bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel Adhesive Silicone‐Based Cell Culture Systems for Differentiation of Human Adipose Tissue‐Derived Mesenchymal Stromal Cells
Authors: Stock Peggy , Meltke Martin , Hildebrand Janis , Böttcher Gunther , Christ Bruno 
Published: April 2020

We established a protocol for the differentiation of hepatocyte‐like cells derived from human adipose tissue mesenchymal stromal cells (hAT‐MSC) in classical two‐dimensional cell culture, yet the physiological functions can be optimized. Classical two‐dimensional cell culture systems exhibit a high modulus of elasticity, yet, without representing the stiffness of human organs. A novel adhesive silicone was utilized to improve the elasticity of the cell culture substratum and to simulate the stiffness of organ tissue.Aim of the study was to analyse the differentiation ability of hAT‐MSC into cells of meso‐ and endodermal lineages on collagen‐coated adhesive silicone layers with low elastic modulus (500 Pa) in comparison with classical 2D cell cultures on polystyrene (103 MPa). hAT‐MSC were differentiated by a two‐step protocol and the progress was verified 8 and 21 days after initiation by the detection of specific histochemical hallmarks and biochemical characteristics.hAT‐MSC attached to the collagen‐coated adhesive silicon and formed cell layers. The formation of spheroids was observed with ongoing culture only in undifferentiated and hepatogenic differentiation, to lower extent also during osteogenic differentiation. Specific stainings on undifferentiated cells on adhesive silicone versus classical cell culture revealed no evident differences in fat content by Oil‐Red‐O staining (adipogenic), alkaline phosphatase by BCIP/NBT staining (osteogenic), and glycogen by PAS staining (hepatogenic). Differentiated cells cultured on adhesive silicone showed more pronounced staining under all three differentiation directions as compared to classically cultured cells. Fat deposit quantification of adipogenic differentiated cells displayed a 2.9‐fold increase in cells cultured on adhesive silicon as compared to the 1.9‐fold increase in cells classically cultured. The urea production rate [μg/10.000 cells*24h] was significantly higher in the culture on adhesive silicone as compared to the classical culture after 21 days of hepatogenic differentiation: 51 ± 13 vs. 0.33 ± 0.04.The results show the appropriateness of adhesive collagen‐coated silicone for the various differentiation directions of hAT‐MSC. The silicone supports the potential of hAT‐MSC to growth as spheroids. Thus, the collagen‐coated adhesive silicone represents an important tool for the development of in vitro culture systems mimicking E‐moduli of tissues and improving physiological functions.Support or Funding InformationThe study was supported by the European Regional Development Fund and the Free State of Saxony (SAB) 100282466 to J.H. and P.S.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Aerobic Physical Training on Thermoregulatory Adjustments During Physical Exercise in Spontaneously Hypertensive Rats
Authors: Drummond Lucas Rios , Campos Helton Oliveira , Rodrigues Quezia Teixeira , Gonçalves Gleisy Kelly , Horta Nayara Abreu , da Costa Monteiro Mateus , Primola‐Gomes Thales Nicolau , Coimbra Candido Celso 
Published: April 2020

This study aimed to investigate the effects of physical training on thermoregulatory adjustments during running exercise and its effects on brain neuronal nitric oxide synthase (n‐NOS) expression. Male spontaneously hypertensive (SHR) and normotensive Wistar rats (NWR) at 8‐weeks age old were divided into four groups: untrained NWR (NWR‐U), trained NWR (NWR‐T), untrained SHR (SHR‐U) and trained SHR (SHR‐T). Experimental procedures were approved by the CEUA‐UFMG (#21/2015). Aerobic exercise training was performed on a treadmill running for 8 weeks. On the day of the experiments, the rats were subjected to a constant‐speed (60% of Vmáx) protocol until fatigue at 25°C. Tail skin temperature (Ts) and abdominal temperature (Ta) were measured every minute throughout the exercise trials. The preoptic area (POA) and paraventricular nuclei (PVN) of hypothalamus was collected to quantification of n‐NOS(ser852) by western blot. During exercise, the SHR‐U group showed greater increase in Ta compared to with NWR‐U group from the 16th min until the 23th min (min 23: 1.62 ± 0.15 °C vs. 1.21 ± 0.12 °C, p<0.05) and lower Ts from the 1th until the 16th min of exercise (min 16: 0.09 ± 0.95 °C vs. 1.39 ± 0.41 °C, p<0.05). Physical training in SHRs group did not change Ta response during exercise (fatigue: SHR‐T: 1.54 ± 0.29 °C vs. SHR‐U: 1.70 ± 0.18 °C, p<0.05) but caused a greater increase in Ts in SHR‐T group (min 12: −0.54 ± 0.93 °C vs. −2.64 ± 0.80 °C, p<0.05). Physical training in NWR group did not change Ta response during exercise (fatigue: NWR‐T: 1.62 ± 0.26 °C vs. NWR‐U: 1.71 ± 0.26 °C, p<0.05) but caused a greater increase in Ts in NWR‐T group (fatigue: 3.65 ± 0.38 °C vs. 1.60 ± 0.47 °C, p<0.05). The p‐nNOS expression in APO was lower in SHR‐U group when compared to the NWR‐U (0.06 ± 0.01 u.a. vs. 0.09 ± 0.01 u.a., p<0.05). Physical training increased the p‐nNOS expression in the APO in the SHR group (0.10 ± 0.01 u.a. vs. 0.06 ± 0.01 u.a., p<0.05). The p‐nNOS expression in PVN was similar between SHR‐U and NWR‐U groups (SHR‐U: 1.10 ± 0.19 u.a vs. NWR NT: 0.94 ± 0.13 u.a., p>0.05). Physical training increased p‐NOS expression in the APO in both SHR group (1.58 ± 0.17 u.a. vs. 1.10 ± 0.19 u.a., p<0.05) and NWR group (1.56 ± 0.25 u.a vs. 0.94 ± 0.13 u.a., p<0.05). We conclude that physical training increases the efficiency of thermoregulatory control during exercise in hypertensive animals, which can be explained, at least in part, to an increase in the activity of neuronal nitric oxide synthase in trained animals.Support or Funding InformationFapemig, Capes, and CNPq.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of aqueous extract of Tamarindus indica Linn. (Leguminosae) pulp on basal and stimulated gastric acid secretion due to histamine, pentagastrin and carbachol in rats
Authors: Onwuchekwa Chinedu , Okon Bassey 
Published: April 2020

Local folklore claims that Tamarindus indica Linn. (Leguminosae) has a gastric beneficial role in humans ( Pankaj et al., 2011). This study was carried out to examine the effects of tamarind on basal and stimulated gastric acid secretion (GAS) in the Wistar rats. Four groups of rats consisting of 6 rats (180 – 210 g) each were treated with aqueous extract of tamarind pulp (1.5 mg/kg, 2.5 mg/kg and 4 mg/kg) orally for 21 days. The ethical guidelines in accordance with NIH Guide for care and use of laboratory animals were followed in the handling of the animals during the studies. At the last day of the treatment, the rats were fasted for 24 hours, anaesthetized with ketamine and cannulated through the femoral vein. GAS (mEq/l−1) induced by histamine, pentagastrin and carbachol administration were assessed by using modified continuous perfusion technique method of Ghosh and Schild (1958) and its acidity determined by titration method. Data were expressed as mean ± SEM. Student's t‐test and one way ANOVA were used to analyze and determine levels of significance at p=0.05.The aqueous extract of tamarind significantly decreased (p<0.05) GAS induced by the histamine, pentagastrin and carbachol at small and medium doses, but not at higher doses over the 80 minute's period in all the treated groups. The gastroprotective action of tamarind may be multifactorial that may include anti‐secretory action.Support or Funding InformationThe financial support of Usmanu Danfodiyo University, Sokoto Nigeria is acknowledgedThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modified team‐based learning as a weekly formative review
Authors: Hartley Rebecca S , McGuire Paul G 
Published: April 2020

Team‐Based Learning is an evidence‐based collaborative learning strategy designed around units of instruction, known as “modules,” that are taught in a three‐step cycle: preparation, in‐class readiness assurance testing, and application‐focused exercise. The Anatomy, Histology and Embryology course at the University of New Mexico School of Medicine modified TBL for use as both summative and formative weekly assessment, ending each week with a TBL module. The individual readiness assessment test (IRAT) served as a weekly “summative” quiz, while the group readiness assessment test (GRAT) served a formative role, allowing students to debrief and learn from each other following the IRAT. The application exercises were clinical cases focused on application of the anatomy, histology and embryology. Instead of additional multiple choice questions, questions were posed in short and long answer formats, giving the students an opportunity to generate their own answers for deeper learning. An addition specific to anatomy, were weekly mini‐practical exams to prepare students for summative lab practical exams. This modified TBL method was well‐received by students and prepared them for summative exams. One drawback was that scores from the GRAT, application exercise, and mini‐practical contributed to passing of some students that did not have a strong enough grasp on the concepts. This method was abandoned when the course was integrated as a thread into the existing organ blocks of the preclinical curriculum. Recently, this form of TBL was modified again to serve a purely formative role in weekly review and consolidation of concepts in the Musculoskeletal, Skin, and Connective Tissue Block of the integrated curriculum. Student feedback and scores strongly support use of this method as a formative review of the weekly material to prepare students for weekly graded quizzes and a summative NBME exam.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Developmental Exposure to Chlorpyrifos Modulates Pulmonary Function in Adult Rats
Authors: Grodzki Ana Cristina , Schelegle Edward , Lein Pamela 
Published: April 2020

Acute OP intoxication is well known to cause bronchoconstriction and increase airway secretions via inhibition of acetylcholinesterase (AChE). However, occupational and environmental exposures to organophosphorus pesticides (OP) at levels not expected to significantly inhibit AChE are associated with increased incidence of asthma and other pulmonary diseases. In support of the epidemiological data, acute exposure of adult guinea pigs and rats to OPs at levels that do not inhibit AChE in the lung, blood or brain have been shown to cause airway hyperreactivity (AHR). More recently, epidemiologic studies have suggested that exposure to OPs during development is also associated with increased risk of asthma. To address whether developmental OP exposures are also causally linked to AHR, we measured respiratory physiology in young adult rats exposed to the OP chlorpyrifos (CPF) at 0, 0.1, 0.3 and 1 mg/kg/d (sc) from postnatal days 1–4. Using a Flexivent mechanical ventilator, airway responsiveness to electrical stimulation of the vagus nerves was quantified. Preliminary findings suggest that adult female rats exposed developmentally to CPF exhibit significantly decreased airway resistance and elastance in response to electrical vagal stimulation relative to vehicle controls. In contrast, male rats exposed developmentally to CPF did not exhibit any changes in airway resistance and elastance in response to electrical vagal stimulation relative to vehicle controls. These results suggest that early life exposure to CPF triggers atypical expansion of the large and small conducting airways in the lungs of female, but not male rats. These findings will significantly advance our understanding of the role of OPs in respiratory diseases, and ultimately, inform risk assessments used to protect human health and safety. The United States Environmental Protection Agency currently uses acetylcholinesterase inhibition as a biomarker of OP toxicity; however, OPs modulate lung function at levels significantly lower than those that inhibit AChE. The findings of this research could potentially provide a scientifically rational basis for reassessment of exposure limits for OPs as well as help direct future research on therapeutic agents for treating/respiratory disorders in OP‐exposed children and adolescents.Support or Funding InformationWork was supported by the NIH (R01 ES017592 to PL and pilot project funding to ACG from P30 ES023513).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assembly of a mechanosensitive complex is required to initiate sprouting angiogenesis
Authors: Bayless Kayla , Duran Camille L. , Abbey Colette A. , Kang Hojin A. 
Published: April 2020

Endothelial cells (ECs) must respond rapidly to a variety of extracellular cues to initiate sprouting events. We undertook the current study to better understand how mechanical forces (shear stress) combine with biochemical cues to promote sprout outgrowth in a defined 3D model of EC invasion, because while the effects of shear stress have been widely studied in atherosclerosis, the molecular events directed by mechanical forces in angiogenic sprouting are incompletely understood. We initially focused on the known mechanosensitive protein Filamin A, which has been implicated in barrier regulation and vascular development. We found that Filamin A is required for EC sprouting in 3D and localizes to junctions with EC activation. Filamin A formed a complex with vascular endothelial (VE)‐cadherin and platelet endothelial cell adhesion molecule‐1 (PECAM‐1), two known members of mechanosensitive complex, following activation with shear stress and pro‐angiogenic factors. Silencing of Filamin A also disrupted the VE‐cadherin and PECAM‐1 complex, which correlates with decreased sprouting. Filamin A co‐localized with SHP2, a phosphatase previously implicated in barrier formation, following EC activation. SHP2 complexed with junctional molecules Filamin A, VE‐cadherin, and PECAM‐1, and complex formation between SHP2, VE‐cadherin, and PECAM‐1 was lost with Filamin A knockdown. In addition, in the absence of Filamin A expression, SHP2 failed to localize to endothelial junctions. Further, SHP2 silencing significantly reduced sprouting, and defects in sprouting were rescued by delivery of wild‐type SHP2. Altogether these data suggest that Filamin A recruits SHP2 to reinforce a transmembrane complex containing VE‐cadherin and PECAM‐1 that is needed to initiate EC sprouting. These data show for the first time that the formation of this mechanosensitive complex containing VE‐cadherin, PECAM‐1, Filamin A, and SHP2 is needed for successful sprout initiation.Support or Funding InformationWork supported by NIH RO1 HL095786, the Department of Molecular & Cellular Medicine, Texas A&M Health Science Center, and AHA Grant‐in‐Aid, Southwest Affiliate.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cannabinoid Inhibition of Carboxylesterase 1 (CES1)
Authors: Qian Yuli , Markowitz John S. 
Published: April 2020

The increasing use of cannabis for both medical indications and on a recreational basis poses a potential for drug interactions between cannabis or one or more of its constituents and conventional therapeutic agents. Although significant efforts have been made to assess the in vitro and in vivo effects of cannabis on cytochrome P450 and UDP‐glucuronosyltransferase enzyme systems, there is no information on the potential influence of cannabis on the major hepatic esterase, carboxylesterase 1 (CES1). In this study, we investigated the in vitro inhibitory effects of the major cannabinoids and metabolites including Δ9‐tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), 11‐nor‐Δ9‐tetrahydrocannabinol‐carboxylic acid (THC‐COOH), and 11‐Hydroxy‐Δ9‐tetrahydrocannabinol (11‐OH‐THC) on CES1 activity.An in vitro system of S9 fractions from human embryonic kidney (HEK 293) cells stably expressing CES1 was utilized to assess the inhibitory effects of the cannabinoids. THC, CBD, and CBN each exhibited substantial inhibitory potency, and were further studied to determine their mechanism of inhibition and kinetic parameters. The inhibition of CES1 by THC, CBD, and CBN was reversible and appears to proceed through a mixed competitive‐noncompetitive mechanism. The Ki values for THC, CBD, and CBN inhibition were 0.541, 0.974, and 0.263 μM (0.170, 0.306, and 0.0817 μg/ml), respectively. Compared to the unbound plasma concentrations achievable through medical or recreational use, the Ki values suggest a potential for clinically significant inhibition of CES1 by THC and CBD. CBN, however, is expected to have limited impact on CES1. Clinical studies are warranted to confirm these in vitro findings.Support or Funding InformationSelf‐fundedThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Hyperinsulinemia in Slipped Capital Femoral Epiphysis
Authors: Montañez-Alvarez Manuel , Flores-Navarro Héctor H. , Cuevas-De Alba Carlos , Arana-Hernández Erika I. , Ramírez-Ruiz Marisa 
Published: April 2020

Background:
Obesity in the prepuberal stage has been directly associated with slipped capital femoral epiphysis (SCFE). Serum insulin level increases in the prepuberal and adolescence stage, to a greater extent in the obese population. The main objective of this article was to analyze the relationship between insulin levels and SCFE.

Methods:
A case-control study was conducted between January 2018 and April 2019. The study group was formed with patients with SCFE and the control group with patients from the pediatric obesity clinic of our hospital selected during their initial evaluation. None were being treated for obesity. Anthropometric measurements of size, weight, waist circumference, and blood pressure were taken. Body mass index (BMI) and waist-height index of all patients were calculated. According to BMI for age, they were classified as normal, overweight, or obese. Serum determinations of glucose, insulin, glycated hemoglobin, lipid profile, and complete blood count were analyzed. Insulin resistance was diagnosed with Homeostatic Model Assessment (HOMA) >3. Insulin levels >13 U/mL for girls and >17 U/mL for boys were considered as hyperinsulinemia.

Results:
We studied 14 patients with SCFE and 23 in the control group. The mean age and BMI in both groups were similar. The elevation of serum insulin was significantly higher in the SCFE group (P=0.001) as was HOMA (P=0.005). Triglycerides and very-low-density lipoprotein were higher in the SCFE group (P=0.037 and 0.009, respectively). Glycemia, glycated hemoglobin, total cholesterol, high-density lipoprotein, low-density lipoprotein, and neutrophils showed no significant difference.

Conclusions:
Patients with SCFE showed elevated levels of insulin, HOMA, triglycerides, and very-low-density lipoprotein, even higher than the control group. Our study demonstrates a significant association between abnormally high serum insulin levels and SCFE. The known effects of insulin on growth cartilage may explain the physeal mechanical insufficiency to support the abnormally high or repetitive loads in accelerated growth stages that lead to SCFE.

Level of Evidence:
Level III—case-control, prognostic study.
Neurologic Manifestations of Severe Respiratory Viral Contagions
Authors: Robinson Christopher P. , Busl Katharina M. 
Published: April 2020

Objectives:
Endemic and pandemic viral respiratory infections have recently emerged as a critical topic of investigation given the recent severe acute respiratory syndrome coronavirus-2 outbreak. Data from such outbreaks indicate that severe systemic comorbidities including acute neurologic illness are associated with illness and lead to significant outcome differences. Herein, we will discuss the neurologic manifestations of severe viral respiratory infections including coronavirus, influenza, respiratory syncytial virus, metapneumovirus, and enterovirus.

Data Sources:
PubMed and EMBASE were searched by two independent investigators up to March 2020.

Study Selection:
Data selection included preclinical and clinical studies detailing neurologic manifestations of viral respiratory infections.

Data Extraction and Synthesis:
Two independent investigators reviewed and extracted the data.

Conclusions:
Neurologic manifestations including seizures, status epilepticus, encephalitis, critical illness neuromyopathy, acute disseminated encephalomyelitis, acute necrotizing encephalitis, Guillan-Barré syndrome, transverse myelitis, and acute flaccid myelitis have all been associated with severe viral respiratory infections. Having an understanding of the direct neurotropism of such viruses is imperative to understanding pathogenesis, clinical presentation, and potential treatment paradigms aimed at improving morbidity and mortality.
Newly Designed Universal Trans-punctal Rigid Lacrimal Microendoscope
Authors: Mimura Masashi , Ko Audrey C. , Alameddine Ramzi M. , Korn Bobby S. , Kikkawa Don O. , Oku Hidehiro , Sato Bunpei , Ikeda Tsunehiko 
Published: April 2020

Purpose:
To report the development and clinical findings of a universal trans-punctal lacrimal microendoscope design.

Methods:
In this study, we examined a unique and original “2-bend” (i.e., double-angle) rigid lacrimal microendoscope designed for universal applications for anatomical variations. The shape of endoscope was initially evaluated in 6 lacrimal systems of cadavers (4 Caucasian-cadaver lacrimal systems and 2 Asian-cadaver lacrimal systems). Second, a prospective clinical study involving 45 consecutive cases of nasolacrimal duct obstruction in Japanese patients was conducted to compare the facility and ease of use between the conventional single-bend type and our original 2-bend type trans-punctal lacrimal microendoscope for examination of the lacrimal drainage system.

Results:
The findings in the cadaver study revealed that original 2-bend lacrimal microendoscope, which was designed double-angled 20°–30° at 10 and 30 mm from the tip of the scope, could more easily used to insert and image all lacrimal systems, while the conventional single-bent lacrimal microendoscope was difficult to insert in subset of patients with a prominent nasal process of the frontal bone. In the clinical trial, our findings showed that 26.7% of lacrimal systems could not be passed using the conventional single-bend design, while using the 2-bend design, all cases could successfully be investigated.

Conclusions:
The original 2-bend-design microendoscope was found to be effective and valuable for universal use in examination and evaluation of the lacrimal passage.