bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the Regulatory Activity of the LSRB3 region of Limb Specific SHH Regulatory Region (LSSRR) in the Chicken Model
Authors: Coy Steven , Pira Charmaine U , Oberg Kerby C
Published: June 2021

During limb development, sonic hedgehog (SHH) from the zone of polarizing activity (ZPA) promoting posterior proliferation and patterning along the anterior‐posterior axis. A 1.8 kb conserved non‐coding sequence, 1 mb upstream of SHH, regulates limb specific SHH expression. We have subdivided the limb specific SHH regulatory region (LSSRR) into two regions, LSRA and LSRB. LSRB is further subdivided into LSRB1, LSRB2, and LSRB3 due to gaps in conservation. We have previously showed that LSRB3 exhibits enhancer activity, but this activity has not been quantified. Our objective was to quantify the enhancer activity of LSRB3. We hypothesize that region LSRB3 contains the minimal enhancer to regulate limb SHH expression. To test our hypothesis, enhancer‐reporter constructs were made by inserting LSSRR or LSRB3 upstream of a minimal HSV‐tk promoter linked to GFP. The constructs were electroporated (EP) into the presumptive limb mesoderm of chicken embryos. After 48 hours, GFP fluorescence was observed and mRNA was evaluated by whole mount in situ hybridization and sqRT‐PCR. GFP was detected in the posterior margin of the limbs EP with LSSRR and LSRB3 indicating enhancer activity. The activity of LSRB3, however, is 40% of LSSRR. These results indicate that the LSRB3 segment of LSSRR contains regulatory sequences sufficient to promote ZPA expression, but that other regions within LSSRR are needed to attain full activity.
The prevalence of bifid tongue congenital anomaly in riverine areas of south east Nigerian population
Authors: Ihentuge Churchill Jonadab , Ugochukwu Chioma Olejuru
Published: June 2021

OBJECTIVE

To determine the occurrence of bifid tongue congenital anomaly in riverine areas of south east Nigerian population

MATERIAL AND METHOD

One thousand six hundred and forty (1640) newborn babies delivered in selected hospitals, health centers, maternity homes and traditional birth homes between November 2009 and October 2012 were used as sample population for this study. 840 were females and 800 males. The subjects were carefully examined within the first week of delivery for bifid tongue congenital anomaly. Consent was also obtained from the subjects mothers and the ethical clearance for the use of human subjects for experimental purpose was also obtained from ethical committee of Imo State University Nigeria.

RESULTS

The result showed that out of the one thousand six hundred and forty (1640) babies examined, sixteen (16) subjects were found with congenital anomalies. The anterior bifid tongue had a prevalence of 0.07% of the studied population and 6.25% of those with congenital lesions.

CONCLUSION

The present study shows that bifid tongue is a very rare congenital anomaly with a prevalence of 0.07%. The maternal health status and familiar history were not contributory to the results of this study but environmental factor may play important role in the etiology which is in support of previous findings. It is therefore imperative that further studies should be conducted to identify the environmental factors that are contributory to the frequency of bifid tongue congenital anomaly.
Maternal immune activation increases neuroinflammation and aggravates EAE in adult offspring
Authors: Zager Adriano , Peron Jean‐Pierre S , Campos‐Rodrigues Sandra , Palermo‐Neto Joao
Published: June 2021

Maternal infection during pregnancy is considered an environmental risk factor in the development of many psychiatric disorders in the adult offspring, including autism and schizophrenia. However, the consequences of this phenomenon in progressive neuroinflammatory diseases, such as Multiple Sclerosis and its mouse model, experimental autoimmune encephalomyelitis (EAE), are still unknown. This study aimed to evaluate the effects of prenatal exposure to lipopolysaccharide (LPS) on the progression of EAE in the adult offspring. LPS (E.coli 0127:B8, 120 ug/kg) was administered IP to pregnant Swiss mice on gestational day 17. Adult female offspring (8 weeks, N=8–10/group) were immunized with MOG 35–55 and the clinical course of EAE was followed for 30 days. Another set of animals was sacrificed during the peak of disease (20d p.i.), where brains and spinal cords were collected for mononuclear cell isolation used in flow cytometry analysis. Prenatal exposure to LPS worsened EAE disease severity, which was linked to an increase in CNS infiltrating macrophages and Th1 and Th17 cells and increased expression of MHC II by macrophages and microglia. Our results indicate that maternal immune activation during late gestation could predispose the adult offspring to increased neuroinflammation and potentiate the clinical manifestation of EAE.
CD44/CD24 expression on salivary glands malignant neoplasms (SGMN)
Authors: Soave Danilo Figueiredo , Costa João Paulo Oliveira , Silveira Giorgia Gobbi , Ianez Renata Carolina Fraga , Oliveira Lucinei Roberto , Lourenço Silvia Vanessa , Silva Alfredo Ribeiro
Published: June 2021

Background

CD44 is the binding site for hyaluronic acid, and CD24 is a receptor that interacts with P‐selectin to induce tumor progression. Present study aims to evaluate CD44/CD24 expression on SGMNs and correlated these data with clinicopathologic features.

Methods

Immunohistochemical stains for CD44/CD24 were performed on 69 SGMN samples. CD44/CD24 expression was correlated to patient clinicopathologic features and outcome.

Results

CD44 expression was associated with the primary site of neoplasm (p=0.046). CD24 was associated with clinical stage III/IV (p=0.008), T stage (p=0,27) and lymph node (p=0,001). The CD44/CD24 profiles were associated with the primary site of injury (p=0.005), lymph node (p=0.011) and T stage (p=0.023).

Conclusion

in summary, we have presented some evidence that the analysis of CD44/CD24 expression could give prognostic information associated to clinicopathologic features in SGMN.
Stabilization and Direct Detection of Green Fluorescent Proteins in Histological Preparations for Pathological Evaluation of Infectious Disease
Authors: Lyons Cassandra Corine , Yellayi Sri , Cann Jennifer , Hart Brit , Postnikova Elena
Published: June 2021

Green florescent protein (GFP)‐expressing recombinant viruses have proven to be invaluable tools for the in vivo monitoring of viral kinetics and disease progression in infectious disease models. Histopathological evaluation of the ex vivo tissues by way of immunohistochemical (IHC) staining for viral antigens further assists in the characterization of disease pathology, but can be insensitive, nonspecific, and time consuming. In this study, we determine if the direct detection of GFP signal provides a suitable alternative to IHC staining in GFP‐expressing tissues. By first creating a GFP‐expressing 293T cell line we were able to monitor signal preservation through critical fixation and processing steps. We then applied our protocol to a transgenic rat model that ubiquitously expresses GFP under a chicken beta‐actin promoter. We examined tissue sections from the liver, kidney and brain and found that we were able to preserve GFP fluorescence through histological processing, however, the interference from autofluorescence, both inherent and fixative‐induced, made reliable direct detection of GFP signal difficult. Future experiments will test a variety of autofluorescence‐reducing techniques, including sodium borohydride and Sudan black pretreatments, and multispectral image analysis of collagen and elastin content to increase the signal to noise ratio.
Asymptomatic African Americans with high‐risk APOL1 genotypes have reduced urinary angiogenesis‐promoting cytokines
Authors: Gutierrez Jean Lorraine , Many Gina , Panchapakesan Karuna , Brown Kristy J , Hathout Yetrib , Marathi Ramya , Devaney Joseph , Hoffman Eric P
Published: June 2021

Background

African Americans (AA) having two high‐risk APOL1 alleles, termed G1 and G2, are about 7 times more likely to develop non‐diabetic kidney diseases, and ApoL1 is differentially expressed in glomerular arterial compartments in those with overt kidney disease.

Objective

To compare angiogenic cytokines in the urine of young, asymptomatic AA with high‐risk for CKD (G1 homozygotes) to lower‐risk AA (non‐carriers and heterozygotes).

Methods

Thirty‐seven previously genotyped participants (5 G1 homozygotes, 20 WT homozygotes, and 12 heterozygotes) donated mid‐stream, first morning urine samples. Cytokine concentrations in unprocessed urine samples were analyzed with Milliplex Human Cytokine/Chemokine Panel bead assay. Urine cytokine concentrations were standardized to urinary creatinine, and the two risk groups were compared using the Mann‐Whitney U Test (SPSS Version 21).

Results

Concentrations of the following angiogenic cytokines were significantly lower in high‐risk G1 homozygotes: TGF‐α (P=0.004), PDGF‐AA (P=0.016), IL‐1‐alpha (P=0.032), VEGF (P=0.036).

Conclusions

This pilot data indicates that young, asymptomatic African Americans with high‐risk APOL1 genotypes may have impaired angiogenesis. These results are consistent with prior evidence indicating that vascular disease may underlie ApoL1‐associated kidney diseases. This study was funded by a CTSA pilot award.
Astilbin inhibits high glucose‐induced vascular smooth muscle cells proliferation and migration
Authors: Shen Zhihua , Deng Xiaojiang , Zheng Shaojiang , Jiang Xiaofan , Ding Ranran , Wu Yanxia , Luo Botao , Guo Junli , Jie Wei
Published: June 2021

Astiblin is a flavonoid compound extracted from Glabrous Greenbrier Rhizome, a traditional Chinese medicinal herb, and show activity in antioxidant effect. In this study, we explored the role of astilbin in anti‐diabetes‐induced vascular smooth muscle cells (VSMCs) proliferation and migration. Tunica media of thoracic aortas of SD rats injected with STZ displayed a higher expression of PCNA, MMP2 and OPN, and a lower expression of SM22¦Á and Myocardin at mRNA and protein levels, administration the rats with astilbin reversed the diabetes‐induced aberrant expression of VSMCs marker genes ex vivo. Results in vitro showed that treatment rat aortic VSMCs with high glucose (25 mmol/l) for 48h led to the increase of PCNA, MMP2 and OPN, as well as the decrease of SM22¦Á and Myocardin when compared with VSMCs maintained in normal glucose (5.5 mmol/l) and D‐Mannitol (25 mmol/l). Interestingly, astilbin attenuated high glucose‐induced aberrant expression of VSMCs marker genes in vitro. Furthermore, astilbin inhibited VSMCs migration via suppression of MMP2 expression and activity. Additionally, astiblin abrogated high glucose‐induced NF¦ÊB‐DNA binding activity, but no effects on Myocardin‐SRF binding activity. Thus, our results suggest that astilbin may be beneficial for the diabetes‐induced vascular complications by inhibition of high glucose‐mediated VSMCs proliferation and migration.
The Best Way to Improve HDL is to Lower LDL Levels.
Authors: Vakili Ladan , Khany Mitra , Karimi Nika , Vazirian Samra , Gapele Ania
Published: June 2021

Objective

Analyses of HDL function.

Introduction

Population studies have shown that plasma levels of HDL (high density lipoprotein) inversely correlate with the prevalence of coronary heart disease. Plasma HDL cholesterol levels in an individual however, does not necessarily correlate with the risk for coronary heart disease. Many patients with high HDL, present with coronary events and many individuals with low HDL cholesterol do not develop coronary complications. Our group previously demonstrated that HDL composition and function could be more important than HDL cholesterol levels. Under excessive inflammatory pressure unsaturated fatty acids in HDL can undergo oxidative modification. Protective enzymes including paraoxonase, LCAT and PLTP can be inactivated by oxidative modification reducing HDL's anti inflammatory properties. This results in HDL not being able to protect LDL from oxidative modification.

Methods

Fasting plasma from patients with documented coronary disease were analyzed for their ability to prevent LDL modification.

Results

We demonstrated that among coronary patients over 69 per cent had non protective HDL while among healthy controls 4.5 per cent had dysfunctional HDL.

Conclusion

While there are effective medications for lowering LDL cholesterol (namely statins) there are few or no satisfactory drugs for raising HDL levels.
Development of functionally selective agonists at the kappa opioid receptor (KOR)
Authors: Lovell Kimberly M , Zhou Lei , Frankowski Kevin J , Slauson Stephen R , Streicher John M , Phillips Angela , Prisinzano Thomas E , Aube Jeffrey , Bohn Laura M
Published: June 2021

The G protein‐coupled, kappa opioid receptor (KOR) is a prominent target for drug discovery efforts for the treatment of pain and addiction behaviors. The therapeutic potential of KOR agonists is currently limited because KOR activation can cause dysphoria. Moreover, dynorphin, an endogenous agonist at the KOR, is a major neuropeptide involved in responding to stress. There is evidence to suggest that KOR‐mediated aversive responses result from KOR interactions with beta‐arrestin2 (βarr2). Therefore, we sought to develop KOR agonists that are functionally selective in promoting G protein coupling over βarr2 recruitment. To discern between G protein and βarr2 pathways, proximal measures of G protein coupling ([35S]‐GTPγS binding) and βarr2:KOR enzyme fragment complementation (DiscoveRx PathHunter® assay) were used. Here we describe compounds that potently activate G protein and have little potency to recruit βarr2. It is hopeful that such pharmacological tools will prove useful in defining the complex contribution of KOR signaling to the physiological stress response and may provide insights for the development of novel drugs for treating pain and drug addiction. This work was supported by NIDA grant 5 R01 DA031927–02
Patchouli Alcohol Possesses Anti‐Inflammatory Activity in RAW264.7 and HT‐29 cells
Authors: Jeong Jin Boo , Shin Yong Kyu , Lee Seong‐Ho
Published: June 2021

Patchouli alcohol (PA) is a chemical compound found in essential oil of patchouli which belongs to the genus Pogostemon, herb of mint family. Although it has been reported that PA suppresses the production of inflammatory mediators, the biological mechanisms of PA for anti‐inflammatory activities have not been studied. Thus, in this study, we investigated whether PA decreases the production of inflammatory mediators through downregulation of the NF‐¥êB and ERK pathway. Our data indicated that PA inhibits the expression of iNOS and IL‐6 in protein and mRNA levels in LPS‐stimulated RAW264.7 and TNF‐¥á stimulated HT‐29 cells. PA reduced I¥êB‐¥á degradation and p65 nuclear translocation, and subsequently suppressed transcriptional activity of NF‐¥êB in LPS‐stimulated RAW264.7 and TNF‐¥á‐stimulated HT‐29 cells. In addition, PA ameliorated LPS‐ or TNF‐¥á‐stimulated ERK1/2 activation. These findings suggest that PA possesses anti‐inflammatory activities through suppressing ERK‐mediated NF‐¥êB pathway in mouse macrophage and human colorectal cancer cells.

This work was supported by start‐up funds to S.‐H. Lee from University of Maryland and in part by RSG‐11–133‐01‐CCE to S.‐H. Lee from American Cancer Society.
Anti‐angiogenic effects of curcumin and its novel analogs EF‐31 and UBS‐109 in colorectal cancer
Authors: Ganji Purnachandra Nagaraju , Zhu Shijun , Ko Jasmine E , Shoji Mamoru , El‐Rayes Bassel
Published: June 2021

Hypoxia‐inducible factors (HIFs) play essential roles in cancer cell growth and metastasis by stimulating angiogenesis. Curcumin inhibits the activity several oncogenic transcriptional factors including HIF‐1á, thus we investigated whether curcumin and its analogs EF‐31 and UBS‐109, could disrupt angiogenesis using colorectal cancer cells (CRC). HCT‐116 and HT‐29 cells were used in these experiments. HUVEC tube formation assay, Matrigel plug assay, Western blotting, QRT‐PCR, and VEGF activity assay were carried out to determine the curcumin, EF‐31 and UBS‐109 role in angiogenesis. Conditioned medium from HCT116 or HT29 cells exposed to curcumin, EF‐31 and UBS‐109 in vitro significantly blocked HUVEC tube assembly in comparison to control. In vivo, Curcumin, EF‐31 and UBS‐109 blocked the vascularization of subcutaneous matrigel plugs and the growth of CRC xenografts. We observed significant inhibition of VEGF synthesis and secretion in both colon cell lines treated with curcumin, EF‐31 and UBS‐109 in concert with the loss of HIF‐1á expression, of which transcriptionally regulate VEGF. These results suggesting that curcumin, EF‐31 and UBS‐109 inhibits VEGF production in part through the degradation of HIF‐1á. Taken together, destabilization of HIF‐1á may be important contributing factors to the antiangiogenic action of curcumin, EF‐31 and UBS‐109 in CRC.
Characterization of the Vascular Phenotype of the Equilibrative Nucleoside Transporter 1 Knockout Mouse
Authors: Best Keisha Arielle , Bone Derek , Feng Qingping , Gros Robert , Hammond James
Published: June 2021

Adenosine is important in the endogenous regulation of vascular tone. Equilibrative Nucleoside Transporter 1 (ENT1) is an established regulator of adenosine concentrations in vascular tissues. Thus, changes in ENT1 expression and/or activity may alter vascular tone regulation. We therefore examined the cardiovascular phenotype of ENT1‐knockout (KO) mice as compared to wild type (WT) mice. Heart rates and blood pressures of KO and WT littermate male mice were obtained. Vascular reactivity was assessed in aortic ring segments obtained from 3 and 6 month old KO and WT mice. KCl‐induced depolarization led to significantly greater constriction (~32% increase independent of tension applied or age) in KO rings compared to WT. However, no significant differences between the two genotypes were observed using a series of receptor‐selective constrictors. Morphological assessment of H&E stained vascularized tissues yielded no significant difference in luminal area or arterial wall thickness between WT and KO mice at the age of 3 months. The enhanced effect of KCl in KO aortic rings suggests a physiological difference in the regulation of depolarization‐induced, but not receptor‐mediated, vasoconstriction response in aorta. Further assessment of this model will aid in our understanding of the role of adenosine in vascular regulation.
PALMITOYLATION OF G‐PROTEIN ALPHAQ IN HYPOXIC PULMONARY HYPERTENSION
Authors: Sikarwar Anurag Singh , Hinton Martha , Dakshinamurti Shyamala
Published: June 2021

Introduction

Hyperreactivity of thromboxane prostanoind receptor (TP) is a major contributor to persistent pulmonary hypertension of the newborn (PPHN). We previously reported that in hypoxic pulmonary artery smooth muscle cells (PASMCs) TP couples mainly with G‐protein alphaq (Gαq), which undergoes palmitoylation. Palmitoylation is a dynamic, reversible post‐translational modification of the N‐terminal cysteine residues of Gαq, which is required for efficient receptor‐Gαq interaction.

Objective

To study the role of Gαq palmitoylation in hypoxic pulmonary hypertension.

Methods

PPHN was induced in newborn piglets by exposure to normobaric hypoxia (FiO2 0.10) for 72h; age‐matched normoxic piglets served as controls. Serum deprived primary cultured myocytes from newborn pig (day 0) were placed in hypoxic (10% O2) or normoxic (21% O2) environment for 72h. Palmitoylable cysteine to alanine mutants of Gαq were obtained commercially and transiently transfected in HEK293T cells stably expressing TP. Calcium mobilization to TP stimulation was studied by calcium indicator dyes in hypoxic and normoxic conditions. Metabolic labeling of Gαq by 3H palmitic acid is currently pursued under hypoxia and normoxia.

Results

Pretreatment with 2‐BrP decreased the maximum contractile response and calcium mobilization of hypoxic pulmonary artery rings and PASMCs respectively. 2‐BrP had minimal effect on rings and PASMCs from normoxic piglets.

Conclusions

Palmitoylation of Gαq could play role in hypoxia‐induced hyperreactivity of TP.

Funding:

MHRC‐MICH Graduate studentship
Role of ABCC1 in protecting the heart against doxorubicin (DOX)‐induced oxidative stress
Authors: Deng Jun , Coy Donna , Sunkara Manjula , Morris Andrew , Chaiswing Luksana , Oberley Terry , Vore Mary
Published: June 2021

We investigated the role of multidrug resistance‐associated protein 1 (Mrp1), an ATP‐binding cassette transporter (ABCC1), in mouse heart following DOX‐induced oxidative stress. Wildtype (WT) and Mrp1−/− (KO) C57BL/6 littermates were administered a single dose of 20 mg/kg DOX or Saline (SAL) intravenously. Hearts were collected 72 h later, and ultrastructural damage measured by electron microscopy (EM) (n=6) and content of the glutathione conjugate of 4‐hydroxynonenal (GS‐HNE), a substrate of Mrp1, determined by LC‐MS (n=6–7). The mRNA of Nrf2 responsive genes, Nqo1, GPX1, GPX3, and GPX4 were quantitated by real time quantitative PCR (n=6). Morphometric analysis of EM data showed a trend (p = 0.063) for decreased cytoplasmic + mitochondrial damage in DOX‐treated KO mice vs WT, but increased (p = 0.031) nuclear injury in KO mice. In contrast, SAL induced more cytoplasmic damage in KO mice (p = 0.015). GS‐HNE (nmol/g heart tissue) of WT and KO SAL mice were similar, whereas DOX decreased (p < 0.001) GS‐HNE in WT mice, but had no effect in KO mice. The basal level of GS‐HNE in naïve KO mice was lower (p = 0.013) than in naive WT mice (n=8). DOX increased mRNA of Nqo1 and GPX3 in hearts of WT and KO mice similarly, but had little influence on GPX1 and GPX4 mRNA. Nevertheless, in naïve mice, basal levels of Nqo1, GPX3, and GPX4 mRNA were lower in KO than in WT (p = 0.001, 0.011, 0.022, respectively). These data imply that the elevated GSH level in hearts of KO mice is able to protect against some forms of DOX‐induced injury, but not others. (CA139844)
THE EFFECTS OF HYDROGEN PEROXIDE (6%) IN GASTRIC MUCOSA â “ AN EXPERIMENTAL STUDY
Authors: Paula Anabela Baptista Pereira , Dias Isabel , Ferreira Manuel , Cabrita AntÃ³nio , Carrilho Eunice
Published: June 2021

The tooth whitening therapeutics can be performed at the clinic or ambulatory, with a greatest danger of eating the products of bleaching.

This experimental study analysis the effect of these products on gastric mucosa with or without non‐tumor pathology. We used 50 mice Wistar‐Han. In periods II and III, which took place on 2nd and 3rd weeks, the animals in groups II, III, IV and V were subjected to induction protocol not gastric tumor by administering a chemical compound (50% ethanol), and/or administration of a 6% hydrogen peroxide bleaching agent (Colgate®Visible White™), by gavage.

From the data collected were made statistical analysis of variance (ANOVA) and multiple comparison of pairs of means over time in a study by a t Student test. It was also performed a frequency analysis of the results of macroscopic observation of the gastric mucosa (JMP Version 7, SAS Institute Inc., NC, USA).

There was a decrease in body weight in animals of groups handled during the study period, which was most pronounced in groups IV and V‐A. Changes in the spleen weight relative to body weight revealed no statistically significant changes. The gastric mucosa revealed in all groups manipulated lesions, which were more frequent in groups III and IV.

We can conclude that there seems to be some signs of toxicity in 3 to 4 days after administration of hydrogen peroxide 6%, and induction of gastric lesions and generalized moderate.
Arterioles from Bariatric Patients with Diabetes Have Blunted Responses to Insulin but not Acetylcholine
Authors: Martinez‐Lemus Luis A. , DeLaTorre Roger A. , Galiñanes Edgar L. , Perna Mark J. , Zhao Guiling , Castorena‐Gonzalez Jorge A.
Published: June 2021

Obesity and type‐2 diabetes are associated with cardiovascular disease and endothelial dysfunction of which the mechanisms leading to aberrant insulin‐dependent vasodilation are incompletely understood. As not all obese individuals are diabetic, we hypothesize that reduced insulin‐dependent vasodilation is a specific endothelial dysfunction independent of the degree of obesity that accompanies the presence of overall insulin resistance. We tested this hypothesis in jejunal submucosal arterioles obtained from bariatric patients undergoing Roux‐en‐Y proximal gastric bypass surgery. Patients were 49±2 years old with a body mass index (BMI) of 48.5±2.4 Kg/m2. No differences in age or BMI were found between diabetics (n=22) and non‐diabetics (n=30). Maximal arteriolar responses to KCl, phenylephrine, acetylcholine, or sodium nitroprusside were not significantly different between diabetics and non‐diabetics. In contrast, insulin‐dependent vasodilation was smaller in diabetics (40.4±6.4 vs. 61.9±8.8%, respectively). This suggests that a blunted insulin‐vasodilation response is a form of endothelial dysfunction not correlated with BMI in type‐2 diabetes. The blunted‐response to insulin occurring in the absence of an abnormal acetylcholine‐induced dilation suggest that the mechanism responsible for the blunted insulin response is upstream of nitric oxide synthase activation.
Effects of Exercise During Pregnancy on Childhood Heart Measures
Authors: May Linda E , Drake William B , Suminski Richard R
Published: June 2021

We have found decreased fetal and infant heart rate and increased heart rate variability as a result of regular maternal exercise suggesting exercise during pregnancy influences cardiac development. It is unknown if these affects persist into childhood. We hypothesized that exercise during pregnancy promotes improved functional heart measurements in children. Mothers (n=25; 30.3 ± 5.5 y of age) of children with normal ultrasounds self‐reported exercise they participated in during their 3rd trimester of pregnancy. Ultrasounds were conducted on offspring 1 month to 7 years after birth. Mothers performed between 120 and 5730 min of exercise (M = 1449 ± 14 min). Minutes of exercise were not significantly correlated with any cardiac measures. After controlling for child's age, min of exercise was negatively associated with left ventricular end systolic and diastolic dimensions (p<0.05) and left ventricular end systolic volume relative to body surface area (p<0.05) but positively related to left ventricular ejection fraction (p<0.05). The results indicate exercise during pregnancy has a significant effect on offspring heart function into childhood. Potential mechanisms warrant further study.
Exploration of the role of tylophorine E ring in anti‐Coronavirus activity‐Tylophorine derived dibenzoquinolines impart multi‐biological activities as orally active agents
Authors: Lee Yue‐Zhi , Yang Cheng‐Wei , Hsu Hsing‐Yu , Qiu Ya‐Qi , Yeh Teng‐Kuang , Chao Yu‐Sheng , Lee* Shiow‐Ju
Published: June 2021

Tylophorine compounds, phenanthroindolizidines and phenanthroquinolizidines, posses multiple potent biological activities including anti‐cancer, anti‐inflammation, and anti‐coronaviruses. In order to explore more classes of tylophorine derivatives for more drug‐like compounds, herein we investigate the role of tylophorine E ring in the biological activities through synthesis of a series of compounds with the modification at the E ring either by increasing the C‐number or un‐cyclizing the E ring and forming different N‐substitutes. We have obtained potent derivatives possessing anti‐coronavirus activity in terms of anti‐TGEV and anti‐MHV replication. In addition, the most potent dibenzoquinoline showed improved solubility, in vivo efficacy, and bioavailability as an orally active agent as well as did not showed significant neurotoxicity. The relations of structure and activity of these N‐substitutes were analyzed. (This work was funded by the National Health Research Institutes, Taiwan, R.O.C. and by the National Science Council of Taiwan, R.O.C. [99–2320‐B‐400–009‐MY3].)
The Effect of Acute Creatine Monohydrate Supplementation on Cognitive Function in Fatigued Healthcare Professionals: A Pilot Study
Authors: Skelton Michele , Giamo Dustin
Published: June 2021

A recent study by the National Institute of Medicine showed as many as 98,000 patients die each year due to preventable hospital errors. The majority of these errors with the largest consequences occur in high stress environments, such as emergency departments (ED) and operating rooms. Clinical studies have demonstrated Creatine (Cr) monohydrate's impact on cognitive function. Increased memory and intelligence have been reported in Cr supplementation studies involving elderly individuals and vegetarians. The purpose of this study is to determine if short term supplementation with oral Cr‐monohydrate improves cognitive function among fatigued healthcare professionals. Two, male healthcare professionals (42.5±24.7 years) working 12‐hour shifts in the ED completed 2.5‐weeks of oral Cr‐monohydrate supplementation (5g per day). The subjects completed a battery of computerized cognitive function tests after a 12‐hour shift prior to and after creatine supplementation. Scores on cognitive function tests involving central executive tasks were significantly higher after supplementation than scores prior to supplementation (p ≤ 0.05). Results from this study suggest that acute supplementation of Cr‐monohydrate may positively impact cognitive function in fatigued healthcare professionals.
Increased systemic TNFα reduces skeletal muscle capillarity but does not change skeletal muscle fatigability in a mouse model of COPD
Authors: Audet Gerald N , Mandler W. Kyle , Thapa Dharendra , Stricker Janelle C , Zachwieja Nicole , Olfert I. Mark
Published: June 2021

Increased systemic inflammation is associated with several different pathologies, including chronic obstructive pulmonary disease(COPD). Systemic inflammation may be the link between central pathology and resultant skeletal muscle comorbidity. We hypothesized that a lung‐specific TNFa overexpressing mouse(shown to have increased circulating TNFa)would have decreased capillarity and increased in vivo muscle fatigue. We measured in vivo muscle fatigue by stimulating the tibial nerve and measuring muscle force using as standardized 3 minute fatigue protocol. After this protocol animals were sacrificed and hindlimb muscle removed for morphometric and molecular analysis. We found that TNFα overexpressing mice had no difference in overall muscle fatigue compared to littermate controls. Although circulating TNFa was elevated by 300%, no difference in skeletal muscle TNFα levels were seen. TNFa overexpressing mice exhibited lower skeletal muscle capillarity in the soleus muscle(down 16%) but not in the plantaris, compared to controls. Surprisingly, no differences in the angiogenic proteins VEGF or TSP‐1 were observed in any muscles. These data suggest that there is a muscle type specific response to capillary rarefaction caused by systemically elevated TNFα. Funding:T32NIH5T32HL090610
Myostatin reduces insulin‐stimulated phosphorylation of AS160 in human skeletal muscle cells
Authors: Consitt Leslie , Gallagher Sean
Published: June 2021

Skeletal muscle insulin resistance is a characteristic feature of type 2 diabetes and plays a prominent role in the development of this disease. Despite this knowledge, the cellular mechanism(s) contributing to insulin resistance remain unknown. Myostatin, a member of the transforming growth factor‐β (TGF‐β) superfamily has been linked to insulin resistance; however, the cellular role of myostatin‐induced insulin resistance in skeletal muscle remains to be elucidated. The purpose of the present study was to 1) identify myostatin‐mediated pathways and 2) determine the role of myostatin in distal insulin signaling (AS160) in human skeletal muscle cells. Human myotubes were incubated with myostatin (100 ng/mL) for up to 24 hours. Myostatin incubation of myotubes resulted in the rapid (5–20 min) phosphorylation of extracellular signal‐regulated kinases 1 and 2 (ERK1/2), p38 MAPK, c‐Jun NH2‐terminal kinase (JNK), and mothers against decapentaplegic homolog 2 (SMAD2). In response to insulin, phosphorylation of AS160‐Thr642 declined 23% (P<0.05) after chronic (24 hour) myostatin incubation. In addition, intramyocellular lipid (IMCL) content increased 29%, (P<0.01) in response to 24 hour myostatin incubation. These data demonstrate that myostatin stimulates a number of intracellular metabolic pathways in human skeletal muscle and suggests this protein may contribute to the accumulation of IMCL and insulin resistance.
Central Rho kinase inhibition improves baroreflex gain and cardiac autonomic balance in conscious rabbits with CHF
Authors: Pellegrino Peter Ricci , Haack Karla , Schiller Alicia , Zucker Irving H.
Published: June 2021

Recent data from our lab have shown that central Rho kinase (ROCK) inhibition normalizes disturbances in baroreflex (BR) function and cardiac sympathovagal balance induced by chronic heart failure (CHF). We hypothesized that non‐invasive measures would similarly reflect the autonomic benefits of central ROCK inhibition. Rabbits were either paced into CHF or unpaced (Sham). Intracerebroventricular infusion of either the ROCK inhibitor Fasudil (Fas; 25 ng/kg/min) or vehicle was initiated. Spontaneous BR sensitivity, pulse rate variability standard deviation of normal‐to‐normal intervals (PRV), and pulse rate variability low‐frequency/high‐frequency power spectral density (PRV LF/HF PSD) were computed from steady state parameters of animals in four treatment groups (Sham Veh, Sham Fas, CHF Veh, CHF Fas). BR sensitivity was decreased in CHF Veh rabbits compared to Sham Veh (15.9 ± 1.9 ms/mmHg vs. 4.5 ± 1.3 ms/mmHg, p < .001) while Fas tended to improve BR sensitivity in CHF animals (9.5 ± 1.3 ms/mmHg). CHF Veh rabbits had decreased PRV compared to Sham Veh (12.6 ± 1.0 ms vs. 38.0 ± 2.4 ms, p < .001) which tended to be restored in CHF Fas animals (18.6 ± 1.3 ms). The increased PRV LF/HF PSD in CHF Veh relative to Sham Veh rabbits (1.41 ± .18 vs. 0.53 ± .13, p < .001) tended to be attenuated in CHF Fas rabbits (0.73 ± 0.21). These data provide further evidence that central ROCK inhibition restores autonomic function and balance in CHF.
Abnormal immune cell populations in SHR hypertension
Authors: Singh Madhu Vrut , Whiteis Carol A. , Cicha Michael Z , Chapleau Mark W. , Abboud Francois M.
Published: June 2021

We have found that autonomic modulator of the immune system is abnormally proinflammatory in the spontaneously hypertensive rats (SHR) and may contribute to hypertension. Nicotine activates cholinergic receptors on splenocytes and suppresses IL‐6 release in response to TLR ligands in Wistar‐Kyoto (WKY) controls, but enhances it in pre‐hypertensive SHR. The objective of this study was to test (1) whether immune cell populations change as SHR age and (2) whether nicotine influences immune cell populations in SHR. We performed immuno‐phenotyping of splenocytes from WKY and SHR at ages ranging from 0 days to 38 weeks. In 0‐day‐old SHR splenocytes, a specific subset of cells (CD161+) was significantly higher than in WKY. The CD161+ cell population increased with age in SHR reaching 30% of the lymphocyte population. Infusion of nicotine increased the of CD161+ splenocytes in SHR but not in WKY. CD161+ cells produce IL‐17, a pro‐inflammatory cytokine involved in hypertension. Upon induction by phorbol myristate acetate and ionomycin, expression of IL‐17 mRNA was increased to a greater extent in SHR vs. WKY splenocytes. We conclude: 1) SHR exhibit a developmentally high abundance of CD161+ immune cells that increases with age as hypertension develops, and (2) nicotine selectively increases the number of potentially IL‐17‐producing CD161+ splenocytes in SHR. To our knowledge, this is the first demonstration of an age‐related increase in a specific immune cell population (CD161+) in genetic hypertension. Funded by NIH PPG HL14388.
Effect of Intestinal Oxalobacter Formigenes Numbers and Colonisation on Kidney Stone Formation
Authors: Pence Sadrettin , Ikizceli Ibrahim , Ozbek Emin , Tiryakioglu Ozan
Published: June 2021

Oxalobacter formigenes is unique bacteria that facilitates oxalate as its main energy source. Due to this unusual characteristic it is thought to reduce intestinal oxalate concentration and absorbtion calcium oxalate kidney stone formation. This study was conducted in order to investigate the association between the oxalate degrading bacteria oxalobacter formigenes and calcium oxalate kidney stone formation. The exact numbers of oxalobacter formigenes in calcium oxalate‐forming patients (n:50) and healthy subjects (n:50) were determined by RT‐PCR using species‐specific primers. The Mann‐Whitney U test showed that the number of oxalobacter formigenes in calcium oxalate‐forming patients was significantly lower than in healthy subjects (p=0,000019). This result indicate a strong association between the number of oxalobacter formigenes and calcium oxalate kidney stone formation.
NO‐induced increases in cGMP and inhibition of NKCC2 activity are blunted in thick ascending limbs from angiotensin II‐hypertensive rats
Authors: Ramseyer Vanesa Daniela , Ortiz Pablo A , Garvin Jeffrey L
Published: June 2021

Thick ascending limbs (THALs) absorb 25% of the filtered NaCl mainly via the Na/K/2Cl cotransporter NKCC2. Transport is enhanced in angiotensin II‐induced hypertension (AIH). NO inhibits NKCC2 activity via cGMP. Phosphodiesterase 5 (PDE5), which degrades cGMP, is increased by angiotensin II in smooth muscle cells. We hypothesized that inhibition of NKCC2 by NO is blunted in AIH due to increased PDE5 activity. Male rats were infused with vehicle (control) or 200 ng/kg/min angiotensin II for 5 days. THAL NKCC2 activity and cGMP levels were measured by fluorescence microscopy and radioimmunoassay respectively. 100 μM Spermine NONOate, a NO donor, reduced NKCC2 activity by 35% from 1.7 ± 0.3 to 1.1 ± 0.2 AU/s (p<0.01) in THALs from control but not AIH. NO increased cGMP in control THALs by 2.2 ± 0.4 fmol/μg protein but only by 1.2 ± 0.3 fmol/μg protein in THALs from AIH (p<0.04). Inhibiting PDE5 with 25 nM vardenafil (Vard) restored NO's ability to reduce NKCC2 activity in AIH (Vard: 1.2 ± 0.2; Vard + NO: 0.4 ± 0.1 AU/s, p<0.02). Vard also restored the ability of NO to increase cGMP in AIH (control: 2.1 ± 0.7; AIH: 2.2 ± 0.3 fmol/μg protein). 500 μM db‐cGMP reduced NKCC2 activity to the same extent in THALs from control and AIH suggesting the signaling cascade downstream of cGMP was not altered. We conclude that PDE5 blunts NO‐induced increases in cGMP and inhibition of NKCC2 activity in THALs from angiotensin II‐hypertensive rats.

Sources of funding: NIH HL 028982 to J.L.G. and AHA 11PRE7510005 to V.D.R.
TRPV1‐dependent afferent nerve sensitivity is augmented during spontaneous phasic contractions in aged mouse urinary bladder
Authors: Tykocki Nathan R. , Heppner Thomas J. , Nelson Mark T.
Published: June 2021

TRPV1 channels in sensory nerves are thought to communicate bladder fullness to the central nervous system. The sense of bladder fullness (“urge”) increases with age. We hypothesized that TRPV1 activity is augmented in urinary bladders from aged mice. Bladders were isolated from 3‐ and 18‐month old C57BL6 male mice for isometric contractility or ex vivo afferent nerve activity recording during constant bladder filling (1.8 ml/hr). Electrical field stimulation caused significantly greater contractions (2‐fold at 50 Hz) in bladder strips from aged mice compared to young. Spontaneous phasic contractions (SPCs), measured as pressure increases during filling of isolated bladders, were not different in young and aged. SPCs elicited significant and transient increases in afferent nerve activity. TRPV1 channel inhibition with capsaicin (1 μM) caused a 23.5% greater reduction in afferent peak frequency and a 35.9% greater reduction in afferent area under the curve in aged versus young bladders. These data suggest that aging increases urinary bladder responsiveness to parasympathetic nerve stimulation and increases the contribution of TRPV1‐signaling to afferent nerve outflow. Supported by NIH R37‐DK053832.
Effect of the tolerance to LPS in the cytokine production and activity of epigenetic enzymes in endotoxemic mice
Authors: Rios Ester Correia Sarmento , de Lima Thais Martins , Soriano Francisco Garcia
Published: June 2021

Epigenetic alteration could be associated to pre‐conditioning procedures. Chromatin remodeling in LPS tolerance is achieved by histone acetylation. LPS low doses induce a gene hypo‐transcription in acute immune response. We investigated how the tolerance to LPS could regulate the epigenetic response in the endotoxemia. Male “Balb C” mice were divided in: C group – control, not manipulated; TD group – induced tolerance and challenged ‐1mg/Kg LPS during 5 days before the challenge (5mg/Kg LPS); D group – challenged without tolerance ‐ 100μL of NaCl 0.9% during 5 days before the challenge (5mg/kg LPS). Spleens and plasma were collected 48 hours after the challenge. Activity of Histone Acetyltransferases (HAT) and Histone Deacetylases (HDAC) were assessed by colorimetric kits (abcam and biovision); Acetylation of histones H3 and H4 were analyzed by western blotting; cytokines production was measured by ELISA. Results represent the media of 5 animals. Statistical analysis was performed by ANOVA. Production of IL‐6, IL‐10 and TNF‐α was inhibited by the induction of the tolerance compared to D group. HAT and HDAC activities were increased in the TD group compared to C/D groups (p<0.05). H3 acetylated was reduced in TD compared to D. H4 acetylated was maintained at basal levels in TD and D groups. In conclusion, the reduction of cytokines during the tolerance to LPS is associated to H3 acetylation. Supported by FAPESP.
KLF4 transcription factor positively regulates the transcription of ATP2A3 gene during the differentiation of human colon and gastric cancer cell lines
Authors: Zarain‐Herzberg Angel , Flores‐Peredo Lucía , Rodríguez Gabriela
Published: June 2021

The sarco(endo)plasmic reticulum Ca2+‐ATPase, ATP2A3 gene, is highly expressed in gastrointestinal cells, but is selectively lost in cancer. Previously, was reported that SERCA3 protein is induced when human gastric and colon cancer cells undergo in vitro differentiation. In this work, we induced differentiation of gastric and colon cancer cells in culture, finding increased SERCA3 mRNA expression and increased transcriptional activity of hATP2A3‐gene‐promoter constructs (up to 50‐fold) after induced differentiation, narrowing the responsive elements within ‐ 135 to +142 bp 5’‐region of the human ATP2A3 promoter containing 8 consensus Sp‐factors binding sites, which previously have shown an important role in the transcriptional regulation of the gene. KLF4 transcription factor is decreased in gastric and colon cancer in a similar pattern to that of SERCA3, and both increase after induced differentiation. In EMSA assays, we found that KLF4 binding increases in differentiated cancer cell lines, suggesting that might function as an activator of ATP2A3 expression. In contrast, the results suggest that Sp3 functions as a repressor in tumor cells keeping low SERCA3 expression. By ChIP assays we found that KLF4 binds to the proximal ATP2A3 promoter as part of a co‐activator complex with the histone‐acetyltransferase p300, regulating positively the transcription of ATP2A3. Supported by grant PAPIIT IN213613.
Stress Hormones are Critical in Maintaining Cardiac Gene Expression and Function in Mice
Authors: Cruz‐Topete Diana , Myers Page , Foley Julie , Willis Monte , Cidlowski John
Published: June 2021

Activation of the hypothalamic–pituitary–adrenal axis results in the release of hormones from the adrenal glands‐ catecholamines, glucocorticoids and mineralocorticoids‐ which function to maintain homeostasis in the face of stressful internal or external challenges. Despite numerous observations suggesting that adrenal hormones exert direct effects on the heart, the molecular mechanisms underlying these actions remain unclear. To understand the biological effects of the stress hormones on the heart, we investigated the impact of loss of endogenous adrenal hormones in mice. Male C57BL/6 mice were adrenalectomized (ADX) at four weeks of age and maintained for 6 months after the surgery to evaluate the effects of long‐term adrenalectomy in cardiac health. We compared heart function and gene expression profiles between intact and ADX mice. Echocardiographic evaluation suggested abnormalities in the left ventricular function with decreased ejection fraction and fractional shortening in ADX mice. Histological evaluation showed an increase in thickness of the left ventricular wall of the ADX mice. Supporting these results electrocardiograms showed that ADX mice presented significantly prolonged QT intervals, indicating that adrenal hormones are important to regulate the electrical activity of the heart. Gene expression analysis of ADX‐hearts revealed an aberrant expression of a large cohort of genes associated with cardiac hypertrophy, arrhythmia and heart failure. Our results indicate that the stress hormones play an essential role in maintaining normal heart function, and suggest that selective modulation of their signaling pathways represent an approach for the treatment of cardiac arrhythmia in humans.
Vitamin D Status and Respiratory Tract Infections: A Systematic Review and Meta‐Analysis of Observational Evidence
Authors: Furlong Kayla , Omand Jessica , Pitino Michael , Science Michelle , O'Connor Deborah , Maguire Jonathon , Tran Dat
Published: June 2021

Background

Vitamin D is postulated to have antimicrobial properties, reduce inflammation, and has been implicated in respiratory health. Serum 25‐hydroxyvitamin D (25[OH]D) has been associated with risk and severity of respiratory tract infections (RTIs) in some, but not all, observational studies.

Methods

We searched MEDLINE, EMBASE, CCTR, AMED, and LILACS (through August 1, 2014) to identify observational studies (with the exception of case reports). Odds ratios, dichotomizing serum 25(OH)D at 50 and 75 nmol/L, were pooled using generic inverse variance methods with random‐effects models. Heterogeneity was assessed using Cochrane's Q and quantified by the I2 statistic. Subgroups included age, location of infection, and study design.

Results

We included 19 studies (n = 44, 301) in one or more meta‐analyses. Serum 25(OH)D levels < 50 nmol/L were associated with an increased odds of RTIs (OR = 2.63, 95% CI: 1.45‐4.76). Subgroup analyses revealed that the effect remained significant in children (OR = 2.80, 95% CI: 1.31‐5.96), lower RTIs (OR = 2.06, 95% CI: 1.16‐3.55), and in case‐control studies (OR = 2.25, 95% CI: 1.14‐4.45). Similar results were obtained when serum 25(OH)D was dichotomized at 75 nmol/L. Moderate heterogeneity was observed in all analyses (I2 蠅 42%).

Conclusions

Low serum 25(OH)D was associated with an increased risk of RTIs and remained significant in various subgroup analyses. Future studies are required to elucidate these subgroup effects.
Imaging Functional Dynamic Processes within Integral Membrane Proteins at the Single‐Molecule Scale
Authors: Blanchard Scott , Terry Daniel , Munro James , Akyuz1 Nurunisa , Gregorio Glenn , Kolster Rachel , Zhou Zhou , Zhao Hong , Altman Roger , Juette Manuel , Zheng Qinsi , Jockusch Steffen , Boudker Olga , Mothes Walther , Javitch Jonathan
Published: June 2021

Advances in single‐molecule imaging and fluorescence technologies now enable direct insights into functional dynamic processes in diverse families of integral membrane proteins. These data pave the way for quantitative descriptions of function from the intuitive perspective of time‐dependent changes in structure. Challenges on this road will be discussed in the context of recent advances in the field.