bluuebunny/crossref_metadata_2025 huggingface dataset:
Characteristics of the substrate‐free mitochondria
Authors: Leem Chae Hun , Ho Quynh Mai , Lee Jeong Hoon , Kim In Young , Youm Jae Boum
Published: April 2020

Mitochondria are critical organelle to convert the metabolites to the life‐essential chemicals, ATP. For serving the role, the mitochondria have very peculiar steps from metabolites to ATP formation. Usually the Ψm was regarded as zero without the mitochondrial substrate or in the presence of uncoupler. However, we found there still considerable Ψm was existed even in those conditions. So basic scheme of the generation of Ψm is hampered and the question how to explain this phenomena remains. For attacking these problems, we used a multiparametric microfluorometric system to monitor NADH, FAD, and TMRE fluorescence, simultaneously and developed a measurement method of Ψm quantitatively. The Ψm in substrate free conditions was around −60 mV. The application of ATP generated considerable hyperpolarization. The application of KATP channel opener, diazoxide (DZX), could depolarize Ψm. When K+ was replaced with meglumin, the Ψm was slightly hyperpolarized and the effect of DZX was abolished. The KATP channel blockers did not have any effect. The application of ATP hyperpolarize Ψm, however, ADP or AMP didn’t. Oligomycin blocked the effect of ATP. Interestingly, oligomycin depolarized the resting Ψm considerably. The effect of ATP was not affected by DZX. The replacement K+ with meglumin slowed the ATP‐induced change of Ψm and made it transient. The return to K+ medium recovered the effect of ATP. From these results, the Ψm in substrate‐free conditions was not zero and K+ may particpate in the formation of the resting Ψm. The F1,F0‐ATPase may participate on the formation of the resting Ψm. K+ may be an important modulator for F1,F0‐ATPase acitivity. The F1,F0‐ATPase and K+ flux might contribute the formation of the resting Ψm but it still need further study on them.

Support or Funding Information

This work is supported by NRF‐2015M3A9B6028310 and NRF‐2016M3C1A6936605.
Adipose tissue heparan sulfate proteoglycans – critical regulators of adipocyte metabolism and glucose homeostasis
Authors: Pessentheiner Ariane R. , Quach Anthony , Al-Azzam Norah , Liu Sihao , Downes Michael , Evans Ronald M. , Gordts Philip L.M.S.
Published: April 2020

Heparan sulfate (HS) proteoglycans are important components of cells and the extracellular microenvironment. Interaction between growth factors and receptors is often modulated by the amount and degree of HS sulfation. We hypothesize that natural variation in HS composition on adipocytes serves as a rheostat for adipocyte function and impacts susceptibility to develop insulin resistance, type‐2 diabetes and obesity.

We use mice with adipose tissue (AT) specific inactivation of the HS biosynthetic enzymes N‐acetylglucosamine‐N‐deacetylase‐N‐sulfotransferase 1 (Ndst1‐AKO) and Exostosin 1 (Ext1‐AKO) mediated by the Adipoq‐Cre driver. These models allow us to study the impact of HS sulfation and chain length, respectively, on adipocyte metabolism and systemic glucose homeostasis in the context of diet‐induced obesity.

On a high fat diet both Ndst1‐AKO and Ext1‐AKO mice gain more weight compared to WT littermates and develop significant glucose intolerance and insulin resistance. A pyruvate tolerance test indicates that disturbed adipocyte‐HS significantly affects liver gluconeogenesis. Moreover, hepatic inflammation is elevated in Ndst1‐AKO mice compared to WT controls, indicating that adipocyte‐HS also influences the organ cross‐talk between AT and liver. Previously, it has been shown that fibroblast growth factor 1 (FGF1) is a potent insulin sensitizer with great therapeutic potential. FGF1 signaling depends on the presence of HS on the plasma membrane. We show that the insulin‐sensitizing effect of FGF1 is diminished in Ndst1‐AKO and Ext1‐AKO mice. However, the insulin sensitization response to FGF21, another metabolic active FGF without HS‐dependency, is unaffected by HS alterations in AT.

Our data suggest that HS composition in AT regulates FGF1 sequestration and signaling, leading to metabolic complications upon diet‐induced obesity. In this context, our results underline the importance of HS composition in AT for signaling processes that are critical for glucose homeostasis.
p23 Co‐chaperone Protects Aryl Hydrocarbon Receptor (AHR) from the Autophagy‐Mediated Degradation
Authors: Yang Yujie , Chan William K.
Published: April 2020

The aryl hydrocarbon receptor (AHR) is first discovered as a transcription factor in response to toxic compounds such as dioxins. It also plays important roles in organ development, immune responses, metabolisms and carcinogenesis. Without ligand activation, AHR resides in the cytosol and is accompanied by Hsp90, p23 and XAP2. AHR is degraded via the 26S proteasome after ligand activation. However, it is still unclear how basal AHR protein levels are regulated. We previously discovered that AHR protein levels are lower in p23 stable knockdown (p23KD) cell lines when there is no ligand treatment. Here we report that treatment of proteasome inhibitor MG132 could not restore AHR in p23KD HeLa cells while the autophagy inhibitor chloroquine increased AHR proteins levels to 2.5 to 3‐fold in p23KD HeLa cells and 2‐fold in WT HeLa cells. Similar results were observed when cells were treated with other autophagy inhibitors (bafilomycin A1 and 3MA). There was higher autophagic flux in p23KD HeLa cells than in WT HeLa cells. Results from the co‐immunoprecipitation experiment and proximity ligation assay showed interaction between AHR and LC3B (LC3B‐II), which is an important protein in macroautophagy. These results support that the AHR protein levels are regulated by autophagy and p23 can protect AHR from the autophagy‐mediated degradation. Additional data will be presented to address the mechanisms of basal AHR protein degradation via autophagy.

Support or Funding Information

NIH R15ES023104
IL‐33/ST2L mediated ADAMTS10 activation regulate retinal endothelial cell sprouting and neovascularization in a murine model of hypoxia‐induced retinopathy
Authors: Singh Nikhlesh Kumar
Published: April 2020

Pathological retinal neovascularization is a clinical manifestation of various proliferative retinopathies, including retinopathy of prematurity (ROP), diabetic retinopathy, and the wet form of macular degeneration. A characteristic feature of these conditions is the presence of hypoxic areas and over‐expression of the proangiogenic vascular endothelial growth factor (VEGF). Treatments of these neovascular diseases using laser or anti‐VEGF therapies not only induce various local and systemic side effects but also are only partially effective. Here, using a murine model of oxygen‐induced retinopathy, we found that either intravitreal injections of IL‐33 siRNA or genetic deletion of IL‐33 resulted in reduced neovascularization with no significant changes in the avascular area. These findings suggest a role for IL‐33 in pathological neovascularization with little or no effect on the normal retinal repair. Interestingly, we observed that along with IL‐33 activation, hypoxia also induces expression of ST2 transmembrane receptor (ST2L), both at mRNA and protein levels, with little or no effect on secretory ST2 (sST2) levels. Our in vitro experiments using human retinal microvascular endothelial cells as a model suggest the importance of IL‐33/ST2L‐signaling on endothelial cell migration, tube formation, and sprouting. Besides it, our in vitro and in vivo findings provide evidence that IL‐33/ST2L‐signaling via ADAMTS10 activation facilitates sprout formation and invasion of connective tissue through controlled degradation of extracellular matrix proteins (ECMs). These results strongly suggest that IL‐33/ST2L‐signaling plays a key role in retinal neovascularization and thus represents a new pharmacological target for the treatment of diseases where excessive neovascularization is the underlying pathology.

Support or Funding Information

The research is supported by NIH EY029709 grant..
Acute High Phosphate Intake Impairs Endothelial‐Dependent Vasodilation in Young Premenopausal Women
Authors: Stephens Brandi , Nandadeva Damsara , Kaur Jasdeep , Barbosa Thales C. , Fadel Paul J.
Published: April 2020

In the United States, the growing abundance of processed foods has led to Americans consuming more than double the recommended dietary intake of inorganic phosphate (Pi). Chronic consumption of Pi is associated with a higher risk of cardiovascular disease in the general population; however, studies investigating underlying mechanisms are limited. Recently, our lab has reported an impairment in brachial artery flow‐mediated dilation (FMD) following acute high Pi consumption in young, healthy men. To our knowledge, the effect of acute high Pi on vascular function in women remains unknown. This becomes important, as previous longitudinal work suggests that women may be protected from the greater cardiovascular risk associated with high serum phosphorus. Therefore, the aim of this study was to determine the effect of acute high Pi consumption on vascular function in young, healthy women. Eight healthy premenopausal women ingested a high‐Pi drink containing 2,000 mg of phosphorus (23 ± 2 yrs; mean ± SEM). Heart rate (HR) (ECG) and arterial blood pressure (BP) (finger photoplethysmography and automated sphygmomanometer) were measured during a 10 min baseline. FMD was used to assess conduit artery endothelial function and microvascular function was quantified as reactive hyperemic velocity AUC following cuff release. In addition, Pulse Wave Velocity (PWV) was used to assess large artery stiffness. All measures were performed before (pre) and 60 min after (post) Pi drink consumption (a time period our lab has shown to elicit peak elevations in serum phosphate following Pi consumption). Resting mean arterial BP and HR were not different pre‐ vs. post‐ Pi consumption (BP= pre, 80 ± 1; post, 81 ± 2 mmHg; p=0.28; HR= pre, 62 ± 4; post, 62 ± 4 beats/min; p=0.64). In contrast, FMD was significantly attenuated post‐Pi consumption, (pre, 6.7 ± 0.5%; post, 4.8 ± 0.8%; p=0.04), whereas, reactive hyperemic velocity AUC remained unchanged (pre, 2590 ± 488 arbitrary units; post, 2056 ± 221 arbitrary units; p=0.30), indicating that acute high Pi consumption had no effect on microvascular function. In addition, PWV was also unaffected by acute high Pi (p=0.18). Collectively, our preliminary findings suggest that, similar to men, exposure to acute high Pi impairs conduit artery endothelial function, but not microvascular function, in young healthy women.

Support or Funding Information

University of Texas at Arlington College of Nursing and Health Innovation
miR‐23a and miR‐23b Are Putative Regulators of KDM6A Protein Expression in Epithelial Mesenchymal Transition
Authors: Kim Joelle , Johnson Kelsey , Taube Joseph
Published: April 2020

Cancer cell metastasis is the leading cause of cancer patient mortality. Tumor progression of epithelial cancers is facilitated by epithelial‐mesenchymal transition (EMT) and mesenchymal‐epithelial transition (MET), reverse processes that enable invasive cellular behavior. A key regulator of EMT/MET is lysine (K)‐specific demethylase 6A (KDM6A), an enzyme involved in epithelial gene expression that is abundant in epithelial cell phenotypes. We have previously determined that KDM6A is not dynamically regulated at the transcription level; we now aim to investigate whether post‐transcriptional mechanisms modulate KDM6A expression patterns. KDM6A mRNA transcripts are potentially regulated by specific miRNAs that inhibit expression of KDM6A protein, influencing EMT and downstream cellular morphology.

To identify miRNAs that likely target KDM6A, eight databases were utilized to compile a list of ~160 potential miRNAs predicted to bind at the KDM6A 3′ untranslated region (UTR). Top miRNA candidates were then selected by number of deep sequencing reads, observed amounts in breast cancer cell lines, and frequency in patient data. miR‐23a, ‐23b, ‐145, and ‐199a were determined as potential regulators of KDM6A. The miRNAs were quantified by performing miRNA‐specific RT‐qPCR on RNA collected from TGFβ‐induced EMT and MET at various timepoints. Of the four miRNA candidates, miR‐23a and ‐23b were identified as likely post‐transcriptional regulators of KDM6A, as their relative quantification levels follow expected patterns opposing KDM6A expression throughout EMT/MET.

Future directions aim to determine whether experimental manipulation of miR‐23a/b alters KDM6A protein expression and whether predicted binding sites at the KDM6A 3′ UTR are required. Cloning the gene for miR‐23a/b into a mammalian expression vector to observe miRNA‐overexpression effects on KDM6A expression can clarify KDM6A’s potential role in metastatic activity and can introduce new targets for research in clinical applications.

Support or Funding Information

Investigation of KDM6A function is supported through funding from Susan G. Komen and Jack G. and Norma Jean Folmar.
Effects of Gravidity on Locust Jump Performance
Authors: Valachovic Abigail Corrine , Kirkton Scott David
Published: April 2020

Among life‐history trade‐offs, the cost of reproduction has been well‐studied. An increase in body mass due to gravidity has been shown to negatively affect survival in vertebrates, but it has not been widely studied in invertebrates. Insects, especially American locusts (Schistocerca americana), are good models to study how life‐history changes affect locomotion. For our research, we collected non‐gravid and gravid female grasshoppers and conducted a five minute endurance jumping test. Non‐gravid females (n=20, mean weight= 2.25 g) had an average maximum jump distance of 189 cm over the first minute. Over the five minute trial, non‐gravid female jump frequency decreased by 7% and mean distance per jump decreased by 46%. In contrast, gravid females (n=12, mean weight= 2.7 g) in the first minute had an average maximum jump distance of 169 cm. Over the five minute trial, gravid female jump frequency decreased by 8% and the mean distance per jump decreased by 52%. Next, we added weights to additional groups of non‐gravid and gravid females to determine how additional mass affected jump performance. The weights were either 20% or 40% of the average group body mass (n=12 for weighted non‐gravid, n=8–10 for weighted gravid). During the five minute trial, jump frequency significantly decreased in weighted grasshoppers regardless of whether they were gravid or not. In addition, the mean distance per jump significantly decreased with weights in both non‐gravid (p<0.008) and gravid females (p<0.0405). Therefore, increased body mass reduced frequency and mean distance per jump during repeated jumping. These results support a trade‐off between gravidity and locomotory performance in American locusts.

Support or Funding Information

Union College Student and Faculty Research Fund
REVERSAL OF UNILATERAL URETERAL OBSTRUCTION LEADS TO SALT‐SENSITIVE HYPERTENSION
Authors: Khan Md Abdul Hye , Stavniichuk Anna , Savchuk Olexiy , Jankiewicz Wojciech K. , Imig John D.
Published: April 2020

Hypertension is a major cardiovascular disease and 50% of the hypertensive population has salt‐sensitive hypertension. There is mounting evidence that a long‐term consequence of acute kidney injury is increased risk for salt‐sensitive hypertension and chronic kidney disease (CKD). Animal models to study the pathophysiological basis for salt‐sensitive hypertension that follows occurrence of kidney injury are lacking. In the current study, we used a reversal unilateral ureteral obstruction (RUUO) model to study long‐term kidney injury consequences to cause salt‐sensitive hypertension. In this model, we removed the ureteral obstruction 10 days following UUO surgery in 8–10 week old male C57BL/6J mice and divided them into four groups (n=5–6/group): Sham‐Normal Salt Diet, Sham‐8% NaCl Salt Diet, RUUO‐Normal Salt Diet, and RUUO‐8% NaCl Diet. We removed the clip form the ureter day10 day which is a time point the UUO mice demonstrate marked renal fibrosis, inflammation, tubular injury, and lower vascular density. At day 10, the mice were fed a normal or 8% NaCl diet for 4 weeks. Weekly blood pressure was measured till the end of the protocol followed by terminal urine and kidney tissue collection for biochemical, histological, immunohistological, and gene expression studies. There was a progressive and steady rise in systolic blood pressure to 130±5 mmHg in RUUO‐8% NaCl diet group compared to other groups that averaged 113–118 mmHg. Interestingly, RUUO‐8% NaCl group had 50% higher albuminuria compared to Sham‐8% NaCl Salt Diet group. We also demonstrated significant increases in renal mRNA expression of adhesion molecules ICAM and VCAM in RUUO‐8% NaCl diet group compared Sham‐8% NaCl Salt Diet. Most importantly, we demonstrated that RUUO‐8% NaCl diet group had 30% lower vascular density in the outer medullary area and 16% lower cortical vascular density compared to RUUO‐8% NaCl diet group. Overall, these findings present a new animal model to study kidney injury to salt‐sensitive hypertension. In addition, development of salt‐sensitive hypertension and kidney injury in RUUO mice is associated with decreased renal vascular density.
Metabolic RNA‐seq profiles from sporadic Alzheimer’s disease patients: Analysis of glycolytic and ketolytic pathways
Authors: Saito Erin , Miller Justin , Kauwe John S. , Bikman Benjamin T.
Published: April 2020

Sporadic Alzheimer’s disease (AD) is a widespread neurodegenerative disorder that progressively impairs memory and behavior. Although much of AD research has centered on the aggregation of amyloid plaques and neurofibrillary tangles, recent research revealing a strong and consistent correlation with impaired glucose metabolism has placed the relevance of these classic AD phenotypes in disease etiology and progression into question. Here we determine whether impairments in glycolytic gene expression might be responsible for AD glucose metabolism dysfunction and examine ketolysis through differential gene expression in AD patients. RNA‐seq profiles of postmortem AD brains from several available datasets were used to compare glycolytic and ketolytic gene expression between AD patients and cognitive‐normal controls. Glycolytic and ketolytic RNA‐seq profiles were also analyzed from progressive supranuclear palsy (PSP) patients compared to cognitive‐normal controls to determine whether these impairments in metabolic function are specific to AD or common to tauopathies. Our analyses show widespread disruption of both glycolytic and ketolytic pathways in several regions of the brain relevant to memory formation in AD brains. These analyses provide further evidence that metabolism is impaired at multiple levels (i.e. substrate transport, glycolysis, ketolysis, etc.) in AD and that these impairments are characteristic of AD, not PSP.

Support or Funding Information

R01AG11380, R01AG21136, R01AG31272, R01AG042611, RF1AG054052
Two weeks of severe food restriction has long‐term adverse effects on cardiac function three months after refeeding in female Fischer rats
Authors: De Souza Aline M.A. , De Almeida Jônathas F.Q. , West Crystal , Ji Hong , Sandberg Kathryn
Published: April 2020

Introduction

Severe food restriction (sFR) is associated with acute cardiovascular dysfunction including long QT intervals, arrhythmias, hypotension, bradycardia and sometimes sudden cardiac death. Little, however, is known regarding the long‐term effects of sFR after these individuals have recovered their body weight (BW).

Methods

Three‐month‐old female Fischer rats were divided into 2 groups and maintained on a control (CT; regular chow ad libitum, n=8) or sFR (60% reduction of daily food intake, n=8) diet. After 2 weeks on the sFR diet, all rats received regular chow ad libitum for three months (‐Refed). Thereafter, cardiac function was assessed in isolated hearts using Langendorff method before and after 30 minutes of ischemia‐reperfusion (I/R).

Results

After 14 days of the sFR diet, rats lost 18% of their BW. Mean arterial pressure (MAP) (p<0.05) and heart rate (HR) (p<0.05) were also reduced compared to CT rats. Although no differences in BW were detected between the CT‐Refed and sFR‐Refed rats, perfusion pressure was increased by approximately 17 mm Hg (p<0.05 by 2‐way ANOVA), left ventricular systolic and diastolic pressures were increased by approximately 17 and 15 mm Hg, respectively (p<0.05 by 2‐way‐ANOVA), and HR was reduced by approximately 81 bpm (p<0.05 by 2 way‐ANOVA) in sFR‐Refed compared to CT‐Refed hearts during I/R. In addition, the sFR‐Refed had 2‐fold more arrhythmias (%: CT‐Refed, 34 ± 8 vs sFR‐Refed, 68 ± 9; p<0.05) during I/R than CT‐Refed hearts.

Conclusion

Despite normalization of BW and basal MAP 3 months after refeeding, isolated hearts from animals subjected to sFR and refeeding had higher perfusion, left ventricular systolic and diastolic pressures and lower HR and were more susceptible to I/R‐induced arrhythmias than hearts from CT‐Refed rats. This study has clinical ramifications for individuals who engage in short term very low calorie dieting or have experienced inadequate food intake from other causes (e.g., very low food security, war, natural disasters) who may be at increased risk of developing cardiovascular disease.

Support or Funding Information

NIH 1R01HL119380 (KS); AHA 19POST34380744 (AS)
mTORC1 is required for the lipidation and ER‐to‐Golgi transport of prechylomicrons in Caco‐2 cells
Authors: Moreau Regis , Wang Zhigang
Published: April 2020

Dietary fats are efficiently digested and absorbed by enterocytes, assembled into triacylglycerol‐rich chylomicrons and finally secreted in a multistep process. The role of mTORC1 (mechanistic target of rapamycin complex 1) in the fate of absorbed lipids, their intracellular trafficking, triacylglycerol packaging in chylomicrons by enterocytes has not been previously studied. On the basis of our previous work in hepatocytes showing that mTORC1 positively regulate VLDL production, we hypothesized that intestinal mTORC1 supports the production and secretion of chylomicrons. Lentivirus mediated shRNA technology was used to knockdown (>90% down) mTORC1 signaling‐associated proteins, Raptor and TSC2, to create three Caco‐2 cell lines that exhibit basal (shScramble cells), low (shRaptor cells) or high (shTSC2 cells) mTORC1 activity. Chylomicron production was primed with the addition of 1.2 mM oleate/1.5% BSA complex to the apical side of ThinCert™ cell culture inserts. We found that knocking down Raptor disrupts triacylglycerol synthesis and chylomicron assembly. Knockdown of Raptor decreased the mRNA levels of APOB (−87%), MTTP (−94%), FASN (−76%), DGAT1 (−77%) and DGAT2 (−81%) compared with control shScramble cells. The cellular protein abundance of apoB (−89%), MTTP (−63%) and FASN (−80%), and the secretion of apoB100 (−51%), apoB48 (−66%) and triacylglycerols (−69%) were decreased compared with control shScramble cells. Rapamycin (a mTORC1 inhibitor) lowered the secretion of apoB100 and apoB48 in shScramble cells by 47% and 75%, respectively. Knockdown of Raptor decreased the gene expression of prechylomicron transport vesicle‐associated proteins VAMP7 (vesicle‐associated membrane protein 7) and SAR1B (secretion associated Ras related GTPase 1B) by 85% and 70%, respectively, compared with shScramble cells, suggesting that prechylomicron trafficking from the ER to the Golgi was repressed in the absence of Raptor. In conclusion, our studies revealed the critical role of mTORC1 in chylomicron metabolism. These findings have broad implications for the absorption of essential fatty acids and lipid‐soluble vitamins, for chylomicron retention disease, chylomicron overproduction and associated hypertriglyceridemia.

Support or Funding Information

USDA Hatch Act, USDA‐NIFA
Effect of exposure to Epigallocatechin gallate‐3‐gallate and Epicatechin in the proliferation in the Breast Cancer cell line MDA‐MB 468
Authors: Mahadeo Arianna
Published: April 2020

Breast cancer is the most commonly diagnosed cancer in women, with more than 252,710 diagnosed in the United States each year. Diet is considered an important factor in breast cancer prevention and some studies suggest that green tea consumption has the potential to reduce breast cancer risk. Specifically catechins, a group of polyphenols that are found in plants, have been shown to have anti‐tumor effects such as promoting the decrease in tumor size and overall cancer growth. Many studies have investigated epigallocatechin‐3‐gallate (EGCG), one of the most abundant catechins in green tea, but less is known about the effect of epicatechin in breast cancer cell growth. Here, we investigated the inhibition of cell proliferation in the breast cancer cell lines, MDA‐MB 468 after exposure to varying concentrations of EGCG and epicatechin. Changes in proliferation were determined using the 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide (MTT) assay after exposing cells for 24 and 48 hours to concentrations of catechins ranging from 1μM to 500μM. We found that after 24‐hr exposure, the cells significantly decreased in growth with a lower percent of cells remaining after exposure to 500μM EGCG. Percentage of cells was higher at 1μM when compared to 500μM (85.3 ± 1.9% vs. 44.0 ± 13.01% respectively (p=0.023)). However, when the cells were exposed to epicatechin in that same time period, there was no significant difference in proliferation between these concentrations (113.83 ± 17.87% vs. 87.94 ± 34.78% respectively (p=0.224)). Similar results were found at 48 hours. We found that as time increased from 24 to 48 hours, EGCG significantly inhibited the growth of cancer cells at 500μM (44.0 ± 13.01% vs. 8.58 ± 0.34%, respectively (p=0.031)). No effect was observed for epicatechin in growth as time increased. At an exposure of 500μM epicatechin, a higher percentage of cells were alive at 24 hours when compared to 48 hours but average values among experiments were not statistically significant (87.94 ± 34.78% vs. 40.47 ± 42.98%, respectively (p=0.174)). Our findings suggest that EGCG is a more potent inhibitor of triple negative breast cancer cell growth than epicatechin. Additional experiments at higher doses and longer exposures are needed to reach final conclusions, and we will be conducting those experiments next.
Effect of Silk Peptide on the B1REL and B2 Cell Population in Old C57BL/6 Mice
Authors: Chei Sungwoo , Oh Hyun-Ji , Lee Kippeum , Jin Heegu , Oh Jae-Hoon , Lee Boo-Yong
Published: April 2020

Silk peptide derived from the silkworm, Bombyx mori, suppresses the inflammation and enhances the anti‐inflammatory activity. However, the alteration of innate‐like B cell population remains unknown by Silk peptide. Changes with age contribute to increasing elderly people’s chances of developing metabolic disease, infectious diseases or cancer in the immune system. Immunosenescence is considered by the enhancement of cell‐mediated immune function, as well as the increasing of moral immune response. Innate‐like B cell is heterogeneous populations of non‐conventional B cells with innate detection and response characteristics. Innate‐like B cell involves of B1REL cell, marginal zone B cell, and other cells related with B cell in mice. In this paper, we assessed the efficacy of silk peptide against innate‐like B cell according to aging. We examined the population of B1REL cell and B2 cell as known as B cell subset in splenocyte of young and old C57BL/6 mice. The population of B1REL cells had an increase in the aged mouse control group than in the young mouse control group. Population of the B1REL cell suppresses in the aged mouse with silk peptide group than in aged mouse control group. In contrast to B1REL, the population of B2 cell decreased compared to the young mouse control group in the aged mouse control group, and the aged mouse group with silk peptide group increased than aged mouse control group. As result, the silk peptide moderates the increase of B1REL population and the decrease of B2 cell population by aging in C57BL/6 mice.
Trehalose Does Not Improve Alcohol‐Impaired TFEB and Liver Injury in Mice
Authors: CHAO XIAOJUAN , Ni Hong-Min , Wang Shaogui , Ding Wen-Xing
Published: April 2020

Alcoholic liver disease (ALD) is one of the major causes of chronic liver disease worldwide. We previously demonstrated that chronic plus acute alcohol binge (“Gao‐binge”) impaired hepatic transcription factor EB (TFEB), a master regulator of lysosomal biogenesis, resulted in insufficient autophagy that contributes to Gao‐binge alcohol‐induced liver injury in mice. Trehalose, a disaccharide, has been shown to activate TFEB and protects against diet‐induced non‐alcoholic fatty liver disease in mice. The aim of the present study was to investigate whether trehalose would reverse impaired TFEB and insufficient autophagy induced by Gao‐binge alcohol and in turn protect against Gao‐binge alcohol‐induced liver injury. We found that trehalose increased TFEB nuclear translocation, elevated levels of LC3‐II and lysosomal proteins in cultured AML12 cells and in mouse livers, confirming the activation of TFEB by trehalose. Surprisingly, trehalose failed to activate TFEB in Gao‐binge alcohol‐treated mouse livers. As a result, trehalose failed to protect against Gao‐binge alcohol‐induced liver injury and steatosis. However, Gao‐binge alcohol increased infiltration of hepatic neutrophils and levels of inflammatory cytokine gene expression, which were attenuated by trehalose, suggesting trehalose inhibits alcohol‐induced inflammation independent of TFEB activation. In conclusion, our results indicate that trehalose fails to improve the impaired TFEB and liver injury but ameliorates hepatic inflammation induced by Gao‐binge alcohol.

Support or Funding Information

NIH R01 AA020518, R01 DK102142, U01 AA024733 AND P30GM118247 (W.X.D).
Silk Peptide Regulates the Level of Immunoglobulin from Young and Aged C57BL/6 Mice Serum
Authors: Oh Hyun-ji , Chei Sungwoo , Lee Kippeum , Jin Heegu , Oh Jae-Hoon , Lee Boo-Yong
Published: April 2020

Immune system contributes to increasing old people chanced of developing metabolic disease in aged‐related change. Age‐associated changes of the immune system include the enhancement of a pre‐inflammatory cytokine and immunoglobulin production induces dysfunctional immune response. It is important to maintain proper amount of cytokine and immunoglobulin within the serum, but the elderly fail to maintain constancy in immune response. In particular, the immunoglobulin secreted by plasma B cells play a key role in the defense against pathogens in adaptive immunity. According to the homeostasis of immunoglobulin is very important. Aging increases levels of serum IgG, IgA and IgM, which reduces sensitivity to several antigen and causes a reaction to self‐antigen. We suggested regulate the homeostasis of immunoglobulin in aged mice serum by effective natural compound. Silk peptide is well known as bioactive component and a natural water‐soluble glycoprotein extracted from raw silk cocoons. However, remains unknown whether it is effective for immune response in aged mice. In this study, aged and young female C57BL/6 mice were orally administered the Silk peptide daily, and total serum were analyzed from aged and young mice. In conclusion, the increased amount of immunoglobulin in aged mouse serum was reduced as much as the young mouse by Silk peptide. This novel result determines that Silk peptide has potent to use in immune therapies to recover a humoral response of aging society.
Cardiomyocyte Specific Knock‐out of Erythropoietin Has Adaptive Non‐Erythropoietic Effects on Cardiac Function: A Novel Cardiac Myokine
Authors: Alshamali Razan , Edgett Brittany , Romanova Nadya , Farquharson Laura , Savinova Iryna , Marrow Jade , Allwood Melissa , Brunt Keith , Simpson Jeremy
Published: April 2020

Introduction

Erythropoietin (EPO), in response to hypoxia, is produced by the kidney, which stimulates erythropoiesis in the bone marrow. While the kidney is regarded as the primary source of EPO, mainly in vitro expression has been shown from cells or tissues of the brain, liver, and reproductive organs. However, whether these organs are an endogenous extra‐renal source and their physiological significance is unknown. Interestingly, recombinant human EPO (rhEPO) confers cardiac cytoprotection and increases contractility. Albeit, this occurs using supra‐physiological doses of rhEPO. Whether the kidney is responsible for these effects remains to be elucidated. Therefore, the
primary objective of this research is to investigate the physiological relevance of endogenous cardiomyocyte EPO production. We
hypothesized that: 1) adult cardiomyoctes are a source of EPO production and 2) the loss of cardiomyocyte EPO expression leads to a decrease in contractility and an increase in susceptibility to ischemia‐reperfusion injury.

Methods

Cardiomyocyte‐specific deletion of EPO (EPOfl/fl‐CM) was obtained by injecting 8‐week old α‐MHC‐MerCreMer+/−: EPO fl/fl male mice with 20 mg/kg tamoxifen daily for 5 days. Left ventricular (LV) EPO expression was determined using qPCR analysis. Cardiac structure and function were assessed by echocardiography, invasive hemodynamics and the Langendorff, respectively. Hematocrit was measured from blood collected via saphenous vein. Data was obtained 8‐weeks post‐tamoxifen.

Results

Paradoxically, EPOfl/fl‐CM mice exhibited a dramatic increase in whole heart EPO expression. Further, cardiac‐derived EPO confers inotropic, lusitropic and hypertrophic effects. Furthermore, in response to ischemia‐reperfusion injury, EPOfl/fl‐CM mice show profound cytoprotection. Importantly, we show that endogenous cardiac‐derived EPO is capable of improving cardiac function without affecting whole body Hct levels.

Conclusion

The knock‐out of EPO from the cardiomyocyte leads to a reciprocal increase in LV EPO expression, whereby an alternate cell in the heart is over‐compensating for the loss of cardiomyocyte‐derived EPO. This study furthers our fundamental understanding of EPO biology and establishes it as a novel cardiac myokine, capable of increasing cardiac function and driving hypertrophy.

Support or Funding Information

Heart and Stroke Foundation

Canadian Institutes of Health Research
Collecting Duct NOS1 is Necessary to Maintain Sodium Balance in Male, But Not Female Mice
Authors: Sedaka Randee S. , Dunaway Luke S , Allan J. Miller , Holliday Alexis D. , Almutlaq Rawan , Hyndman Kelly A. , Pollock Jennifer S.
Published: April 2020

Nitric oxide synthase 1 (NOS1) is a well‐established regulator of fluid and electrolyte balance in the collecting duct (CD). We previously reported that a loss of CDNOS1 in male mice results in reduced sodium excretion, urine volume, and nitric oxide (NO) excretion as well as increased plasma sodium compared to flox control mice after being challenged with 1 day of high salt diet. However, it is not yet known if this genotype difference is conserved in female mice. Therefore, we investigated whether collecting duct NOS1 is critical in maintaining homeostasis in female mice. Female CDNOS1KO and flox control mice were acclimated to metabolic cages and placed on a sodium deficient (LS, 1 week) then challenged with a high sodium (HS, 1 day) diet and urine collected. Female CDNOS1KO mice and flox control mice had similar sodium excretion (Flox: 3.24±0.32, KO: 2.62±0.31mmol/day; n=8–12/group, p>0.05), urine volume (Flox: 7.69±0.75, KO: 6.42±0.82mL/day; n=8–12/group, p>0.05), and NOx excretion (Flox: 1099±205, KO: 1339±389 μmol/day; n=8–12/group, p>0.05) in addition to comparable plasma sodium levels (Flox: 146±1.0, KO: 147±0.5mmol/L; n=6/group, p>0.05) after one day of a high salt challenge. Furthermore, urinary prostaglandin E metabolite excretion was similar between genotypes after a high salt challenge (Flox: 3187±382, KO: 2744±291pg/day; n=8–10/group, p>0.05), suggesting that female mice are able to compensate for a loss of NOS1‐derived NO in the collecting duct. Additionally, we performed RT‐PCR on male and female renal vessels and found that unlike males, female CDNOS1KO mice have significantly elevated vascular endothelin 1 mRNA expression (Male‐ flox: 1.00±0.09, KO: 0.98±0.03; n=6/group, p>0.05; Female‐ flox: 1.00±0.18, KO: 1.75±0.24; n=5–6/group, p=0.03). In conclusion, female mice do not rely on collecting duct NOS1‐derived NO to maintain fluid‐electrolyte balance. We propose that female mice utilize endothelin‐1 and/or NO production in other renal segments.

Support or Funding Information

F31DK111067 to RSS, F31HL149235 to LSD, and P01HL136267, AHA 19SRG34930023 to JSP
Survival After In-Hospital Cardiac Arrest In Critically Ill Patients Implications For The Covid-19 Pandemic?
Authors: Girotra Saket , Tang Yuanyuan , Chan Paul S. , Nallamothu Brahmajee K.
Published: April 2020

The coronavirus disease 2019 (COVID-19) outbreak is placing a considerable strain on U.S. healthcare systems. Due to presumptions of poor outcomes in such critically ill patients, many hospitals have started considering a universal do-not-resuscitate order in patients with confirmed Covid-19 given a limited supply of intensive care unit (ICU) beds and the potential risk of transmission of infection to healthcare workers during resuscitation. However, empirical data on survival of cardiac arrest in Covid-19 patients are unavailable at this time.

To inform this debate, we report survival outcomes following cardiopulmonary resuscitation in a cohort of similar critically ill patients with pneumonia or sepsis who were receiving mechanical ventilation in an ICU at the time of arrest. The probability of survival without severe neurological disability (CPC of 1 or 2) ranged from less than 3% to over 22% across key patient subgroups, For patients with an initial rhythm of asystole or PEA, who were also receiving vasopressors at the time of arrest, fewer than 10% were discharged without severe neurological disability (CPC of 1 or 2), and this number dropped to less than 3% in patients over 80 years old. In contrast, survival rates were much higher in younger patients, patients with an initial rhythm of VF or pulseless VT, and in patients receiving ventilatory support without vasopressors.

Our findings suggest caution in universal resuscitation policies. Even in a cohort of critically ill patients on mechanical ventilation, survival outcomes following in-hospital resuscitation were not uniformly poor and varied markedly depending on age, co-morbidities and illness severity. We believe that these data can help inform discussions among patients, providers and hospital leaders regarding resuscitation policies and goals of care in the context of the COVID-19 pandemic.
An experimental trial of recombinant human interferon alpha nasal drops to prevent COVID-19 in medical staff in an epidemic area
Authors: Meng Zhongji , Wang Tongyu , Chen Li , Chen Xinhe , Li Longti , Qin Xueqin , Li Hai , Luo Jie
Published: April 2020

Objective

To investigate the efficacy and safety of recombinant human interferon alpha (rhIFN-α) nasal drops in healthy medical staff to prevent coronavirus disease 2019 (COVID-19).

Methods

A prospective, open-label study was conducted in January 21, 2020at Taihe Hospital in Shiyan City, Hubei Province. Totally, 2944 medical staff members were recruited and allocated into low-risk group or high-risk group according to whether they were directly exposed to COVID-19 patients. Participants in the low-risk group received rhIFN-α nasal drops (2–3 drops/nostril/time, 4 times/day) for 28 days with first-level protection; those in the high-risk group received identical rhIFN-α nasal drops combined with thymosin-α1 (1.6 mg, hypodermic injection, once a week) along with secondary-level or third-level protection. The primary outcome was new-onset COVID-19 over 28 days. The secondary outcome was new-onset fever or respiratory symptoms but with negative pulmonary images. The results were compared with new-onset COVID-19 in medical staff in Hubei Province (including Wuhan) during the same period. Adverse reactions to interferon nasal drops were also observed.

Results

Among the 2944 subjects in our study, 2415 were included in the low-risk group, including 997 doctors and 1418 nurses with average ages of 37.38 and 33.56 years, respectively; 529 were included in the high-risk group, including 122 doctors and 407 nurses with average ages of 35.24 and 32.16 years, respectively. The 28-day incidence of COVID-19 was zero in both the high and low-risk groups. The 28-day incidence of new-onset clinical symptoms with negative images for pneumonia was also zero in both the high and low-risk groups. As control, a total of 2035 medical personnel with confirmed COVID-19 from the same area (Hubei Province) was observed between January 21 to February 23, 2020. No serious adverse events were observed in our trial during the intervention period.

Conclusion

In this investigator-initiated open-label study, we observed that rhIFN-α nasal drops may effectively prevent COVID-19 in medical staff, as an enhancement protection on the basis of standard physical isolation. Our results also indicate that rhIFN-α nasal drops have potential promise for protecting susceptible healthy people during the coronavirus pandemic.
Learning as We Go: An Examination of the Statistical Accuracy of COVID19 Daily Death Count Predictions
Authors: Marchant Roman , Samia Noelle I. , Rosen Ori , Tanner Martin A. , Cripps Sally
Published: April 2020

A recent model developed at the Institute for Health Metrics and Evaluation (IHME) provides forecasts for ventilator use and hospital beds required for the care of COVID19 patients on a state-by-state basis throughout the United States over the period March 2020 through August 2020 (See the related website https://covid19.healthdata.org/projections for interactive data visualizations). In addition, the manuscript and associated website provide projections of deaths per day and total deaths throughout this period for the entire US, as well as for the District of Columbia. This research has received extensive attention in social media, as well as in the mass media. Moreover, this work has influenced policy makers at the highest levels of the United States government, having been mentioned at White House Press conferences, including March 31, 2020.

In this paper, we evaluate the predictive validity of model forecasts for COVID19 outcomes as data become sequentially available, using the IHME prediction of daily deaths. We have found that the predictions for daily number of deaths provided by the IHME model have been highly inaccurate. The model has been found to perform poorly even when attempting to predict the number of next day deaths. In particular, the true number of next day deaths has been outside the IHME prediction intervals as much as 70% of the time.
Modelling and analysis of COVID-19 epidemic in India
Authors: Tiwari Alok
Published: April 2020

COVID-19 epidemic is declared as the public health emergency of international concern by the World Health Organisation in the second week of March 2020. This disease originated from China in December 2019 has already caused havoc around the world, including India. The first case in India was reported on 30thJanuary 2020, with the cases crossing 6000 on the day paper was written. Complete lockdown of the nation for 21 days and immediate isolation of infected cases are the proactive steps taken by the authorities. For a better understanding of the evolution of COVID-19 in the country, Susceptible-Infectious-Quarantined-Recovered (SIQR) model is used in this paper. It is predicted that actual infectious population is ten times the reported positive case (quarantined) in the country. Also, a single case can infect 1.55 more individuals of the population. Epidemic doubling time is estimated to be around 4.1 days. All indicators are compared with Brazil and Italy as well. SIQR model has also predicted that India will see the peak with 22,000 active cases during the last week of April followed by reduction in active cases. It may take complete July for India to get over with COVID-19.
Social decision making is influenced by size of shoal but not personality or familiarity in Deccan Mahseer (Tor khudree)
Authors: Varma Vishwanath , Singh Abhishek , Vijayan Jintu , Binoy VV
Published: April 2020

Shoals formed by many piscine species are fission-fusion societies where decisions to leave or join a group can have consequences on the fitness of individuals. Some important factors that determine shoal choice are shoal size, familiarity and species composition. However, individuals and species often exhibit distinct shoaling preferences. Individual differences in shoaling preferences may also be related to personality traits such as boldness and sociability. In this study, we examined the link between shoaling decisions and personality traits in a hatchery reared population of an endangered megafish, the Deccan Mahseer (Tor khudree). We found that this fish exhibits a distinct preference for larger shoals at ratios of 1:2 or greater. However, they did not prefer to associate with an isolated familiar individual over unfamiliar ones or with a member of their own species over an invasive species. Moreover, shoaling preferences in individuals did not correlate with their boldness or sociability. These results suggest that hatchery reared mahseers which are reintroduced into natural habitats may shoal with invasive species, negatively affecting their viability. Modifying social behaviour of mahseers by amending rearing practices may be a useful strategy to improve outcomes of restocking interventions.
Machine Learning Analysis of Chest CT Scan Images as a Complementary Digital Test of Coronavirus (COVID-19) Patients
Authors: Al-Karawi Dhurgham , Al-Zaidi Shakir , Polus Nisreen , Jassim Sabah
Published: April 2020

This paper reports on the development and performance of machine learning schemes for the analysis of Chest CT Scan images of Coronavirus COVID-19 patients and demonstrates significant success in efficiently and automatically testing for COVID-19 infection. In particular, an innovative frequency domain algorithm, to be called FFT-Gabor scheme, will be shown to predict in almost real-time the state of the patient with an average accuracy of 95.37%, sensitivity 95.99% and specificity 94.76%. The FFT-Gabor scheme is adequately informative in that clinicians can visually examine the FFT-Gabor feature to support their final diagnostic.

Key Strengths

The proposed FFT-Gabor scheme is an automatic machine learning scheme that works in real time and achieves significantly high accuracy with very low false negative, and can provide supporting evidences of the predicted decision by visually displaying the final features upon which decision is made. This scheme will be most beneficial when used in addition to the RT-PCR swab test of non-symptomatic cases.