) was evaluated using the electronic version of the Von Frey before and after the injection of carrageenan (300 μg/paw‐Cg) or LP (0.5, 5 and 50 mg/kg, iv, 30 min before Cg application) into the mice paw (1, 3 and 5 hours after Cg). c) After 3 h of Cg injection, subplantar tissue was collected for measurement of: myeloperoxidase activity (MPO); cytokines (TNF‐α and IL‐1β) and chemokine (KC) production; prostaglandin E2 (PGE2) production. d) ANOVA/Bonferroni was used as statistical tests. P<0.05 was accepted. e) Ethics Committee 61/11.

Results

LP 5 mg/kg reduced MH in 25%, 55% and 46% and LP 50 mg/kg in 39%, 64% and 60% in the 1st, 3rd and 5th hours. LP at 5 and 50 mg/kg decreased MPO concentration in 80% and 94% respectively. LP at 5mg/kg reduced the concentration of TNF‐ α, IL‐1β, KC and PGE2 at 71%, 81%, 72% and 72% respectively.

Conclusion

LP has antinociceptive properties in animal model of acute pain and this may be associated with the inhibition of neutrophil migration, cytokine and chemokine production. CAPES and CNPq.
Cerebro‐Vasculoprotective Effects of Azilsartan Medoxomil in Diabetes
Authors: Abdelsaid Mohammed A , Coucha Maha M , Ergul Adviye
Published: June 2021

We have shown that glycemic control prevents diabetes‐mediated remodeling of the cerebrovasculature. However, whether the remodeling can be reversed is unknown. Given that angiotensin II Type 1 receptor blockers (ARBs) reverse pathological vascular remodeling and function independent of changes in blood pressure in other vascular beds, we hypothesized that azilsartan medoxomil, a new ARB, reverses cerebrovascular remodeling in diabetes independent of blood pressure.

Methods

Control Wistar and diabetic Goto‐Kakizaki rats (n=20/group), were treated with vehicle (water) or Azilsartan medoxomil (3 mg/kg/day) from 18 to 22 weeks of age. Blood glucose and blood pressure was monitored and middle cerebral artery structure and function was evaluated using pressurized arteriography.

Results

Blood glucose was higher in the GK compared to Wistar rats. Azilsartan treatment lowered blood glucose. There was no difference in blood pressure among the groups. Diabetic animals exhibited lower myogenic tone, increased wall thickness, and cross sectional area compared to controls which were corrected by azilsartan treatment.

Conclusion

Azilsartan medoxomil restores diabetes‐induced cerebrovascular remodeling independent of blood pressure. This effect may be partially mediated by improvement of blood glucose. Disclosure The project is supported by Takeda Pharmaceuticals North America Inc.
The influence of ethanol on the hyperglycemia and renal activities of glycolytic and gluconeogenic enzymes of diabetic rats
Authors: Patel Sanket N. , Parikh Mitul C. , Ashby Charles R. , Lau‐Cam Cesar A.
Published: June 2021

The role of ethanol (EtOH) on the blood glucose (BG) and activities of glycolytic and gluconeogenic enzymes in diabetes was investigated in the kidney of normal and streptozotocin (STZ)‐ treated rats as a function of EtOH concentration (5% and 15%, v/v) and timing of treatment with STZ relative to the start of EtOH intake. In rats receiving 5% EtOH in the drinking water for 28 days and STZ (60 mg/kg i.p.) on day 15, the BG rose to a level below that of STZ alone. A change to 15% EtOH led to a BG level that peaked by day 17 and then declined to below the value seen with 5% EtOH. From the renal activities of glycolytic and gluconeogenic enzymes, it was apparent that diabetes inhibits glycolysis but stimulates gluconeogenesis. In these rats, consumption of 5% or15% EtOH led to a concentration‐dependent reduction of the biochemical alterations induced by STZ (p≤0.01). A treatment with STZ 14 days before EtOH intake for another 14 days had a smaller, although significant (p≤0.01), reducing effect on the BG than when given later. With this treatment schedule, the effects of EtOH on enzyme activities were rather similar in pattern and magnitude to those observed with EtOH preceding STZ. In short, chronic consumption of EtOH is capable of attenuating changes in BG and renal glycolytic and gluconeogenic enzyme activities associated with diabetes in a concentration‐related manner and independently of the time of induction of diabetes.
The effect of a DPP4 inhibitor on morphometry of pancreatic beta cells after lactation on female wistar rats.
Authors: Suen Vivian , Ribas‐Filho Durval , Cersosimo Eugenio , Martins Alex , Marchini Julio Sergio , Azoubel Reinaldo
Published: June 2021

Objective

To evaluate through cariometry and estereology if vildagliptin causes morphological changes in the endocrine pancreas of Wistar rats.

Methods

Sixteen male rats were selected and divided into two groups, control and treated, which received respectively physiological and vildagliptin solution through nasogastric intubation for 10 days. After the sacrifice of all animals and pancreas dissection, histological slices were obtained for cariometric and stereological studies. Computarized programs were used to analyse nuclear parameters of pancreatic islet cells, where insulin is secreted: Major Diameter and Minor Diameter. Mann‐Whitney U Test was applied for statistical analysis.

Results and Conclusion

Vildagliptina causes significant cariometric and stereological alterations in pancreatic islet cells, with nuclear hyperplasia, without hypertrophy, and an increase in metabolism, with tissue hyperplasia.
Effect of L‐tryptophan on cytoprotection against long term postprandial hyperglycemia‐induced esophageal damage in rats
Authors: Hrytsevych Nazar Romanovych , Zayachkivska Oksana , Yaschenko Antonina
Published: June 2021

Beneficial effects of L‐tryptophan (L‐Try) have already been described by many activities in kynurenine and serotonin synthesis, redox reactions in all living cells which play key role for cancer invasion and proliferation. Recently we revealed that postprandial hyperglycemia (PPH) which characterized by hyperglycemic spikes leads to nitro‐oxidative stress and may predispose to foregut disorders. Nevertheless, the effect of L‐Try on PPH is unknown. In this study, we elaluated effect of L‐Try on NO/NOS expression in OM during PPH and abnormal PG/COX activity.

Rats were used with without/with L‐Try treatment (400 mg/kg) and indomethacin (Indo, 5 mg/kg) during PPH (Kozar, 2009); oesophagus (OM) and lower oesophageal sphincter (LOS) lesions were investigated by histology score index (HIS); esophageal NO2, cNOS and iNOS via bioanalysis.

PPH caused destructive lesions in the OM and LOS, accompanied by the up‐regulation of iNOS and down‐regulation of cNOS expressions, excessive NO2‐ while Indo significantly aggravated severity of these lesions; L‐Try prevented ulcerogenic response to PPH with potent up‐regulation of cNOS but did not affect synthesis NO2‐.

These results suggest that L‐Try prevents L‐Try‐induced OM and LOS damage but oesophagoprotective‐promoting effect may be associated with the stimulation of endothelial metabolism, but not through an amelioration of PG/COX activity.
Angiotensin AT2R activation prevents adiposity under high‐fat diet‐condition in ovariectomized mice
Authors: Nag Sourashish , Hussain Tahir
Published: June 2021

Recently we demonstrated that the genetic deletion of angiotensin AT2 receptor (AT2R) increases adiposity in female mice on high‐fat diet (HFD). Further these changes were associated with decrease in the urinary 17,β‐estradiol (E2), a known regulator of adiposity. This study was designed to test whether AT2R activation prevents adiposity dependent on estrogen. Female C57BL/6 ovariectomized (Ovx) female mice (8‐weeks old) were supplemented with E2 pellets and/or pretreated with AT2R agonist C21 (0.3 mg/kg, daily i.p.) for 4 days. Thereafter the mice were placed on normal diet (ND) or HFD and the drugs treatment continued for next 10 days. Ovx mice had increased body weight (BW), gonadal adipose tissue weight (gATW), plasma free fatty acid (pFFA) compared to mice with intact ovary on ND. High‐fat diet (HFD) in Ovx mice further increased BW, gATW, pFFA. As expected, E2 supplementation prevented the above mentioned parameters under HFD in Ovx mice. Interestingly, C21 treatment alone in Ovx mice significantly prevented BW gain (19%), gATW (42%), pFFA (55%) despite similar energy intake compared to Ovx mice on HFD. We conclude that the pharmacological activation of AT2R alone prevents adiposity in ovariectomized mice indicating estrogen‐independent role of AT2R in preventing HFD‐induced adiposity in females.

Grant Funding Source: NIH RO1 DK‐61578
JNK1 phosphorylation of HNF4α represses miR‐122, which causes PTP1B induction
Authors: Yang Yoon Mee , Seo So Yeon , Kim Tae Hyun , Kim Sang Geon
Published: June 2021

MicroRNAs have emerged as key regulators of energy metabolism. The miR‐122 is the most abundant microRNA in the liver, and may affect cholesterol metabolism. Nevertheless, the role of miR‐122 in insulin sensitivity and blood glucose control had not been studied. This study investigated whether dysregulation of miR‐122 contributes to hepatic insulin resistance using animal and cell models. The levels of miR‐122 were down‐regulated in the liver of high‐fat diet (HFD)‐fed mice. miR‐122 was also repressed in hepatocytes treated with either TNFα or palmitate. We identified protein tyrosine phosphatase 1B (PTP1B), a key negative regulator of insulin signaling, as a novel target of miR‐122; decrease of miR‐122 caused PTP1B induction, as indicated by the results of miR‐122 mimic and its inhibitor experiments. JNK1 facilitated serine/threonine phosphorylation of HNF4α (i.e., HNF4α inhibition), which was responsible for miR‐122 down‐regulation in hepatocytes. Consistently, functional inhibition of JNK by licorice flavonoid treatment caused not only the restoration of miR‐122 levels, but also the repression of PTP1B. Moreover, JNK1 inhibition enabled cells to restore tyrosine‐phosphorylation of IR or IRS1/2, lowering blood glucose contents in HFD‐fed or ob/ob mice. In conclusion, miR‐122 dysregulation may increase insulin resistance through PTP1B induction, which depends on HNF4α phosphorylation by JNK1.