bluuebunny/crossref_metadata_2025 huggingface dataset:
The distinct in vivo roles of the long‐form and short‐form of glutathione peroxidase 4: the short‐form is essential for somatic cell functions and the long‐form is important for male fertility
Authors: Liang Hanyu , Yoo Si‐Eun , Na Ren , Walter Christi , Richardson Arlan , Ran Qitao
Published: June 2021

Glutathione Peroxidase 4 (Gpx4) is an essential antioxidant enzyme that protects cells against membrane lipid peroxidation. The Gpx4 gene encodes for three isoforms: a long‐form (lGpx4), a short form (sGpx4) and a nuclear form. Genetic studies indicate that the nuclear form of Gpx4 plays a role in sperm chromatin condensation. However, the in vivo roles of lGpx4 and sGpx4, which are distinguishable by the presence or lack of a mitochondrial targeting signal peptide at the N‐terminal of the Gpx4 protein, remain unclear. In this study, we generated transgenic mice using mutated human GPX4 genes encoding either lGPX4 [Tg(lGPX4) mice] or sGPX4 [Tg(sGPX4) mice]. Tg(sGPX4) mice had increased total Gpx4 proteins in all tissues whereas Tg(lGPX4) mice had increased total Gpx4 proteins only in the testes. Tg(sGPX4) mice, but not the Tg(lGPX4) mice, were protected against diquat‐induced liver apoptosis. By cross‐breeding the Tg(sGPX4) mice with the Gpx4 knockout mice, we showed that the sGPX4 transgene was able to rescue the lethal phenotype of mouse Gpx4 null mutation. In contrast, the lGPX4 transgene failed to rescue Gpx4 null mice. Gpx4 protein expression data from the sGPX4‐rescued Gpx4 null mice indicated that sGPX4 was the major isoform in the somatic tissue subcellular fractions including mitochondria whereas lGPX4 was predominant in testes. The male sGPX4‐rescued mice were infertile and exhibited low sperm count and reduced sperm mobility, suggesting lGPX4 plays a role in male fertility. Together, our results indicate that the sGpx4 is the major isoform in somatic tissues and it is essential for survival and protection against apoptosis; the lGpx4 is predominant in the testes and it is important in regulating male fertility.
Excitation of the genioglossus in REM sleep
Authors: fraigne jimmy , Orem John
Published: June 2021

Although attention has focused on atonia of the genioglossus (GG) in rapid eye movement (REM) sleep, the fact is that, as shown here in the rat, the overall level of activity of the muscle is greater in REM than in NREM sleep. Three Wistar rats were anesthetized and electrodes were implanted to record electroencephalogram (EEG) and electromyograms (EMG) of the diaphragm, GG and neck muscle. After recovery and adaptation, the animals were recorded during REM sleep. GG activity emerged on an atonic background an average of 20s (±2 SEM) after the onset of REM sleep and was present for 71% (±6 SEM) of the REM duration. This activity developed to a peak 2‐3 fold higher than the average level of activity present during the preceding NREM period. The profile of GG activity, obtain from 88 REM periods, was unimodal with a peak in the second half of the REM period. GG activity correlated with an increased diaphragmatic muscle activity. However, nuchal activity did not present a similar pattern of activity. These results indicate that there is an excitatory drive affecting specifically the respiratory system, including the genioglossus muscle, during REM sleep in rat.
A SUMMER ELECTIVE COURSE IN DISSECTION IN GROSS ANATOMY AS A METHOD OF INTEGRATING CLINICAL AND BASIC SCIENCE EDUCATION
Authors: Zill Sasha N
Published: June 2021

Following a curriculum revision, we have taught Gross Anatomy to medical students in a prosection‐based course. The opportunity for experience in cadaver dissection has been offered in a summer elective course (six weeks duration). The only course requirement is the complete dissection of one body region and grading is pass/fail. Review of didactic material is self‐paced and self‐directed. In addition, clinical faculty (Emergency medicine, Orthopedics, Surgery) have given lectures on the anatomical basis of procedures and surgical approaches. These lectures are accompanied by laboratory sessions in which 1) students perform procedures (ex. thoracotomies) or 2) physicians demonstrate surgical approaches on cadaver specimens. The response of students has been enthusiastic and approximately half the class has attended the course. Specimens generated in the dissection course have also been used as prosections in our main course in Gross Anatomy. This approach is advantageous in 1) it provides a review of material presented in a compressed format 2) it allows students to appreciate the three dimensionality of body structure and 3) it provides experiences requisite for subsequent clinical training. The summer dissection course, thus, provides a direct method of integrating clinical and basic science education.
SNAP‐tag, A Multipurpose Visualization Tool for Studying Protein Dynamics in Living Cells
Authors: Sun Luo , Zhang Aihua , Ghosh Inca , Xu Ming‐Qun
Published: June 2021

The creation of cell lines expressing proteins fused to intrinsically fluorescent tags (e.g., GFP) and their use in cell based studies of protein localization have become routine. The fluorescent signal from such tags cannot be routinely switched on at will, precluding some time‐resolved studies of expression, localization and degradation.

A complementary and more flexible imaging system based on a family of small, self‐labeling protein tags is being introduced by New England Biolabs. SNAP‐tag and CLIP‐tag, orthogonal tags with a variety of fluorescent substrates, are suitable for imaging intracellular and cell surface proteins in live cells. These tags can also be used for time‐resolved dual labeling, highly efficient covalent pull‐downs and other biochemical assays. Furthermore, a collection of non‐fluorescent substrates that block SNAP‐ and CLIP‐tag reactivity enable pulse‐chase studies and an examination of the temporal dynamics of nascent protein synthesis and complex formation in live cells.

In this report, several studies of protein localization, turnover and receptor internalization using SNAP‐ and CLIP‐tags in living cells are demonstrated. Using two fluorescent dyes in pulse‐chase experiments, we examined the turnover of several intracellular proteins including the Cytochrome c oxidase subunit 8A (COX8A) and the Transferrin Receptor. We demonstrate that fluorescent protein labeling using SNAP‐tag and CLIP‐tag provides a powerful tool to study protein localization, turnover and internalization.
Identification and characterization of the human N‐acetyltransferase complexes hNatB and hNatC.
Authors: Starheim Kristian Kobbenes , Arnesen Thomas , Gromyko Darina , Ryningen Anita , Evjenth Rune , Varhaug Jan , Lillehaug Johan
Published: June 2021

Protein N‐alpha‐terminal acetylation is a conserved and widespread protein modification in eukaryotes. Several studies have linked it to normal cell function and cancer development, but nevertheless, little is known about its biological function. In yeast, three major complexes, NatA, NatB and NatC catalyse near all N‐alpha‐terminal acetylation, acetylating specific subsets of proteins. In humans, the NatA complex is the most extensively studied. We here present the human NatB and NatC complexes (hNatB and hNatC). They are conserved from yeast with respect to subunit composition, substrate specificity and ribosome binding. Knockdown of hNatB subunits disrupts normal cell cycle progression, and induces growth inhibition in HeLa cells and the thyroid cancer cell line CAL‐62. Knockdown of hNatC subunits results in p53‐dependent cell death and reduced growth in human cell lines. Taken together, these studies emphasize the biological importance of N‐alpha‐terminal acetylation.
Phosphatidylethanolamine Is Required for Normal Cell Morphology and Cytokinesis in the Fission Yeast, Schizosaccharomyces pombe
Authors: Marcus Stevan , Luo Jun , Matsuo Yasuhiro , Gulis Galina , Patton‐Vogt Jana
Published: June 2021

We describe the characterization of three predicted genes, designated psd1‐3, in the fission yeast, Schizosaccharomyces pombe, encoding phosphatidylserine decarboxylases (PSDs), which catalyze the conversion of phosphatidylserine to phosphatidylethanolamine (PE). While S. pombe mutants carrying deletions in any one or two psd genes are viable in both rich and minimal media, psd1‐3δ mutants carrying deletions in all three psd genes grow slowly in rich medium and are inviable in minimal medium, indicating that the psd1‐3 gene products share overlapping essential cellular functions. Supplementation of growth media with ethanolamine, which can be converted to PE by the Kennedy pathway, restores growth to psd1‐3δ cells in minimal medium, indicating that PE is essential for S. pombe cell growth. psd1‐3δ cells appear morphologically indistinguishable from wild type S. pombe cells in medium supplemented with ethanolamine but when cultured in non‐supplemented medium produce high frequencies of abnormally shaped cells as well as cells exhibiting severe septation defects, including multiple, mispositioned, deformed, and misoriented septa. Our results demonstrate that PE is essential for cell growth and for normal morphology and cytokinesis in S. pombe and illustrate the usefulness of this model eukaryote for investigating potentially conserved biological and molecular functions of PE.
Effects of oligomeric state on catalysis by UDP‐glucose dehydrogenase
Authors: Neely Willis Melanie , Easley Katherine E , Barycki Joseph J
Published: June 2021

Human UDP‐glucose dehydrogenase (UGDH) is an enzyme that catalyzes the conversion of UDP‐glucose to UDP‐glucuronate by two successive oxidation reactions. Previous studies have highlighted the importance of several conserved active site residues and many of the mechanistic details have been examined. Recent studies indicate that wild‐type UGDH exists as a hexamer, composed of a trimer of catalytically active dimers. Examination of the available UGDH crystal structure suggests that disruption of its oligomeric state may impact its enzymatic activity. To test this hypothesis, we have designed, expressed, and purified two UGDH mutants, E110A and T325A. Both mutants were found primarily in the dimeric state by gel filtration and sedimentation velocity experiments. Importantly, these engineered dimer mutants have kinetic constants comparable to wild‐type enzyme. The E110A and T325A mutants were subsequently used to examine the importance of the hexameric structure using cell culture models. Results of these studies suggest that the intact hexamer is required for optimal UGDH activity.
Non‐canonical Wnt signaling regulates the stability of xSyndecan4
Authors: Escobedo Noelia A. , Carvallo Loreto , Munoz Rosana , Larrain Juan
Published: June 2021

The non‐canonical or beta‐catenin independent Wnt signaling pathway regulates convergence and extension movements in vertebrate embryos and planar cell polarity. Recently in our laboratory, we have demonstrated that a cell surface heparan sulphate proteoglycan, Syndecan4 (xSyn4), is critical for Xenopus laevis gastrulation and is a component of the non‐canonical Wnt signalling pathway. We have found that xSyn4 is expressed in the same tissues and interacts functionally and biochemically with Frizzled‐7 (xFz7) and Dishevelled (xDsh). Furthermore, xSyn4 is necessary and sufficient to activate translocation of xDsh to the plasma membrane. In this work we have used methodologies such as GST‐pulldown and immunoprecipitation in HEK293T cells in order to investigate the interaction between xSyn4, xDsh and xFz7. We have observed that xDsh and xFz7 interact with xSyn4 cytoplasmic domain. Also, we have found that xSyn4 stability is regulated by activation of the non‐canonical Wnt pathway and that the cytoplasmic domain is required for such effect.

This work is supported by FONDECYT Post‐Doctoral Fellowship 3070015 to L.C. and the Center for Cell Regulation and Pathology (FONDAP).
Role of S100A8 and S100A9 proteins on breast cancer aggressivity
Authors: Cormier Kevin A , Fournier Sebastien , O'Brien Jeannick , Tessier Philippe , Robichaud Gilles A
Published: June 2021

Women are more affected by breast cancers than any other type of cancer. Studies show that proteins from the S100 family are differentially expressed in cancers and autoimmune disease. The S100 protein family is the largest family of calcium binding proteins where most members are overexpressed in certain types of cancers. More specifically, S100A8 and S100A9 seem to be overexpressed in mammary ductal carcinomas. Nevertheless, little is known on the cellular effects of S100 proteins and their roles in breast cancer pathogenesis.

In this study, we assessed cancerous biological processes (proliferation, apoptosis, cell adherence and ECM invasion) in mammary cells following: 1) the treatment with various concentrations of recombinant and purified S100A8 and S100A9 proteins and, 2) the knocked‐up or knocked‐down expression of the S100A8 and S100A9 genes. We found that extracellular treatments with S100 proteins affected growth rates of cancer cells. We also observed that the conditioned expression of S100 protein in breast cancer cells affected cancer processes in a cell type dependent manner. Details from these results will be discussed here.

These results will aid to a better understand the roles of S100A8 and S100A9 proteins in epithelial cell biology and breast cancer pathogenesis. The outcome of these experiments could potentially lead to a better diagnostics or more specialized treatments of breast cancers.
Effect of the Ala391Glu mutation on FGFR3 activation in cellular membranes
Authors: Chen Fenghao , Horton William , Hristova Kalina
Published: June 2021

Fibroblast growth factor 3 (FGFR3), a member of the receptor tyrosine kinase (RTK) family, is an important mediator of cellular signaling. Signal transduction is achieved by receptor dimerization, phosphorylation and stimulation of downstream signaling cascades. Single amino acid mutations can affect the signaling and lead to overactivation and pathologies. Ala391Glu is one FGFR3 pathogenic mutation, causing crouzon syndrome with acanthosis nigricans. We investigate the effect of this mutation in the context of full length FGFR3 in 293T cell, and in the presence of the ligand acidic fibroblast growth factor (FGF1). By fitting the Western blot data to a thermodynamic model describing an apparent equilibrium between inactive monomers and active dimers, we find that the Ala391Glu mutation increases the activation prospensity by about ‐2 kcal/mole.
Rho Residue 43 is a Structural Determinant of GEF Activity and Selectivity
Authors: Goetzinger Chris J. , Sloan Christina M. , Quinn Clancy V. , Ellerbroek Shawn M.
Published: June 2021

RhoA, RhoB, and RhoC GTPases share over 80% sequence identity but diverge in cellular function. In previous work, we hypothesized that variation at amino acid residue 43 between RhoA/B (valine) and RhoC (isoleucine) was a structural impediment to XPLN activation of RhoC. In this work, we have systematically studied a putative role of Rho residue 43 as a structural determinant of guanine nucleotide exchange factor (GEF) activity. Specifically, we performed in vitro analysis of GEF activity (VAV2, XPLN, Dbs, GEFT, and Dbl) against both normal RhoA/B/C proteins and those mutated at residue 43. Substitution of val43 with an isoleucine in RhoA reduced 40‐50% of GEF catalyzed exchange, except for GEFT (reduced only 10‐20%). Substitution of val43 with an isoleucine in RhoB had variable effect on exchange, with Vav2 and GEFT modestly sensitive (10% decrease) and XPLN‐stimulated exchange reduced 80%. On the other hand, substitution of ile43 with a valine in RhoC promoted all GEF catalyzed exchange (1.3‐4.0 fold increases). Substitution of a polar threonine at residue 43 significantly reduced all RhoA GEF exchange 60‐80%, nearly eliminated RhoB GEF exchange (80‐100% decrease), but had little to no affect on RhoC exchange, with the exception of GEFT (reduced 75%). In support, RhoA V43T displayed a reduced phenotype when expressed in NIH3T3 fibroblasts, whereas constitutively active RhoA V43T, 63L was phenotypical equivalent to RhoA 63L. Together, these findings support the hypothesis that a divergence at residue 43 between RhoA/B and RhoC is a structural determinant of GEF activity and selectivity. This work was supported with funds from the RJ McElroy Trust.
A Targeted Therapeutic Rescues Botulinum‐A Poisoned Neurons
Authors: Zhang Peng , Ray Radharaman , Singh Bal Ram , Li Dan , Adler Michael , Ray Prabhati
Published: June 2021

Botulinum neurotoxin (BoNT) is the most poisonous poison known to mankind for which currently no antidote is available to rescue poisoned nerves. An effective medical countermeasure strategy will be required and would include its efficient delivery specifically to presynaptic nerve terminals. Here we present the report of rescue of botulinum poisoned nerve cells by Mastoparan‐7, a peptide constituent of bee venom that was delivered through a drug delivery vehicle (DDV) constructed from the non‐toxic fragment of botulinum neurotoxin itself. We found that mastoparan‐7 that was delivered into spinal cord cells rescued BoNT/A poisoned cells by restoring over 40% neurotransmitter release. The rescue of the cell poisoning does not seem to occur from inhibition of the endopeptidase activity of BoNT/A against its well known substrate, SNAP‐25, mechanistically involved in the exocytosis process. Rather, Mastoparan‐7 induces a physiological host response apparently unrelated to SNAP‐25 but linked to the phospholipase signal transduction pathway. Our results open new avenues to examine mechanism of exocytosis process, and also to examine alternative cellular mechanism of botulinum neurotoxin action. Successful demonstration of BoNT‐based DDV opens door for its utility for drug delivery against many neuronal and neuron‐muscular disorders.

This work was supported JSTO‐CBD/DTRA #3.10003_05_WR_B
The Novel Role of Guanylyl Cyclase C Signaling in Appetite Suppression and Body Weight Regulation
Authors: Valentino Michael , Lin Jieru , Li Peng , Snook Adam , Pitari Giovanni , Schulz Stephanie , Waldman Scott
Published: June 2021

Guanylyl cyclase C (GCC) is an intestine‐specific transmembrane receptor that synthesizes cGMP as its proximal effector. Recently, GCC has emerged as an important intermediary in signaling programs controlling appetite and body weight. C57/BL6 mice in which GCC signaling was eliminated exhibited excess body weight which was associated with adipocyte hypertrophy, an increase in subcutaneous and visceral adipose mass, and obesity‐related co‐morbidities including hepatic steatosis, hyperinsulinemia, and glucose intolerance. Furthermore, GCC‐deficient mice exhibited hyperphagia, and their excess weight gain was eliminated by restricting food intake to levels consumed by wild‐type mice. The observed hyperphagia correlates with a defect in gut pathways regulating food intake, as GCC‐deficient mice exhibit ~50% reduction in the enteroendocrine satiety hormones cholecystokinin and glucagon‐like peptide within the jejunum and ileum, respectively. These observations demonstrate a previously unrecognized role for GCC signaling in regulating appetite and body weight by modulating the function of enteroendocrine cells. This highlights a novel therapeutic paradigm in which oral hormone supplementation with GCC ligands could enhance enteroendocrine hormone levels and amplify nutrient satiety responses, thereby restricting appetite and defending against obesity. Supported by the PhRMA Foundation
Therapeutic Effect of Ginkgo Biloba Extract on Rat Heart Failure Induced by Isoprenaline
Authors: Yi Hongwei , Wang Yongqing , Ding Fuyun , Meng Ling , Wu Xudong
Published: June 2021

Objective

To investigate the effect of Ginkgo biloba extract (GBE) on rat heart failure induced by isoprenaline and its mechanism.

Methods

The SD rat heart failure was induced by isoprenaline administrated subcutaneouly for 11 days. 100 mg/kg/d GBE and 10 mg/kg/d Metoprolol were pretreated 3 days before isoprenaline. The index of haemodynamics and serum contents were detected.

Results

Isoprenaline increased the heart index and left ventricle index significantly, but GBE and Metoprolol could alleviate rat myocardial hypertrophy. GBE had stronger effect on inhibition of the increase in AST and LDH than Metoprolol; on the contrary, Metoprolol was more potential for decreasing creatine kinase, MB isoenzyme of creatine kinase and hydroxybutyrate dehydrogenase level. Isoprenaline decreased the level of LVSP, LVEDP, LV +dp/dt max and LV ‐dp/dt max and increased heart rate. Metoprolol and GBE blocked the effects of isoprenaline respectively, but there is not significant difference between GBE and Metoprolol. GBE and Metoprolol inhibited the increase in mRNA levels of AMP‐activated protein kinase alpha 1, 2 and ryanodine receptor 1 induced by isoprenaline significantly.

Conclusion

GBE attenuated the damage of cardiac muscle, improved the dysfunction of rat heart induced by isoprenaline. Not like Metoprolol, a beta receptor blocker, GBE exerts a different mechanism in curing heart failure.
Role of mouse CYP2A5 in nicotine clearance and testosterone homeostasis
Authors: Ding Xinxin , Zhou Xin , Xie Fang
Published: June 2021

CYP2A5, a mouse P450 enzyme, is expressed abundantly in the olfactory mucosa (OM), and in other tissues, including the liver and the lateral nasal gland (LNG). To determine the biological and pharmacological functions of CYP2A5 in vivo, we have generated a Cyp2a5‐null mouse. Homozygous Cyp2a5‐null mice are viable and fertile. OM microsomes from Cyp2a5‐null mice had significantly lower activities in the metabolism of testosterone (T) than did OM microsomes from wild‐type (WT) mice. In contrast, hepatic microsomes from Cyp2a5‐null mice showed only small decreases in the overall rates of formation of T metabolites, compared to hepatic microsomes from WT mice. Serum T levels were not altered by the loss of CYP2A5 function in male mice, a result confirming that CYP2A5 does not play a major role in systemic T clearance in vivo. Nevertheless, loss of CYP2A5 was accompanied by significant increases in tissue levels of T in the LNG, a gland important for nasal mucus secretion and production of odorant binding proteins. The Cyp2a5‐null mice also showed decreased clearance of nicotine, with significant increases in the half life and AUC values for serum nicotine, compared with WT mice. These findings indicate that CYP2A5 is the major nicotine oxidase in the mouse liver, and that CYP2A5 plays an important role in the regulation of T levels in the LNG of male mice. (Supported in part by NIH grant ES‐07462)
Reduction of the trans fatty acid absorption in Caco‐2 cell by grain extracts.
Authors: Lee Sung‐Joon , Jeun Jungae , Cho Hee‐Jun , Shin Ki‐Yong
Published: June 2021

Reduction of the trans fatty acid absorption in Caco‐2 cell by grain extracts. Jungae Jeun, Hee‐Jun Cho, Ki‐Yong Shin, Sung‐Joon Lee

Intake of trans‐fatty acids could increase LDL‐cholesterol and reduce HDL‐cholesterol levels thus may increase the risk for coronary diseases. We investigated the inhibition of grain extracts on the elaidic acid (t18:1) uptake, predominant trans‐fatty acid in the processed foods. Water, ethyl acetate, and hexane extracts from white and black soybeans, barley, brown rice, and wheat powder were used in the study. The composition profile of each extract was analysed by GC‐MS analysis and the effects of each extracts on the elaidic acid absorption on Caco‐2 cells were measured by GC analysis. RT‐PCR and western blot were performed to assess gene expression in the cellular fatty acid uptake. Hexane extracts of black soybean was the most effective and inhibited 15% of elaidic acid uptake, and down regulated the expression of CD36 and FATP2 by 54% and 49%, respectively. ACS1, FABP, NPC1L1 were marginally altered. These suggest that soybean may reduce trans‐fatty acid uptake by altering the expression of fatty acid transporters in the small intestine.
Statins Improve Hemodynamic Status in STZ‐Diabetic Rats.
Authors: Quidgley Jose , Cruz Nildris , Creagh Osmar , Torres Hector , Crespo Maria J.
Published: June 2021

Statin therapy has been associated with improved cardiac status in normocholesterolemic diabetic patients. Whether this effect is drug‐specific or class‐dependent remains unknown. In this study, we compared the effects of the daily administration of pravastatin (PVT, 10 mg/kg/day) and simvastatin (SVT, 10 mg/kg/day) on cardiac function in streptozotocin (STZ)‐diabetic rats. We administered these drugs over a 4‐week period. Systolic blood pressure (SBP) and echocardiographic parameters were evaluated. SBP was significantly higher in diabetic rats than in age‐matched non‐diabetic rats. SVT decreased SBP in diabetic rats by 21% (P<0.05), whereas PVT did not modify this parameter. SVT increased stroke volume (from 0.17 ml to 0.33 ml, n=5, P<0.05) and cardiac output (n=5, P<0.05). PVT increased ejection fraction by 46% (n=5, P<0.05) and reduced LVESV from 0.18 ml to 0.06 ml (n=5, P<0.05), but did not improve cardiac output (P>0.05). These results suggest SVT and PVT have a positive hemodynamic effect in diabetic rats. The underlying mechanisms of action, however, are different; whereas SVT improves heart function by reducing afterload, PVT improves cardiovascular status by increasing systolic function.
Osteologic exploration of the clavicle: a new approach
Authors: Bortier Hilde Elisa , Bernat Amit , Huysmans Toon , Van Glabbeek Francis , sijbers jan , Pinho Romulo , Gielen Jan , Hubens Guy
Published: June 2021

Introduction

Clavicles have a complex osteologic structure which makes a morphometric analysis extremely difficult. Our analysis shows the exact measurements and variations of the clavicle.

Materials and Methods

90 clavicles were dissected, CAT scanned and reconstructed. All measurements were automatically performed. The length and curvatures were calculated around the central line and for each cross‐section the average, sagittal and axial diameter was calculated.

Results

The average length is 163±11 mm. For the length, there is a 9% difference between the gender and 1.2% between the left and right clavicle. Between the genders there is a volume difference of 36%. The extremities show the biggest diameter, this decrease as approaching to the inflexion point, which is the smallest average diameter. In the axial view the acromial curvature is shorter and more curved than the sternal one. In males, the maximum acromial curvature has a difference of 18% and the maximum sternal curvature a difference of 4% compared with the females. In the coronal view there is a concave curvature with a maximum of 6 mm. In females the acromial end bends more posteroinferiorly.

Discussion

This is the first 3D analysis performed on the clavicle. Females have a smaller clavicle with a shorter and less curved acromial curvature and a posteroinferior bending. The right clavicle is slightly shorter, thicker and more robust.
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).

Methods

Thirty 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.

Results

BP 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.

Conclusion

Aldosterone 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

Objective

In 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.

Methods

We 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.

Results

All 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.

Conclusion

MRF 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.
Elevated O‐GlcNAcylation of ATP synthase β subunits in cardiomyopathic hearts in Syrian hamsters
Authors: Suzuki Osamu , Koura Minako , Noguchi Yoko , Uchio‐Yamada Kozue , Matsuda Junichiro
Published: June 2021

In recent years, the posttranslational modification of serine and threonine residues of heart proteins by O‐linked attachment of the monosaccharide β‐N‐acetylglucosamine (O‐GlcNAc) has received a lot of attention as a key regulator in the cardiovascular function. To study the possible involvement of the O‐GlcNAcylation in the cardiomyopathy, we examined the O‐GlcNAcylation status of ventricular proteins from both normal (J2N‐n strain) and cardiomyopathic (J2N‐k strain) Syrian hamsters. Soluble fractions of heart proteins were extracted from left ventricles of both strains using a ReadyPrep sequential extraction kit (Bio‐Rad). Western blot analysis of the soluble fractions with a mouse monoclonal antibody against O‐GlcNAc (clone CTD110.6) indicated one J2N‐k specific band (~52 kDa). After isolation by monoclonal antibody columns and SDS‐PAGE, the band was suggested to be an ATP synthase β subunit (ATP5B) by mass spectrometry. These results suggest that the energy production machinery is modulated by O‐GlcNAcylation in the cardiomyopathy. This work was supported by a grant from the Ministry of Health, Labor, and Welfare of Japan.
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.
Enhanced expression of phospholipase C by cholesterol increases the production of amyloidogenic peptide
Authors: Chung Sungkwon , Chun Yoon Sun , Park Myoung Kyu
Published: June 2021

Cerebral elevation and accumulation of amyloid β‐peptide (Aβ) are necessary steps in the pathogenesis of Alzheimer's disease (AD). More amyloidogenic 42‐residue Aβ (Aβ42) is considered as a pathogenic agent. Even though cholesterol has been suggested as the primary cause for AD, the role of cholesterol in the pathogenesis of AD is not clear yet. We have reported that mutations in familial AD (FAD)‐linked presenilin (PS) down‐regulate the level of phosphatidylinositol 4,5‐bisphosphate (PIP2), and that the modulation of cellular PIP2 levels closely correlates with Aβ42 biogenesis. Recently, we showed that cholesterol enrichment down‐regulates PIP2 level via activation of phospholipase C (PLC). In this study, we found that cholesterol enrichment decreased PIP2 level, and increased the production of Aβ42. Supplying PIP2 by using PIPn‐carrier system blocked the effect of cholesterol on Aβ42 production. We also found that cholesterol increased the expression of PLCβ1 and PLCβ3. Inhibiting new protein synthesis or silencing the expression of PLCβ1 by siRNA method prevented the effects of cholesterol on PIP2 level as well as on Aβ42 production. These results suggest that cholesterol enrichment increased Aβ42 production by down‐regulating PIP2 level via enhancing the expression of PLCβ1. Thus, these results may indicate a new link between cholesterol and the pathogenesis of AD.
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.

Methods

100 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.

Results

90/100 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%.

Conclusion

Eosinophils 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.
Anticoagulation of Heparin Compromised Population with a Direct Synthetic Anti Xa agent Otamixaban
Authors: Fareed Jawed , Thethi Indermohan , Walenga Jeanine Marie , Hoppensteadt Debra , Lewis Bruce
Published: June 2021

Otamixaban is a synthetic anti‐Xa agent with potent anticoagulation activity. This study investigated the cross reactivity of otamixaban with HIT antibodies and its modulatory actions on anticoagualtion. The cross reactivity studies with antibodies were carried out in the platelet aggregation and 14C serotonin release assays. In contrast to the unfractionated heparin (UFH), and low molecular weight heparins (LMWHs), otamixaban did not exhibit any cross reactivity with sera from HIT patients (N=250). In the 14C serotonin release assays, otamixaban did not exhibit any cross reactivity with HIT antisera in contrast to UFH and enoxaparin. There results suggest that otamixaban can be used as an effective anticoagulant in Heparin compromised patients.
Detection of early infection with flu viruses in patients with bronchial asthma and chronic obstructive pulmonary disease (COPD) in winter season in the state of Qatar: A One year prospective study
Authors: Al‐Thani Asma Ahmed , Sattar Hisham A. , Singh Rajvir , Majeed Ehssan Mohd A. , Shaath Noor Jawdat , Khanji Moza Al
Published: June 2021

Flu infections are an important cause of morbidity in patients with various medical conditions which require inpatient and outpatient care. To reduce the impact of Flu infections in Qatar; continued studies, intense surveillance, increased use of influenza vaccine, and anti‐virals with the potential for broader protection were required. The objective of this study is to understand the viral etiologies of respiratory tract infection during the winter season associated with asthma and COPD and to evaluate a new diagnostic‐ respiratory detection by Real‐time PCR to detect 15 virus at the same time (influenza A (FluA) and B (FluB); corona viruses NL63 (Cor63), 229E (Cor229) and OC43 (Cor43), parainfluenza 1, 2, 3 and 4 (para1, para2, para3, para4), human metapneumo virus A and B (Hump A and B), rhinovirus (rhino), respiratory syncytial viruses A and B (RSVA and B) and adenovirus (AV). Nasal swabs from 200 patients above 18 years old with asthma and COPD attending the chest clinic at HMC during winter season between 1st of Oct. 2008 and 31st March 2009 were collected. Among the sample population it was found that, 190 patients had asthma and 10 had COPD. Only 36 patients had viruses related to the respiratory tract infection; 31 of which associated with asthma and 5 with COPD. The most predominant respiratory virus was Rhino virus 17 %. Additionally, it was found that the new diagnostic respiratory detection by Real‐time PCR machine was effective in detection of the fifteen viruses.
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.
Localization of the Dopamine‐ and cAMP‐regulated Phosphoprotein, Mr 32 kDa (DARPP‐32) in the Ventral Tegmental Area (VTA)
Authors: Gouty Shawn Christian , Cox Brian , Nugent Fereshteh
Published: June 2021

The VTA is a critical component of the brain reward circuitry and necessary for processing reward as well as drug reward. DARPP‐32, first discovered in the striatum represents an exciting target for drug‐induced neuroplasticity in the brain, yet the cellular localization of DARPP‐32 in the VTA has not been confirmed. We explored the possibility whether VTA GABAergic neurons, the main local inhibitor of VTA dopamine neurons, express DARPP‐32. To confirm whether a subset of these DARPP‐32‐immunoreactive terminals arise from local GABAergic interneurons, we used a double immunoflurescence confocal microscopy technique using antibodies against mu opioid receptors (as the marker for GABAergic neurons) and DARPP‐32 to visualize GABAergic neurons expressing DARPP‐32. Our preliminary experiments confirmed that some subset of mu opioid receptor‐expressing GABA neurons also expresses DARPP‐32, suggesting that the source of DARPP‐32 in the VTA may arise from some subset of local GABA neurons in the VTA. To our knowledge, this is the first demonstration of the cellular localization of DARPP‐32 in the VTA. Supported by R075OU‐DOD/USUHS and PhRMA Research Starter Grants.
Behavioral and transcriptional effects of acrylamide‐induced neurotoxicity in rat pups
Authors: El‐Alfy Abir T , Seale Suzanne , Feng Q , Mark M , Baerson Scott , Agarwal Ameeta
Published: June 2021

The goal of this project was to assess both behavioral effect and gene expression changes induced by acrylamide neurotoxicity in rats when administered during early postnatal life. Three week old male Wistar rats were administered acrylamide daily (30 mg/kg, p.o.) for twenty one days. Neurobehavioral effects were assessed using locomotor activity, weight, hind‐limb heel splay, fore‐limb and hind‐limb grip strength. Acrylamide treatment induced significant characteristic neurotoxic symptoms: increased heel splay, decrease in grip strength, and decrease in locomotor activity. Acrylamide caused significant loss of weight gain starting at day fourteen (p<0.001) and a significant reduction in hind‐limb grip strength (p<0.001). Similarly, on day fourteen hind limb heel splay was significantly increased (p<0.01). Reduction in locomotor activity began in the treated rats on day sixteen (p<0.01). On the other hand, fore‐limb grip strength was not affected. Gene expression changes were evaluated in the cerebellum, spinal cord, and sciatic nerve of acrylamide‐treated rats. At a 2 fold cutoff, one gene was up‐regulated and fifteen genes were down‐regulated among the three tissues. RT‐PCR was conducted to validate the microarray data. The identified differentially expressed genes play a role in neuronal development, muscle contraction, and control of motor function.
Ketamine disturbs normal neurogenesis in cultured rat cortical neural stem progenitor cells
Authors: Dong Chaoxuan , Rovnaghi Cynthia S , Anand K.J.S.
Published: June 2021

Ketamine, an N‐Methyl‐D‐aspartate (NMDA) receptor antagonist, is widely used for anesthesia, analgesia or sedation in pediatric clinical settings. Fate of neural stem stem/progenitor cells (NSPCs) determines normal development of embryonic brains. Current studies indicate that ketamine damages the development of developing brains through inducing neuronal death. However, effects of ketamine on NSPCs have not been detected systemically. This study explored time‐ and dose‐dependent effects of ketamine on cell death and neurogenesis of cultured rat cortical NSPCs. In this study, NSPCs were isolated from embryos of timed‐pregnant SD rats, were treated by varying concentrations of ketamine (0, 1, 10, 20, 20, 50, and 100 μM) for 24 hours or were exposed to 10 μM of ketamine for different durations (0, 1/2, 1, 2, 4, 6, 8, 10, 12, 18, 24, and 48 hrs). Treated cultures were immunostained to detect apoptosis (anti‐cleaved caspase‐3), proliferation (anti‐BrdU & anti‐Ki67), and neuronal differentiation (anti‐Tuj‐1). Additionally, LDH assay was utilized to detect necrosis in cultures. Results showed that ketamine (<100 μM) did not induce apoptosis and necrosis in NSPCs critically. But, ketamine significantly reduced proliferation and promoted neuronal differentiation in rat cortical NSPCs in dose/time dependent manners. This study suggests that ketamine exposure may result in potential abnormalities of cortical structure and functions in developing rat brains.
Mechanisms underlying the neuroprotective effects of fisetin and ibuprofen in the MPTP model of Parkinson's disease
Authors: Patel Mital Yogeshbhai , Benzeroual Kenza E.
Published: June 2021

Several studies mentioned the association of oxidative stress and neuroinflammation with Parkinson's disease (PD). In the MPTP‐injured PC12 cells, a rat model of PD, MPTP induces cell death through complex I inhibition in the mitochondria, increases the production of nitric oxide (NO) and pro‐inflammatory mediators such as IL‐1beta and TNF‐alpha. This study, investigated the mechanisms underlying the neuroprotective effect of two anti‐inflammatory drugs, Fisetin and Ibuprofen in MPTP‐induced PC12. Cell viability was measured by MTT assay, IL‐1beta levels were quantified by ELISA, TNF‐alpha expression was assessed by western blot, and NO levels were also evaluated. PC12 cells were pretreated with varying doses of fisetin, ibuprofen, or both drugs in combination for 2 hours, prior to MPTP treatment. Pretreatment with fisetin produced a dose‐dependent increase in cell viability, and a dose‐dependent decrease in the expression of TNF‐alpha. Ibuprofen at 2 μg/ml also decreased TNF‐alpha expression in MPTP treated PC12 cells. The combination of both drugs showed a synergistic effect in the inhibition of TNF‐alpha expression. Fisetin also suppressed the production of NO and IL‐1beta in a dose‐dependent manner, and that both drugs combined showed a synergistic effect in suppressing the NO and IL‐1beta production. The present study indicates that fisetin and ibuprofen may provide a promising approach for the treatment of neuroinflammation associated with PD.