bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of L‐arginine supplementation on renal morphology of rats submitted to unilateral ureteral obstruction.
Authors: Gomes Guiomar Nascimento , Pletiskaitz Thais Maria , Carraro‐Lacroix Luciene Regina , Gil Frida Zaladek 
Published: June 2021

This work was designed to study the effect of L‐Arginine (Arg) treatment on renal morphology changes after unilateral ureteral obstruction (UO). Ureteral obstruction was performed by ligation of the left ureter of Male Wistar rats, 3 month old. Four experimental groups were studied: control‐sham operated: C; obstructed: UO; control treated with Arg: CA; obstructed treated with Arg: UOA. Groups CA, and UOA received as drinking water a solution containing arginine 1% and sucrose 1%, since 24 hours before surgery until 14 days of obstruction, when the animals were studied. The groups C and UO received 1% sucrose solution. We studied: glomerular morphology, alpha smooth muscle actin (α‐SMA) expression, macrophage infiltration (ED1), proliferating cell nuclear antigen (PCNA) expression and fibrosis. We observed that 14 days after UO, α‐SMA expression and macrophage infiltration (ED1) were increased in UO kidney. In OUA, both α‐SMA expression and ED1 presence were reduced. Glomerular hypertrophy was observed in contralateral kidney from UO rats. The treatment with Arg also reduced this response. These results indicate that L‐arginine supplementation has a beneficial effect on renal morphological changes caused by ureteral obstruction.Supported by FAPESP, CAPES, CNPq.
A calcium‐induced calcium‐release mechanism supports Luteinizing Hormone (LH) induced testosterone secretion in mice Leydig cells
Authors: Varanda Wamberto Antonio , Franci Celso Rodrigues , Costa Roberta Ribeiro 
Published: June 2021

Binding of LH to a G Protein Coupled Receptor in the plasma membrane, results in a primary increase in cAMP and in the intracellular calcium concentration ([Ca2+]i in Leydig cells. Immunocytochemical labeling with specific antibodies show the presence of three distinct isoforms of both ryanodine receptors (RYRs) and inositol 1,4,5‐trisphosphate receptors (IP3Rs). Measurements of intracellular calcium activity, using the fluorescent calcium sensitive dye Fluo3 and confocal microscopy, demonstrate that these receptors are involved in a calcium induced calcium release mechanism (CICR), responsible for the amplification of the initial Ca2+ influx through plasma membrane T‐type calcium channels (CaV3), induced by LH. The RYRs and IP3Rs are functional, as judged both from their activation by caffeine and IP3 and block by ryanodine and 2‐APB, respectively. Nevertheless, the intracellular global calcium changes resulting from treatment of the cells with LH are readily blocked 100 μM ryanodine but not by 2‐APB or xestospongin C. Besides this, blockage of the ryanodine receptor inhibits both the hormone‐induced [Ca2+]i transients and steroid secretion by Leydig cells. These results not only broaden the understanding of the role played by calcium in Leydig cells but also show, for the first time, that ryanodine receptors have a crucial role in determining secretion of testosterone by the testis.Financial Support: FAPESP, CNPq
Phosphorylation of TonEBP/OREBP by cyclin dependent kinase 5 (CDK5) mediates its NaCl‐induced nuclear translocation
Authors: Gallazzini Morgan , Heussler Gary E , Williams Chester K , Burg Maurice B , Ferraris Joan D 
Published: June 2021

High NaCl increases TonEBP/OREBP nuclear localization. We find that high NaCl increases phosphorylation of TonEBP/OREBP‐T135, located in its Auxiliary Export Domain (amino acids 132–156). Mutation of T135 to alanine decreases high NaCl‐induced nuclear localization of TonEBP/OREBP. T135 is in a consensus site for phosphorylation by protein kinase CDK5. High NaCl increases phosphorylation of CDK5‐Y15, indicating increased activity. Inhibitors of CDK5 and Kinase Dead (KD) dominant negative CDK5 reduce high NaCl‐induced nuclear localization of TonEBP/OREBP. Furthermore, CDK5 associates with TonEBP/OREBP (coimmunoprecipitation) and directly phosphorylates T135 in vitro. Over expression of CDK5 KD strongly decreases phosphorylation of T135. We do not find any regulation of TonEBP/OREBP transactivating activity by CDK5. We conclude that high NaCl stimulates CDK5 kinase activity which in turn contributes to nuclear localization of TonEBP by phosphorylating it on T135.This research was supported by the Intramural Research Program of NHLBI.
PP2R2C is involved in the resensitization of the dopamine D1 receptor in human renal proximal tubule cells.
Authors: Zhang Yanrong , Yang Yu , Villar Van Anthony , Pascua Annabelle , Armando Ines , Jose Pedro A 
Published: June 2021

Dopamine is important in the regulation of blood pressure, sodium balance, and renal function acting through D1‐like and D2‐like dopamine receptors. Protein phosphatase 2A (PP2A) is a heterotrimeric serine/threonine phosphatase. There is tissue specific expression of the B subunits of PP2A; human renal proximal tubule cells express B55γ (PP2R2C, γ isoform, B regulatory subunit) and PP2A B56 γ (□ isoform, B′ regulatory subunit). We have reported that dopamine D1 receptor (D1R) dephosphorylation, recycling to the cell surface membrane, and resensitization are dependent on PP2A activity (Kidney Int. 2006;70:1072–9). We aimed to determine the role of PP2R2C in the spatial and temporal events involved in the resensitization of D1R in human renal proximal tubule cells. PP2A B (entire B regulatory subunit), PP2A C (catalytic subunit) and PP2R2C subunits were found to be expressed in the cytosol of immortalized human renal proximal tubular cells by confocal microscopy (n=3). Co‐immunoprecipitation studies demonstrated that the D1R interacts with PP2A B, PP2A C and PP2R2C subunits (n=3). Treatment with the D1‐like agonist fenoldopam (FEN, 1 □M) decreased expression of PP2A B (15%; n=3) and PP2A B56γ (30%; n=3) but increased that of PP2R2C (25%: n=3). Treatment with FEN for 15 min increased total phosphatase activity (18%; n=2). Upon FEN stimulation the D1R internalized and co‐localized with of PP2R2C (n=3). These results suggest that PP2A and PP2R2C are involved in the regulation of the resensitization and trafficking of the D1R.This work was supported in part by grants from the National Institutes of Health, HL68686, HL23081, HL074940, HL092196 and DK39308
Effects of hindlimb unweighting on estrogen replete and deficient mice
Authors: Crissey Jacqueline Marie , Ferreira J Andries , Brown Marybeth 
Published: June 2021

Skeletal muscle atrophy is often debilitating. In women the decline in lean muscle mass accelerates at the onset of menopause due in part to lower estrogen (E2) levels. We evaluated the role of E2 to regulate skeletal muscle mass and maintain contractile function over 14 days of hindlimb unloading (HU) and 3 days of reloading (REC) in E2 replete (estrogen receptor‐α knock‐out, ERαKO) and E2 deficient (aromatase knockout, ARKO) mice, compared to wild type (WT) animals. Mice were separated into 3 groups: 1) CON: ambulatory controls, 2) HU: 14days of HU, 3) REC: 14 days of HU + 3 days REC. We observed differential effects on body weight (BW) after HU and REC. BW significantly decreased in ARKO mice (−10.9%) and ERαKO (−10.5%) after HU, while BW of WT mice was not different. Neither ARKO nor ERαKO recovered to pre‐HU BW values following REC. Expected declines in muscle mass of soleus (SOL), plantaris (PLAN), gastrocnemius (GAS), and tibialis anterior (TA) in WT mice after HU (Table 1) were observed. ARKO mice trended towards losing more muscle mass during HU and recovered slower than WT.
Percent atrophy of muscle mass loss in HU & REC compared to CON.

WT
SOL
PLAN
GAS
TA

HU
−38
−20
−15
−11

REC
−18
−6
−6
−2

ARKO
SOL
PLAN
GAS
TA

HU
−43
−30
−23
−12

REC
−34
−17
−19
−15

ERαKO
SOL
PLAN
GAS
TA

HU
−46
−9
−15
−4

REC
−25
−7
−7
−8

The ERαKO muscle mass profile resembled that of WT mice, suggesting that the presence of E2 rather than the absence of ERα mediates estrogen effects during HU and REC. These data suggest that estrogen deficiency slows skeletal muscle mass recovery. Support: HD‐058834 & University of MO Life Sciences Fellowship.
Substrate‐dependent localization of transglutaminase 2
Authors: van den Akker Jeroen , Afink Gijs B , van Weert Angela , de Vos Judith , Bakker Erik NTP , vanBavel Ed 
Published: June 2021

Transglutaminase 2 (TG2) is known to be important for both cross‐linking of matrix proteins as well as cell adhesion, cytoskeletal organization and signaling. However, its cellular distribution and mechanisms of secretion and activation during matrix remodeling remain unclear. We tagged TG2 at the C‐terminal with eGFP. TG2/eGFP was transfected into HEK/293T cells using Effectene. Western blot and in vitro activity assay showed a >100‐fold increase in active TG2. No difference was observed in TG2 activity between TG2/eGFP and its non‐fluorescent control.TG2 distribution in HEK cells strongly depended on the underlying substrate. When seeded on collagen type I, TG2 appeared mainly as intense intracellular and membrane‐bound spots, as well as in microparticles at distances up to 100 micron from the cell. However, on a fibronectin coating, TG2 was rather homogenously distributed throughout the cytosol. In HEK cells plated on plastic, TG2 localized strongly along the cell membrane. Here, transfection induced a large increase in fibronectin staining, which colocalized with TG2 fluorescence. TG2 was almost absent in the nuclei of living cells.In conclusion, we developed a construct of green fluorescing TG2. HEK cells expressing TG2/eGFP showed distinct localization of TG2, depending on its ECM. Only TG2‐transfected cells were able to produce a membrane‐bound fibronectin matrix.Supported by NHF grant 2005.B080
Albumingel for cartilage and intervertebral disc regeneration prevents blood vessel formation
Authors: Scholz Beate , Kinzelmann Claudia , Benz Karin , Baumer Yvonne , Mollenhauer Jürgen , Schlosshauer Burkhard 
Published: June 2021

Healthy cartilage is devoid of blood vessels, thus an implant matrix for autologous chondrocyte implantation should exhibit antiangiogenic properties. The purpose of the work was to investigate the influence of an albumin gel (AG) for cartilage and intervertebral disc (IVD) regeneration on blood vessel formation. The AG is composed of activated sheep serum albumin, hyaluronic acid and a polyethylene glycol spacer. Primary human endothelial cells (EC) cultured on the AG did not adhere on the surface and did not proliferate. Life‐dead staining displayed an increase in dead cells within 5 days of culture on the AG, while extracts of the AG showed no cytotoxic effect. Controversely, the vast majority of human primary chondrocytes cultured within the AG were vital after 5 days. EC stimulated by vascular endothelial growth factor did hardly invade through AG coated transwell filter. Blood vessels of the chick chorioallantoic membrane did not grow into the implanted AG. Additionally, IVD cells enclosed in the AG were implanted subcutaneously in immunodeficient mice. After 2 weeks there was no evidence of blood vessel formation in the implant.The results indicate that the AG is a suitable 3D‐matrix for chondrocytes exhibiting nonpermissive properties for EC and therefore could be an ideal implant for cartilage and IVD regeneration.All procedures were approved by the local Ethics and the local Animal Care and Use Committee.
Rotational Stress Impairs Cutaneous Wound Healing In Mice
Authors: Monte‐Alto‐Costa Andréa , Romana‐Souza Bruna 
Published: June 2021

ObjectiveThe aim of this study was to investigate the effects of the rotational stress on cutaneous wound healing in mice.MethodsMale mice were spun at 45rpm for 15minutes every hour from three days before wounding until euthanasia. Control animals were not submitted to stress. A full‐thickness lesion was created and measured. Fourteen days after, animals were killed and lesions were collected. Sections were stained with HE and immunostained against F4/80 and alpha‐smooth muscle actin to quantify macrophages and myofibroblasts, respectively. Data are expressed as mean±S.E.M and analyzed by Student's t test.ResultsFourteen days after wounding, wound area was greater in the stress group than in the control group (p<0.05), while re‐epithelialized wound area was smaller in the stress group than in the control group (p<0.05). Stressed mice presented less organized collagenous granulation tissue (rich in inflammatory cells) when compared with control mice. The macrophages number and myofibroblasts density was greater in the stress group than in the control group (p<0.05).ConclusionThe rotational stress delays the cutaneous wound healing in mice.
3D‐X: making spatial sense of cross‐sectional anatomy in an online learning environment
Authors: Bhattacharyya Siddhartha , Dorosh Kyle , Wilson Tim , Johnson Marjorie 
Published: June 2021

Residency programs are demanding an increased knowledge of cross‐sectional anatomy from entering physicians, while medical schools are decreasing time allotted to teaching anatomy. The increasing use of e‐learning resources in medical education led us to create a web‐based tool to facilitate the learning of cross‐sectional anatomy.A 3D model of the abdomen was constructed in Amira 5.0, by segmenting 600 images from the Visible Male cryo‐section data. A number of cross‐sectional images were imported into Flash CS4 along with five reconstructed models, and interactive controls for the 3D models and dynamic labels for the cross‐sections were programmed using the ActionScript 3.0 engine in CS4. The application, titled 3D‐X, was exported in a web‐friendly flash format and published online.In 3D‐X, users can view seven key cross‐sections, while the planes of origin are indicated on the 3D model. The 3D model can be rotated and modified to view various structures from different angles. Moving the cursor over an anatomical structure on a 2D cross‐section causes the structure's textual label to appear and colors the structure to match its counterpart on the 3D model. Clicking a structure on a 2D cross‐section modifies the 3D model to create an unobstructed view of the structure and may help users correlate 2D and 3D information.Based on a research study, 3D‐X co‐administered with cadaveric anatomy improved CT interpretation when compared with learning solely through cadaveric anatomy. 3D‐X therefore shows promise as a tool for enhanced learning of cross‐sectional anatomy by providing a familiar 3D context to aid with spatial orientation.
Cadaver Computed Tomography as a Tool for Teaching Anatomy: an Update
Authors: Ellozy Sharif Hamed , Lerner Susan , Reidenberg Joy S. , Weintraub Joshua , Marin Michael L. , Laitman Jeffrey T 
Published: June 2021

Cadaver dissection has been the mainstay of anatomy instruction. However, advances in clinical imaging have made familiarity with cross‐sectional anatomy crucial. Additionally, clinical relevance in the preclinical curriculum is key in generating student enthusiasm. In order to facilitate these goals, high resolution computed tomography was performed on all cadavers used in gross anatomy.Multiplanar and 3D reconstructions were performed on 50 cadavers over two years. Specific reconstructions of skeletal, aerodigestive, and vascular systems were done, as well as any anatomic variants or postsurgical changes. The CT scans were read by a radiologist. Students were given access to all of the studies and instructed on how to use a DICOM viewer. CT images were included on the examinations. Student groups were required to identify a structure of interest in their cadaver, research the clinical significance of this finding, and illustrate the finding with a CT image.Introducing the principles of imaging early in medical school may facilitate the transfer of knowledge from the dissection laboratory to its clinical application. The ability to identify anatomic variants and pathology a priori allows for focused dissection in select cadavers. We have found cadaver computed tomography to be a valuable addition to traditional dissection in teaching anatomy.
Diagnosis of Pediatric Intussusception using Point of Care Ultrasound in the Pediatric Emergency Department
Authors: Halm Brunhild Minette , Boychuk Rodney B , Franke Adrian A 
Published: June 2021

Over the past decade Point‐of‐Care Ultrasound (POC US) is increasingly performed in emergency medicine for a variety of indications. Pediatric emergency medicine physicians have been slower to understand the importance of POC US in the diagnosis of some specific pediatric conditions.The use of POC US and knowledge of the sonographic characteristics of intussusception can help any emergency physician in the prompt diagnosis of this condition, thereby reducing morbidity that may result from a delay in diagnosis. This report illustrates the use of POC US in a case of a 10 month old male that presented to the emergency department (ED) at midnight with fussiness as the only symptom. A diagnosis of intussusception was made using real‐time POC US by a pediatric emergency medicine attending.In some pediatric hospitals, like our, the radiologists and the ultrasound technicians are not readily available after hours and need to be called in. The goal is to diagnose and treat intussusception as early as possible since a delay in diagnosis may result in the need for surgical reduction or even intestinal resection. The diagnosis of intussusception is best performed by an experienced sonographer, such as the radiologist. However, if a delay for this study is anticipated POC US by a pediatric emergency physician trained in POC US can be crucial in reducing time to diagnosis and definitive treatment.
Medical gross anatomy laboratory tours have a positive impact on participant interest in the health care professions
Authors: Eckel Christine M. , Pratt Rebecca L , Hermey Donna , Hanna Jandy B 
Published: June 2021

To encourage HS and allied health students in West Virginia who have an interest in the health care professions, WVSOM has a laboratory tour program for students enrolled in human anatomy courses at the HS or undergraduate level. Each tour begins with a presentation covering proper laboratory etiquette, the role of the Human Gift Registry, and scientific aspects of cadaver embalming and plastination. Participants are then given a tour of the lab and demonstrations of human anatomy emphasizing clinical applications using cadaveric specimens, radiographs, and/or plastinated specimens. To assess the value of the tours, we ask all participants to complete pre and post tour questionnaires. In the spring of 2009, 255 tour participants returned questionnaires; a response rate of 90.4%. 82% of participants indicated a strong or very strong interest in pursuing a career within the field of health care, and 27% said attending the tour changed their opinion about the health care professions. Of those whose interest changed, 78% indicated they were more interested in a health related career as a result of the tour. We conclude that our anatomy tour program positively influences the interest of participants in the health care professions.Grant Funding Source: NA
A Genome‐wide RNAi screen identifies novel factors involved in the processing of snRNA
Authors: Wagner Eric J , Albrecht Todd R. , Chen Jiandong , Ezzeddine Nader 
Published: June 2021

Proper 3′ end formation of snRNA depends on a conserved element termed the 3′ box and is thought to be tightly coupled to transcription. Moreover, there is a poorly understood complex of proteins called the “Integrator Complex” that is thought to mediate the processing of snRNAs. This complex contains at least 12 unique polypeptides that are likely loaded onto RNA polymerase II at the promoter and are involved in 3′ end formation.We are using the Drosophila U7 snRNA gene as a model substrate to study pre‐snRNA processing. We designed a cell‐based reporter by cloning the U7 snRNA gene upstream of the GFP ORF. In the event that an essential processing factor is depleted, the polymerase will transcribe past the processing site resulting in expression of GFP.Using this reporter, we performed a genome‐wide RNAi screen in S2 cells to identify factors required for the 3′ end formation of snRNA. We found that nearly all of the Integrator proteins identified biochemically are functionally required for the 3′ end processing of snRNA. Two of these Integrator proteins are indeed required for processing of endogenous U7snRNA in mutant flies and we also found that U4 snRNA biosynthesis is only sensitive to a subset of the Integrator proteins leading to the potential that there are distinct complexes associated at different snRNA genes. We also identified several novel genes that had not been previously implicated in snRNA biogenesis.
Use of RNA affinity chromatography to investigate factors involved in selenoprotein synthesis and nonsense decay
Authors: Berry Marla J , Seale Lucia , Gilman Christy , Seyedali Ali 
Published: June 2021

Selenocysteine, the 21st amino acid, is the defining component of selenoproteins, a family whose members function in cellular oxidative defenses, male fertility, and thyroid function. Selenium deficiency has been linked to the pathogenesis of cancers, diabetes, cardiovascular disease, neurodegenerative diseases and other diseases in which cumulative oxidative damage is implicated. Selenoprotein mRNAs recode UGA, which typically signals termination, to instead specify insertion of selenocysteine. This recoding process involves assembly of complexes at specific secondary structures in the 3' untranslated regions of these mRNAs termed SECIS elements. When selenium is limiting, UGA is recognized as a termination codon and can trigger nonsense‐mediated decay (NMD) of the corresponding mRNAs. Different selenoprotein mRNAs exhibit differential sensitivity to NMD, and the mechanistic basis for this is not known. We are using glutathione‐RNA (GRNA) chromatography to isolate RNA‐binding proteins that bind selenoprotein mRNAs, to further elucidate the mechanism of selenoprotein synthesis and the factors involved in conferring sensitivity or resistance to NMD. Using GRNA with w.t. and mutant SECIS elements, we have identified three new SECIS‐binding proteins and are characterizing the physiological functions of these proteins in selenoprotein synthesis and NMD.Supported by NIH grant DK‐47320 to MJB.