bluuebunny/crossref_metadata_2025_split huggingface dataset:
Physiological Assessment Of The PEPCK‐Cmus Mouse Heart. A Mouse Model Of An Athletic Heart Without Imposed Training?
Authors: Flores Saul , Hakimi Parvin , Liner Anna , Yu Xin , Hanson Richard W. , Snyder Christopher S. , Hoit Brian D. , Watanabe Michiko 
Published: June 2021

BackgroundCardiac hypertrophy in athletes is adaptive and distinct from pathological hypertrophy. Mice that over‐express phosphoenolpyruvate carboxykinase (PEPCK‐C) in skeletal muscle are more active and live longer compared to controls. We hypothesized that PEPCK‐Cmus mice would have an athletic cardiac phenotype in the absence of an imposed exercise regimen.MethodsAnalysis of activity level, cardiac structure with conventional (M‐mode, Doppler) and deformational (midwall strain, strain rate) echocardiographic parameters, and cardiac MRI for torsion and time to peak strain (Tpk) in PEPCK‐Cmus and control mice.ResultsThe heart rate, end diastolic dimension, LV fraction shortening and ventricular torsion were similar between groups.

Measurement
Unit
Control (n=5)
PEPCK‐Cmus (n=6)

Activity

Distance moved
cm
61999 +25.2
85830 +33.5*

Echocardiography

RWT
cm
0.56 +0.13
0.89 +0.16*

LVMI
mg/weight
3 +0.11
5+0.11*

MPI (LV)

0.45 +0.02
0.35 +0.07*

CO
cc
0.09 +0.02
0.24 +0.06*

CS
%
−27.4 +5.7
−49.1+3.1*

CS rate
1/s
−7.7 +1.8
−17.5+1.9*

RS
%
+16.9 +1.2
+37.386 +5.9*

RS rate
1/s
+4.6 +1.1
+7.8 +1.6*

Cardiac MRI

Tpk
%
60 +1.2
40 +2.1*

P‐value <0.05,
RWT=relative wall thickness; LVMI=LV mass index; MPI=myocardial performance index; CO=cardiac output; CS=circumferential strain; RS=radial strain

ConclusionsUntrained PEPCK‐Cmus mice have greater activity and their hearts have phenotypical changes characteristic of the athlete's heart, and are a novel model for non‐pathological cardiac hypertrophy that may be useful for comparison with other mice models of hypertrophy.The radial and circumferential strain measurements were distinctly different in the PEPCK‐Cmus mice indicating these parameters should be considered when clinically assessing hypertrophy.Our future plan is to analyze molecular changes (VEGF, PECAM) by Western blots in PEPCK‐Cmus and control mice.Grant Funding Source: N/A
Role of Sca‐1 after ischemia/reperfusion injury in adult mouse kidney
Authors: Camarata Troy , Vasilyev Aleksandr , Arnaout Amin 
Published: June 2021

Acute kidney injury (AKI) is a common clinical problem with significant consequences on healthcare. It is of great importance to understand the genetic and cellular changes induced by AKI. Stem cell antigen‐1 (Sca‐1, Ly6a), is a GPI‐anchored protein expressed on progenitor cell as well as differentiated cell populations. Although commonly used as a marker to enrich for stem/progenitor cells, little is known about the function of Sca‐1. Recent work has suggested Sca‐1 may play role in Smad activation downstream of TGFβ receptor 1, a pathway known to function in the progression of kidney disease. We sought to determine the role of Sca‐1 in the adult kidney using ischemia/reperfusion injury (IRI) as a model for AKI. Sca‐1 is expressed in epithelial cells of the proximal tubule, loop‐of‐Henle, and distal tubule as well as in specific interstitial and endothelial cells. After IRI, Sca‐1 knockout mice displayed significantly increased kidney injury molecule 1 expression and prolonged Smad2 phosphorylation compared to wild‐type controls. Interestingly, uninjured Sca‐1 knock‐out mice also had increased Smad2 activation. In addition, loss of Sca‐1 resulted in increased cytokine and collagen expression after IRI. These results suggest Sca‐1 is required to inhibit Smad activation by the TGFβ pathway and may protect from AKI induced interstitial fibrosis.Grant Funding Source: N/A
DNA repair choice defines a common pathway for recruitment of chromatin regulators
Authors: Bennett Gwendolyn , Papamichos‐Chronakis Manolis , Peterson Craig 
Published: June 2021

DNA double‐strand break (DSB) repair is essential for maintenance of genome stability. Recent work has implicated a host of chromatin regulators in the DNA damage response, but their functional roles remain unclear. In budding yeast, DSB recruitment of chromatin regulators has been monitored primarily in asynchronous cell populations, and thus it is unclear if these events are linked to non‐homologous end joining (NHEJ) or homologous recombination (HR). By using synchronized cell populations, we surprisingly find that subunits of the INO80, SWR‐C, NuA4, SWI/SNF, and RSC enzymes, as monitored by chromatin immunoprecipitation (ChIP), are primarily recruited outside of the G1 cell phase. Furthermore, recruitment is inhibited by the NHEJ machinery, and requires early steps of HR, strongly supporting a role for these enzymes in the HR repair pathway. Strikingly, and in contrast to previous reports, we also find no significant role for H2A.X phosphorylation (γH2AX) in the recruitment of chromatin regulators, but rather their recruitment coincides with reduced levels of γH2AX.This work was supported by a grant from the NIH (GM54096).
A unique acquired resistance mechanism to agents that target GLI (GANT61) in the Hedgehog (HH) signaling pathway
Authors: Agyeman Akwasi , Mazunder Tapati , Houghton Janet 
Published: June 2021

Canonical HH signaling engages the transmembrane receptor PTCH, the intermediary signaling molecule SMO, and the HH signaling response molecules, GLI1 and GLI2. In colon cancers, GLI is aberrantly regulated, driving cell survival and genomic instability in oncogenesis, progression and metastasis. Following inhibition of GLI by a protracted exposure of HT29 to GANT61, we have identified a previously undescribed, novel mechanism by which cancer cells evade cell death. Stepwise selection did not produce a proliferative colon cancer cell line resistant to GANT61, confirming the critical importance of GLI genes to cell survival. In contrast, in HT29 cells, by inhibiting GLI1/GLI2 by prolonged exposure to semi‐lethal concentrations of GANT61 (10 μM; G10), we observed reduced proliferation, decreased appearance of γH2AX nuclear foci, and increased methylation of histone H3K9 (H3K9me1–3) within the promoters of the GLI1 and GLI2 genes. Increased H3K9 methylation was also demonstrated in HT29(G10) cells in the promoter regions of FoxM1 and CDC6, a G1/S and PreReplication Complex assembly regulatory genes, respectively. Data suggest that the GLI2‐>;GLI1‐>;FOXM1‐>;CDC6 promoters are coordinately regulated by H3K9 methylation in a pathway to promote gene silencing, reduce gene transcription, and reduce cell cycle transition at G1/S, constituting a unique mechanism of acquired resistance to drugs that target GLI.This work is supported by NCI awards RO1 CA 32613 (J.A. Houghton) and NCI RO1 108929 (J.A. Houghton)
CTLA4 Polymorphisms are associated with Severe Pulmonary Tuberculosis in a Chinese Han population
Authors: Wang Chong , Jiang Tingting , Wei Liliang , Zhong Jianping , Liu Jiyan , Zhang Xing , Xu Dandan , Chen Zhongliang , Li Zhongjie , Li Ji‐Cheng 
Published: June 2021

The cytotoxic T lymphocyte antigen‐4 (CTLA4) is a key negative regulator in T lymphocyte activation. Polymorphisms on CTLA4 are associated with susceptibility to many autoimmune diseases, resulting in down‐regulation of the inhibition of cellular immune response. The present study aimed to investigate potential associations of CTLA4 +49A>;G (rs231775), +6230G>;A (rs3087243), and 11430G>;A (rs11571319) polymorphisms with pulmonary tuberculosis (PTB) in a Chinese Han population. A case‐control study was performed, and the three SNPs were genotyped by PCR and DNA sequencing methods. We found the frequency of CTLA4 +49AG genotype in 274 PTB cases was significantly lower than that of the 265 controls (Pcor=0.025, 0R 0.613, 95% CI 0.401–0.937). However, no associations were found between the other 2 SNPs (P>;0.05). After divided PTB cases into two groups: with or without cavities, we found a novel relation of +6230AG genotype to cavities (Pcor=0.046, 0R 2.677, 95% CI 1.029–6.965). Lately haplotype analysis showed that haplotype AGG was significantly associated with healthy controls (P=0.002, OR 0.286, 95% CI 0.124–0.657), while haplotype AAG was related to PTB with cavities (P=0.047, OR 2.053, 95% CI 1.000–4.215). Our results suggest the CTLA4 polymorphisms can be associated with PTB in a Chinese Han population, and +49AG may reduce the risk of being PTB, while +6230AG may contribute to cavities formation.This work was supported by grants from National Natural Science Foundation of China (No.81072724, No.81273882), National Special Sci‐Tech Projects (No.2012ZX10005001–006, No.2008ZX10005–010), and Zhejiang Province Special Sci‐Tech Projects (No.2009C03011–3).
Purification and Analysis of Rho‐1D4‐tagged Peripheral Cannabinoid Receptor CB2
Authors: Locatelli‐Hoops Silvia Cristina , Gorshkova Inna , Gawrisch Klaus , Yeliseev Alexei A 
Published: June 2021

Human peripheral cannabinoid receptor CB2 is involved in regulation of the immune response. For studies requiring surface immobilization, pure and functional receptor with a reliable tag is required. The goal of this project is to test the usefulness of the C‐terminal epitope of rhodopsin (rho‐1D4‐tag) for purification and surface immobilization of recombinant CB2 using the monoclonal antibody 1D4.Expression of rho‐1D4‐tagged CB2 as a fusion with maltose binding protein (MBP) in E coli was evaluated in terms of protein levels, accessibility of the rho‐1D4 tag and activity. Highly pure tagged CB2 was obtained by rho‐1D4 and Ni‐NTA affinity chromatography. The reconstituted protein was functionally active by G protein activation assay. The interaction of rho‐1D4‐tagged CB2 with 1D4 antibody was characterized by surface plasmon resonance (SPR). Either purified or fusion CB2 from crude extracts was captured on the 1D4 antibody‐coated surface in a specific and quantitative fashion, and its presence confirmed by biding of an anti‐CB2 monoclonal antibody. Ligand binding to purified receptor in detergents was studied upon immobilization of fusion CB2 on 1D4 antibody resin. This methodology was also used to study thermal stability of the receptor in micelles.In conclusion, the rho‐1D4 tag was successfully utilized for purification, immobilization, biochemical and biophysical studies of functional CB2.
TGF‐β regulated miRNAs: therapeutic targets for cardiac hypertrophy
Authors: Yang Xiao , Wang Jian 
Published: June 2021

Heart failure (HF) is one of the most frequent causes of death worldwide. The underlying causes of HF are diverse but often relate to cardiac hypertrophy. The role of TGF‐β signaling in cardiac hypertrophy has been extremely contradictory. We have previously showed that targeted deletion of Smad4 unexpectedly leads to cardiac hypertrophy, demonstrating that the endogenous cardiomyocyte Smad4‐dependent TGF‐β pathway protects heart from cardiac hypertrophy and fibrosis.Recently, we have revealed that the function of endogenous TGF‐β/Smad signaling in maintaining cardiac homeostasis involves the downregulation of miRNAs inducing cardiac hypertrophy. We show that TGF‐β1 inhibits the expression of miR‐23a/miR‐27a/miR‐24–2 and miR‐23b/miR‐27b/miR‐24–1 clusters at the transcriptional level. Transgenic mice with cardiomyocyte‐specific overexpression of miR‐27b exhibit cardiac hypertrophy and dysfunction by directly targeting PPAR‐¦Ã. Most importantly, in vivo silencing of miR‐27b in a pressure‐overload‐induced mouse model of HF attenuates cardiac hypertrophy and dysfunction. The function and mechanisms of other miRNAs regulated by TGF‐β/Smad signaling in cardiac hypertrophy have also been investigated. All these results provide critical genetic evidence showing that TGF‐β‐regulated miRNAs might serve as efficient therapeutic targets for cardiac diseases.
Adaptation of a compact rapid vitamin A assay device (iCHECK™) to simulated field conditions and relevant substrates in Guatemala
Authors: Zambo Zsofia Magdolna , Sanchez‐Mena Jose David , Orozco Monica N , Solomons Noel W 
Published: June 2021

BackgroundGuatemala has mandated fortifying granulated table sugar with vitamin A at 10–20 ppm. Children in the interior commonly continue partial breast‐feeding through the first 2 y of life.ObjectiveTo adapt the preparation of samples of sugar and bovine milk for a rapid assessment of vitamin A using nonlaboratory items for mixing and measuring.MethodsSugar samples were mixed for homogenization in plastic bags, with 4 g delivered into a 20 mL syringe to prepare a 20 w/v% solution. Heavy cream was diluted in plastic bags. Samples were injected into iEX™ extraction vials and measured by fluorometry in an iCHECK™(BioAnalyt, Telbow, Germany) rapid assay unit, providing digital read‐outs in μ/L.ResultsIntra‐sample CVs ranged from 0.3–7.8% and intraobserver CVs ranged from 2.1– 15.0% for dairy. The r value for the inter‐observer assay association for 12 sugar specimens was 0.86 (p<0.001). Inter‐observer accord on median vitamin A values was excellent for both substrates. Plausible values for sugar vitamin A, with a median of 11 ppm, were found for 58 samples of sugar gathered from across 4 regions.ConclusionThe iCHECK™ system provides plausible values for vitamin A content of fortified sugar and bovine milk and cream. It offers important promise for a rapid, on‐site, field‐level evaluation of materials for timely public health responses. Supported by a donation from Sight & Life
Fructose reduction improves CVD risk in adolescents with NAFLD
Authors: Vos Miriam , Jin Ran , Welsh Jean , Le Ngoc‐Anh 
Published: June 2021

ObjectiveNonalcoholic fatty liver disease is associated with increased CVD risk. We have previously reported abnormal post‐prandial TG response after fructose‐enriched meals in adolescents with NAFLD suggesting that fructose may contribute to the increased CVD risk. We hypothesized that fructose reduction would improve CVD risk factors in adolescents with NAFLD.MethodsOverweight, Hispanic adolescents who self‐reported high consumption of sugar beverages (>;24oz/day) were recruited for hepatic fat screening by MRS. 24 adolescents had high hepatic fat (HHF, ≥5%) and were randomized to either 4 weeks of fructose reduction (home beverages replaced with 3 glucose study provided beverages per day) or 4 weeks of continued high fructose (home beverages replaced with 3 fructose study provided beverages per day). MRS, fasting labs, and anthropometrics were repeated at baseline, 2 and 4 weeks.ResultsAfter 4‐weeks of glucose beverages (fructose reduction), subjects had improved adipose IR (p=0.034), hs‐c reactive protien (p=0.026), circulating oxidized LDL (p=0.056), and LDL oxidation susceptibility (p=0.009); which were not observed in fructose beverage group. No improvement was seen in BW, hepatic fat, and fasting TG.ConclusionReduction of fructose containing beverages for 4 weeks improved CVD risk profile in adolescents with HHF despite a lack of change in hepatic fat.
Effects of topically applied Platycarya strobilacea S. et Z. extract on hair growth in C57BL/6 mice
Authors: Lee Bog‐Hieu , Kim Eun Jin , Choi Joo‐Yeon , Yu Miri 
Published: June 2021

The study was conducted to evaluate the effect of Platycarya strobilacea S. et Z. extract on hair growth. Four‐week‐old male C57BL/6 mice were assigned to three experimental groups (n=5/group); the dimethyl sulfoxide (DMSO) group (NC), the minoxidil group (PC), and the Platycarya strobilacea S. et Z. extract group (PSE). Hairs of animals were shaved on the dorsal skin before the study and 100 μℓ of test samples were topically applied for 3 weeks everyday, while the dorsal skin was taken pictures on the 0th, 7th, 14th, 21st day. We observed morphology, length of hair follicle, and no. of mast cells and stem cell factors (SCF) on skin tissues. PSE group markedly promoted the hair growth than did NC and PC groups. Depth of hair follicle in PSE group(843.33±84.33 μm) was the deepest the deepest in PSE group, while those of NC or PC group were 758.33±150.05 μm and 655.00±145.45 μm, respectively. In case of number of mast cells, PSE group have shown the lowest in number(1.50±0.29) in comparison to NC(6.33±0.88) and PC(7.00±0.71)(p<0.001). Also, SCF staining in follicles, mast cells, skin layer indicated that PSE‐treated group have shown the strongest immune response than did the NC and PC groups. From the findings, topical application of PSE had a positive effect on hair growth and improved hair follicle length, mast cell, and immune response. Further research should be conducted to clarify the action mechanism related to Platycarya strobilacea S. et Z. extract.Grant Funding Source: This research was supported by Basic Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education, Science and Technology(2011–0013949)
A Synopsis of Chromosomal Microarray Cases at Dartmouth‐Hitchcock Medical Center
Authors: Tayoun Ahmad N Abou , Tsongalis Gregory J. , Lefferts Joel A. 
Published: June 2021

BackgroundChromosomal microarrays (CMA) used to assess DNA copy number are currently considered first‐tier clinical diagnostic tests for postnatal evaluation of individuals with intellectual disability (ID), developmental delay (DD), autism spectrum disorders (ASD), and/or multiple congenital anomalies (MCA). Here, we provide a summary of the CMA cases performed at Dartmouth‐Hitchcock Medical Center during the first month that clinical CMA testing was offered.MethodsWe used the CytoScan®HD microarray containing 750,000 SNP probes and 1.9 million non‐polymorphic probes with an average intragenic spacing of 880 base pairs and 384 base pairs for 340 genes recommended by the International Standards for Cytogenomic Array (ISCA) consortium. Several databases of copy number variants were used during interpretation including ISCA, OMIM, DECIPHER and the Database of Genomic Variants (DGV).Results16 cases (9 females and 5 males) with an average age of ~7yrs were analyzed and reported. The clinical indication for testing in 14/16 cases (87.5%) was ID, DD or ASD. Clinically significant copy number variants were detected and reported in 3/16 cases, a diagnostic yield of 18.75%. Additionally, two variants of uncertain significance (VOUS) were also reported in two cases with unexplained developmental delay.ConclusionsDespite the small number of cases, our diagnostic yield matches the well‐established CMA yield (15–20%) for genetic testing of individuals with ID, DD, ASD or MCA. This further emphasizes the fact that CMA provides higher sensitivity compared to G‐banded karyotyping albeit with the slight disadvantage of increased VOUS detection.
Do pericytes represent the cellular switch between kidney health and disease?
Authors: Peppiatt‐Wildman Claire M 
Published: June 2021

Pericytes are pluripotent cells with numerous functional roles. We focus on the role of pericytes in kidney (patho)physiology and describe data identifying pericytes as being pivotal in both settings. Using the live kidney slice model [1], we have demonstrated i) that pericytes respond to various endogenous vasoactive mediators to regulate vasa recta diameter and thus medullary blood flow (MBF) [1], ii) neuro‐vascular coupling between pericytes and sympathetic nerves and iii) hypotonicity‐induced regulation of vessel diameter by pericytes, which is indicative of pericyte‐mediated tubulovascular crosstalk [1]. Collectively these represent novel mechanisms by which pericytes match MBF with the high metabolic demand of the tissue.Additionally, we describe a critical role for pericytes in drug‐induced nephrotoxicity that is established for various commonly prescribed drugs. We demonstrate pericyte‐mediated vasoconstriction in response to immunosuppressant antibiotics and non‐steroidal anti‐inflammatory drugs, which may cause exacerbated local ischemia responsible for the acute kidney injury observed clinically. Our data, in combination with studies highlighting a key role for pericytes in renal diseases, suggests pericytes are the cellular switch underpinning the onset/progression of renal disease.
Gating modulation of the sodium‐activated potassium channel KCa4.1 (Slack) by bithionol and niclosamide
Authors: Fleming Matthew R. , Singh Vikrant , Jenkins David Paul , Wulff Heike , Kaczmarek Leonard K. 
Published: June 2021

The Na+‐activated K+ channel Slack (also known as KCNT1, Slo2.2, or KCa4.1) is widely expressed throughout the central nervous system, where it contributes to the slow afterhyperpolarization and is involved in controlling bursting behavior and spike frequency adaptation. Gain‐of‐function mutations in Slack have recently been shown to cause malignant migrating partial seizures of infancy. KNa channels further have long been known to protect cells from hypoxic injury. Slack activators therefore should be therapeutically useful for reducing neuronal excitability in epilepsy and ataxia as well as for the prevention of hypoxia associated neuronal damage in stroke and cerebral palsy. Slack channel activity is increased by the antiparasitic compounds bithionol and niclosamide. However, while bithionol is a positive gating‐modulator that requires intracellular sodium to be elevated to exert an effect, niclosamide is a true opener that activates Slack currents in whole‐cell patch‐clamp experiments even with a KF containing intracellular solution in the nominal absence of both Na+ and Ca2+. Both types of Slack channel activators can be detected by resonance wavelength grating optical biosensors (the SRU Biosystems BIND), a high‐throughput assay measuring changes in mass at the plasma membrane. We are currently using this assay to screen for both Slack channel activators and inhibitors.Supported by R21 NS073943
Small arteries of gestational rats are less sensitive to relaxation induced by inhibition of rho A‐associated kinase (ROK).
Authors: Chi Mei , Zielnik Barbara , Davidge Sandra , Mitchell Bryan 
Published: June 2021

Inhibition of ROK has been proposed as a method to arrest uterine contractions during preterm labor. Because of concern about vascular side effects, we studied the effects of the ROK inhibitor g‐H1152 (g‐H) on relaxation of mesenteric and uterine arteries.Arteries from non‐pregnant (n=5) and pregnant (gestation day17–21; n=6) rats were studied using wire myography. Concentration response curves (CRC) to calculate IC50 for g‐H were obtained following phenylephrine (PE) or endothelin‐1 (ET‐1) induced constriction. Using vessels from pregnant rats, CRC for PE were performed before and after a 20‐min incubation with g‐H at 300 nM, 1 and 3 μM.Mesenteric arteries were more sensitive than uterine arteries to g‐H‐induced relaxation in both non‐pregnant and pregnant rats. Both arteries were significantly less sensitive to g‐H‐induced relaxation during pregnancy. Sensitivity to PE was not changed with any dose of g‐H. However, using 2‐factor ANOVA, the maximal PE‐induced tension was significantly reduced by the two highest concentrations of g‐H in both vessels from pregnant rats but to a greater extent in mesenteric vessels (34±5.6%, 18±4.1%) compared to uterine (14±4%, 7±2.7%).The ROK pathway plays a significant role in agonist‐induced constriction of mesenteric and uterine vessels, though less so during gestation. If ROK inhibitors are used during pregnancy in vivo, maternal blood pressure and uterine blood flow will be assessed.