bluuebunny/crossref_metadata_2025_split huggingface dataset:
Spironolactone increases myocardial performance and reduces right ventricular and atrial weights in spontaneously hypertensive rats
Authors: Cezar Marcelo Diarcadia Mariano , Damatto Ricardo , Martinez Paula , Lima Aline , Rosa Camila , Bonomo Camila , Guizoni Daniele , Blotta Daniella , Gimenes Rodrigo , Cicogna Antonio , Campana Alvaro , Okoshi Marina , Okoshi Katashi 
Published: June 2021

The purpose of this study was to evaluate the effects of spironolactone (SPR), started before clinical evidence of heart failure, on cardiac remodeling in spontaneously hypertensive rats (SHR).MethodsThirty six 13 month old SHR were divided into control and SPR (20 mg/kg/day for six months) groups. Systolic blood pressure (BP) was measured by tail‐cuff method. Echocardiogram (ECHO) was performed to evaluate in vivo cardiac structures and left ventricular (LV) function. Myocardial function was analyzed in LV papillary muscle during isometric contractions at 1.25 mM calcium concentration. Right ventricle, LV, and atria were weighed. Statistics: Student's t test and Fisher exact test.ResultsBP was not different between groups at the end of the experiment (control: 184±24; SPR: 197±22 mmHg; p>0.05). Mortality rate was lower in SPR group but without statistical difference (12.5% vs 25%, p>0.05). ECHO did not show differences in cardiac structures or LV systolic and diastolic function between groups. Myocardial function evaluated in vitro showed better functional performance in SPR group. Right ventricular and atrial weights were lower in SPR versus control groups.ConclusionAldosterone blockade started before clinical evidence of heart failure shows benefitial effects on myocardial performance and right ventricular and atrial weights in spontaneously hypertensive rats. Support: FAPESP and CNPq.
Myosin heavy chain changes in heart failure‐rat diaphragm muscle are not related to myogenic regulatory factors expression
Authors: Lima Aline Regina Ruiz , Martinez Paula F , Damatto Ricardo L , Cezar Marcelo D M , Guizoni Daniele M , Bonomo Camila , Zornoff Leonardo A M , Rosa Camila M , Campana Alvaro O , Silva Maeli Dal‐Pai , Okoshi Katashi , Okoshi Marina P 
Published: June 2021

ObjectiveIn this study we tested the hypothesis that diaphragm myosin heavy chain (MyHC) isoform changes are related to alterations of myogenic regulatory factors (MRF) expression in chronic heart failure.MethodsWe studied Sham (n=10) and infarcted rats with heart failure (MI/HF, n=10). Cardiac structures and ventricular function were assessed by echocardiogram. MyHC isoforms were analyzed by electrophoresis. MRF myogenin, MyoD, and MRF4 gene expression was evaluated by real time RT‐PCR and protein expression was analyzed by Western‐blot. Statistical analysis was performed by Student's t test and Pearson correlation.ResultsAll MI/HF rats had pulmonary congestion and right ventricular hypertrophy. In echocardiographic evaluation, MI/HF presented left cardiac chambers dilation and left ventricular systolic and diastolic dysfunction. The percentage of MyHC I isoform was higher and IIb isoform was lower in MI/HF. Left atrial diameter‐to‐body weight ratio positively correlated with MyHC I and negatively correlated with MyHC IIb isoform percentage. No differences were observed in MRF gene and protein expressions between groups.ConclusionMRF expression does not modulate diaphragm MyHC isoforms changes in rats with chronic heart failure. Diaphragm MyHC changes are related to ventricular dysfunction severity. Financial support: Fapesp.
Ghrelin Immunexpression in Pituitary Adenomas
Authors: Rotondo Fabio , Cusimano Michael , Scheithauer Bernd W , Rotondo Angelo , Syro Luis V , Latta Eleanor , Kovacs Kalman 
Published: June 2021

Ghrelin, an orexigenic hormone occurs mainly in stomach, and in several tumor types. We investigated ghrelin immunopositivity using the streptavidin‐biotin‐peroxidase complex method in the full spectrum of pituitary adenoma subtypes. Quantification was undertaken by analyzing both the percentage of tumor cells stained for ghrelin and the staining intensity.Cytoplasmic ghrelin immunopositivity was identified in several adenoma subtypes. Not every cell was immunopositive; the intensity of staining differed among cells within the given lesion and among the various tumors. The highest scores were noted in GH adenomas exposed to somatostatin analogues. In decreasing order, lower scores were encountered in ACTH‐producing, silent subtype 1 and 2 adenomas in Cushing disease, silent subtype 3 adenomas, untreated GH adenomas, dopamine agonist‐treated PRL adenomas, gonadotroph adenomas, and ACTH adenomas in Nelson syndrome. No immunoreactivity was found in TSH, untreated PRL‐producing, and null cell adenomas. The high immunoexpression of ghrelin in GH adenomas exposed to somatostatin analogues, may be due to increased ghrelin synthesis or to suppression of its release.
Concurrent plasma cells and eosinophils in endometrial biopsy specimens: Chronic endometritis or simply inflammatory response?
Authors: Symons Wendy A. , Hartman Sarah K. , Yeh I‐Tien 
Published: June 2021

Chronic endometritis is commonly diagnosed by identification of plasma cells that may be difficult to distinguish from stromal cells or lymphocytes. We correlate the concurrent presence of eosinophils and plasma cells by H&E and CD138 stains in endometrial biopsies.Methods100 consecutive endometrial biopsies were examined for eosinophils and plasma cells. Cases were stained with anti‐CD138 (mouse monoclonal, clone B‐A38; Cell Marque, Rocklin, CA). H&E and CD138 slides were reviewed, and the number of plasma cells and eosinophils/10 hpfs were recorded. The numbers of plasma cells and eosinophils were recorded again after re‐review of H&Es in areas with high numbers of CD138+ cells.Results90/100 cases were + for plasma cells by CD138. 57% of the CD138 immunostained subset contained eosinophils by H&E. Utilizing the CD138 stain to map the area of positivity, eosinophils were identified in the area as the plasma cells in 92% of cases and ranged from 1–61/10 hpf. Of the 10 cases lacking plasma cells by CD138, eosinophils were present in 10%.ConclusionEosinophils are a surrogate marker for the presence of plasma cells in endometrial biopsies. However, plasma cells are more frequently found than eosinophils: 52% contain eosinophils and 90% contain plasma cells. Both may be present as a result of a variety of inflammatory states and are often seen. Clinical correlation is critical for diagnosis of chronic endometritis.
Immunohistochemical Demonstration of Ghrelin in the Nontumorous Human Hypophysis
Authors: Rotondo Angelo , Rotondo Fabio , Scheithauer Bernd W , Sharma Soniya , Syro Luis V , Cusimano Michael , Kovacs Kalman 
Published: June 2021

Ghrelin, a 28 amino acid‐containing orexigenic peptide, is an endogenous ligand of the GH secretagogue receptor. It was extracted from the rat stomach, but is present in other organs. The aim of this work was to localize ghrelin by immunohistochemistry in autopsy‐derived non‐tumorous human pituitaries. Immunohistochemistry was performed using the streptavidin‐biotin‐peroxidase complex method and the results were analyzed independently by 3 of the authors. Double Immunostaining was also undertaken to determine which cell type was expressing both the adenohypophysial hormone and ghrelin.Results showed that ghrelin can be localized in the adenohypophysis. It's immunoexpression being cytoplasmic, weak to moderate, and present in only a subset of cells. The neurohypophysis is immunonegative. Double immunostaining showed that ghrelin is co‐localized with GH, ACTH and LH and is present to 51–90% of such cells. PRL, TSH and FSH cells are consistently non‐reactive. The role of ghrelin in adenohypophysial endocrine activity is not clear. It may well be that ghrelin exerts an autocrine‐paracrine effect and can modulate hormone synthesis and release.
Molecular Mechanisms of Antidepressant‐Induced Neurogenesis
Authors: Lakshmikuttyamma Ashakumary , Undieh Ashiwel S 
Published: June 2021

While induction of neurogenesis probably contributes to the clinical efficacy of antidepressant agents, it remains unclear whether antidepressants induce neural cell proliferation, promote neuronal differentiation, and/or enhance survival of newly differentiated neurons. This study addressed these mechanisms using mouse fetal cortical neuronal precursor cells and the RenCell human neural progenitor cell line. The antidepressants imipramine and fluoxetine concentration‐dependently increased proliferation of mouse and human precursor cells. Differentiation of precursor cells into neuronal phenotype was also enhanced at nanomolar drug concentrations. Flow cytometric analysis showed that 0.3uM fluoxetine significantly increased the proportion of precursor cells in the S phase of the cell cycle. Moreover, on examining the activities of genes regulating cell cycle progression from G1 to S phase, we observed that fluoxetine enhanced the expression levels of cyclin D1and cyclin E, while markedly decreasing the expression of the cyclin‐dependent kinase inhibitors p21 and p27. These results suggest that fluoxetine and other antidepressants may induce neuronal precursor cell proliferation and subsequent differentiation by suppressing cyclin‐dependent kinase inhibitors and concomitantly activating mitogenic signals.Supported by Thomas Jefferson University Foundation, Philadelphia, Pennsylvania.
Endothelial TRPC1 and TRPC3 channels contribute to ATP‐mediated relaxation in mouse cerebral arteries
Authors: Kochukov Mikhail Y , Noel Rebecca C , Abramowitz Joel , Birnbaumer Lutz , Marrelli Sean P 
Published: June 2021

Canonical transient receptor potential (TRPC) channels in endothelial cells (EC) may play an important role in regulation of vascular tone by mediating Ca2+ dependent relaxation. We tested the role of TRPC1 and C3 in ATP‐mediated Ca2+ signaling and vasorelaxation in mouse posterior cerebral artery (PCA).Experiments were performed on wild type (WT), TRPC1, and TRPC3 knockout (KO) mice. Isolated PCA were placed in a pressurized artery chamber and monitored for changes in tone by video‐microscopy. ATP was delivered to the endothelium through the artery lumen. Intracellular calcium ([Ca2+]i) was measured by Fura 2 in primary cultures of aorta EC.PCA from TRPC1 and C3 KO mice developed tone and constricted to endothelin‐1 comparably to WT, but demonstrated impaired vasodilation responses to ATP. In aorta EC, ATP produced an increase in [Ca2+]i insensitive to P2X antagonist NF279 but susceptible to inhibition by TRPC modulator La3+ or PLC blocker U73122. ATP‐induced changes in [Ca2+]i were similar in WT and C3 KO but reduced in C1 KO EC. Pyr3, a TRPC3 blocker, inhibited the [Ca2+]i response significantly in WT but not C3 KO, demonstrating a role of TRPC3 in EC.Our data support a critical role for TRPC1 and TRPC3 in endothelium‐dependent relaxation to ATP in cerebral arteries. TRPC1 and C3 both appear to be functionally important to ATP‐mediated Ca signaling in aorta EC.Supported by NIEHS Z01‐ES‐101684 and NIH R01 HL088435
The effects of structurally disparate Kv7 activators in the rat vasculature
Authors: Jepps Thomas Andrew , Hansen Rie Schultz , Olesen Søren Peter , Greenwood Iain Andrew 
Published: June 2021

Kv7 voltage‐gated potassium channels, encoded by KCNQ1–5 genes, are present functionally in vascular and non‐vascular smooth muscle cells. The aim of the present study was to compare the effect of structurally different activators of Kv7.2–7.5 channels: S‐1, BMS‐204352, ICA27243 and retigabine in the rat mesenteric artery, thoracic aorta and coronary circulation. Isometric tension recording was performed on segments of mesenteric artery and thoracic aorta. In addition the coronary circulation was studied using the Langendorff heart preparation. In mesenteric arteries the Kv7 activators demonstrated EC50 values of: 1.9 μM (BMS204352), 2.5 μM (S‐1), 5.4 μM (ICA27243), and 14.3 μM (retigabine). Application of all Kv7 activators at 10 μM relaxed the thoracic aorta albeit with varying efficacies. In the isolated heart increasing concentrations of Kv7 activators produced a concentration‐dependent increase in coronary flow. This study is the first to fully characterise the effects of various Kv7 channel activators in the rat mesenteric artery, thoracic aorta and coronary artery.Work was funded by BBSRC‐CASE studentship with NeuroSearch A/S.
Ferula hermonis root extract increases fiber size and myonuclear number within tibialis anterior muscle of exercised rats
Authors: Allouh Mohammed Zohair 
Published: June 2021

Ferula hermonis Boiss. is an aphrodisiac plant that grows in the Mediterranean region. It has been reported that acute treatment with the extract from the root of this plant significantly increases serum testosterone in the rat. This study investigates the effects of F. hermonis extract alone or combined with exercise on rat skeletal muscle fibers. Adult male rats were divided into four groups: control sedentary (CS) that had no treatment or exercise; ferula sedentary (FS) that orally treated with ferula extract at a dose of 60 mg/kg/rat every other day over a period of 20 days; control exercised (CE) that trained by swimming for 40 minutes every other day; and ferula exercised (FE) that received combined treatment and exercise. At the end of experiments, the left tibialis anterior muscle was dissected for study from each rat. Fiber diameter, myonuclear number and myonuclear domain size were measured by using immunofluorescent techniques and software analyses. FE group showed significant (P<0.05) increases in muscle weight, fiber size and myonuclear number compared to the remaining groups. However, no significant variations were found among CS, FS and CE groups. This study concludes that exercise combined with administration of F. hermonis extract is more effective in enhancing the growth of skeletal muscle fibers than exercise alone. Funds for this study were provided by the Deanship of Research at JUST (34/2010).Grant Funding Source: JUST
Calcium signaling regulates gravin‐dependent compartmentalization of PKA
Authors: Schott Micah Bruce , Grove Bryon 
Published: June 2021

Gravin (AKAP12) is a multivalent scaffolding protein that anchors PKA to the plasma membrane, but undergoes redistribution to an intracellular compartment in response to PKC activation or increased [Ca2+]i. We postulate that through this mechanism gravin may mediate cross‐talk between Ca2+‐dependent and cAMP‐dependent signaling pathways. To assess this, we examined the effect of Ca2+‐elevating agents on gravin and gravin‐dependent PKA distribution in cells lacking endogenous gravin but expressing EGFP/ EYFP‐gravin constructs. Ionomycin and thapsigargin initiated rapid gravin redistribution into the cytosol, an event which was inhibited by Ca2+ chelation using BAPTA‐AM. Treatment of cells coexpressing gravin‐EYFP and PKA RII‐ECFP with Ca2+‐elevating agents also resulted in PKA redistribution, but only when gravin was present. Consistent with this, treatment with lysophosphatidic acid (LPA) also resulted in rapid gravin redistribution in concert with a rise in [Ca2+]i and resulted in PKA redistribution in cells coexpressing gravin and PKA constructs. However, inhibition studies using BAPTA‐AM and bisindolylmaleimide suggest that in addition to changes in [Ca2+]i, LPA‐mediated gravin dynamics may involve other mechanisms. Studies to further elucidate the mechanism behind LPA‐mediated gravin dynamics are currently underway. This work was supported by funding from UNDSMHS and NIH 5P20RR017699.Grant Funding Source: Institutional/NIH
Ethanol‐induced oxidative stress in oteoblast‐like cells (UMR 106‐01 BSP)
Authors: Joseph Eugene E , Garcia Beronica 
Published: June 2021

Ethanol is known to induce oxidative stress which has been associated with cancer, neurodegenerative and cardiovascular diseases and osteoporosis. Oxidative stress adversely affects bone cells leading to decreased mineralization of bone. Many studies have been done documenting the effect of ethanol on bone. However, the cellular mechanisms leading to the effects of ethanol‐induced oxidative stress and demineralization in bone cell cultures are incompletely understood. In this study, it was hypothesized that ethanol decreases mineralization in UMR cells by inducing oxidative stress. UMR cells were treated with varying concentrations of ethanol and incubated at 37 °C for 60 hours. The cell cultures were treated with 7mM beta‐glycerophosphate for 24 hours to stimulate mineralization. Cell cultures were stained with 40 mM alizarin red to assess mineralization. Mineral content was extracted with 10 mM HCl/70% ethanol and spectrophotometrically quantified using a microplate reader (Spectramax 190). Vector Red staining was also done to test for alkaline phosphatase (ALP). Lactate dehydrogenaes (LDH) concentrations were measured colorimetrically using an LDH bioassay kit. F test ANOVA was used to compare variances and all values in the results were expressed as means ±SD. In all cases, p values of <0.05 were considered statistically significant. The results showed an inverse correlation of ethanol concentrations to both mineral content and alkaline phosphatase concentrations, while a concomitant increase of LDH concentrations was noted with the increasing ethanol concentrations. These results suggest that ethanol causes induced oxidative stress by affecting ALP and LDH activities, leading to an overall demineralization in UMR cell cultures.(Supported by the Dean's Office of the College of Arts and Sciences of La Sierra University, Riverside, CA).
Distribution of mitochondrial chaperonins in lung cells
Authors: Cappello Francesco , Corrao Simona , Anzalone Rita , Magno Francesca , Loria Tiziana , Iacono Melania Lo , Farina Felicia , Zummo Giovanni , de Macario Everly Conway , Macario Alberto JL , La Rocca Giampiero 
Published: June 2021

Hsp60 and Hsp10 are considered typical mitochondrial chaperonins, residing inside the organelle, but works in the last years showed they can also be outside mitochondria and cells. The exact extramitochondrial locations of the chaperonins in the various normal cells and tissues under diverse conditions remain to be ascertained. Consequently, we undertook to examine the subcellular distribution of Hsp60 and Hsp10 in lung cells in order to provide a baseline for comparison between normal and pathological tissues. We used a battery of methods (light and electron microscopies, immunocytochemistry, subcellular fractionation‐purification plus electrophoresis, Western blotting) to study two cell types, lung fibroblasts and epithelial cells, unstressed and exposed to the stressor cigarette smoke extract, which is pertinent to lung pathology. Here we focus on Hsp10, which was found in the mitochondria, cytosol, and nucleus, with their levels changing after stress. Nuclear Hsp10 has not been reported before and was demonstrated in various ways, including immunoelectromicroscopy with specific antibodies.Grant Funding Source: University of Palermo
Novel transcript variants arise from alternate promoter usage and alternative splicing in 5′ regions of cAMP‐dependent Protein kinase A subunit genes in mouse
Authors: Banday Abdul Rouf , Azim Shafquat , Tabish Mohammad 
Published: June 2021

Protein kinase A (PKA) is composed of four membered structure with two regulatory (R) and two catalytic (C) subunits. There are four different R subunits (Iα and Iβ, IIα and IIβ) and two different C subunits (Cα and Cβ) in mouse, each encoded by separate gene. Earlier, few alternative spliced variants have been identified with tissue specific function. The objective of this study was to analyze splicing events in 5′ region of R and C subunit genes of PKA in mouse.Analysis was done with the help of combinatorial approach of bioinformatics tools and molecular biology techniques like gene and exon prediction programmes, ORF finders, EST searches, alignment tools, 5′ RACE, RT‐PCR, sub‐cloning, sequencing etc. We were able to sequence two types of cDNA clones from mouse tissues, one type having alternative 5′ untranslated exon (s) and other type having alternative 5′ first coding exon. In former the variants arise either from alternate promoter usage or splicing of 5′ untranslated exons and in later new exon replaces the canonical first coding exon to exhibit different N‐terminal to translated protein. However, alternate promoter usage is mostly linked to regulatory subunit genes.The numbers of new transcript variants sequenced were four in Cα, two in Cβ, three in R Iα and four in R Iβ. Their expression profiles showed variation across the tissues together at different development stages. The research was funded by DBT, India.
ERAD and autophagy cooperatively facilitate clearance of aggregation‐prone mutant Surfactant Protein C
Authors: Maguire Jean Ann , MacNabb C. Marshall , Guttentag Susan H. , Beers Michael F. 
Published: June 2021

We have previously shown that mutations within the BRICHOS domain of Surfactant Protein C (SP‐C), a conserved C‐terminal region of ~100 amino acids found in a number of degenerative disease‐related proteins, lead to UPR activation, cytosolic aggregation, and activation of caspase‐mediated apoptotic pathways. We hypothesized that in addition to upregulation of quality control mechanisms, SP‐C BRICHOS mutants would induce autophagy as a means of cytoprotection. To study this, A549 and HEK293 cells were transfected with human SP‐CΔexon4 mutant. Expression induced autophagosome formation, assessed by detection of LC3 isoform conversion, and by alteration of the intracellular localization of GFP‐tagged LC3. These results were also observed in primary human alveolar type II cells. Inhibition of either the 26S proteasome with MG‐132, or autophagy by 3‐methyladenine resulted in a significant increase in expression levels of mutant SP‐C, suggesting both pathways play a prominent role in mutant clearance. Inhibition of autophagy in cells expressing SP‐CΔexon4 also led to pronounced Caspase 3 activation, while augmentation of autophagy through rapamycin treatment enhanced the clearance of mutant protein. These results indicate that macroautophagy is important for lung epithelial cell homeostasis via its participation in quality control responses activated by aggregation‐prone BRICHOS mutants.Funding: HL19737
Folding of individual subunits of AcrB before trimerization
Authors: Zhong Meng , Lu Wei , Wei Yinan 
Published: June 2021

AcrB and its homologous are the inner membrane components of the major multidrug transporter in Gram‐negative bacteria. In Escherichia coli, AcrB recognizes and exports a broad range of functionally and structurally unrelated substrates. The pumps confer elevated drug tolerance to bacteria, thereby increasing the difficulty to cure infections caused by these bacteria. AcrB exists and operates as a homotrimer. To investigate to what extent each individual subunit folds before they oligomerize, we created a reporter platform to detect subtle conformational changes in AcrB in vivo. We expect this thiol‐trapping based method to be generally useful in the study of many other inner membrane proteins that contain periplasmic domain. Taking advantage of this reporter platform, we created an AcrB mutant that existd as a monomer in the cell membrane. The mutant AcrB was not functional, but folded into a conformation highly resembling that of the wild type AcrB. The success in the creation of a well‐folded AcrB monomer suggests that after translation, each individual subunit folds to a conformation very similar to its final state before trimerization occurs.The work was supported by faculty start‐up fund to Wei Y. and RCTF fellowship from University of Kentucky.