scholarly journals Diagnostic efficacy of Brucella abortus strain RB51 in experimentally inoculated Sprague-Dawley rats using western blot assay

2008 ◽  
Vol 2 (05) ◽  
Author(s):  
Siddiqur Rahman ◽  
Byeong Kirl Baek
Keyword(s):  
Western Blot ◽  
Brucella Abortus ◽  
Sprague Dawley ◽  
Western Blot Assay ◽  
Diagnostic Efficacy ◽  
Sprague Dawley Rats

Glycosaminoglycan polymerization may enable osmotically inactive Na+ storage in the skin

2004 ◽  
Vol 287 (1) ◽  
pp. H203-H208 ◽  
Author(s):  
Jens Titze ◽  
Mehdi Shakibaei ◽  
Markus Schafflhuber ◽  
Gundula Schulze-Tanzil ◽  
Markus Porst ◽  
...  
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Dermatan Sulfate ◽  
Active Process ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Dry Ashing ◽  
Mrna Content ◽  
Water Accumulation ◽  
Chain Polymerization

Osmotically inactive skin Na+ storage is characterized by Na+ accumulation without water accumulation in the skin. Negatively charged glycosaminoglycans (GAGs) may be important in skin Na+ storage. We investigated changes in skin GAG content and key enzymes of GAG chain polymerization during osmotically inactive skin Na+ storage. Female Sprague-Dawley rats were fed a 0.1% or 8% NaCl diet for 8 wk. Skin GAG content was measured by Western blot analysis. mRNA content of key dermatan sulfate polymerization enzymes was measured by real-time PCR. The Na+ concentration in skin was determined by dry ashing. Skin Na+ concentration during osmotically inactive Na+ storage was 180–190 mmol/l. Increasing skin Na+ coincided with increasing GAG content in cartilage and skin. Dietary NaCl loading coincided with increased chondroitin synthase mRNA content in the skin, whereas xylosyl transferase, biglycan, and decorin content were unchanged. We conclude that osmotically inactive skin Na+ storage is an active process characterized by an increased GAG content in the reservoir tissue. Inhibition or disinhibition of GAG chain polymerization may regulate osmotically inactive Na+ storage.

scholarly journals Effect of metformin on the improvement of prostate cancer in diabetic rats

2019 ◽  
Vol 17 ◽  
pp. 205873921985855
Author(s):  
Kaijian Hou ◽  
Wansheng Ke ◽  
Jianping Xiong
Keyword(s):  
Prostate Cancer ◽  
Western Blot ◽  
Intervention Group ◽  
Diabetic Rats ◽  
Pathological Examination ◽  
Control Group ◽  
Nuclear Staining ◽  
Sprague Dawley ◽  
Western Blot Assay ◽  
E Cadherin

This study was designed to investigate the effect of metformin on the improvement of prostate cancer in diabetic rats. A total of 20 Sprague Dawley (SD) rats were equally divided into control and intervention groups. The intervention group received intragastric metformin 200 mg/kg, while the control group was given intragastric drinking water for 4 weeks. Tumor volumes were compared, all tumor specimens underwent routine pathological examination, immunohistochemical detection of E-cadherin and N-cadherin, and western blot assay. The tumor volume of control and intervention group was 462.15 ± 45.67 and 23.46 ± 5.32 mm3, respectively. Hematoxylin and eosin (HE) staining showed partial visible glandular structure with deepened nuclear staining in the intervention group. Immunohistochemistry showed high expression (6.5 ± 0.28 vs 3.8 ± 0.26, P < 0.05) of E-cadherin and low expression (3.4 ± 0.12 vs 7.8 ± 0.34, P < 0.05) of N-cadherin in the intervention group. Western blot assay showed higher expression of E-cadherin, while low N-cadherin in the intervention group. Metformin can effectively alleviate lesion extent of prostate cancer and mechanism may be related to upregulation of E-cadherin and downregulation of N-cadherin expression.

scholarly journals 2251

2017 ◽  
Vol 1 (S1) ◽  
pp. 60-60
Author(s):  
Andrea Lee Frump ◽  
Margie Albrecht ◽  
Sandra Breuils-Bonnet ◽  
Bakhtiyor Yakubov ◽  
Mary Beth Brown ◽  
...  
Keyword(s):  
Western Blot ◽  
Knockout Mice ◽  
Sprague Dawley ◽  
Protein Receptor ◽  
Morphogenetic Protein ◽  
Sprague Dawley Rats ◽  
Tnf Α ◽  
Study Population ◽  
Direct Binding ◽  
17Β Estradiol

OBJECTIVES/SPECIFIC AIMS: Women with pulmonary arterial hypertension (PAH) exhibit superior right ventricular (RV) function and survival compared with men, a phenomenon attributed to poorly understood cardioprotective effects of 17β-estradiol (E2). We hypothesize that E2, through ERα, attenuates PH-induced RV dysfunction by upregulating the pro-contractile and pro-angiogenic peptide apelin. This ERα-mediated increase in apelin is mediated by the myocardial remodeling effector bone morphogenetic protein receptor 2 (BMPR2). METHODS/STUDY POPULATION: ERα, BMPR2, and apelin were measured (western blot) in RVs from patients with PAH-induced RV failure and in RV homogenates from male or female Sprague-Dawley rats with sugen/hypoxia (SuHx) or monocrotaline (MCT)-induced PH. H9c2 rat cardiomyoblasts and cardiac endothelial cells were stressed with TNF-α (10 ng/mL) or staurosporine (50 nM)±E2 (100 nM; 24 h). ERα-, BMPR2-, and apelin-dependence were evaluated by siRNA (5 pM). Downstream apelin target and pro-survival factor ERK1/2 expression was measured (western blot). p<0.05 by ANOVA was considered significant. RESULTS/ANTICIPATED RESULTS: ERα correlated positively with BMPR2 and apelin expression in SuHx-RVs and human RVs. Treatment of SuHx-PH rats with E2 or ERα agonist increased RV BMPR2 and apelin, whereas RV apelin was decreased in E2-treated hypoxic ERα knockout mice (p<0.05), but not in ERβ knockout mice. In H9c2 cells, E2 or ERα agonist attenuated TNF-α- or staurosporine-induced decreases in BMPR2, apelin, and phospho-ERK1/2 (p<0.05 for all endpoints). E2 protection was lost in presence of siRNA directed against ERα, BMPR2, or apelin (p<0.05). ERα was necessary for E2-mediated increases in BMPR2, apelin, and ERK1/2, and BMPR2 was required for the E2-mediated increase in apelin (p<0.05 for siRNA vs. scramble). ERα, BMPR2, and apelin protein was increased in decompensated human RVs but downstream phospho-ERK signaling was disrupted. DISCUSSION/SIGNIFICANCE OF IMPACT: E2, via ERα, increases BMPR2 and apelin in the failing RV and in stressed rat cardiomyoblasts. The E2-mediated increase in apelin is BMPR2-dependent and likely occurs through direct binding of ERα to the BMPR2 promoter. Harnessing this E2-ERα-BMPR2-apelin axis during RV compensation may lead to novel, RV-targeted therapies for PAH patients of either sex.

scholarly journals Repetitive Treatment with Volatile Anesthetics Does Not Affect the In Vivo Plasma Concentration and Composition of Extracellular Vesicles in Rats

2021 ◽  
Vol 43 (3) ◽  
pp. 1997-2010
Author(s):  
Christian Bleilevens ◽  
Christian Beckers ◽  
Alexander Theissen ◽  
Tamara Fechter ◽  
Eva Miriam Buhl ◽  
...  
Keyword(s):  
Cell Migration ◽  
Western Blot ◽  
Volatile Anesthetics ◽  
Extracellular Vesicle ◽  
Sprague Dawley ◽  
Scratch Assay ◽  
Transmission Electron ◽  
Sprague Dawley Rats

Background: Anesthetic-induced preconditioning (AIP) with volatile anesthetics is a well-known experimental technique to protect tissues from ischemic injury or oxidative stress. Additionally, plasmatic extracellular vesicle (EV) populations and their cargo are known to be affected by AIP in vitro, and to provide organ protective properties via their cargo. We investigated whether AIP would affect the generation of EVs in an in vivo rat model. Methods: Twenty male Sprague Dawley rats received a repetitive treatment with either isoflurane or with sevoflurane for a duration of 4 or 8 weeks. EVs from blood plasma were characterized by nanoparticle tracking analysis, transmission electron microscopy (TEM) and Western blot. A scratch assay (H9C2 cardiomyoblast cell line) was performed to investigate the protective capabilities of the isolated EVs. Results: TEM images as well as Western blot analysis indicated that EVs were successfully isolated. The AIP changed the flotillin and CD63 expression on the EV surface, but not the EV concentration. The scratch assay did not show increased cell migration and/or proliferation after EV treatment. Conclusion: AIP in rats changed the cargo of EVs but had no effect on EV concentration or cell migration/proliferation. Future studies are needed to investigate the cargo on a miRNA level and to investigate the properties of these EVs in additional functional experiments.

Overexpression of miR-146a inhibits the apoptosis of hippocampal neurons of rats with cerebral hemorrhage by regulating autophagy

2020 ◽  
Vol 39 (9) ◽  
pp. 1178-1189
Author(s):  
S Huan ◽  
J Jin ◽  
C-x Shi ◽  
T Li ◽  
Z Dai ◽  
...  
Keyword(s):  
Western Blot ◽  
Hippocampal Neurons ◽  
Caspase 3 ◽  
Nuclear Factor Κb ◽  
Beclin 1 ◽  
Sprague Dawley ◽  
Related Factors ◽  
Expression Levels ◽  
Transmission Electron ◽  
Sprague Dawley Rats

In this study, to investigate the effect of overexpression of miR-146a on autophagy of hippocampal neurons in rats with intracerebral hemorrhage (ICH), 72 Sprague-Dawley rats were randomly divided into the sham, ICH, miR-146a agomir, and miR-146a agomir control groups. The ICH model was constructed by injection of collagenase VII. The apoptosis of hippocampal neurons was measured by TUNEL assay. The levels of LC3 and Beclin 1 were analyzed by immunohistochemistry. Mitochondrial autophagy was examined by transmission electron microscopy. The levels of LC3A, LC3B, Beclin 1, Bax, Bcl-2, and cleaved caspase 3 were examined by Western blot. Western blot was also used to evaluate the expression of nuclear factor κB signaling pathway-related factors. To examine the effect of autophagy inhibitor (3-methyladenine (3-MA)) on miR-146a-regulated apoptotic protein expression, 30 rats were further divided into the sham, ICH, miR-146a agomir, 3-MA, and miR-146a + 3-MA groups. The levels of Bax, Bcl-2, and cleaved caspase 3 were examined by Western blot. Compared with the sham group, the nerve function scores, brain water content, the percentage of apoptotic cells, and the expression levels of LC3, Beclin 1, Bax, cleaved caspase 3, and p-P65 in the hippocampus of rats in the ICH group were all significantly increased ( p < 0.05), whereas the expression levels of miR-146a, Bcl-2, and p-IκBα were markedly decreased ( p < 0.05). Mitochondrial autophagy was also evident. Furthermore, compared with the ICH group, the results of the abovementioned tests in the miR-146a agomir group were reversed. The overexpression of miR-146a inhibited the autophagy of hippocampal neurons in rats with ICH.

scholarly journals Plate and Tube Agglutination Tests for Diagnosis of Brucella abortus Biotype 1 Infection in Sprague-Dawley Rats

1970 ◽  
Vol 2 (1) ◽  
pp. 63-67 ◽  
Author(s):  
MS Rahman
Keyword(s):  
Antibody Titer ◽  
Brucella Abortus ◽  
Post Infection ◽  
Physiological Saline ◽  
Potential Candidate ◽  
Sprague Dawley ◽  
Sd Rats ◽  
Sprague Dawley Rats ◽  
Group A ◽  
Group B

The plate and tube agglutination tests were evaluated for the diagnosis of experimentally induced Brucella abortus biotype 1 infection in 45 female, 6 to 10 months old Sprague- Dawley (SD) rats during the period from 2001 to 2002. These 45 rats were divided into two groups A and B, of which group A consisting of 27 rats used for experimental infection, whereas 18 rats of group B served as uninfected control. Each rat of group A was injected subcutaneously @ 1.0×109 colony forming units (CFU) in 500 µl of bovine pathogenic strain of B. abortus biotype 1 suspension in physiological saline. The SD rats were monitored at regular intervals by serological and bacteriological methods. The reciprocal antibody titer was 1:400 through tube agglutination test (TAT) whereas it was 1:800 through plate agglutination (PAT) at first week of post-infection. There was no reciprocal antibody titer in sera of 24 weeks of post-infection both through PAT and TAT despite the presence of bacteremia and these tests were evaluated for the first time using sera from rat with brucellosis. PAT using B. abortus strain 1119-3 (S1119-3) whole cell antigen was a potential candidate as an improved diagnostic method for field diagnosis of brucellosis in wild animals. Key words: B. abortus biotype 1; plate and tube agglutination tests; Sprague-Dawley rats doi: 10.3329/bjvm.v2i1.1938 Bangl. J. Vet. Med. (2004). 2 (1) : 63-67

scholarly journals Regulations of expressions of rat/human sulfotransferases (SULTs) by anti-cancer drug, nolatrexed and micronutrients

2020 ◽  
Author(s):  
Smarajit Maiti ◽  
Sangita MaitiDutta ◽  
Guangping Chen
Keyword(s):  
Folic Acid ◽  
Western Blot ◽  
Folinic Acid ◽  
Hepg2 Cells ◽  
Clinical Importance ◽  
Cancer Drug ◽  
Sprague Dawley ◽  
Physiological Processes ◽  
Sprague Dawley Rats ◽  
Level Of Significance

AbstractCancer is a disease related to cellular proliferative-state. Drastically increase in cell-cycle regulations augments cellular folate-pool and folate-metabolism. So, this pathway is targeted therapeutically. A number of drugs are involved in this metabolism i.e. folic-acid/folinic-acid/nolatrexed(NT)/ methotrexate(MTX) for the research and treatment of cancer. Our previous study showed that MTX significantly modulated rat/human SULTs. Present study was an attempt to study the effect of NT (widely used in different cancers) and these micronutrients on the expressions of rat/human SULTs. Male Sprague-Dawley rats were treated with NT (01,10 or 100 mg/Kg) or both sexes were treated to folic acid (100,200 or 400 mg/kg) for 2-weeks and their AST-IV (2-napthol sulfation) and STa (DHEA-sulfation) activities, protein-expression (Western-Blot) and mRNA-expression (RT-PCR) were tested. In cultured HepG2 cells NT (1nM-1.2mM) or folonic-acid (10nM-10μM) were applied for 10 days. Folic acid (0-10μM) was treated to human hepatocarcinoma (HepG2) cells. PPST (phenol-catalyzing), MPST (dopamine) DHEAST (dehydroepiandrosterone,DHEA) and EST (estradiol-sulfating) protein-expressions (Western-blot) were tested in all HepG2 study. Present results suggest NT significantly increased SULTs expressions in rat (protein/mRNA/activity) and HepG2 cells. Folic acid increased SULTs activity/protein in sex-dependant manner (ASTIV in female/ STa in male). Both folic and folinic acid increased several hSULTs isoforms with varied level of significance (least or no increase at highest-dose) in HepG2 cells pointing its dose-dependent multi-phasic responses. The clinical importance of this study may be furthered in the verification of sulfation-metabolism of several exogenous/endogenous molecules, drug-drug interaction and their influences on the patho-physiological processes. Further studies are necessary in this regard.

Activation of local aldosterone system within podocytes is involved in apoptosis under diabetic conditions

2009 ◽  
Vol 297 (5) ◽  
pp. F1381-F1390 ◽  
Author(s):  
Sun Ha Lee ◽  
Tae-Hyun Yoo ◽  
Bo-Young Nam ◽  
Dong Ki Kim ◽  
Jin Ji Li ◽  
...  
Keyword(s):  
Western Blot ◽  
Tunel Assay ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Sprague Dawley ◽  
Hoechst 33342 ◽  
Sprague Dawley Rats ◽  
Mrna And Protein Expression ◽  
Related Proteins

Previous studies have shown that mineralocorticoid receptor (MCR) blocker reduces proteinuria in diabetic nephropathy (DN), but the role of aldosterone in podocyte injury has never been explored in DN. This study was undertaken to elucidate whether a local aldosterone system existed in podocytes and to examine its role in podocyte apoptosis under diabetic conditions. In vitro, immortalized podocytes were exposed to 5.6 mM glucose (NG), NG + 24.4 mM mannitol, and 30 mM glucose (HG) with or without 10−7 M spironolactone (SPR). In vivo, 32 Sprague-Dawley rats were injected with diluent (C, n = 16) or streptozotocin intraperitoneally [diabetes mellitus (DM), n = 16], and 8 rats from each group were treated with SPR for 3 mo. Aldosterone synthase (CYP11B2) and MCR mRNA and protein expression were determined by real-time PCR and Western blot, respectively, and aldosterone levels by radioimmunoassay. Western blot for apoptosis-related molecules, Hoechst 33342 staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were performed to determine apoptosis. CYP11B2 and MCR expression were significantly higher in HG-stimulated podocytes and DM glomeruli compared with NG cells and C glomeruli, respectively, along with increased aldosterone levels. Western blot analysis revealed that cleaved caspase-3 and Bax expression was significantly increased, whereas Bcl-2 expression was significantly decreased in HG-stimulated podocytes and in DM glomeruli. Apoptosis determined by Hoechst 33342 staining and TUNEL assay were also significantly increased in podocytes under diabetic conditions. These changes in the expression of apoptosis-related proteins and the increase in apoptotic cells were inhibited by SPR treatment. These findings suggest that a local aldosterone system is activated and is involved in podocyte apoptosis under diabetic conditions.

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