bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of early candidate urine biomarkers for measure Escitalopram treatment response from major depressive disorder
Authors: Huan Yuhang , Wei Jing , Zhou Jingjing , Liu Min , Yang Jian , Gao Youhe 
Published: April 2020

Major depressive disorder (MDD) is a common mental disorder that can cause substantial impairments in quality of life. Clinical treatment is usually built on trial and error method, which lasts about 12 weeks to evaluate whether the treatment is efficient or not, leading some inefficient treatment measures. We therefore identify early candidate urine biomarkers to predict efficient treatment response in MDD patients. In this study, urine samples were collected from 10 respondent and 10 non‐respondent to escitalopram MDD patients with 0‐, 2‐, and 12‐week treatment. Differential urinary proteins were then analyzed by liquid chromatography coupled with tandem mass spectrometry (LC‐MS/MS). On week 2, there were 14 differential proteins identified in response group when compared with week 0 and these proteins can’t be enriched by random allocation analysis, 10 among which have been associated with the mechanisms of MDD. While in non‐response group, there were 35 differential proteins identified on week 2, and 18 among which have been associated with the mechanisms of MDD. In addition, differential urinary proteins on week 2 between respondent and non‐response group can be distinguished clearly by using Orthogonal Partial Least Squares Discrimination Analysis (OPLS‐DA). Our small pilot results indicated that the urine proteome can reflect early changes between respondent and non‐respondent group to escitalopram therapy since on week 2, which will provide potential early candidate urine biomarkers to predict efficient treatment measures in MDD patients.Support or Funding InformationThis research was supported by the National Key Research and Development Program of China (2018YFC0910202, 2016 YFC 1306300), the Key Basic Research Program of the Ministry of Science and Technology of China (2013FY114100), the Beijing Natural Science Foundation (7173264, 7172076), the Beijing cooperative construction project (110651103), the Beijing Normal University (11100704), the Peking Union Medical College Hospital (2016–2.27), and Beijing Biobank of Clinical Resources‐‐Mental Disorders, BBCR‐MD.
Deletion of Elavl1 in renal collecting duct cells is associated with a specific gene expression pattern resulting in major functional changes
Authors: Bornkessel Nicola Marcia , Chernyakov Dmitry , Groß Alexander , Gallwitz Lisa , Becker Franziska , Fischer Annika , Edemir Bayram 
Published: April 2020

IntroductionThe action of vasopressin and aldosterone on water and sodium reabsorption in the renal collecting duct includes the action of the RNA binding protein human antigen R also known as Embryonic Lethal Abnormal Vision Like 1 (Elavl1). Elavl1 modifies the protein expression of the epithelial sodium channel EnaC by stabilizing the mRNA. Under hypertonic conditions it down regulates the expression of the mineralocorticoid receptor (MR), the endogenous target of aldosterone. So far, the role of Elavl1 on the expression of other factors has not been studied, nor whether hypertonicity has an effect on Elavl1 mediated gene expression. Within this study we identified hundreds of genes that show differences in expression upon Elavl1 knock out.MethodsWe used the CRISPR‐Cas9 technology to delete Elavl1. First we cloned specific guideRNAs targeting Elavl1 and scrambled gRNA into the lentiV2‐vector and used them to produce virus particles in HEK293T‐cells. With those particles we transduced a mouse cortical collecting duct cell line (mpkccd). The successful induction of mutation in the Elavl1 locus was verified by Sanger‐Sequencing. Single cell clones were selected and the deletion on protein level was confirmed by immunofluorescence analysis. The Elavl1‐deficient cells were used for functional analysis like cell proliferation, migration and colony forming capacity. Using next generation sequencing we analyzed the gene expression profile comparing Elavl1 deficient to control cells and identified signaling pathways, which were deregulated by deletion of Elavl1.ResultsThe successful deletion of Elavl1 was functionally associated with significantly slower proliferation, migration and lower colony forming capacity. As expected, Elavl1 deficiencies lead to down regulation of the MR gene. Beside MR the expression of more than 440 genes is significantly (p<0,001) up and more than 2700 genes are down regulated. Classification by gene ontology terms showed that gene members of the ontology categories like “enzyme binding”, „rna binding”,„cell stress” and „cell death” are significantly enriched. KEGG pathway analysis showed that the differentially expressed genes are involved in metabolic and cancer associated pathways, such as „PI3K‐AKT‐Signaling pathway” or „HIF1A Signaling pathway”.DiscussionWithin this study we showed a comprehensive analysis of Elval1 function in renal collecting duct cells. Beside mayor physiological differences, Elavl1 deficiency is associated with massive changes in gene expression. Further analyses are needed to investigate its role for in vivo renal function.
Role of Microglia in Ventilatory and Blood Pressure Responses to Acute Hypoxia
Authors: Okada Yasumasa , Yoshizawa Masashi , Fukushi Isato , Takeda Kotaro , Kono Yosuke , Hasebe Yohei , Koizumi Keiichi , Toda Takako 
Published: April 2020

Microglia induces neuroinflammation in response to intermittent and chronic hypoxia, and enhances ventilatory augmentation and blood pressure elevation during hypoxic exposure. However, the role of microglia in ventilatory and blood pressure responses to acute hypoxia, especially to brief period of hypoxic exposure, has not been well investigated. In the present study we aimed to clarify the role of microglia in ventilatory and blood pressure responses to short duration acute hypoxia. To investigate this issue we conducted experiments using adult rats. A blood pressure telemeter was implanted intraperitoneally with a pressure transducer tip in the abdominal aorta. After at least one week recovery period, ventilatory and blood pressure responses to hypoxia were measured in an unanesthetized freely moving condition by whole body plethysmography and blood pressure telemetry, respectively. Measurement was first conducted without pretreatment with minocycline which is an inhibitor of microglial activation. Then, in more than one week after this control measurement, the animal was pretreated with minocycline for three consecutive days, and ventilatory and blood pressure responses to hypoxia were measured under the suppression of microglial activation. To test hypoxic responses, mild (13% O2, 4 min) and severe (7% O2, 5 min) hypoxia was loaded with a sufficient interval. In results we did not find differences ether in ventilatory or blood pressure response to either level of hypoxia between the conditions before and after pretreatment with minocycline. These results indicate that microglial activation does not affect ventilatory or blood pressure response to short duration acute hypoxia, which is not in agreement with previous reports, and this disagreement may be attributed to the different period of hypoxic loading between the previous reports and our present study, which could you too short to induce neuroinflammatory action by activated microglia.Support or Funding InformationThis work was supported by JSPS KAKENHI Grant Numbers 17K08559, 17H05540, 18K17783, 19K17386 and 19K17620 and a Grant‐in‐aid of the Cardiovascular Research Fund, Tokyo, Japan.
Arterial Baroreflex Function in Patients with Parkinson’s Disease: a Pharmacological Approach
Authors: Sabino-Carvalho Jeann L. , Cartafina Roberta A. , Guimarães Gabriel M. N. , Brandão Pedro R. P. , Vianna Lauro C. 
Published: April 2020

Parkinson’s disease (PD) is a neurodegenerative disorder that is known for its motor dysfunction. Moreover, PD also causes non‐motor impairments, and approximately 40% of the patients experience coexistent abnormalities, like orthostatic hypotension (OH). The OH induces incapacitating symptoms that can contribute to an increased risk of falls, which have a great impact on patients’ quality of life. The arterial baroreflex system plays a pivotal role in counteracting transient falls in blood pressure during the execution of activities of daily living. Although a plethora of studies have indicated that an impaired baroreflex function is a clinical feature in patients with PD, none of the human studies have used the modified oxford method, which is considered the gold standard approach for assessing arterial baroreflex sensitivity (BRS). Additionally, since the OH and arterial BRS have an age‐dependent effect, studies with younger patients with PD is lacking. In this sense, the aim of the present study was to test the arterial baroreflex function in patients with PD with and without OH and aged‐matched control during rapid changes in blood pressure induced by sequential boluses of vasoactive drugs (Modified‐Oxford Technique). Beat‐to‐beat HR (electrocardiography) and BP (finger photoplethysmography) were obtained during 10‐min of supine rest preceding a modified‐Oxford [bolus injection of 65–100μg sodium nitroprusside (NTP) followed by 65–100μg phenylephrine hydrochloride (PHE) 1 min afterwards] in five patients with PD (50±6yr; Hohen and Yhar: <3a.u.; diagnose time: 8.8±3.7yr) and seven age‐matched controls (48±7yr). Arterial BRS was analyzed using the slopes of the linear portions of the R‐R interval‐systolic BP relationship. Patients with PD had lower BP at rest compared to controls (82±7 vs. 94±5mmHg, P=0.01) and a trend for higher HR (72±13 vs. 59±8bpm, P=0.06). The average of falling (ΔSBP: −37±0.65 vs. −31±0.43, P=0.26) and rising systolic BP (ΔSBP: −27±0.76 vs. −20±0.58, P=0.38) was similar between patients with PD and controls. Our preliminary results showed that arterial BRS was lower in patients with PD compared to controls for falling pressure (NTP trial: 4.52±0.34 vs. 9.81±0.23ms/mmHg, P=0.01) and for rising pressure (PHE trial: 5.08±0.27 vs. 13.60±0.55ms/mmHg, P=0.03). Of note, only one of the patients presented OH, and the baroreflex function was characterized by a marked blunted arterial BRS for failing pressure (NTP trial: 1.83ms/mmHg) and an averaged BRS for rising in pressure. Our preliminary results indicated that the ups and downs of baroreflex function are impaired in patients with PD, and the downs appear to be further impaired in patients with OH. These findings provide a better understanding of the neural control of blood pressure in patients with PD, and future studies should focus on OH.Support or Funding InformationCNPq, FAPDF and CAPES.
Early Life Stress Induces Anxiety‐like Hyperactivity but Not Altered Ethanol Drinking in Outbred Heterogeneous Stock Rats
Authors: Deal Aaron , Cooper Nicholas , Weiner Jeffrey , Woods Leah Solberg 
Published: April 2020

Early life stress (ELS) during adolescence is known to have lasting effects impacting vulnerability to psychopathological disorders in adulthood, including post‐traumatic stress disorder (PTSD) and alcohol use disorder (AUD). The mechanisms by which ELS affects an individual’s susceptibility to these psychopathological outcomes are not fully understood. There is evidence suggesting that genetics may play a role in susceptibility and resistance to ELS‐induced disorders, though few specific genes have been identified. Therefore, we investigated the outbred Heterogeneous Stock (HS) rat as a model to study the genetics contributing to ELS‐induced PTSD and AUD‐like behavior. The HS population has high genetic diversity and more closely resembles the range of phenotypes expected within a human population than inbred rodent strains. We hypothesized that adolescent social isolation (ASI) of HS rats would increase anxiety‐like behavior, induce hyperactivity in response to a novel environment, and increase ethanol self‐administration.In this study, we isolated (n=32) or group‐housed (n=32) male HS rats at 4 weeks of age. After 5 weeks of isolation (ISO) or group‐housing (GH), behavioral outcomes were measured via elevated plus maze (EPM), open field test (OFT), and forced swim test (FST). Following behavioral testing, all rats were individually housed and assessed for voluntary ethanol self‐administration.We found no differences between ISO and GH rats in EPM anxiety‐like behavior as measured via open arm time or closed arm time. However, we did find ISO increased hyperactivity in response to a novel environment compared to GH in the OFT, including increased movement time and distance, increased beam breaks, and increased rearing. Interestingly, we found that ISO rats demonstrated decreased despair‐like behavior in the FST, including decreased immobility and increased wall climbing. When presented with 20% ethanol, we found no difference in self‐administration between ISO and GH animals. At increasing concentrations of ethanol, GH rats appeared to consume more ethanol than ISO, though results did not reach significance. We observed high variability in individual propensity to self‐administer ethanol in both ISO and GH groups which may be playing a larger role than ELS in this population.We have shown that HS rats acquire hyperactivity in response to a novel environment following ELS but are resistant to ELS‐induced despair‐like behavior and alcohol consumption. There is literature to support ELS as a protective event in certain environmental situations. These results support the use of the HS rat as a model to investigate the genetics involved in resistance to ELS‐induced AUD.Support or Funding InformationFunding for this study was provided by the Wake Forest Center for Research on Substance Use and Addiction Pilot Grant with support from the Wake Forest Alcohol Center NIH/NIAAA award P50 AA26117.
A Novel Hybrid Silk Fibroin/Polyurethane Arteriovenous Graft for Hemodialysis: Proof‐of‐principle Animal Study in an Ovine Model
Authors: Acocella Fabio , Tozzi Matteo , Bagardi Mara , Ravasio Giuliano , Cigalino Giorgio , Crippa Luca , Piccolo Solange , Nahal Emma Amal , Spandri Mattia , Catto Valentina Ilaria Maria , Tironi Matteo , Remuzzi Andrea , Greco Francesco Giovanni , Riboldi Stefania Adele 
Published: April 2020

BACKGROUNDIn an attempt at answering the need for a valuable and durable hemodialysis vascular access, a novel semi‐degradable hybrid vascular graft, manufactured by electrospinning using silk fibroin and polyurethane (Silkothane®), has been developed and characterized in vitro according to standards.OBJECTIVESIn this proof‐of‐principle animal study, we aimed at evaluating the performances of Silkothane® grafts in a sheep model of arteriovenous shunt, with respect to patency and short‐term graft remodeling.MATERIALS AND METHODSNine Silkothane® grafts (6mm inner diameter, ≈7cm long, ≈400μm thick) were surgically placed into the left cervical fossa of nine female sheep, between the common carotid artery and the external jugular vein. During the housing period the animals were treated with an inhibitor of platelets aggregation. Grafts were examined by palpation three times per week, and by duplex ultrasound every two weeks to exclude significant stenosis, dilation, presence of thromboses, neointimal proliferation or other complicating features. The sheep were euthanized at 30, 60 and 90 days (N=3 for each group). At sacrifice, grafts were harvested, fixed, and submitted for histologic, immunohistochemical and Scanning Electron Microscopy examination.RESULTSNo cases of graft‐related complications (e.g. seroma formation, transgraft bleeding, aneurismal dilation, calcification, structural yielding) were recorded in this study. One graft failed due to a thrombotic event likely related to a technical error (11%). One sheep had post‐operative wound infection, surgically resolved (11%). Four out of nine sheep (44%) showed post‐operative edema, usually disappearing in 7 days. Eight of nine sheep (89%) showed 100% primary unassisted patency at the respective time of sacrifice, as confirmed by palpation and ultrasound evaluation (average flow rate 1.76 ± 0.61 l/min). Histological, immunoistochemical and SEM analysis evidenced signs of inflammation and formation of neointimal hyperplasia at the venous anastomosis, as it typically happens with arteriovenous fistulae and synthetic grafts.CONCLUSIONSIn our ovine model of arteriovenous shunt, Silkothane® grafts provided evidences of safety and efficacy in granting 100% patency up to 90 days. Further studied need to be conducted in order to directly compare the novel grafts with commercially available synthetic grafts.
Sex differences in cholinergic regulation of behavioral responses to rewarding and aversive stimuli
Authors: Tat Jennifer , Brady Lillian J. , Zachry Jennifer E. , Kutlu Munir Gunes , Calipari Erin S. 
Published: April 2020

Substance use disorder (SUD) is a major public health concern, characterized by dysregulation in drug taking, drug seeking, and compulsive behavior, as well as deficits in the valuation of drugs and alternative reinforcers. Determining neurobiological factors that contribute to individual variability of SUD is critical for identification of at‐risk populations as well as the use of preventative measures and personalized medicine in treatment. Women represent a particularly vulnerable population when it comes to the development and trajectory of SUD, therefore understanding the sex‐specific mechanisms underlying the differential control of reward and motivation will be critical for improving treatment outcomes in women. At the core of this SUD vulnerability is an enhancement in reward processing and motivation for both natural and drug rewards. Nicotinic acetylcholine receptors (nAChR) have been directly linked to the control of reward‐related behavior for both natural and drug reinforcers, thus, here we aimed to understand how these receptor populations differentially control motivated behavior for positive and negative stimuli in a sex‐specific fashion. Mice were trained to acquire positive (response = reward delivery) and negative (response = shock removal) reinforcement in the presence of discrete antecedent auditory cues. We found sex differences in performance at baseline, with females showing reduced rates of acquisition for negative reinforcement and increase motivation for positive reinforcers. Next, the non‐selective nAChR antagonist, mecamylamine, was administered systemically to assess the effect of nAChRs on task performance. Blocking nAChRs reduced discrimination for auditory cues in both sexes regardless of what task type they were predicting, demonstrating that its effects on reinforcement are not stimulus specific. Further, while we saw sex‐differences in reinforcement at baseline, we observed no sex differences following mecamylamine treatment. Together this data shows that nAChR modulation of reward‐related behavior eliminates basal sex differences in reinforcement behavior and suggests that the cholinergic system may be a crucial factor in sex differences in motivation.Support or Funding Information
Funding was provided by startup funds from Vanderbilt University School of Medicine Department of Pharmacology and the National Institutes of Health (NIH) (to E.S.C.) and the Vanderbilt University Academic Pathways Postdoctoral Research Fellowship (to L.J.B.). Funds from the National Institute of Drug Abuse (NIDA) DA042111 and DA048931(E.S.C), the Brain and Behavior Research Foundation (to E.S.C), Whitehall Foundation (to E.S.C), and the Edward J Mallinckrodt Jr. Foundation (to E.S.C) also supported this work.
Reactivation of eNOS in the endothelium rescues eNOS KO mice from endothelial dysfunction and hypertension
Authors: Leo Francesca , Piragine Eugenia , Vornholz Lukas , Suvorava Tatsiana , Isakson Brant , Cortese-Krott Miriam Margherita 
Published: April 2020

Production of nitric oxide (NO) by endothelial NO synthase (eNOS; NOS3) plays a key role in regulation of vascular tone and blood pressure. The aim of this study was to establish “loss” and “gain” of function models for eNOS by using tissue specific Cre‐induced gene inactivation or reactivation. To accomplish this, two independent genotypes were created: 1) a conventional floxed eNOS (eNOSflox/flox) against exon 2 or 2) a Cre‐inducible eNOS knock‐in (KI) gene construct (eNOSinv/inv) allowing targeted removal or reactivation of eNOS expression. Both the eNOSflox/flox and eNOSinv/inv were crossed with endothelial cell (EC)‐specific tamoxifen (TAM)‐inducible Cre mice to create EC‐specific knock‐out (EC eNOS KO) and EC eNOS KI mice. We found that Cre‐specific DNA recombination occurred specifically in endothelium of both EC eNOS KO and EC eNOS KI, as assessed by quantitative polymerase chain reaction (qPCR). TAM treatment fully abolished mRNA and protein expression of eNOS in the aorta of EC eNOS KO (eNOSflox/flox Crepos) mice, but not in Cre negative or untreated mice, as determined by retro transcription qPCR and western blot analysis. Functionally, the EC eNOS KO demonstrated inhibition of Ach‐induced vasodilation of aortic rings ex vivo, as well as flow‐mediated dilation (FMD) of the arteria iliaca in vivo (assessed by ultrasound). Moreover, absence of eNOS in the endothelium of EC eNOS KO was correlated with spontaneous hypertension, assessed by Millar catheter (MAP was 80.37 ± 2.135 mmHg in EC eNOS KO against 67.88 ± 2.723 mmHg in control mice). Next, we tested blood pressure of eNOSinv/inv which fully lacked eNOS expression in all tissues and was hypertensive. In EC eNOS KI (eNOSinv/inv Crepos treated with TAM) the expression of eNOS was completely restored in endothelium, leading to a significant decrease of the blood pressure as compared to eNOSinv/inv Creneg mice. In conclusion, we report the first inducible “loss” and “gain” of function models of eNOS in the vascular endothelium demonstrating that reactivation of eNOS rescues from endothelial dysfunction and hypertension. These mice models can further be applied to non endothelial tissues to further investigate the role of eNOS in different tissues in modulating blood pressure and systemic hemodynamics.Support or Funding InformationThis work was supported by the German Research Council (DFG CO 1305/2‐1 to M.M.C.‐K.; F.L. is a scholar of the IRTG1902 to M.M.C.‐K. and B.E.I. and by a research grant of the Forschungskommission to M.M.C.‐K., Medical Faculty, Heinrich Heine University of Düsseldorf.