bluuebunny/crossref_metadata_2025_split huggingface dataset:
Receptor for advanced glycation end products is involved in remodeling of diabetic coronary arterioles
Authors: Trask Aaron J , Katz Paige S , Stewart James A , Lucchesi Pamela A 
Published: June 2021

The aim of this study was to determine whether the receptor for advanced glycation end products (RAGE) contributes to remodeling of type 2 diabetic coronary arterioles (<150 μm diameter) of leptin‐receptor deficient db/db mice. Coronary arterioles isolated from control Db/db, type 2 diabetic db/db, and RAGE−/− × db/db mice at 16 weeks of age were mounted on a pressure myograph to measure and record wall thickness and diameters over a range of pressures (0–125 mmHg). Coronary arterioles isolated from db/db mice exhibited decreased luminal diameter (Db/db: 118 ± 5 μm vs. db/db: 103 ± 4 μm p < 0.05) with thicker microvascular walls (Db/db: 6.9 ± 0.6 μm vs. db/db: 9.7 ± 0.5 μm p < 0.01), which significantly increased the wall:lumen ratio (Db/db: 6.0 ± 0.8 vs. db/db: 9.5 ± 0.4, p < 0.01). Compared to db/db mice, coronary arterioles isolated from RAGE−/−x db/db mice had similar internal diameter (104 ± 9 μm, p > 0.05), while wall thickness (6.1 ± 0.5 μm, p < 0.001), wall:lumen ratio (6.2 ± 0.8 μm, p < 0.01), and growth index (db/db: 28.1 vs. RAGE−/−x db/db: 22.3, % of control) were reduced. External diameters were not different among groups, although there was a tendency for reduced external diameter in RAGE−/−x db/db. These data suggest that the RAGE receptor is at least partially responsible for inward hypertrophic remodeling observed in coronary arterioles of type 2 diabetic mice.Support: NIH T32HL098039 to AJT; NIH HL056046 to PAL.
Protein O‐GlcNAcylation Promotes Post‐hypoxic Survival of Cardiac Progenitor Cells
Authors: Zafir Ayesha , Li Qianhong , Bolli Roberto , Jones Steven P 
Published: June 2021

BackgroundCardiac progenitor cells (CPCs) represent a viable route to promote post‐infarct cardiac repair. We hypothesized that the auto‐protective stress signal, known as protein O‐GlcNAcylation, would confer a pro‐survival benefit in CPCs.Methods/ResultsWe genetically and pharmacologically augmented O‐GlcNAc levels in CPCs (c‐kit+/lin−) with loss‐ and gain‐of‐function approaches, i.e., adenoviral (Ad) overexpression of OGT (adds the O‐GlcNAc modification to proteins) and inhibition of OGA (removes the O‐GlcNAc modification from proteins) by PUGNAc (0.2 mM). We also used DON (0.05 mM) as an inhibitor of the rate‐limiting enzyme of the hexosamine biosynthetic pathway, GFAT. CPCs were subjected to 3 h hypoxia, followed by a 3 h reoxygenation period. Augmenting O‐GlcNAc levels (via AdOGT or PUGNAc) significantly attenuated post‐hypoxic cell death (according to LDH release), whereas reducing O‐GlcNAc levels (via DON) augmented post‐hypoxic cell death.ConclusionsEnhancing protein O‐GlcNAcylation represents a novel approach to minimize cell death in CPCs. Such approaches could be explored with the goal of improving CPC survival at the time of adoptive transfer to bolster the reparative capacity of CPCs in vivo.Financial support: NIH, AHA.
Exercise ancestry decreases lipogenesis‐related gene expression in skeletal muscle of male offspring
Authors: Guth Lisa M. , Ludlow Andrew T. , Witkowski Sarah , Marshall Mallory R. , Lima Laila , Venezia Andrew C. , Xiao Tao , Lee Mei‐Ling Ting , Spangenburg Espen E. , Roth Stephen M. 
Published: June 2021

The study objective was to examine the influence of exercise (EX) ancestry compared to sedentary (SED) ancestry on offspring skeletal muscle gene expression. C57/Bl6 mice (F0) were exposed to EX (wheel running, N=20) or a SED condition (no wheel access, N=20) for 12 weeks prior to breeding. EX males and EX females, and SED males and SED females were bred to produce F1 pups. F0 EX mice had continued access to the wheel during breeding, pregnancy, and weaning. F1 pups were sacrificed at 8 wk without direct exercise exposure. Microarray analysis of gene expression was performed on pooled gastrocnemius RNA from F1 males of each ancestry (5 animals per array; 4 total arrays). After processing the microarray data to minimize false positives, 109 transcripts were differentially expressed between F1 EX and F1 SED. Three targets identified as upregulated in F1 SED offspring have been previously associated with lipogenesis and were selected for real‐time PCR validation (Adipoq, Scd1, and Cidec). Adipoq and Scd1 were verified as significantly upregulated in F1 SED offspring (p < 0.05) and Cidec tended to be upregulated in F1 SED offspring (p = 0.06). These preliminary findings indicate that exercise ancestry (i.e., parental exercise stimulus) can affect lipogenesis‐related skeletal muscle gene expression in male mouse offspring.Supported by the UMD School of Public Health Research Seed Money Program and NIH AG000268.
Integrative approach reveals specific disturbances in β‐adrenergic signaling in a porcine model of post‐myocardial infarction remodeling
Authors: Kuster Diederik W , Verhoeven Adrie J. , Merkus Daphne , van der Velden Jolanda , Duncker Dirk J. 
Published: June 2021

Loss of myocardial tissue after myocardial infarction (MI) leads to neurohumoral activation that is aimed at restoring cardiac pump function, but eventually leads to cardiac remodeling, which is an independent risk factor for the development of heart failure.We studied the process of post‐MI remodeling in the pig, a physiologically relevant animal model, at the organismal, cellular and molecular level.At 3 weeks after MI, β‐adrenergic sensitivity of the heart was reduced and left ventricular remodeling and dysfunction were apparent. At the cardiomyocyte level, force development was reduced and Ca‐sensitivity increased. Microarray analysis revealed alterations in the β‐adrenergic signaling pathway. Protein analysis showed increased PKA regulatory subunit while PKA catalytic subunit was decreased, which were normalized upon β‐blocker treatment. The β‐agonist induced increase in phosphorylation of the PKA target protein troponin I was attenuated.This integrative approach allowed us to link the blunted response to exercise and the disturbed myofilament function to changes in expression of PKA subunits. β‐blocker treatment restored functional parameters to control levels. These observations are in line with medical practice that treatment with β‐blockers protects MI patients from developing heart failure.Supported by a grant from Netherlands Heart Foundation (NHS2005B234)
Dynamic Imaging of Airways During Bronchoconstriction and Deep Inspiration in Intact Rats
Authors: Rapalo Gabriel , Sinclair Scott E. , Makena Patrudu , Molthen Robert C. , Waters Christopher M. 
Published: June 2021

Airways are continuously subjected to cyclic mechanical stretch during normal respiratory movements, and large distensions occur periodically with deep sighs. However, there is limited information on the levels of mechanical stretch in vivo and how these might change in hyper‐responsive airways. Using a unique microfocal X‐ray imaging system to visualize airways coated with tantalum dust, we imaged dynamic changes in dimensions of rat airways in vivo following exposure to methacholine during mechanical ventilation. Circumferential strain and longitudinal strain were measured based upon changes in diameter and length of segments during a breath cycle relative to dimensions at end expiration. We simultaneously performed measurements of lung resistance and compliance using the Flexivent system. We found significant changes in the diameters of airways of all sizes (~0.2 mm to 2 mm), but there was substantially more heterogeneity in smaller airways. We also found that a deep inspiration caused significant relaxation of airways circumferentially, but deep inspiration had little effect on longitudinal dimensions. These measurements allow us to quantify changes in the levels of mechanical stretch in airways during bronchoconstriction and to assess the time‐dependent response to a deep inspiration.Supported by NIH grant HL094366.
Phrenic Nerve Morphometry in Short Term and Long Term Experimental Diabetes
Authors: Alcântara Adriana Cristina Licursi , Tanaka Natália Massumi , Filho Omar Andrade Rodrigues , Fazan Valéria Paula Sassoli 
Published: June 2021

The aim of the present study was to investigate the morphometric parameters of the phrenic nerve in short term and long term experimental diabetes. Male Wistar rats received a single intravenous injection of streptozotocin (40mg/Kg) 15 days (N=7) or 12 weeks (N=10) before the experiments. Controls rats (N=7 and N=4, respectively) received citrate buffer (vehicle). On the experimental day, rats were perfused with glutaraldehyde 2.5%, had the right and left phrenic nerves dissected and the proximal and distal segments prepared for light microscopy study as described previously (Rodrigues Filho & Fazan, 2006). Morphometry was performed with the aid of computer software and took into consideration the fascicle area and diameter, number of myelinated fibers and their density. Comparison was made between segments, sides and groups and differences were considered significant when p < 0.05. Our results show that the myelinated fiber number is similar between control groups but larger on both diabetic groups, compared to controls, particularly on the distal segments. This is suggestive of a “dying back” neuropathy, with signs of nerve regeneration.Support: CNPq, CAPES, FAPESP and FAEPAGrant Funding Source: CNPq, CAPES, FAPESP and FAEPA
Phrenic Nerve Morphology in Normotensive and Spontaneously Hypertensive Rats with chronically induced diabetes
Authors: da Silva Ferreira Renata , Sanada Luciana Sayuri , Salgado Hélio Cesar , Fazan Valéria Paula Sassoli 
Published: June 2021

The aim of this study was to investigate morphometric alterations in phrenic nerves of spontaneously hypertensive rats (SHR), compared to normotensive rats (WR), both with chronically induced diabetes (SHR+STZ and WR+STZ, respectively). WR+STZ and SHR+STZ received a single intravenous injection of streptozotocin (STZ) while WR and SHR received vehicle. Twelve weeks after injection, rats were anesthetized and had their blood pressure and heart rate recorded. After intracardiac perfusion, right and left phrenic nerves were prepared for light microscopy. Morphometric analysis was performed with the aid of computer software, and took into consideration the fascicle area and diameter, as well as myelinated fiber number, density, area and diameter. No differences in fascicular morphology were observed between groups and the phrenic nerves were statistically symmetrical longitudinally and laterally in all groups studied. However, both diabetic groups showed alteration of the myelinated fibers. Despite little alteration on phrenic nerve morphology in SHR+STZ, this study provides a morphological basis for future neuropathy studies in these models of human diseases.Support: CNPQ, CAPES, FAPESP and FAEPA
Spread of injected dye after ultrasound‐guided transversus abdominis plane block in cadavers
Authors: Morris Laura Ruth , Johnson Marjorie , Ganapathy Sugantha 
Published: June 2021

Transverse abdominis plane (TAP) block is done before abdominal surgery to relieve post surgery pain. A TAP block deposits local anaesthetic between transversus abdominis and internal oblique muscles, anesthetising the thoracolumbar nerves. Difficulty encountered when performing this procedure is consistency reaching T7‐L1 nerves. In previous TAP block studies, only some of these nerves were covered by dye, mimicking anaesthetic migration, in embalmed cadavers. The goal of this study is to explore techniques that enable TAP blocking of the T7 to L1 nerves. Different injected substances tested were methylene blue dye, green paint, red latex solution, gelatine and paint solution, and methylcellulose and paint solution in embalmed cadavers using ultrasound as a guide. Using methylcellulose and paint solution, T7 to L1 nerves were reached in embalmed cadavers with 20cc of methylcellulose/paint solution injected rostral and with 10 cc of methylcellulose/paint solution injected caudal to the umbilicus at the anterior axillary line. These clinical findings may provide anaesthesiologists with information to more consistently relieve pain and decrease recovery time.Grant Funding Source: Deparmental
Supra and infralevator autonomic‐somatic communications: Computer‐assisted anatomic dissection in male and female foetuses
Authors: alsaid bayan , MOSZKOWICZ David , PESCHAUD Frederique , BESSEDE Thomas , ZAITOUNA Mazen , KARAM Ibrahim , BENOIT Gérard , DROUPY Stéphane 
Published: June 2021

Objectiveto identify the location and the nature of communications between supra and infralevator pathways.Materials and Methods6 male and 6 female human fetal pelvic specimens, 15 to 32 wk of gestation, were studied using the 3D computer‐assisted anatomic dissection (CAAD) with neural, nitrergic and myelin sheath markers.ResultsSupra and infra levator neural pathways cross each other distally in spatial X shape area. They yield dual innervation in 5 directions: 1‐anal sphincter, 2‐levator ani muscle, 3‐urethral sphincter, 4‐corpus spongiosum and perineal muscles, and 5‐corpora cavernosa. The communications between the two pathways exist in 3 levels: proximal supralevator, intermediary intralevator and distal infralevator. Distal communications exist when autonomic nitrergic cavernous nerves interact with some somatic nitrergic branches of the dorsal penis/clitoris nerve and carried the nitrergic activity in its distal portion.ConclusionsThe pelvic structures responsible for sphincters continence and sexual function receive a dual innervation from the autonomic supralevator and the somatic infralevator pathways. Distal communication takes over the nitrergic activity to the distal part of cavernous bodies in both gender and represents an important structure which may impact erectile function, especially during reconstructive surgery.Grant Funding Source: non
Anti‐Fertility Effects of Amlodipine in Adult Male Wistar Rats
Authors: Akinlolu Adelaja Abdulazeez , Ghazali Olaide , Adefule Adebayo Kehinde 
Published: June 2021

Amlodipine is a long‐acting calcium channel blocker used in the treatment of hypertension and angina. In adult Man, the treatment regimen is 5 or 10mg daily. This study tested the hypothesis that prolonged oral administration of Amlodipine impairs fertility in adult male rats. Thirty‐three normotensive adult male wistar rats (140 – 250g) comprising of seven groups were employed in the study; Control Groups I and II received 4mls of Normal Saline while experimental Groups III – VII received 5, 10, 15, 20 and 40mg/kg BW of Amlodipine respectively for 56 – 65 days. Gross morphology, testicular histology, testosterone levels, sperm cells morphology, count and motility were examined in all rats. Gross morphology evaluations showed reduced bodyweight, hair loss and loss of locomotion and balance in rats treated with Amlodipine in weeks 4 – 9 in a dose‐dependent manner. No adverse effects were observed on testosterone levels in all rats. Dose ‐ dependent anomalies of sperm morphology such as sperms with no head and double tail as well as significantly reduced sperm cells' count and motility were observed in rats of the experimental group compared to rats of the control group. Histological analyses showed scanty seminiferous tubules, wider tubular lumen and reduced interstitial cells in rats of the experimental groups. This study observed impaired fertility in adult male rats treated with 5mg/kg BW or higher doses of Amlodipine.Grant Funding Source: UNIVERSITY OF ILORIN, NIGERIA
The spectrin cytoskeleton is an integral target and crucial component of adherent, invasive triggering and invasive zippering bacterial diseases
Authors: Ruetz Tyson Joel , Cornick Steve , Guttman Julian Andrew 
Published: June 2021

Entropathogenic Escherichia coli (EPEC), Salmonella Typhimurium (S.Typhimurium), and Listeria monocytogenes (L. monocytogenes) all manipulate the host cell cytoskeleton at crucial stages of their disease processes. Although the actin cytoskeleton has been previously identified as a ubiquitous target of these pathogens, the examination of the spectrin cytoskeleton has been largely overlooked. Here we show that spectrin and the spectrin‐associated proteins adducin and protein 4.1 are recruited to sites of EPEC pedestals as well as sites of S. Typhimurium and L. monocytogenes invasion. Individual siRNA knockdowns of each spectrin cytoskeletal protein significantly impeded the ability of EPEC to form pedestals and inhibited the abilities of both S.Typhimurium and L.monocytogenes to invade host cells. Further studies identified spectrin cytoskeleton recruitment to later time‐points of S.Typhimurium and L. monocytogenes infections during the intercellular stage of pathogenesis, revealing novel cytoplasmic roles for the spectrin cytoskeleton. This work demonstrates that the spectrin cytoskeletal system is crucial for bacterial pathogenesis and represents a potential target for therapeutic treatment for a broad list of infections.Grant Funding Source: CIHR
Beta1 integrin plays a role in lens fiber cell structure and interaction with the cytoskeleton
Authors: Scheiblin David A. , Simirskii Vladimir N. , Czymmek Kirk J. , Duncan Melinda K. 
Published: June 2021

A conditional knockout for β1 integrin of lens fiber cells was produced. Scanning electron microscopy was used to assess the ultrastructure. Immunofluorescence of sectioned and whole lens was used to determine the expression of β1 integrin and its crosstalk with different cytoskeletal components.At 14.5 dpc (days post coitum) expression of β1 integrin is lost in lens fiber cells. Mice lacking β1 integrin in lens fiber cells have variable phenotypes of clear and cataract lens. The ultrastructure of the lenses was analyzed from two different areas. Cortical fiber cells are newly differentiated cells that contain a highly regular structurally repeated ball and socket pattern along the membranes of fiber cells in wildtype mice. Lens lacking β1 integrin have ball and socket structures that are disorganized exhibiting variable width and extension from the cell. Nuclear fiber cells are older cells that contain ball and sockets as well as a random arrangement of membrane furrows in wildtype mice. Lens lacking β1 integrin have a membrane furrow pattern that is highly organized into a lattice like geometric pattern, no ball and sockets and a defect of Aquaporin‐0. Lens lacking β1 integrin also have a modified f‐actin network. The lack of the f‐actin network disrupts the hexagonal packing arrangement and radial cell columns of these fiber cells.β1 integrin is needed for proper structure of lens fiber cells. Funded by NEI EY015279.Grant Funding Source: NEI EY015279
Multi‐tasking focal adhesion protein zyxin is present at tubulobulbar complexes
Authors: Young J'Nelle Sarah , Vogl Wayne 
Published: June 2021

Tubulobulbar complexes (TBCs) are actin‐related double‐membrane projections that form at intercellular junctions in the seminiferous epithelium of the mammalian testis. They form at basal junction complexes between neighboring Sertoli cells and apical junctions between Sertoli cells and germ cells. They are proposed to internalize intact junctions during sperm release and during the translocation of spermatocytes through basal junction complexes between neighboring Sertoli cells. Although TBCs are testis‐specific, they morphologically resemble podosomes in osteoclasts. Previously, we have reported that a complex of N‐WASp, Arp2/3, cortactin and dynamin that is found at podosomes also is present at TBCs. Here we demonstrate that zyxin, a focal adhesion protein known to be present at podosomes also is present at apical TBCs. A rabbit polyclonal anti‐zyxin antibody (B71) [generous gift from M.C. Beckerle, Huntsman Cancer Institute] was used to label fixed fragments and frozen sections of testis. Zyxin staining was intense in Sertoli cell regions known to contain tubular regions of TBCs. Zyxin is comprised of a proline rich N‐terminal domain and three LIM domains in its C‐terminal half. Although known to interact with the cytoplasmic domain of integrins, zyxin may be part of the mechanism of tubulobulbar complex formation. Supported by an NSERC Discovery Grant to AWV.Grant Funding Source: NSERC
Purine biosynthesis and the case of the “missing” genes
Authors: Sarisky Catherine Ann , Smee Kevin M , Brown Anne M 
Published: June 2021

This work is a survey of known and suspected purine biosynthesis genes in sequenced archaea. We searched existing sequence databases for genes for the 11 steps (17 distinct genes) of de novo purine biosynthesis in 65 sequenced archaea, and carefully inspected each hit for the presence of active site residues and other residues known or suspected to be required for function. We found a number of problems with existing annotations, such as missing active site residues, fusions, and incorrect initiation sites. There is also evidence that the pathway has been shaped by horizontal gene transfer, duplication, and gene loss. Two groups of archaea lack a gene normally required for purine biosynthesis, suggesting that there are at least two more variants in this pathway to discover. We propose the existence of a gene encoding a GAR transformylase enzyme that is structurally unrelated to either known GAR transformylase, expected to be present in seven of the 65 archaea studied. A second proposed gene encodes a novel IMP cyclohydrolase function and is expected to be present in the Crenarchaea and Pyrococci.Functional characterization of some purine biosynthesis enzymes will also be reported.
Enzyme Kinetics of Native and Recombinant Secondary Alcohol Dehydrogenase originated from Micrococcus leuteus WIUJH20
Authors: Huang Jenq‐Kuen , Youngblut Nick D , Shoger Nicholas J , Takkalpalli Vamshi , Lin Chi‐Tsai , Wen Lisa 
Published: June 2021

A novel secondary alcohol dehydrogenase (2°‐ADH) from M. luteus WIUJH20 has been characterized and its gene cloned & expressed in this lab previously. This 2°‐ADH belongs to NAD+‐dependent short‐chain L‐3‐hydroxyacyl‐CoA dehydrogenase (SCADH) based on amino acid sequence homology. The native and recombinant 2°‐ADH have been purified to homogeneity. Both forms of the enzyme were soluble and were stable at −20°C which facilitated our kinetic studies toward various substrates. Our results showed that the 2°‐ADH has low substrate specificity: it catalyzed either the hydroxyl fatty acids or straight chain aliphatic secondary alcohols with hydroxyl group in either D or L form on a number of carbon positions; hydroxyl fatty esters were poor substrates; and cyclic aliphatic secondary alcohols and hydroxyacyl‐CoA were not substrates for both forms of 2°‐ADHs. To address the substrate specificities between the 2°‐ADH and SCADH, we have created three amino acids substitution mutants by site‐directed mutagenesis. The kinetic properties of the mutant proteins toward various substrates are compared to that of the wild‐type 2°‐ADH.This research is supported in part by Western Illinois University (WIU) Foundation and the College of Arts and Sciences.