bluuebunny/crossref_metadata_2025_split huggingface dataset:
Proteome Analysis of Plasma Exosomes in a Mouse Model of Dilated Cardiomyopathy (4C30 strain)
Authors: Suzuki Osamu 
Published: June 2021

AimThe role of exosomes is focused on in various diseases. In this study, I examined protein profiles in plasma exosomes of a mouse model of dilated cardiomyopathy (4C30 strain) to discover possible protein biomarkers for cardiomyopathy in plasma exosomes.MethodsPlasma samples were collected from 15‐week‐old male mice of cardiomyopathic 4C30 and normal C57BL/6NCr mice (five mice/strain). Plasma exosomes were collected from 1 mL of plasma, pooled from the five mice (0.2 mL/mice) in each strain using Exosome extraction kit for plasma (Invitrogen). Exosomal proteins were solubilized, cleaned up, reduced and alkylated. Approximately 50 µg of exosomal proteins were separated by isoelectric focusing and then SDS‐PAGE. After SyproRuby staining, two‐dimension electropherograms of exosomal proteins were compared between two strains. Some protein spots were punched out and their protein identities were determined by mass spectrometry.Results and DiscussionOnly a few differences were found in exosomal proteins between strains. Of proteins preferentially present in the 4C30 exosome, three spots were picked up and examined by mass spectrometry. One of them was fibrinogen gamma chain (FGG), which was thought to be a contaminant from plasma soluble components. The other two spots were under examination. Our results suggest that cardiomyopathy induces only a subtle change in exosomal proteins. This work was supported by a grant from the Ministry of Health, Labour and Welfare of Japan.
Anti‐arthritic effects of melatonin through blockade of cytonuclear shift of MMP‐13
Authors: Hong Yunkyung , Hong Yonggeun 
Published: June 2021

No curative treatments for osteoarthritic cartilage have been reported. The aim of the present study was to determine the therapeutic effects of melatonin on osteoarthritic chondrocytes. Chondrocytes were isolated from the cartilage specimens of the neonatal rats, cultured in monolayer, and treated with TNF‐α (10 ng/mL) repeatedly. We added DMSO combined with or without melatonin to medium. Caspase‐3 cleavage was induced by TNF‐α within 15 min, which is consistent with reduced cell viability. Co‐treatment of TNF‐α with DMSO induced not only increases of MMP‐13, both full length pro‐ and active proteins, but also shift of cytosolic MMP‐13 into the nucleus. Also there were metabolic disruption of COL2A1, proteoglycan loss, their atypical distribution, and excessive Ca2+ deposit. Melatonin, which was treated at the concentration of 1 nM, restored aberrant levels of COL2A1. Cytonuclear translocation of MMP‐13 was blocked by nanomolar concentration of melatonin partially, although its level was still remained high. Melatonin induced increase of the amount of proteoglycan with well‐distribution in chondrocytes. Moreover, the number of cells containing Ca2+ crystals decreased by melatonin treatment. Based on these results, we suggest that melatonin might be able to inhibit turning on of catabolic switch through regulation of MMP‐13 localization.Funding: 2012R1A1A2005089, 2013R1A2A2A01067169, KGM4611512
Building a Better Mouse Model to Understand Infant Respiratory Syncytial Virus Disease
Authors: You Dahui , Siefker David , Shrestha Bishwas , Saravia Jordy , Cormier Stephania 
Published: June 2021

Respiratory syncytial virus (RSV) is the number one cause of lower respiratory tract infection in infants; and there are no vaccines or specific antiviral drugs against RSV. The pathogenesis of RSV remains elusive. This is partly due to the fact that the mouse is semi‐permissive for human RSV. The present study sought to determine if a better neonatal mouse model of RSV infection could be obtained using a recombinant virus in which the F protein of standard laboratory strain A2 was replaced with that from the line 19 isolate (rA2‐19F).Five‐day‐old pups were infected with A2 or rA2‐19F. rA2‐19F caused significantly more lung inflammation and mucus than A2. More type II T helper (Th2) cells and less type I (Th1) cells and effector CD8+ T cells (Tc1) were observed in the rA2‐19F infected lungs compared to A2. A cohort of infected neonates were allowed to mature to adulthood and reinfected with the same strain at 4 wks post primary infection. Similarly, secondary infection with rA2‐19F caused exaggerated Th2 and reduced Th1 and Tc1 responses. These immune responses during reinfection were associated with increased airway hyperreactivity, mucus hyperproduction and eosinophilia in rA2‐19F vs. A2 infected mice. The mechanisms are unknown, but are associated with higher pulmonary viral loads in rA2‐19F vs. A2 infected mice during primary infection.Therefore, rA2‐19F caused more severe lung pathologies and Th2 immune responses compared to A2, possibly due to higher lung titers during primary infection. The rA2‐19F strain represents a better neonatal mouse model of RSV infection.This work is supported by U.S. National Institutes of Health.
Factor XIII A Clone EP3372 Provides Superior Staining on FFPE Human Tissue Sections Compared to Clone AC‐1A1
Authors: Liu Haiping , Muralitharan Sharmini 
Published: June 2021

Factor XIII is a plasma proenzyme essential for normal clot formation. Its 'A' subunit (Factor XIII A) exists intracellularly in cells such as dermal dendritic cells, histiocytic reticulum cells as well as megakaryocytes, macrophages and blood monocytes. Detection of Factor XIII A (FXIIIA) by Immunohistochemistry (IHC) is useful in the distinction between dermatofibroma (DF) and dermatofibrosarcoma protuberans, with DFs mostly being FXIIIA positive. However, the frequently used FXIIIA antibody, clone AC‐1A1, is associated with considerable non‐specific staining in IHC. The aim of the study is to find a good IHC assay for FXIIIA. Clone AC‐1A1 concentrate antibody was optimized with two IHC platforms on FFPE human tissue sections. However, the non‐specific staining was lingering. A Rabbit monoclonal antibody clone EP3372 was subsequently obtained and optimized. The optimized protocols for clone EP3372 and clone AC‐1A1 were compared. It was found that clone EP3372 not only produced stronger staining on tumor cells of dermatofibroma than clone AC‐1A1, but also devoid of any non‐specific staining. To further characterize clone EP3372, multi normal tissue arrays and tumor arrays were stained. In the normal tissues interstitial macrophages and stromal fibroblasts showed strong cytoplasmic staining. They were found in abundance in skin, gastrointestinal tracts, bladder, lymphoid tissues; and scattered in other tissues. Strong staining was found in dermatofibroma, capillary hemangioblastoma and fibrosarcoma. In addition, FXIIIA was also diffusely positive in 5 of 13 lymphomas. This study suggests that FXIIIA clone EP3372 performs superior to clone AC‐1A1. The finding that some lymphomas are FXIIIA positive needs further investigation.
Pharmacology Specialty and Brain Drain are Factors for Consideration in Improving Pharmacology Education in Nigeria ‐ Lagos State University College of Medicine (LASUCOM) as Example
Authors: John Theresa , Agaga Luther 
Published: June 2021

INTRODUCTIONIn Nigeria, pharmacology departments teach medical students didactically towards direct health care delivery. Is there a premise for expanding pharmacology departments' programs to include science degree curricula using LASUCOM as example?METHODSMedical students of LASUCOM participated in different 18‐question surveys two months apart.RESULTSRespondents associated pharmacology with medicine (61.3%), science (24.9%), industry (16.8 %), and government (11.1%); 32.8% want to know clinical pharmacology, 7.1% basic pharmacology; 45.8% prefer to study lecturers' notes, 26.7% textbooks, 9.8% the Internet, and 2.7% journals;primarily to be able to treat patients (40%), obtain MBBS degree (39.1%) and 8.9% to know this subject, 3.1% to make money; 0.4% would definitely and 33.8% would probably become pharmacologists, while 27.1% are uninterested and 8.4% would never be pharmacologists. Regarding Nigeria as not yet developed (51%), most would visit a developed country (61%) after MBBS (52%); some to spend professional life (59.52%); highest interests are in USA (46.7%), Canada (44.8%) and any European country (40.48%); based on scientific and technical advancement (60%). Some 23.8% consider it important for self fulfillment; 32.9%, 26.2%, and 21.9% for opportunity, bright future, and job satisfaction; 63% would necessarily return to Nigeria for roots (39%), to bring expertise (51.9%) and experience (53.3%).CONCLUSIONRespondents relate pharmacology to MBBS requirements; pharmacology departments could include science degree programs, collaborating with advanced countries, to contribute to local scientific and technical advancement, lack of which promotes brain drain.
Assessing the Safety and Tolerability of Long‐Term Multi Day IVM Treatment for Alcohol Use Disorder
Authors: Arabian Natalie , Huynh Nhat , Lieu Dustin , Thuy Jamie , Patel Neel , Rodgers Kathleen , Davies Daryl 
Published: June 2021

Alcohol use disorder (AUD) is a health crisis of unmet need. We propose that ivermectin (IVM), an FDA approved drug, can be repurposed to address this problem. Our group recently reported that long‐term (30 day) oral administration of IVM (10 mg/kg) significantly reduces murine ethanol intake without causing any overt signs of toxicity, suggesting that IVM represents a safe, tolerable, and effective agent for the treatment of AUDs. This first longer term study focused on an alcohol drinking paradigm that resulted in levels of alcohol intake that would more closely mimic the levels one would expect from someone drinking at a moderate level (i.e., social drinker). The current study extends this investigation and further assesses the safety and utility of multi‐day oral administration of IVM using a Drinking in the Dark (DID) paradigm, where animals in a very short period of time reach blood ethanol levels that are associated with “binge” drinking in humans. This was accomplished by investigating the effects of orally delivered IVM (10 mg/kg/day) in male alcohol preferring C57BL/6 over a 6 week (30 day, M‐F drinking) treatment period. We found that IVM significantly reduces ethanol intake and preference over the 6 week study without exerting any significant signs of toxicity, which was assessed using aspartate aminotransferase (AST) and alanine aminotransferase (ALT) liver enzyme level measurements. Histopathology assessments are currently ongoing. Taken together, findings, to date, continue to provide support for the development of of IVM for the treatment of AUD.Suport: Research grants SC CTSI NIH/NCRR/NCATS –UL1TR000130 (D.L. D.)
Sestrin2 Regulates TLR‐mediated Proinflammatory Signaling in Macrophages
Authors: Ki Sung Hwan , Yang Ji Hye 
Published: June 2021

Sestrins (Sesns) are conserved antioxidant proteins that accumulate in cells in response to various stresses. However, the regulatory roles of Sesn2 in the immune system and in inflammatory responses remain obscure. In the present study, we investigated whether Sesn2 regulates TLR‐mediated inflammatory signaling and sought to identify the molecular mechanism responsible. In cells expressing Sesn2, it was found that Sesn2 almost completely inhibited lipopolysaccharide (LPS)‐induced NO release and iNOS expression. A gene knockdown experiment confirmed the role of Sesn2 in LPS‐activated RAW264.7 cells. Consistently, proinflammatory cytokine release and expression were inhibited in Sesn2‐expressing cells. Furthermore, Sesn2 prevented LPS‐elicited cell death and ROS production via inhibition of NADPH oxidase. NF‐κB and AP‐1 are redox‐sensitive transcription factors that regulate the expressions of diverse inflammatory genes. Surprisingly, Sesn2 specifically inhibited AP‐1 luciferase activity and its DNA binding, but not those of NF‐κB. AP‐1 inhibition by Sesn2 was found to be due to a lack of JNK, p38, and c‐Jun phosphorylation. Next, we investigated whether Sesn2 protects galactosamine (Gal)/LPS‐induced liver injury in mice infected with a recombinant adenovirus Sesn2 (Ad‐Sesn2). Ad‐Sesn2 present less severe hepatic injury as supported by decreases in the ALT, AST, and hepatocyte degeneration. Moreover, Ad‐Sesn2 attenuated Gal/LPS‐induced proinflammatory gene expression in mice. The study shows that Sesn2 inhibits TLR‐induced proinflammatory signaling and protects cells by inhibiting JNK‐ or p38‐mediated c‐Jun phosphorylation.This study was supported by the National Research Foundation (NRF) funded by the Korea government (MOSIFP) (Grant No. R13‐2008‐010‐00000‐0)
Neuropeptide‐interactions in Gastric Mucosal Defense Initiated Centrally in the Rat.
Authors: Gyires Klara , Németh József , Zádori Zoltán 
Published: June 2021

BackgroundSeveral neuropeptides were shown to elicit gastric mucosal protective effect given centrally (intracerebroventricularly /i.c.v./, intracisternally or into discrete brain areas), e.g. TRH, opioid peptides, ghrelin, nociceptin, agmatine, substance P (SP), angiotensin II (Ang II) etc. However, numerous data suggest interactions between neuropeptides or with other endogenous mediators. Aim: To analyse if additional neuropeptides are involved in the gastroprotective effect of Ang II or agmatine (endogenous ligand of imidazoline receptors). Methods: Gastric mucosal lesion was elicited by oral administration of acidified ethanol. The compounds were given i.c.v. 10 min prior to ethanol and the lesions were determined 60 min later. The mucosal somatostatin and CGRP contents were determined by RIA. Results: 1. Both Ang II (0.012‐0.191 nmol) and agmatine (0.88‐4.4.nmol) induced significant reduction of the gastric lesions. 2. Parallel, the dramatic decrease of mucosal CGRP and somatostatin levels of gastric mucosa following ethanol administration was restored by both compounds. 3. The gastroprotective effect of Ang II was inhibited both by candesartan and L 733 060, selective NK1 receptor antagonist. Moreover, the protective effect of agmatine was inhibited both by alpha2/imidazoline I1 receptor antagonists idazoxan/efaroxan and the selective I1 receptor ligand, AGN 192403 as well as the opioid receptor antagonist, naloxone.ConclusionSP and opioid peptides might be involved in the gastroprotive effect of Ang II and agmatine, respectively.Supported by OTKA 75965
Lung Progenitor Cells Expressing PW1 Gene Participate in Vascular Remodeling During Pulmonary Arterial Hypertension
Authors: NADAUD Sophie , DIERICK France , HERY Tiphaine , MONCEAU Virginie , MOUGENOT Nathalie , CRISAN Mihaela , DORFMULLER Peter , MARODON Gilles , BESSON Vanessa , MARAZZI Giovanna , SASSOON David , HULOT Jean‐Sébastien , SOUBRIER Florent 
Published: June 2021

AIMPW1+ progenitors were identified in adult tissues and can differentiate in smooth muscle cells (SMC) in vitro. Our hypothesis is that PW1+ progenitors are recruited to participate in the vascular remodeling during pulmonary hypertension (PAH).METHODSPW1IRESnLacZ+/‐ mice express the β‐galactosidase as a reporter gene for PW1 expression allowing to follow the lineage of PW1+ cells. These mice were exposed to chronic hypoxia (CH) to induce PAH and lung vessels neomuscularisation.RESULTSIn rodent and human lung, PW1+ cells were adjacent to small vessels. They express c‐Kit, PDGFRα and pericyte markers and differentiate in vitro into SMC. After 4 days of CH, proliferating lung PW1+ cells were increased (x3, p<0.01) as well as PDGFR‐α+/PW1+ and c‐Kit+/PW1+ cells (x2 and x8, p<0.001). In small pulmonary vessels, the proportion of β‐Gal+ vascular SMC derived from PW1+ cells was increased (64% vs 35%, p<0.05) suggesting a recruitment and differentiation of PW1+ cells into lung vascular SMC. Moreover, WT mice irradiated and engrafted with GFP+/β‐Gal+ bone marrow cells did not show any increase in GFP+ or β‐gal+ SMC in lung vessels after 4 days of CH indicating that the lung PW1+ progenitors are not derived from bone marrow. In the human PAH lung, PW1+ cells were also observed in the media of remodeled vessels.CONCLUSIONThese results suggest that lung resident PW1+ progenitors are recruited to participate in the vascular remodeling of small pulmonary vessels in experimental and human PAH.
Targeted delivery of microRNAs by microparticles to inflamed endothelium ameliorates endothelial inflammation and atherosclerosis
Authors: Ma Shuangtao , Tian Xiao Yu , Mu Chaofeng , Shen Haifa , Bismuth Jean , Wong Wing Tak 
Published: June 2021

RationaleThe E‐selectin in endothelium promotes the growth of atherosclerotic plaques. We identified an E‐selectin‐targeting thioaptamer (ESTA) and conjugated it onto a multistage vector (MSV) to form an ESTA‐MSV microparticle. We tested E‐selectin‐targeted delivery of microRNAs (miRs) for the treatment of atherosclerosis.Methods and ResultsThe E‐selectin was up‐regulated, while the miR‐146a and miR‐181b were down‐regulated, in TNF‐α‐treated human microvascular endothelial cells (HMVECs). MiR‐146a and miR‐181b mimics were packaged into the ESTA‐MSV. Both miR‐146a and miR‐181b attenuated TNF‐α‐induced endothelial inflammation. Moreover, the monocyte adhesion to HMVECs was also attenuated by miR‐146a or miR‐181b. The ApoE‐/‐ mice were injected with ESTA‐MSV‐packaged miR‐146a or miR‐181b biweekly through tail vein for 12 weeks. Injection of miR mimics resulted in a 4.7‐ and 5.7‐fold overexpression of miRs in mouse aortas. The immunofluorescence staining showed the co‐localization of E‐selectin and miRs in aortic endothelium. MiR‐146a and miR‐181b significantly inhibited atherosclerotic lesion size, lipids deposition, macrophage and lymphocyte infiltration in plaques, and expression of adhesion molecules in aortas.ConclusionsE‐selectin‐targeting delivery of miR‐146a and miR‐181b to inflamed endothelium inhibits atherosclerosis. These findings provide a novel delivery system for miR‐based therapy of atherosclerosis.
Perivascular Adipose Does Not Affect Endothelium‐Dependent Relaxation or Contractionin Spontaneously Hypertensive and Wistar Kyoto Rat Aorta
Authors: Wood Laura , Denniss Steven , Holder Peter , Rush James 
Published: June 2021

This study examined the effect of the presence or absence of naturally adhering perivascular adipose tissue (PVAT+ or PVAT‐, respectively) on acetylcholine (Ach)‐stimulated endothelium‐dependent vasorelaxation inisolated phenylephrine‐precontractedaortic rings, and onAch‐stimulated endothelium‐dependent vasocontraction in isolated quiescent (non‐precontracted) L‐NG‐Nitroarginine methyl ester‐preincubated aortic rings from 25‐30 wk old Spontaneously Hypertensive rats (SHR; n=4‐8) and Wistar Kyoto rats (WKY; n=6‐8). Ach from ‐10.0 to ‐5.0 LogM resulted in a dose‐dependent relaxation response in PVAT‐ rings from both WKY (Maximum Amplitude (Max Amp): 85.6±5.8%; EC50: ‐7.72±0.08LogM; Area Under the Curve (AUC): 236±18) and SHR (Max Amp: 87.3±6.1%; EC50: ‐7.90±0.07LogM; AUC: 255±23), with no statistically significant differences found between strains in any dose‐response curve‐fit parameter (p<0.05). In both WKY and SHR, the curve‐fit parameters of the Ach‐stimulated relaxation response in PVAT+ rings were found not to be statistically different from thoseof the PVAT‐ rings.Inquiescent rings, Ach resulted in a dose‐dependent vasocontractile response in PVAT‐ rings from both WKY (Max Amp: 16.5±1.6%; EC50: ‐6.32±0.10LogM; AUC: 28±4) and SHR (Max Amp: 43.7±7.0%; EC50: ‐6.25±0.11LogM; AUC: 59±10), with a greater Max Amp and AUC in SHR (p<0.01). In both WKY and SHR, the curve‐fit parameters of the Ach‐stimulated contractile response in PVAT+ rings were found not to be statistically different from those of the PVAT‐ rings. These preliminary data suggest that, under the conditions examined, the presence of PVAT does not affect endothelium‐dependent relaxation or contraction.Funded by NSERC Canada.
Effect of Altered Endogenous Melatonin Regulated by Conditional Light Exposure on Neural Reorganization by Regulation of ER Stress Response and Autophagy after Spinal Cord Injury
Authors: Kim Joo‐Heon , Jin Yunho , Park Sookyoung , Hong Yonggeun 
Published: June 2021

Spinal cord injury causes unfolded protein response (UPR) to recover from endoplasmic reticulum (ER) stress. Melatonin is known to delay ER stress‐induced apoptosis. Furthermore, it has been suggested that melatonin‐mediated autophagy can induce neuroprotection.The purpose of this study was to elucidate the effect of endogenous melatonin on neural reorganization and locomotor recovery after spinal cord injury. To diversify the concentration of endogenous melatonin, differentiated light exposure time was needed. Thus, experimental animals were divided into 5 groups; group1 (12‐hr light/12‐hr dark), group2 (6‐hr light/18‐hr dark), group3 (24‐hr dark), group4 (18‐hr light/6‐hr dark), group5 (24‐hr light). 7 days after injury, animals were engaged in locomotor assessment.The BBB score of experimental animals was increased inversely proportional to light exposure time. Similarly, the more the rats were blocked from light, the more neural cells were reorganized. Furthermore, axonal sprouting showed a tendency to relatively increase in accordance with light blockade.In conclusion, UPR and autophagy might interact to play a neuroprotective role and, this effect is intensified by endogenous melatonin.Funding : 2012R1A1A2005089, 2013R1A2A2A01067169, KGM4611512, 2014R1A1A3051724
Synthesis of type I collagen in an animal model of skin injury via TGF‐β1 and IGF‐1R upregulation
Authors: Choi Jeonghyun , Hong Yonggeun 
Published: June 2021

Many approaches that accelerate the wound healing process have been developed. However, improving the speed and quality of healing remain a challenging task faced by clinicians. Collagens are an important component in all phage of wound healing. They impart integrity and strength to all tissue. Transforming growth factor‐beta1 (TGF‐β1) is important in inflammation, angiogenesis, reepithelialization, and connective tissue regeneration. This play a role in wound contraction by facilitating fibroblast contraction of the collagen matrix. Insulin‐like growth factor‐1 (IGF‐1) is a cytokine that participates in the cellular granulation process during wound healing. IGF‐1 has also described as an agent that acts anabolically and anti‐catabolically in a wide variety of cell type. The aim of this study was to evaluate whether upregulation of TGF‐β1 and IGF‐1R related to tissue repair process of wound in an animal model. Skin injury were created 1.5㎠ on dorsal right and left. Animal were housed in ad libitum and controlled with a photoperiod of 12h. On postoperative 3h, 15h, 2d, 7d, and 10d the complete wound with a 0.5 ㎝ margin was carefully removed. TGF‐β1 gene expression was increased in 2d, 7d, and 10d. IGF‐1R gene expression was showed in the same manner. Synthesis of type I collagen was growing as time dependent. These finding suggest that typeIcollagen synthesize was influenced by TGF‐β1 and IGF‐1R.Funding: 2012R1A1A2005089, 2013R1A2A2A01067169, KGM4611512