bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of Pre‐exercise and PM2.5 on Lung Pulmonary Function and Inflammation in Aged Rats
Authors: Qin Fei , Zhao Jiexiu , Xu Minxiao , Wang Zhongwei 
Published: April 2020

PURPOSEExposure of particulate matter of less than 2.5 μm (PM2.5) has been associated with adverse respiratory and the risk of inflammation. While regular physical activity (PA) reduces the risk of many adverse health effects. This study aimed to examine the protection of pre‐exercise on adverse health effects of Pulmonary Function and inflammatory induced by PM2.5 exposures in aged rats.METHODS24 male wistar rats, aged 16 months, were randomly divided into 4 groups: Sedentary (S), Exercise (E), Sedentary+PM2.5 exposures (S+PM), and Exercise+ PM2.5 exposures (E+PM).The rats in all E‐related groups went through an 8‐week aerobic treadmill exercise protocol (65–75%VO2max, 30 min) at every other day. The PM‐related groups of aged rats were exposed to concentrated PM2.5 in Beijing(7 days/week, 4 hours/day). After 2‐week PM Exposure, the pulmonary function examined by whole body plethysmography, structure of lung tissues were observed by HE staining, and the counts of leukocyte, sIgA, CRP and TNF‐α concentration were examined.RESULTS1)Compared with S group, following changes occurred in S+PM group after 2‐week PM2.5 exposure: lung tissues were seriously damaged, local bleeding, pus exudation, and inflammatory cell infiltration, as well as the decline of sIgA level (↓45%, ES=0.87, p<0.05) and percentage of neutrophil (↓62%, ES=0.45,p<0.05), while the incline of the Index of constriction (↑76%,ES=3.97, p<0.05), the rejection index, CRP(↑24%, ES=0.83, p<0.05), and TNF‐α(↑38%, ES=0.84, p<0.05) were observed. 2) Compared with S+PM group, the structure of lung tissues and pulmonary function were obviously improved in E+PM group. In addition, the sIgA concentration (↑85%, ES=0.70, p<0.05) were enhanced markedly, while CRP (↑21%, ES=0.76, p<0.05), and TNF‐α level (↑25%, ES=0.74, p<0.05) were decreased in E+PM group compared with S+PM group.CONCLUSION2‐week PM2.5 exposures led to an increased susceptibility of infections, index of constriction and susceptibility of pulmonary function in aged rats. Moderate pre‐exercise has beneficial effects on pulmonary function and reducing the susceptibility to inflammation.Support or Funding Informationthe National Natural Science Foundation of China (11775059 and 31900845), the China Postdoctoral Science Foundation (2018T110076), and the Fundamental Research Foundation of the China Institute of Sport Science (18‐02, 19–22, and 19–34).Effect of Pre‐exercise and PM2.5 on Pulmonary Functions in Aged Rats.*P<.05. All values were presented as Mean ± SD, n= 10 for each group.Figure 1Impact of PM2.5 Exposures and Pre‐exercise on Inflammation in Aged Rats.*P<.05. All values were presented as Mean ± SD, n= 10 for each group.Figure 2
Continuous Glucose Monitoring Implemented in Pregnant Mice
Authors: Wuyts Caroline , Simoens Caroline , Philippaert Koenraad , Pinto Sílvia João Poseiro coutinho , Vennekens Rudi 
Published: April 2020

Pregnancy is a challenge for the female body. Increased insulin resistance of the mother allows appropriate nutrient flow to the fetus. This is counterbalanced by increased insulin secretion from pancreatic β‐cells. Gestational diabetes mellitus (GDM) arises when insulin resistance is increased excessively or pancreatic β‐cells fail to adapt.To study basic mechanisms and novel therapeutic strategies for GDM, it is necessary to test glycaemia in pregnant mice. We investigated the potential of continuous glucose monitoring via telemetry as a reliable and stress‐free approach to follow glycaemia during pregnancy. The transmitter is capable of continuous monitoring the temperature, activity, and plasma glucose levels in the arterial blood during full pregnancy. The telemetric implant had no effect on the pregnancy and the litter. Oral glucose tolerance tests were performed before, during and after pregnancy using intermittent blood sampling and telemetry. With both methods, we observed a similar glucose tolerance profile in wildtype mice during pregnancy. However, the use of telemetry results in a large amount of continuous data, which greatly extends the level of detail in which glycaemia during pregnancy can be monitored. We combined camera surveillance and telemetry to observe glycaemia during spontaneous feeding behavior. Once this behavior was known, glucose excursions and their respective parameters could be determined, e.g. amplitude, area under the curve, and time to peak. In addition, several parameters of glycaemic variability, important in diabetes research, were calculated from this dataset. For example, the standard deviation, interquartile range, and continuous net glycaemic action over 30 min seem to be disturbed specifically around delivery. In general, wildtype C57BL/6N mice are able to cope with the extra metabolic demand placed upon the mother by pregnancy. However, changes in glycaemic variability, average glucose levels and activity could be found throughout pregnancy. To conclude, continuous glucose monitoring is a safe method that provides insight in glycaemic control during pregnancy with unprecedented detail.Support or Funding InformationKU Leuven Bijzonder Onderzoeksfonds: TRP Research platform Leuven (TRPLe)Figure 1
Left Bifid Ureter and Renal Artery Variation in an Adult Female Cadaver
Authors: Sullivan Hannah , Tan Yun , Daly Daniel T. 
Published: April 2020

During the retroperitoneal space dissection of a 77‐year‐old female cadaver, a left duplicated ureter (DU) was observed along with variations of left renal artery branching pattern. The ureter, typically 25–30cm in length, is a continuation of the renal pelvis that receives filtrate from 2 or 3 major calyces before entering the bladder. In this cadaver, two separate ureters that drained upper and lower kidney, with lengths of 12.4 cm and 10.6 cm respectively. The two ureters then joined to form a common ureter with a length of 12.7 cm before entering the bladder. There are two types of DU; this type of DU has been previously described as an incomplete or bifid ureter and is a commonly asymptomatic congenital abnormality of the kidney and urinary tract (CAKUT). However, it can produce symptoms of frequent urinary tract infections, urinary lithiasis, ureteric stenosis and renal colic
(1). The incidence of any type of DU is 1 in 100 to 1 in 500
(2) and is also more common in women than men, with a ratio of 2:1
(1). Unusual blood supply of the left kidney was also observed with an aberrant left renal artery arising from the abdominal aorta 4 cm inferior to the left renal artery, crossing anterior to the DU and supplying the lower pole of the left kidney. The ovarian artery was observed arising from this aberrant left renal artery. The incidence of the gonadal artery arising from a source other than the abdominal aorta is approximately 20%
(3). The incidence of an aberrant renal artery is observed in 20–30% of the population and has the potential to result in hydronephrosis
(4).Support or Funding Information
Impact of Common Polymorphic Variant A503V in POR on Drug Metabolism: Implications for POR Deficiency
Authors: Velazquez Maria Natalia Rojas , Parween Shaheena , Pandey Amit V. 
Published: April 2020

A broad spectrum of human diseases, including abnormalities in steroidogenesis, is caused by mutations in the NADPH cytochrome P450 oxidoreductase (POR)
[1,
2]. Cytochrome P450 proteins perform several reactions, including the metabolism of steroids, drugs, and other xenobiotics. Therefore, genetic variations in POR can impact many different metabolic pathways by changing the activities of cytochromes P450. In 2004 the first human patients with defects in POR were reported with symptoms of ambiguous genitalia and bone malformations, and over 200 variations in POR are known.By analyzing the POR sequences from genomics databases, we focused on the common variant in POR A503V, which is often present in heterozygous form in many PORD patients. We expressed the A503V variant of POR in bacteria and characterized it by functional studies using purified recombinant proteins. The POR, as well as P450 Proteins, were expressed in E. coli BL21(DE3) and purified by ion‐exchange, affinity, and metal chelate chromatography for activity assays as described in our earlier publications
[3,
4]. Activities of cytochrome P450 enzymes were tested with lipids into which P450 and P450 reductase proteins were embedded and assayed using fluorogenic substrates on a microplate spectrofluorometer.We found a positive effect on many drug‐metabolizing enzyme activities due to the A503V variant in POR. The activity of CYP2C9 was increased to 180% of the WT POR, and CYP2C19 showed 243% of WT activity with the A503V variant of POR. Most remarkable was the impact of the A503V variant on CYP3A5 activity, which was increased by 421% compared to WT POR.Effect on drug metabolism in patients with PORD requires careful considerations of cytochrome P450 mediated activities. While many PORD patients have low or reduced drug metabolism due to defects in POR, the presence of the A503V allele can alter the overall effect on drug metabolism. Significantly, tacrolimus, a drug given during organ transplant, can be metabolized several folds faster due to the presence of the A503V variant of POR. The presence of the A503V allele is likely beneficial in patients with POR deficiency.Support or Funding InformationThis work was supported by the Swiss National Science Foundation (31003A‐134926), the Novartis Foundation for Medical‐Biological Research (18A053), and a grant from Burgergemeiende Bern to AP.
Effect of Macamides N‐(benzyl)linolenamide and N‐(3‐methoxybenzyl)linolenamide on Autophagy in HepG2 and HeLa Cells
Authors: Martínez-Málaga Jimena Alexandra , Gonzales-Urday Ana Lucia , Rondón-Ortiz Alejandro Nico , Böhlke Mark , Pino-Figueroa Alejandro 
Published: April 2020

Autophagy is a conserved catabolic process which channels intracellular proteins and organelles into lysosomes to be degraded. This process can be triggered either by nutrient deprivation or by metabolic stress. Autophagy malfunction has been linked to a variety of human diseases such as neurodegeneration, myopathies, infectious diseases and different cancer types. Therefore, the search for novel autophagy modulators is of great interest to reveal autophagic mechanisms involved in these disease processes. As many natural product‐derived compounds have been used as traditional medicines, they have great potential as lead autophagy modulators. Macamides are secondary metabolites isolated from the Peruvian plant Lepidium meyenii L. (Maca). These metabolites have exhibited various biological activities, including anti‐oxidative, anti‐fatigue, anti‐cancer, anti‐osteoporotic and neuroprotective effects. The purpose of this study was to evaluate the effect of two macamides: N‐(benzyl)linolenamide (Cmpd 1) and N‐(3‐methoxybenzyl)linolenamide (Cmpd 2) on autophagy in both HepG2 and HeLa cancer cell lines. Cell viability in both cell lines was strongly reduced by both Cmpd 1 (HepG2, IC50 = 75.6 μM; HeLa, IC50 = 116 μM) and Cmpd 2 (HepG2, IC50 = 62.8 μM; HeLa, IC50 = 117 μM) as determined by the MTS assay. Subsequently, it was investigated whether macamides inhibit lysosomal acidification, by monitoring the red fluorescence emitted by cells stained with LysoTracker Red DND‐99. Significantly, both Cmpd 1 and Cmpd 2 (50 μM) decreased the red fluorescence in both cell lines. Furthermore, to evaluate autophagic flux in HepG2 and HeLa cells, protein levels of microtubule‐associated protein 1A/1B‐light chain 3 (LC3) and p62 were measured. No significant differences were observed in LC3 levels. Nonetheless, Cmpd 1 was identified as the most effective compound to induce accumulation of p62 in HepG2 cells. Because p62 is degraded in the lysosome during the last stage of autophagy, these results suggest that macamides inhibit autolysosomal degradation. However, a more in‐depth study is needed to determine the inhibitory mechanism and/or if another type of autophagy is involved.Cmpd 1 and Cmpd 2 decreases acidification of lysosomes in HepG2 cells.Figure 1Cmpd 1 and Cmpd 2 decreases acidification of lysosomes in HeLa cells.Figure 2
New Insights into the Role of Intracellular Ca2+ in the Regulation of the Human Kidney Chloride Channel ClC‐Ka
Authors: Carmosino Monica , Gerbino Andrea , De Zio Roberta , Milano Serena , Russo Daniela , Milella Luigi , Procino Giuseppe , Svelto Maria 
Published: April 2020

It has been recognized that the physiological role of ClC‐Ka and its mouse homolog ClC‐K1 in the nephron is to confer high Cl− permeability in the thin Ascending Limb of Henle (tAL), which in turn is essential for establishing the high osmolarity of the renal medulla in the countercurrent system. Accordingly ClC‐K1 KO mice display a severe polyuria resembling a Nephrogenic Diabetes Insipidus (NDI) phenotype. Thus, ClC‐Ka in human nephron could be an interesting target for diuretic and anti‐hypertensive drugs. Unfortunately the regulation of ClC‐Ka in and its electrophysiological properties are still not completely known. We analyzed, by whole‐cell voltage‐clamp, Cl− currents in HEK293 cells expressing GFP‐ClC‐Ka and the accessory m‐Cherry Barttin subunit upon intracellular Ca2+mobilization. The patch clamp protocol consists of a holding potential from 0 mV, which is stepped to various test potentials from −195 and +125 mV for 150 ms. Pulses ended with a tail pulse to −125 mV for 20 ms. In control conditions we found a linear current/voltage relationship over a broad range of membrane voltages except of hyperpolarization below −100 mV, where the channel open probability was reduced, providing the well‐known hook‐shaped appearance of the I‐V curve for ClC‐Ka channel. Interestingly, both 100 μM ATP and 400 μg/ml of dandelion root extract (DRE), both increasing intracellular Ca2+ in HEK293, significantly inhibited Cl− currents in a time dependent way. Pretreatment of cells with either the Ca2+ chelator BAPTA‐AM or the PCK inhibitor Calphostin C, reverted the inhibitory effects of both ATP and DRE. Interestingly, 1μM of Phorbol Meristate Acetate (PMA), a specific PKC activator, mimicked the inhibitory effect of ATP and DRE on Cl− currents. We finally demonstrated that 30 μM Heclin, an inhibitor of E3 ubiquitin ligase, did not revert the DRE‐ and ATP‐ induced Cl− currents inhibition, thus suggesting that the ClC‐Ka inhibition is independent of channel ubiquitination. Accordingly, live confocal analysis showed that all the Ca2+mobilizing compounds tested did not induced neither ClC‐Ka nor Barttin internalization. We hypothesized that phosphorylation in the putative PKC consensus sites present in ClC‐Ka amino acidic sequence may be involved in the inhibition of channel activity. In conclusion, we demonstrated for the first time that the activity of ClC‐Ka in renal cells could by inhibited by either receptor agonists or natural molecules able to activate an intracellular Ca2+/PKC pathway, thus opening venues for the development of new diuretic drugs.Support or Funding InformationGrant n# ARS01_01081 to MC from The National Operational Program (PON) of the Ministry of Education, ITALYFigure 1
Structure‐Function Studies of the Helicase Domain of NS1 Protein of Human Parvovirus B19
Authors: Ghadirian Niloofar , Horton Nancy 
Published: April 2020

Human Parvovirus B19 can cause serious fatal acute infections and is a cause of miscarriages, it has also been suggested it can trigger the development of autoimmune diseases. NS1 protein of B19V has roles in viral replication and modulation of host genes and pathways. The helicase domain of this protein has predicted ATPase activity and is implicated in strand separation at the viral origin of replication. It may also play another role in separating the two strands of DNA following initial strand synthesis of the single‐stranded genome, viral packaging, protein‐protein interactions in various cellular pathways important for viral replication and virus production, and in gene transactivation. Purifying this protein from bacterial sources and characterizing its ATPase and DNA strand separation activity, as well as performing structural investigations (using analytical ultracentrifugation, x‐ray crystallography, and cryo‐electron microscopy) and functional investigation (for DNA‐binding, cleaving, helicasing) are recent works that will be presented. Completion of this work can provide new targets for diagnostics and therapeutics in Human Parvovirus B19 disease.Support or Funding InformationThe University of ArizonaSDS‐PAGE gel after purification of B19V NS1 through Talon resinFigure 1a)Native gels for DNA helicase assay using hishelicase domain of NS1 for Ori‐top26*+Ori‐bot75 construct. b) plot of ds and ssDNA with time for helicase activity.Figure 2
Sulfonated Arylurea Derivatives are Potent Inhibitors of Human Neutrophil Elastase
Authors: Al-Horani Rami A. , Mottamal Madhusoodanan 
Published: April 2020

Human neutrophil elastase (HNE) is a serine protease in the chymotrypsin superfamily of enzymes. HNE has been reported to play important roles in inflammation, innate immune response, and tissue remodeling processes. Therefore, HNE has been actively pursued as a drug target, particularly for the treatment of cardiopulmonary diseases. Although thousands of molecules have been reported to inhibit HNE, yet very few have been tested in early clinical trials and with only marginal success. We report here a novel chemotype of heparin‐like sulfonated arylurea derivatives as potent and noncompetitive inhibitors of HNE. Using a chromogenic substrate hydrolysis assay, 14 heparin‐like sulfonated molecules were tested for their inhibitory activity against HNE. Only 12 molecules inhibited HNE with IC50 values of 0.22 – 88.3 μM. The inhibition of HNE by these molecules was NaCl‐dependent. Interestingly, a specific hexa‐sulfonated arylurea derivative inhibited HNE with an IC50 value of 0.22 μM via noncompetitive mechanism as demonstrated by Michaelis‐Menten kinetics. The hexa‐sulfonated derivative demonstrated 455‐, 221‐, 21‐fold selectivity indices over other heparin‐binding serine proteases including thrombin, factor Xa, and factor XIa, respectively. The molecule also demonstrated 381‐fold selectivity over factor XIIIa, a transglutaminase clotting factor with a potential heparin‐binding domain. The molecule did not show heparin‐like effects on clotting times of human plasma as determined by partial activated thromboplastin time and prothrombin time assays. A computational approach was exploited to identify the putative binding site for this novel class of HNE inhibitors. Overall, reported sulfonated arylurea derivatives serve as a new platform to develop potent, selective, and noncompetitive inhibitors of HNE for therapeutic purposes.Support or Funding InformationThis work was supported by NIGMS/NIH under award number SC3GM131986 and by IDeA program from NIGMS/NIH under grant number P20 GM103424 to RAAH. The content is solely the responsibility of the authors and does not necessarily represent the official views of NIH.
Intermittent Functioning of Baroreflex Contributes to the Etiology of Hypertension in Spontaneously Hypertensive Rats
Authors: Gu Feng , Michele Daniel E. , Carlson Brian E. , Whitesall Steven , Fink Gegory D. , Beard Daniel A 
Published: April 2020

The potential roles of the baroreflex in long‐term arterial blood pressure (BP) regulation and in the etiology of primary hypertension have been long debated. We have hypothesized that aging‐associated stiffening of large arteries desensitizes baroreceptors in the aortic arch and carotid sinus, and thereby may contribute to a mechano/neurogenic etiology. To test that hypothesis, we obtained long time‐course continuous arterial pressure data in spontaneously hypertensive (SHR) and control Wistar‐Kyoto (WKY) rats at various ages (7–15 weeks) during the development of the hypertensive phenotype in the SHR animals. Rather than the expected simple proportional relationship between arterial vessel distensibility and baroreflex gain, analysis of data from these animals reveals a markedly intermittent functioning of the baroreflex arc, with long periods (up to several minutes long) with arterial pressure fluctuations coupled to corresponding physiological responses in heart rate and with similarly long periods where the pressure and heart rate fluctuations are not coupled. Several potentially important trends are observed to be associated with the intermittent apparent baroreflex “on” and “off” time periods: (1.) during normal dark‐cycle recording the fraction of time for which the baroreflex is operating in the “on” state ranges from 45–70% for both strains and all ages observed; (2.) the fraction of time in the “on” state decreases with age in both the SHR and WKY strains; (3.) the mean arterial pressure is tightly correlated with fractional “on” time in all rats (see figure
); (4.) mean pressures during “off” states are higher than during “on” states for both strains, both sexes, and at all ages studied; and (5.) mean pressure tends to decrease during “on” states and rise during “off” states in WKY rats, while mean pressure remains elevated at all times in SHR rats. From these observations we hypothesize that impaired function of the baroreflex in the SHR strain contributes to the SHR hypertensive phenotype. This impaired function is reflected in less frequently coupled “on” function in the SHR compared to the WKY and the inability of the functioning baroreflex to reduce pressure during “on” states in the SHR.Support or Funding InformationSupported by NIH grant HL139813Correlation between mean arterial pressure and fractional baroreflex “on” time in SHR and WKY rats.Figure 1
Morphometric Analysis of Efnb1 Dependent Palate Growth
Authors: O'Neill Marisol Ann , Percival Christopher J. , Bush Jeffrey O. 
Published: April 2020

Congenital craniofacial anomalies represent one of the most prevalent types of structural birth defects and include common dysmorphology such as cleft palate. While morphometric analysis of surfaces generated from micro computed tomography (micro‐CT) and magnetic resonance imaging (MRI) is a well‐established method, it has not yet been applied to understanding the three‐dimensional changes in palatal shelf morphology that result in a cleft palate phenotype. EFNB1 mutations result in craniofrontonasal syndrome, an X linked condition, consisting of a variety of craniofacial anomalies, including cleft palate. Mouse models for loss of Efnb1, have demonstrated its importance during normal development of the anterior secondary palate, with loss of Efnb1 resulting in changes in secondary palatal shelf shape including apparently reduced anterior palatal shelf extension, leading to a cleft secondary palate. Palatal shelf shape appears more disrupted in Efnb1+/− embryos, which are mosaic for EPHRIN‐B1 expression, but how this mosaicism contributes to palate dysmorphology has been difficult to understand in two dimensions. This study aims to establish a method of quantifying secondary palatal shelf outgrowth and shape that is coupled with an ability to quantify gene expression changes through the use of morphometrics. We performed whole‐mount immunofluorescent staining of E‐CADHERIN, EPHRIN‐B1, and DAPI followed by confocal microscopy to generate 3D renderings that we quantify using landmark based morphometrics. This method allows a deeper understanding of changes in tissue shape as they relate to gene expression.Support or Funding InformationThis study was supported by NIH grant R01DE023337 from NIDCR