bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Short‐term Warm Water Bathing on Cardiac Baroreflex Sensitivity in Healthy Men
Authors: Sugawara Jun , Tomoto Tsubasa
Published: April 2020

Bath‐related fatal (sudden cardiac arrest) and non‐fatal accident (dizziness and disturbance of consciousness) have been reported frequently in Japan. Cardiac baroreflex sensitivity (BRS) may relate to these symptoms. However, underlying mechanism(s) is poorly understood. With this information as background, we determined the effects of short‐term warm water immersion on cardiac BRS in healthy men. Twelve apparent healthy men underwent the single sit‐stand test (e.g., 5‐min sitting on the floor of bathtub and then standing up and remained standing for 5 min quietly) twice (e.g., without and with 41 °C warm water at the heart level). Beat‐to‐beat blood pressure was continuously recorded through the protocol (via Finometer). BRS was evaluated by the sequence method from heart rate response to the standing‐induced abrupt drop of mean arterial pressure (MAP). Additionally, after the each test, brachial‐ankle pulse wave velocity (baPWV: an index of arterial stiffness) was measured in the supine position. During the bathing, heart rate and cardiac output (estimated by Modelflow method) were significantly increased but MAP, stroke volume, total peripheral resistance, and tympanic temperature did not change significantly (vs. pre‐bathing). Regarding the single sit‐stand test, there were no significant changes in the baseline (sitting) MAP (from 73±16 to 77±16 mmHg, P=0.322) and the standing‐induced decreases in MAP (from −33±12 to −33±13 mmHg, P=0.970) between pre‐ and post‐bathing, but MAP‐HR slope was blunted after the bathing (from −0.47±0.26 to −0.35±0.25 bpm/mmHg, P=0.024, Figure
1). The baPWV was significantly decreased after the bathing (vs. pre‐bathing (from 12.2±1.2 to 11.8±1.2 m/sec, P=0.002). Change in MAP‐HR slope correlated with corresponding change in baPWV (r=0.610, P=0.036, Figure
2). Our results indicate that: (1) even a short‐term warm water bathing deteriorates cardiac BRS although it did not induce greater orthostatic hypotension, and (2) the change in cardiac BRS with the short‐term bathing is associated with the change in arterial stiffness. Further study to determine the impact of prolonged warm water bathing is needed.

Support or Funding Information

This study was supported by Japan Health & Research Institute Research Grant.

Cardiac barorefex sensitivity (BRS) gain and brachial‐ankle pulse wave velocity (baPWV) before and after 5‐min warm‐water bathing. Data are mean and SD. Thin lines indicate individual data.

Figure 1

Relationship between 5‐min bathing induced changes in baPWV and cardiac BRS gain.

Figure 2
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

PURPOSE

Exposure 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.

METHODS

24 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.

RESULTS

1)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.

CONCLUSION

2‐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 Information

the 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 1

Impact 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 Information

KU 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 Information

This 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 1

Cmpd 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 Information

Grant n# ARS01_01081 to MC from The National Operational Program (PON) of the Ministry of Education, ITALY

Figure 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 Information

The University of Arizona

SDS‐PAGE gel after purification of B19V NS1 through Talon resin

Figure 1

a)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 Information

This 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 Information

Supported by NIH grant HL139813

Correlation 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 Information

This study was supported by NIH grant R01DE023337 from NIDCR
Down‐regulation of LRRc17 secreted by BMSCs alleviates age‐related bone aging through autophagy enhancement
Authors: Liu Fei , Yuan Yujia , Li Yuanmin , Zhang Jie , Lu Yanrong , Chen Younan , Cheng Jingqiu
Published: April 2020

Backgrounds

The impaired function of endogenous bone marrow mesenchymal stem cells (BMSCs) is a determinant in the development of osteoporosis (OP). BMSCs have been described the potential relevance of autophagy to the pathogenesis of OP. However, the underlying mechanism remains unclear. Our previous researches have found leucine‐rich repeat‐containing 17 (LRRc17) secreted by MSCs, a member of the LRR superfamily, was markedly enriched in the age‐related pathways. This study aims to investigate the effects of LRRc17 in aging process of MSCs.

Methods

Hydrogen peroxide (H2O2) induced aging MSCs and natural senescent cell cultivated from old and young C57BL/6 mice were used to evaluate the effects of LRRc17 on MSCs. Western blot, flow cytometry was used to detect autophagy, apoptosis and cell cycle respectively. Furthermore, mitochondrial function was measured by Mitosox, JC‐1, ATP. The mitochondrial morphology was detected by Mitotracker. Moreover, co‐culture was performed to detect the effects of LRRc17 secreted by MSCs on osteoblast (OB) and osteoclast (OC).

Results

MSCs exhibited a senescent phenotype after treatment with hydrogen peroxide. This result was improved after the addition of rapamycin. Interestingly, compared with the H2O2‐treated group, MSCs transfected with shLRRc17 showed increased DNA synthesis phase, and decreased apoptosis ratio, ROS (Fig.
1A, B, C). Autophagy related protein LC3 II, Beclin1 were up‐upregulated and P62 was down‐regulated (Fig.
1G). And mitochondrial morphology was also significantly improved (Fig.
1D, E). More importantly, LRRc17 reversed the loss of Ψm (Fig.
2A) and enhanced ATP content (Fig.
1F). Co‐culture showed that silenced LRRc17 in MSCs enhanced the osteogenic ability of OB and decreased the bone resorption capacity of OC, and the opposite result was obtained after overexpression (Fig.
2B).

Conclusions

LRRc17 enhances characterizations of BMSCs through attenuating MSC senescence during OP, and which could lead to novel therapies to target altered bone formation.

Support or Funding Information

This work was supported by the Chengdu Giant Panda Breeding Research Foundation (2016cdspt001).

Figure 1Figure 2
Co‐Delivery of Anti‐Inflammatory and Proliferative Agents by Injectable Hydrogel to Promote Tissue Repair after Acute Kidney Injury
Authors: Wang Yizhuo , Liu Shuyun , Zhao Meng , Wang Chengshi , Yuan Yujia , Li Lan , Liao Guangneng , Chen Younan , Cheng Jingqiu , Lu Yanrong , Liu Jingping
Published: April 2020

Object

Ischemia‐reperfusion (I/R)‐induced acute renal injury has become a serious issue in clinical. The increasingly mortality, induced by chronic fibrosis and further renal dysfunction which caused by poor recovery of acute injury, has made it urgent to address kidney tissue repair. By using a renal I/R injury model, we aimed to investigate the role of a hydrogel‐based dual‐drug delivery platform on promoting tissue repair.

Methods

An injectable, self‐assembling peptide/heparin (SAP/Hep) hydrogel was employed to co‐deliver TNF‐α neutralizing antibody (anti‐TNF‐α) and hepatocyte growth factor (HGF). We analyzed the microstructure and controlled release properties of KLD2R/Hep hydrogel in vivo. The effects of the drug‐loaded hydrogel (SAP‐drug) on renal injury were evaluated in a rodent renal I/R injury model.

Results

In vitro, the SAP/Hep hydrogel exhibited a faster release of anti‐TNF‐α with a sustained release of HGF, and both drugs could maintain better bioactivities after release. In vivo, combined anti‐TNF‐α/HGF showed better renal protective potential than anti‐TNF‐α or HGF alone. Compared with free‐drug (anti‐TNF‐α/HGF in solution) or SAP alone , SAP‐drug treatment significantly reduced the levels of serum creatinine (Scr)/blood urea nitrogen (BUN), tubular apoptosis, renal inflammatory factors and macrophage infiltration. Moreover, the SAP‐drug group exhibited better efficacy on promoting tubular cell proliferation and dedifferentiation than SAP or free‐drug alone, and thus reduced chronic renal fibrosis in I/R injury mice.

Conclusions

We confirmed that SAP could sequentially deliver anti‐TNF‐α and HGF to achieve anti‐inflammatory and pro‐proliferative effects simultaneously with only once injection, and thus can be a promising delivery platform for tissue repair.

Support or Funding Information

This work was supported by grants from National Natural Science Foundation of China (81571808, 31871001, 31200754) and Sichuan Science and Technology Program (2019YJ0069)

Design of dual‐drug delivery KLD2R/Hep hydrogel for enhancing renal repair. The formation of the cationic KLD2R/anionic heparin hybrid hydrogel via electrostatic interaction, and heparin further had high affinity to HGF. The encapsulated anti‐TNF‐α and HGF were released from hydrogel in a sequential manner, and thus synergistically promoted tissue repair.

Figure 1
Electrophysiological measurements of peripheral nerves in rats using the Qtrac approach
Authors: Pedersen Michael , Skov Martin Nors , Larsen Yrsa Marie Damgaard , Tankisi Hatice , Matchkov Vladimir
Published: April 2020

Introduction

The aim of this study was to build an implant and benchmark it for further improvement, specifically aimed to measure periphery nerves in rats. The nerve algorithm was based on the Qtrac approach for measurements of excitability measurements on motor and sensory nerves. This design was designed for enhanced measurements of nerve function related to diabetic neuropathy; a common and sometime a debilitating condition that eventually may result in foot ulcers and limb amputation. In order to better treat and prevent complications it is of great importance to recognize the sensory and autonomic dysfunction as early as possible. At the moment, it is unclear how and when the autonomic changes develop in relation to the somatosensory signs. The pathology of DN is primarily characterized by loss of nerve fibers. A typical course of the disease often starts with small‐fiber neuropathy/dysfunction in toes and feet, pain and paresthesia. Development of the proposed device addresses this problem.

Methods

We created a working pre‐amp and filter prototype and used conventional Qtrac algorithms. The performance consisted of multiple iterations to determine the best configuration for the amplification, signal‐to‐noise, filtration, space occupied on the PCB and battery use. The Qtrac package includes an automated stimulation<−>response threshold tracking software and hardware setup with specialized algorithms to determine multiple variables in nerve excitability studies. The device was measured in the nerves: sciatic, ulna and tail.

Results

Eight rats were examined in 3 nerves (Fig
1A–1C). These data showed no significant difference between a regular Qtrac setup and the newly‐designed Qtrac setup. We showed a measurable difference between the nerves in question as shown in Figure
1D, showing 5 different variables obtained from the Qtrac protocol from the different nerves. The data is shown as a mean value of all rats in the study. At the same time, the difficulty in precisely measure in the same spot on different rats, became clear. Especially the Ulnar nerve proved too hard to use for this purpose, and we are reluctant to use this nerve in further studies.

Conclusion

The study demonstrated a novel Qtrac design for measurements of periphery nerves in rats. The study revealed reliable data, showing significant difference in several excitability parameters for the three nerves. Potentially, explained by different amounts of fibers in the examined nerves, different fiber sizes and compositions. Further studies should include telemetric add‐on and measurements in rats suffering from DN. .

needle placement in rats. A. Sciatic, B. Ulna, C. Tail. Graphs (D) show 5 different variables obtained from the Qtrac protocol from the different nerves.

Figure 1
IQE.DYN‐Integrated Quantum Energy Dynamics by G.P. Einstein Physio‐Neuro Function
Authors: Sainvil Frantz , Tulp Orien Lee , Einstein George P.
Published: April 2020

In a mathematical context, a dynamic system is a set of deterministic partial differential equations for the time neuro‐physiological processes with multiple variables. After long times a dynamic system displays one of a few possible types of dynamic states such as a steady state, oscillations or chaos. The dynamic equations typically depend on parameters that qualitatively influence the dynamic states of the biological system.

The implications of the neuronal dynamics we studied experimentally and applied G. P Einstein Integrated Quantum Energy Dynamic of Physio‐Neuro Mathematical Function Model.

G. P. Einstein Mathematical Model of Effective DYNAMICS Neuronal stimulation by Sigma Photonics Quantum Energy Therapy can be presented:

d ∑ IQBET/dt1 > d ∑ IQBPDE/dt2

where: d ∑ I Q B E T ‐ derivative ∑ Integrated Quantum Bioelectromagnetic Energy Therapy d ∑ I Q B P D ‐ derivative ∑ Integrated Quantum

In neuronal stimulation, function is supported by Infrared Quantum Dynamics. It is the process of using an external source to activate ion channels causing depolarization of the neural membrane and evoking an action potential to propagate down the axon of a stimulated neuron.

The Quantum Dynamics generate action potentials in both the central nervous system (CNS) and peripheral nervous system (PNS). Quantum Dynamics in Neuronal Stimulation by Sigma Photonics Quantum Energy was applied. The application of 11 basics mathematical function therapies is supported by specific integrated quanta energy matrixes. Each mathematical quantum energy function contains the specific matrix quanta energy therapy. By applying a controlled dose of Integrated Quantum Dynamics Physio‐Neuro stimulation by Each mathematical quantum energy function we can alter the properties of the tissue and the internal organs and bring their characteristics specific matrix quantum energy treatment. With careful choice of frequencies, which depend on each individual patient, his/her condition, degree of pathology and the time‐dependent properties of the treatment, we can treat a wide range of tumor cancers.

The neuronal stimulation function was derived from neuro‐physio model as presented in figures
1 and
2.

Support or Funding Information

Supported by Institutional Resources of the University of Science Arts & Technology [USAT], Montserrat and the Einstein Institute USA.

Figure 1Figure 2
Changes in Steroids, Xenobiotics, and Drug‐Metabolism in Humans by Genetic Variations in NADPH Cytochrome P450 Oxidoreductase
Authors: Pandey Amit V.
Published: April 2020

A broad spectrum of human diseases, including disordered steroid metabolism, are caused by mutations in the NADPH cytochrome P450 oxidoreductase (POR)
[1,
2]. Human POR is a diflavin reductase that transfers electrons from NADPH to small molecules, non‐P450 redox partners, and cytochrome P450 proteins in the endoplasmic reticulum
[2]. Cytochrome P450 proteins perform an extensive range of reactions, including the metabolism of steroids, drugs, and other xenobiotics. Therefore, genetic variations in POR change several metabolic pathways by changing the activities cytochromes P450
[2]. Due to this unique role of POR in metabolism, it was believed for a long time that genetic defects in POR are unlikely, and indeed a POR knockout mouse is embryonically lethal. However, in 2004, the first human patients with defects in POR were reported, and since then, over 200 different variations in POR have been found
[3,
4].

Recently, we have characterized POR variations from non‐clinical samples
[5]. By analyzing the POR sequences from the 1000 genome and other sequencing projects, we identified potentially disease‐causing variations and characterized these by functional studies using recombinant proteins produced in bacteria, yeast, and mammalian cells
[6,
7].

Identification of severe effects of POR mutations on both the drug and the steroid metabolizing cytochromes P450 indicates that likely pathogenic mutations may be found in non‐clinical population. Their combination as compound heterozygotes or homozygous may severely impact both steroid and drug metabolism by modifications of cytochrome P450 activities. Variations in POR need to be evaluated individually. Changes in drug and steroid metabolism due to genetic variations can be addressed using personalized metabolic profiling and supplementation using modified dosages of drugs and steroids.

Support or Funding Information

Supported by grants from the Swiss National Science Foundation (31003A‐134926) & Novartis Foundation for Medical‐Biological Research (18A053) to AVP.
The Extent of Metaplastic Hard Tissue Within the Appendicular Skeleton of the Nine‐Banded Armadillo (Dasypus novemcinctus)
Authors: Heck Christian , Woodward Holly N.
Published: April 2020

The developmental and functional advantages of vertebrate metaplastic hard tissue is still relatively unclear. Metaplastic tissue forms from the direct transformation of one cell type to another. Specifically, mineralization of preformed dense connective tissue results in intratendinous metaplastic tissue, and has been described in a variety of taxa and elements including turtle shell, reptilian osteoderms, sturgeon scutes, dinosaur cranial and vertebral elements, ossified tendons, portions of bird skeletons, and tendinous entheses in mammalian bone. However, histological analysis of the extent of metaplastic hard tissues, both across Mammalia and within individual mammalian bones, is lacking. Here we serially sectioned both tibiae of a nine‐banded armadillo, Dasypus novemcinctus [Xenarthra: Cingulata], transversely and longitudinally to identify the existence and possible patterns of metaplastic hard tissue. All skeletal elements were processed using standard procedure for the preparation of undecalcified bone. We found evidence of metaplastic hard tissue in the tibial crest, as well as the medial side of the diaphysis and the proximal lateral side of the tibia. The metaplastic tissue in the tibial crest appears interstitially within a primary woven bone matrix. Along the lateral side, metaplastic tissue forms a thin patch in the mid‐cortex superficial to compact coarse cancellous bone. The presence of metaplastic hard tissue seems consistent with tendinous insertions through fibrous entheses along the length of the tibia and tibial crest. The entrapment of metaplastic tissue within the tibia is likely due to appositional bone growth at the enthesis. However, the presence of metaplastic hard tissue on the medial side of the mid‐diaphyseal and proximal portion of the tibia does not appear to be associated with any tendinous insertions. Previously, unassociated metaplastic tissue was found extensively in osteoderms, ankylosaur tail clubs, and dinosaur cranial ornamentation such as the parietal frill of Triceratops. Study of metaplastic hard tissue in modern bone, independent of tendinous insertions, can improve our understanding of its potential biomechanical advantages and thereby possibly provide clues to its use in ornamentation and within the appendicular skeleton. Metaplastic hard tissues have likely been misidentified and understudied in previous investigations of modern hard tissue microstructure. Additionally, because Xenarthra is proposed to be one of the four original clades of placental mammals, analysis of metaplastic tissue in the skeleton of D. novemcinctus could have implications for early mammalian bone development patterns.

Support or Funding Information

Partial funding was provided by Montana State University Undergraduate Scholar’s Program

Figure 1
BDNF/TrkB Signaling Increases Autophagy Flux in Cervical Spinal Cord
Authors: Ducaji Emily C. , Pareja-Cajiao Miguel , Gransee Heather M. , Sieck Gary C. , Mantilla Carlos B.
Published: April 2020

Autophagy is a homeostatic process for the elimination of damaged, unnecessary proteins or organelles within cells. Our overarching hypothesis is that autophagy contributes to neuromuscular dysfunction, specifically increased NMTF seen with aging. Understanding the mechanisms regulating autophagy flux is essential to discovering therapeutic agents which may mitigate the effects of neuromuscular dysfunction across conditions including old age. In recent studies, we found that in 18 month old mice autophagy in the cervical spinal cord is accelerated and neuromuscular transmission failure (NMTF) in diaphragm muscle‐phrenic nerve preparations is increased. Neurotrophin activity, specifically brain‐derived neurotrophic factor (BDNF) signaling through its high affinity receptor tropomyosin‐related kinase receptor subtype B (TrkB) decreases NMTF. Thus, we predict BDNF/TrkB signaling suppresses autophagy in phrenic motor neurons located in the cervical spinal cord. We hypothesize that inhibiting TrkB kinase activity accelerates autophagy in phrenic motor neurons. TrkBF616A mice at 18 months of age were randomized to receive 1 week treatment with vehicle (n=4) or 1NMPP1 (TrkB kinase inhibitor; n=6). Additionally, 18 month old wild type C57BL/6 mice (n=2) were studied. Whole cervical spinal cord (CSC) was harvested for measurements of autophagy marker protein expression analyzed by Western blotting. Data for the vehicle‐treated TrkBF616A and wild type mice was combined given lack of differences for any measurement. p62, a marker associated with the initiation of autophagy, was significantly decreased in the 1NMPP1‐treated group compared to vehicle‐treated or wild type mice. LC3, a protein marking the autophagosome membrane elongation phase, was not significantly changed across groups indicating that LC3‐tagged autophagosomes proceed to lysosomal degradation and do not accumulate. These results support our hypothesis that inhibiting BDNF/TrkB signaling accelerates autophagy flux in the CSC and reveal trophic effects of BDNF/TrkB signaling in the neuromuscular system. Accelerated autophagy may underlie age‐related neuromuscular dysfunction in a BDNF/TrkB signaling dependent manner.

Support or Funding Information

Support: NIH R01 AG057052
Is Chemotherapy‐Induced Cardiomyopathy Caused by Myocardial Ischemia?
Authors: Davidian Anahis , Mistry Maahi , Pucci Thomas , Kolz Christopher L , Shockling Lindsay , Enrick Molly , Yin Liya , Chilian William , Ohanyan Vahagn
Published: April 2020

Over the last 50 years progress has been made in treating childhood and adult solid and hematological tumors. One commonly used chemotherapeutic agent is doxorubicin (DOX), an anthracycline antitumor antibiotic, which was discovered in early 1960s. Anthracycline (ANT) chemotherapy alone or in combination with other chemotherapeutic agents markedly improves the survival rate of cancer patients, particularly among children, where survival rates have more than doubled over 5 decades. However, the use of ANT chemotherapy is a double‐edged sword in that it can produce chemotherapy induced cardiac dysfunction (CRCD), which can cause death in 50% of the patients with who present with the cardiomyopathy. We hypothesized, that one of the causes for CRGD is a reduction in myocardial blood flow (MBF) that leads to cardiac myocyte hypoxia, producing small areas of ischemia and myocyte damage. We also hypothesize that increasing MBF by the coronary vasodilator chromonar prevents cardiomyocytes from hypoxic damage.

To test these hypotheses, we treated mice genetically engineered to show cardiac myocyte fluorescence if the cells are hypoxic (hypoxia fate mapping [HFM]) with DOX for 6 weeks (group 1, N=8) and Dox + Chromonar for 6 weeks (group 2 N=8). Five weeks after treatment we started tamoxifen for 5 days to activate the HFM protocol. Tamoxifen induces Cre in cardiac myocytes and hypoxia activates a program that results in the expression of td‐tomato only in hypoxic cardiac myocytes. In tissue sections, hypoxic cardiac myocytes are visualized by fluorescence microscopy. Cardiac function was measured by ultrasound and MBF was measured with contrast echocardiography.

Six weeks after DOX treatment many hypoxic myocytes were observed in the myocardium. Cardiac function was significantly decreased at the end of DOX treatment (% ejection fraction EF=39±3) compared to baseline (62±5.8%). Simultaneous administration of chromonar prevents the cardiomyocytes from hypoxic damage and prevents cardiac dysfunction development. %EF was 58±6 at the end of the chemotherapy. Chromonar also significantly increased MBF in group 2 mice compared to group 1 (P<0.05).

Based on these data CRCD may be a type of ischemic heart disease where the treatment reduces MBF, which leads cardiomyocyte hypoxia, and causes progressive, accumulative cardiac injury. Moreover, increasing myocardial blood flow by chromonar prevents the cardiomyocytes from hypoxic damage and development of CRCD.

Support or Funding Information

NIH, NHLBI

Panel A shows the relationship between myocardial blood flow and cardiac work 6 weeks after DOX and DOX + Chromonar treatment. Panel B shows ejection fraction changes in DOX +Chromonar (Group 2 mice)

Figure 1