bluuebunny/crossref_metadata_2025_split huggingface dataset:
Klotho Gene Delivery Suppresses Endothelin‐1 Production but Upregulates ETB Receptors in Kidneys in SHR
Authors: Wang Yuhong , Sun Zhongjie 
Published: June 2021

Klotho is a newly‐discovered anti‐aging gene. We recently reported that klotho gene delivery prevents progression of spontaneous hypertension and renal damage in spontaneously hypertensive rats (SHRs). The purpose of this experiment was to further determine the effect of klotho gene delivery on the production of ET‐1 and regulation of SOD in kidneys of SHRs. AAV carrying mouse klotho full‐length cDNA (AAV.mKL) was constructed for in vivo expression of klotho. Four groups of male SHRs (5 rats/group) and 1 group of sex‐ and age‐matched WKY rats were used. Briefly, 4 groups of SHRs received IV injection of AAV.mKL, AAV.LacZ, AAV.GFP, and PBS, respectively. The WKY group received PBS and served as a control. AAV delivery of klotho gene significantly attenuated spontaneous hypertension. All animals were sacrificed at the end of week 12 after gene delivery. AAV.mKL decreased the ET‐1 levels in plasma and kidneys. Interestingly, klotho gene delivery increased ETB receptor expression in renal distal tubule and decreased the ETA/ETB receptor ratio, suggesting, for the first time, that klotho may up‐regulate renal ETB receptors. Klotho gene delivery also enhanced superoxide dismutase (SOD) expression in kidneys.ConclusionsKlotho gene delivery suppressed the ET‐1 production and up‐regulated ETB and SOD in kidneys of SHRs. These findings may offer new insights into the relationship between renal klotho and the ET system in hypertension.
CFTR and ClC‐5 Chloride Channels Modulate Plasma Membrane H+‐ATPase in Proximal Tubule Cells
Authors: Carraro‐Lacroix Luciene Regina , Lessa Lucília M. , Pessoa Thaíssa D. , Nogueira Camila , Souza‐Menezes Jackson , Morales Marcelo M. , Girardi Adriana C. , Malnic Gerhard 
Published: June 2021

H+ transport across membranes by H‐ATPase creates a lumen‐positive voltage, generating an electrochemical potential gradient that limits pump activity. This DP is normally dissipated by a Cl‐ movement, which provides an electric shunt compensating for the transferred positive charge. This work was performed to identify the associated Cl−‐conducting proteins involved in the regulation of plasma membrane (PM) H‐ATPase from rat kidney proximal tubule (PT). Through in vivo stationary microperfusion, it was observed that ATPase‐mediated HCO3− reabsorption was significantly reduced in the presence of the Cl− channels inhibitor NPPB. This effect was confirmed in vitro by measuring the cell pH recovery rates after NH4Cl pulse in a cultured cell line derived from PT, IRPTC. In these cells, even after abolishing the membrane potential with valinomycin, ATPase activity was seen to be still dependent on Cl−. siRNA‐mediated CFTR and ClC‐5 channels knockdown significantly reduced H‐ATPase activity. These results indicate that PM H‐ATPase activity from PT depends on both CFTR and ClC‐5 chloride channels.Supported by FAPESP
NBCe1‐A residues mutated in patients with pRTA: A Topological Analysis
Authors: Zhu Quansheng , Kao Liyo , Newman Debra , Liu Weixin , Azimov Rustam , Kurtz Ira 
Published: June 2021

Eight missense mutations in Na+ coupled HCO3− cotransporter 1 (NBCe1‐A) (R298S, S427L, T485S, G486R, R510H, L522P, A799V and R881C) are reported to cause proximal renal tubular acidosis (pRTA). In this study, we explored location of the pRTA causing mutations and their structural importance in NBCe1‐A by substituted cysteine accessibility method (SCAM). The 8 residues were individually substituted with cysteine (cys) in a modified NBCe1‐A backbone with no free reactive cys (NBCe1‐A‐5C−). Locations of the introduced cys were determined by whole cell labeling with two cysteine specific reagents: membrane permeant biotin maleimide and a membrane impermeant MTS‐TAMRA. Results showed none of the 8 residues is on the surface of NBCe1‐A. Selective mutation of T485 revealed it resides in a unique position that is important for normal function. Further topologic analysis of NBCe1‐A by SCAM on 56 individually introduced cys in the NBCe1‐A‐5C− revealed that the first half of the transmembrane region of NBCe1‐A contains 8 transmembrane segments (TM) highly homologous to anion exchanger 1 (AE1), whereas the remainder of the cotransporter folds tightly unlike AE1. On the basis of these results, we propose a new topologic model of NBCe1‐A which allows the assignment of pRTA causing mutations to the following regions: S427L, TM1; T485S and G486R, TM2; R510H and L522P, TM3; A799V, TM10; R881C, TM12.Source of Research Support: NIH
TP53 polymorphisms and association with endometriosis in Brazilian population
Authors: Carvalho Cristina Valletta , Camargo‐Kosugi Cintia Meirelles , Kosugi Eduardo Macoto , D'Amora Paulo , Rojas Jéssica Vidal , Schor Eduardo , da Silva Ismael Dale Cotrim Guerreiro 
Published: June 2021

IntroductionEndometriosis is a benign gynaecologycal disease, defined as the presence of functional endometrial implants out of uterus, mainly in pelvis. Cell cycle and apoptosis disturbs could be associated with this disease as well as alterations in the genes that control.AimTo investigate the possible association between the presence of the polymorphisms in the genes TP53: codons 11; 72; 248 with endometriosis.MethodsCross sectional case–control study was performed. Genomic DNA was extracted from cells collected from buccal swabs. The TP53: codons 11; 72; 248 polymorphisms were investigated using the polymerase chain reaction restriction fragment length polymorphism (PCR‐RFLP) method in a hospital‐based Brazilian population.ResultsThe P53 gene, codon 11 (P53*11) polymorphism study showed that mostly of women did not present the allelic variance (p=0.255). The P53 gene, codon 72 (P53*72) polymorphism study presented significant differences between endometriosis and control groups (P=0.01; OR=2.19; 95% IC=1.196–4.039). The P53 gene, codon 248 polymorphism study showed that only one woman from endometriosis group presented heterozygosis to variate allele.ConclusionsP53*72 polymorphisms studies could be associated with endometriosis.
Role of caveolin ‐ 1 in the entry of HCMV into human extravillous trophoblast cells
Authors: Wang Lin , Zhou Li 
Published: June 2021

ObjectiveTo establish the cell model of HCMV infect human extravillous trophoblast cells (EVTs)and use this valuable cell model to study the role of caveolin‐1 in the entry of HCMV into EVTs.MethodsEVTs separated from early gestation villous tissue (5–6 weeks gestation) was used to establish the cell model of HCMV infection. The dynamic change of caveolin‐1 in EVTs after infection of HCMV was characterized by using flow cytometry and immunocytochemistry and RT‐PCR.ResultsThe positive products of HCMV immediate early protein can be observed in the nucleuses of infected cells using immunocytochemistry method, mRNA from HCMV IE gene was detected in the EVT cells 24 and 36 hours after infection. The result of flow cytometry shows that the percentage of EVTs apoptosis does not increase at early period, but improve 48 hours later after infection; The expression of caveolin‐1 in uninfected EVT cells is higher, began to reduce at 12 hours after infection, the lowest expression of caveolin‐1 occurs at 24 hours after infection. The expression of caveolin‐1 began to increase at 48 hours, with peak expression occurring at 96 hours after infection.ConclusionEstablished a cell model of HCMV infect EVT cells successfully. The dynamic change of caveolin‐1 in EVTs imply that HCMV enters EVT cells via unique endocytic pathway that involves caveolae/caveolin‐1.
Nexilin is a component of Enteropathogenic Escherichia coli (EPEC) pedestals
Authors: Law H T , Jackson Jeremy S , Guttman Julian A 
Published: June 2021

Enteropathogenic Escherichia coli (EPEC) are extracellular bacteria that colonize intestinal epithelial cells, leading to severe diarrheal disease and possibly death. Following bacterial attachment to the cell surface these microbes utilize a syringe‐like apparatus, called a Type III Secretion System (T3SS), to inject various pathogenic bacterial proteins into the host cell to cause disease. One of these proteins, Tir (Transmembrane intimin receptor), inserts itself into the host plasma membrane where it functions to recruit actin beneath the attached bacteria, generating structures called “pedestals”. These pedestals are common a characteristic of EPEC pathogenesis and allow the bacteria to ‘surf’ atop infected cells. Here we report the discovery of nexilin, an actin binding protein known to localize to focal adhesions, at EPEC pedestals. We demonstrate that nexilin recruitment beneath the attached bacteria requires a functional T3SS and specifically the bacterial protein Tir. Our discovery sets the foundation for investigating its contribution during these infections and is a method of exploiting EPEC to tease out the cellular function of novel host proteins.Grant Funding Source: NSERC
Aging causes accumulation of non‐proliferative adipose derived stem cells that is attenuated by caloric restriction in mice
Authors: Schmuck Eric , Mulligan Jacob D , Saupe Kurt W 
Published: June 2021

While white adipose tissue (WAT) is a promising source of mesenchymal stem cells, the effects of WAT remodeling factors such as aging and diet on these cells are unknown. To address this adipose‐derived stem cells (ADSC), defined as the non‐adipocyte cell fraction that is Lin‐/CD34+/CD31‐, were isolated from epididymal fat pads of adult (10–12 months, n=12) and aged (>24months, n=12) C57BL/6 mice fed ad lib or a calorie restricted (CR) diet known to retard aging and tumor development. ADSC abundance, single cell clonality, expression of stem cell associated genes and enzymatic activities were quantified. Aging increased ADSC abundance 3.2 fold and expression of the genes Cdkn2a 9.8 fold and Isl1 60.6 fold. However, clonality was decreased 33% suggesting that aging causes accumulation of non‐replicative ADSC. CR attenuated the age‐associated increase in ADCS abundance by greater than 50%, but decreased clonality to 10% and 7% of levels in adult and aged ad lib groups respectively. Therefore, CR appears to have anti‐proliferative effects on ADSC that may be advantageous from the perspective of cancer, but disadvantageous for regenerative medicine applications.Grant Funding Source: 1R21HL092477‐01 Proposal Title: Impact of chronic caloric restriction on resident progenitor cells in the heart
Continuous Measurement of Negative Pressure Effects on Cell Migration in Wounded Monolayer Cell with Electrical Wound‐Healing Assay
Authors: Hsu Chih‐Chin , Tsai Wen‐Chung , Wang Jong‐Shyan 
Published: June 2021

BackgroundNegative pressure wound therapy has recently gained popularity in chronic wound care. This study attempted to explore effects of different negative pressures on cell migration in wound healing process.MethodsWe used the electric cell‐substrate impedance sensing technique to create a 5×10−4 cm2 wound in the monolayer Madin‐Darby Canine Kidney cells and continuously monitor cell migration activities. The wounded cells were cultured in a self‐constructed negative pressure incubator at ambient pressure and negative pressures of 75 mmHg, 125 mmHg and 175 mmHg. The effective time, complete wound healing time, healing rate, cells radius and wound area over time were evaluated.FindingsThe effective time (1.25±0.27 hrs), complete healing time (1.76±0.32 hrs), healing rate (2.94±0.62 cm2/hr), cell radius (11.5±0.2 μm) at complete wound healing of cells treated at negative pressure of 125 mmHg were significantly different from those at other three pressure conditions.InterpretationThe study has established a cellular model for observing cell migration at different negative pressures uninterruptedly. Results have shown that the negative pressure of 125 mmHg can enhance cell migration activites.
Effects of Negative Pressure on Cell‐Cell Junctions and Cell Migration
Authors: Lu Yun‐Mei , Hsu Chih‐Chin , Wang Jong‐Shyan 
Published: June 2021

BackgroundNegative pressure wound therapy has been developed for treatment of chronic wounds. This study attempted to highlight effects of negative pressure of 125 mmHg on cell migration in wound healing process.MethodsA monolayer of Madin‐Darby Canine Kidney cells were grown in the chambered coverglass system. We created a scratching wound on the well with a pipette tip. Then the prepared traditional wound‐healing assays were harvested in a self‐constructed negative pressure incubator at ambient pressure and negative pressure 125 mmHg for three hours separately. Immunocytochemical stain of cells vitality, tight junctions and actins were performed later.FindingsNo differences of cell viabilities between cells cultured at ambient pressure and negative pressure of 125 mmHg. While compared with cells at ambient pressure, nearly total loss of tight junction fluorescent intensities and lamellipodium together with filopodia could be identified in cells treated with negative pressure.InterpretationLoosening of cell‐cell junctions may induce actin assembly to enhance cell migration at negative pressure of 125 mmHg.
Upregulation of Sca‐1 in satellite cells during diaphragmatic skeletal muscle regeneration: Potential novel mechanism of skeletal muscle regeneration
Authors: Rezk Bashir Mahmoud , Yoshida Tadashi , Semprun‐Prieto Laura , Sukhanov Sergiy , Delafontane Patrice 
Published: June 2021

Stem cell antigen‐1 (Sca‐1) is an 18kDa glycosyl phosphatidylinositol‐anchored cell surface protein of the Ly‐6 family that is involved in cell activation, proliferation and differentiation. Regeneration of skeletal muscle is a multifactorial process which includes proliferation and differentiation of satellite cells into new muscle fibers. Mechanisms of skeletal muscle regeneration, however, are poorly understood. We previously reported that angiotensin II (Ang II) is implicated in the induction of skeletal muscle atrophy that is characteristic of advanced heart failure. This study was designed to investigate the effects of Ang II on diaphragmatic skeletal muscle and determine mechanism(s) of skeletal muscle regeneration. FVB mice were infused with 1μg/Kg/min Ang II or sham‐infused for one week. Immunostaining for embryonic myosin heavy chain revealed multiple areas of regenerating muscle fibers and Sca‐1 expression was markedly upregulated in the satellite cells of these injured areas. Both proliferation markers PCNA and Ki67 were upregulated as detected by Western blot for PCNA and immunostaining for Ki67. The myogenic factor, M‐cadherin was also increased in the injured muscle. In conclusion, our results demonstrate that Ang II induced injury of diaphragmatic skeletal muscle is accompanied by increased proliferation and myogenesis that is potentially mediated, at least in part, by increased activity of Sca‐1 in muscle satellite cells.Grant Funding Source: NIH
Integrating Anatomical and Clinical Illustrations in a Teaching Format
Authors: Cui Dongmei , Lynch James C , Yang Gongchao , Haines Duane E 
Published: June 2021

Integrating anatomical structures with appropriate clinical examples can enhance the understanding of basic medical sciences and speed up the learning process. We have integrated illustrations of normal tissues with illustrations of diseased to highlight the differences between normal and abnormal tissues, and to demonstrate the processes in which normal change into abnormal tissues. Examples include: (1) Connective tissue cells in normal tissue compared to the cells in the tissue during an allergic reaction. (2) Normal and abnormal endothelium in atherosclerosis in blood vessels. (3) Normal stratified columnar cells of the conjunctiva of the eye are compared to cells with inflammatory granulations of the conjunctiva. (4) A normal optic nerve papilla is compared with an optic nerve papilla that exhibits papilledema. (5) Comparison of normal dentinal tissue and abnormal dentin tissue in a case of dentinogenesis imperfecta. Clinical correlations with well‐designed illustrations benefit students in learning clinical concepts, enhance the student's understanding of basic medical sciences, and help the students acquire clinical skills. This learning process paves the road for medical students as well as dental students to develop critical thinking skills while they are still in their early years of medical education, thus guiding them in their future study.Grant Funding Source: University of Mississippi Medical center
Experience of true life: A qualitative inquiry of first‐year medical students’ experience of cadaver dissection.
Authors: Skinner Miki D , Morrow Sue , Morton David A 
Published: June 2021

This research project explored the experience of first‐year medical students studying anatomy through full cadaver dissection. Literature suggests that cadaver dissection is a learning experience comprised of multiple variables that do not lend themselves to quantitative analysis. The goal of this research was to gain a greater understanding of the meaning that medical students take from cadaver dissection. The study was qualitative in nature. Perspectives of 16 first‐year medical students were drawn together through individual interviews and one focus group to produce a grounded theory hierarchy of the meaning‐making process associated with cadaver dissection. Themes emerging from the data were congruent with those identified in the literature. However the predominant theme of Balancing Respect is not reflected in the current scholarly literature regarding the use of cadaver dissection as an educational tool. These results suggest that the cadaver lab is the initial training ground for doctors to learn how to balance personal values connected with the human body with the invasive procedures they are required to do in order to be competent physicians.Grant Funding Source: AAA Student Travel Award
Testing knowledge of clinically‐relevant surface anatomy
Authors: Hankin Mark , Baptista Carlos A.C. , Lane Richard D. , Morse Dennis E. , Bennett‐Clarke Carol A. 
Published: June 2021

INTRODUCTIONA thorough knowledge of surface anatomy is essential for diagnosis and management of patients. For many students, “instruction” in this subject is limited to an introductory lecture. While surface anatomy is available for study on cadavers, dissection unavoidably removes skin making advanced study and learning difficult. Therefore, students’ knowledge of surface anatomy is rarely assessed on practical exams but instead measured in a limited way on written exams.METHODSThis report describes the use of undissected cadavers in practical exams given to physician assistant students (N=34) to assess their knowledge of surface anatomy on written and practical exams. Four pairs of subject‐matched, multiple‐choice surface anatomy questions on three exams were used to assess the association between performance on the subject‐matched questions and exam scores.RESULTSSpearman non‐parametric analysis showed strong correlations (0.79≤ r≤ 0.88, p<0.001) between overall performance on written and practical exams. While there was no correlation between performance on subject‐matched questions for Exam 1, significant correlations were observed for Exam 2 (r=0.37 p=0.3) and Exam 3 (r=0.39, p=0.02). Performance on subject‐matched questions also correlated with overall performance for Exams 2 and 3, but only for practical surface anatomy questions in Exam 1.CONCLUSIONThis data suggests that it is possible to accurately assess clinically‐relevant surface anatomy on a practical exam. Since physical examination is an important clinical application of surface anatomy, it is essential that we have an accurate way to assess our students' ability to recognize essential surface landmarks.
CORRELATING NUMERICAL COURSE GRADES BETWEEN GROSS ANATOMY AND CLINICAL COURSES WITH STRONG ORTHOPEDIC CONTENT IN THE PHYSICAL THERAPY CURRICULUM
Authors: Bukoskey Thomas Patrick , Barredo Ronald 
Published: June 2021

ObjectiveThe purpose of this study is to determine the presence, strength, and significance of correlations between the Gross Anatomy course and the Orthopedics course in the Doctor of Physical Therapy (DPT) curriculum.SubjectsNumerical course grades of students in the first DPT class were used to determine the correlations.MethodsCorrelations were conducted using Pearson r. Numerical course grades in the Gross Anatomy course (Year 1, Semester 1) were correlated with those from the Orthopedics course (Year 2, Semester 2). In addition, correlations were determined between the Gross Anatomy course and courses with strong orthopedic content which were taken prior to the actual Orthopedics course, including: Tests and Measurements (Year 1, Semester 2), Biomechanics and Movement Science I (Year 1, Semester 2), and Biomechanics and Movement Science II (Year 1, Semester 3).ResultsCorrelation significant at the 0.05 level was identified between Gross Anatomy and Orthopedics. Strong correlations significant at the 0.01 level were identified between the Gross Anatomy course and the following courses: Tests & Measurements, Biomechanics and Movement Science I, and Biomechanics and Movement Science II. Regression analysis between and among these courses revealed significance at the 0.00 level.ConclusionsStrong correlations existed between and among the courses foundational to the Orthopedics course.RelevanceThis study underscored the foundational role of the Gross Anatomy course in courses with strong orthopedic content. A strong background in Gross Anatomy was important when taking courses with strong orthopedic content.
Protocadherin of the liver, kidney and colon associates with detergent‐resistant membranes during cellular differentiation
Authors: Rizk Sandra , Krahn Michael P. , Alfalah Marwan , Behrendt Marc , Naim Hassan Y. 
Published: June 2021

Protocadherin LKC (PLKC) is a member of the heterogeneous subgroup of protocadherins, which was identified and described as a potential tumor‐suppressor gene involved in contact inhibition. Several aspects of the structure, posttranslational processing, targeting and function of this new protocadherin are still not known. Here, we generated MDCK cell lines that stably express PLKC and investigated its cellular distribution, biosynthesis, glycosylation and association with detergent‐resistant membranes, particularly during cellular differentiation. We demonstrate that PLKC is localized at the apical membrane and adherens junctions, but it does not colocalize with the tight junctions proteins ZO‐1 and occludin. Unlike E‐cadherin, PLKC is not redistributed from the cell membrane to submembranous and vesicular structures upon Ca2+ removal. Interestingly, PLKC associates during differentiation with detergent‐resistant membranes that trigger its redistribution from intracellular membranes to the cell surface. This association occurs concomitant with alterations in the O‐glycosylation pattern that is increased in postconfluent, i.e. well differentiated MDCK cells. Finally, we demonstrate that PLKC is directly involved in the establishment of a proper epithelial cell polarity.This work has been supported by grants from the German Research Council, DFG (SFB 621) to HYN.