scholarly journals 2350

2017 ◽  
Vol 1 (S1) ◽  
pp. 8-8
Author(s):  
Dimitri Koutzoumis ◽  
Jose Antonio Pino ◽  
Sharonda S. Harris ◽  
Marisol Quiroz ◽  
Mansour Mohamadzadeh ◽  
...  
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Mobile Phase ◽  
Hplc Analysis ◽  
Treated Group ◽  
Control Group ◽  
Tissue Preparation ◽  
Western Blots ◽  
The Impact

OBJECTIVES/SPECIFIC AIMS: Several clinical studies have established a correlation between changes in relative bacterial populations in the gut and Parkinson disease. However, few published experiments have been able to parse out whether these associations are causative or correlative. Our aim is to determine how bacteria in the gut may impact the health and resilience of dopaminergic signaling. Our experiment is designed to serve as a proof-of-principle that controlled alterations to the gut microbiome alters mechanisms in dopamine homeostasis in the midbrain. METHODS/STUDY POPULATION: Bacterial inoculation 8–10-week-old germ-free male mice (C57BL/6) were exclusively used in this experiment. Mice were orally gavaged every 3 days (D0, 3, 6, and 9) with 100 µL novel bacterial suspension (~108 CFU resuspended in PBS with 1.5% NaHCO3) or vehicle and were sacrificed on D11. Tissue preparation—brains were quickly extracted and the striatum was isolated and homogenized in either RIPA buffer with protease inhibitors (for Western blot analysis) or in 0.1 N HClO4 (for HPLC processing). The homogenates were processed through fractional centrifugation to remove cellular debris. Lysate samples were frozen at −80°C until ready for analysis. Protein expression quantification—expression of proteins were measured using intensity of bands from Western blots. Lysates were denatured prior to loading with LB with 10% β-mercaptoethanol and 30-minute incubation at 37°C. All immunoblots were normalized to immunoreactivity to α-tubulin. Immunoblot intensity was determined using the ImageJ software. Dopamine/dopamine metabolite quantification HPLC analysis was used to determine dopamine and dopamine metabolite concentration. Aliquots of the lysate were injected onto a C18 column using a mobile phase consisting of 50 mM H2NaO4P·H2O, 0.72 mM sodium octyl sulfate, 75 µM Na2 EDTA, and 10% acetonitrile (pH 3.0). The mobile phase was pumped through the system at 0.3 mL/minute. RESULTS/ANTICIPATED RESULTS: Measured total dopamine concentration through HPLC analysis in the striatum showed no significant differences in the bacteria-treated group relative to the control group. The metabolites DOPAC and HVA had an elevated measured concentration in the bacteria-treated group relative to the control group. Western blot analysis showed decreased immunoreactivity for DAT and TH in the bacteria-treated group compared with the control group. There was no significance difference in the immunoreactivity for VMAT2. DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrates that dopamine signaling dynamics in the midbrain can be altered by changes in the gut flora in mice. These results further substantiate the impact of the gut-brain axis and may even point to a potential avenue of bolstering the resilience of dopaminergic neurons in preventing the onset of PD. Further experiments must be performed to understand the mechanism of the observed changes and to determine if these changes have any salutary effect.

Structural and Functional Interactions of KCl Cotransport Proteins KCC1 and KCC3 in Sickle and Normal Erythrocyte Membranes

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2474-2474
Author(s):  
Mary Risinger ◽  
Jesse Rinehart ◽  
Scott Crable ◽  
Anna Ottlinger ◽  
Richard Winkelmann ◽  
...  
Keyword(s):  
Western Blot ◽  
Specific Antibody ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Erythrocyte Membranes ◽  
Western Blots ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
In Cells

Abstract The KCl cotransporter (KCC) mediates volume reduction in normal reticulocytes and exaggerated KCC activity in sickle red blood cells (SS RBC) (Joiner et al, Blood109:1728, 2007) contributes to pathological dehydration that potentiates sickling. Three separate genes (KCC1, KCC3, KCC4) are expressed in RBC (Crable et al, Exp. Hem.33:624, 2005). KCC1 and KCC3 proteins have been shown to interact in ex vivo expression systems (Simard et al, JBC282(25):18083, 2007), and co-expression of an N-terminal truncation of KCC1 reduces KCC activity mediated by full-length KCC1 or KCC3 (Casula et al. JBC276:41870, 2001), suggesting functional interaction. We show here via western blot analysis that SS RBC membranes contain more KCC1 protein (relative to KCC3) than AA RBC, independent of the reticulocytosis of sickle blood. Immunoprecipitation of solubilized SS RBC membranes with KCC3-specific antibody yielded a band at 125 kD on SDS PAGE which contained KCC1, as identified by western blotting with KCC1-specific antibody and by TOF mass spectroscopy. The effect of co-expression of KCC1 and KCC3 on KCC activity was assessed by measuring NEM-stimulated, Cl-dependent, (ouabain + bumetanide)-insensitive Rb uptake in HEK 293 cells. The Flip-In T-rex HEK 293 cell line (Invitrogen) containing a tetracycline-response promoter was transfected with a pcDNA5a plasmid containing KCC3a cDNA. Recombination of the plasmid with the integrated tet-promoter construct inserts the KCC3a gene under control of a tetracycline-responsive promoter. These cells were subsequently transduced with a retroviral vector (SF-91. Hildinger et at, Gene Ther. 5:1575, 1998) containing KCC1 cDNA linked to a GFP cassette. Control cells contained SF-91 vector lacking KCC1. Cells were selected for GFP expression and grown in the absence (un-induced, no KCC3a expression) or presence of tetracycline (induced, KCC3a expression). From this binary matrix, four types of cells were obtained: Cells with no additional KCC expression, representing endogenous KCC activity; cells with only KCC1 or KCC3a expression; cells with both KCC1 and KCC3a expression. Western blots indicated similar KCC1 expression in cells with KCC1 only and [KCC1 + KCC3] and similar KCC3 expression in cells with KCC3 only and [KCC1 + KCC3]. Thus, the expression of neither isoform was affected by the presence of the other. KCC activity in cells overexpressing KCC1 only was similar to endogenous activity in HEK 293 cells; i.e., transport activity of KCC1 alone was minimal. Cells overexpressing KCC3 only had a 5-fold increase in KCC activity over endogenous levels. When KCC1 was co-expressed with KCC3 in [KCC1 + KCC3] cells, an additional 50% increase in KCC activity was observed (p < 0.05 by paired t-test, N=4), despite similar levels of KCC3 expression by western blot analysis. This synergistic effect was dependent on the cytoplasmic N-terminus of KCC1, as it was not seen when the first 39 amino acids of KCC1 were removed. Interestingly, removal of the entire cytoplasmic N-terminal domain (117 aa) produced an inhibitory effect when co-expressed with KCC3a in HEK cells, as previously reported in Xenopus oocytes (Casula et al.). These data indicate that KCC1 and KCC3 interact structurally and functionally in RBC membranes, and provide another potential mechanism for regulation of KCC activity via multimeric associations between KCC isoforms. Thus, KCC activity could be modulated not only by transcriptional mechanisms and post-translational modification (phosphorylation), but also by altering the ratios of KCC isoforms or the kinetics of their association. We speculate that higher levels of KCC1 protein relative to KCC3 in SS RBC membranes could account for higher KCC activity in these cells relative to AA RBC.

Lidocaine Inhibits Ovarian Cancer Cell Proliferation and Induces Apoptosis: An In Vitro Study

2020 ◽  
Vol 10 (5) ◽  
pp. 724-729
Author(s):  
Yaping Xu ◽  
Xiaoqin Fang ◽  
Xianjiang Wei
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Protein Expression ◽  
Western Blot ◽  
Cancer Cell ◽  
Blot Analysis ◽  
Group Treatment ◽  
Western Blot Analysis ◽  
Ovarian Cancer Cell ◽  
Control Group

Objective: The present study aimed to explore the effects and related mechanism of lidocaine on human ovarian cancer cell lines. Methods: Human ovarian cancer cell lines (SKOV3 and ES-2) were treated with different concentrations of lidocaine for different time. We treated SKOV3 and ES-2 cells using lidocaine then used MTT assay and flow cytometry to detect the cell proliferation and cell apoptosis. In addition, we used western blot analysis to explore the protein expression of Bax and Bcl-2 in SKOV3 and ES-2 cells. Western blot analysis and qRT-PCR were performed for the detection of EMT markers (E-cadherin, N-cadherin). The protein expression levels of TRAF3 and p-p65 in SKOV3 and ES-2 cells were determined by Western blot analysis. Results: Compared to the control group, 0.5, 1, 5, and 10 mM of lidocaine significantly inhibited ovarian cancer cell proliferation at different time points, while 0.1 mM of lidocaine had no significant effect. 1, 5 mM of lidocaine induced the cell apoptosis, and observably reduced expression of Bcl-2 protein, but improved Bax expression markedly compared with the control group. Treatment of lidocaine increased E-cadherin expression, but decreased N-cadherin expression when compared with control group. Treatment of lidocaine increased TRAF3 protein expression, but decreased p-p65 protein expression in ES-2 and SKOV3 cells. Conclusion: We demonstrated that lidocaine inhibited cell proliferation, induced apoptosis, and inhibited EMT in ovarian cancer cells via regulating TRAF3/NF-κB pathway.

scholarly journals Is Oxidative Stress in Mice Brain Regions Diminished by 2-[(2,6-Dichlorobenzylidene)amino]-5,6-dihydro-4H-cyclopenta[b]thiophene-3-carbonitrile?

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
A. C. Fortes ◽  
A. A. C. Almeida ◽  
G. A. L. Oliveira ◽  
P. S. Santos ◽  
W. De Lucca Junior ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Superoxide Dismutase ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Brain Regions ◽  
Saline Control ◽  
Control Group ◽  
Nitrite Content

2-[(2,6-Dichlorobenzylidene)amino]-5,6-dihydro-4H-cyclopenta[b]thiophene-3-carbonitrile, 5TIO1, is a new 2-aminothiophene derivative with promising pharmacological activities. The aim of this study was to evaluate its antioxidant activity in different areas of mice central nervous system. Male Swiss adult mice were intraperitoneally treated with Tween 80 dissolved in 0.9% saline (control group) and 5TIO1 (0.1, 1, and 10 mg kg−1). Brain homogenates—hippocampus, striatum, frontal cortex, and cerebellum—were obtained after 24 h of observation. Superoxide dismutase and catalase activities, lipid peroxidation and nitrite content were measured using spectrophotometrical methods. To clarify the 5TIO1’s mechanism on oxidative stress, western blot analysis of superoxide dismutase and catalase was also performed. 5TIO1 decreased lipid peroxidation and nitrite content in all brain areas and increased the antioxidant enzymatic activities, specially, in cerebellum. The data of Western blot analysis did not demonstrate evidence of the upregulation of these enzymes after the administration of this compound. Our findings strongly support that 5TIO1 can protect the brain against neuronal damages regularly observed during neuropathologies.

scholarly journals Emerging techniques of western blotting for purification and analysis of protein

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Krishna Kumar Singh ◽  
Anshika Gupta ◽  
Charu Bharti ◽  
Himanchal Sharma
Keyword(s):  
Capillary Electrophoresis ◽  
Western Blot ◽  
Western Blotting ◽  
Blot Analysis ◽  
Clinical Relevance ◽  
Western Blot Analysis ◽  
Molecular Techniques ◽  
Clinical Implications ◽  
Western Blots ◽  
Main Text

Abstract Background Western blotting is frequently employed in molecular techniques like Proteomics and Biology. Because it is a sequential framework, differences and inaccuracies could even take place at any stage, decreasing this particular method's reproducibility and reliability. Main text New approaches, like automated microfluid western blotting, DigiWest, single cell resolution, microchip electrophoresis, and capillary electrophoresis, were all implemented to reduce the future conflicts linked with the western blot analysis approach. Discovery of new in devices and higher susceptibility for western blots gives innovative opportunities to expand Western blot’s clinical relevance. The advancements in various region of west blotting included in this analysis of transfer of protein and validation of antibody are described. Conclusion This paper describes another very developed strategy available as well as demonstrated the correlation among Western blotting techniques of the next generation and their clinical implications. In this review, the different techniques of western blotting and their improvement in different stages have been discussed.

Carboplatin-induced Senescence in Ovarian Cancer Cells Is Reversed Through EGFR and NF-κB Signaling

2021 ◽  
Author(s):  
Yue Li ◽  
Mingxu Fu ◽  
Ling Guo ◽  
Xiaoxiao Sun ◽  
Yuhang Chen ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Western Blot ◽  
Cancer Cells ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Cell Counting ◽  
Blue Staining ◽  
Ovarian Cancer Cells ◽  
Positive Staining ◽  
The Impact

Abstract Background: Metastases and recurrence of ovarian cancer after surgery and chemotherapy account for most cancer-related deaths, yet the mechanism underlying metastases and recurrence remains poorly understood. Recent evidence demonstrates that although long-lasting cells were considered tumor suppressors, senescent cancer cells, can induce the metastases and recurrence. In this study, we focused on the fate of ovarian cancer cells treated with carboplatin and explored the mechanism underlying ovarian cancer cell recovery from chemotherapy-induced senescence. Methods: SÁ-β-galactosidase staining was used to detect the impact of carboplatin on senescence of ovarian cancer cells. Cell proliferation was determined using direct cell counting, clone formation assay and 3D tumor spheroid formation assay. Lentivirus-mediated transduction was used to silence or upregulate EGFR expression. Quantitative real-time PCR and western blot analysis validated the efficacy of the knockdown or overexpression effect. Immunofluorescence staining and western blot analysis were used to examined the expression of EGFR and NF-KB. Cell death was determined using trypan blue staining assay. Results: Ovarian cancer cells treated by carboplatin exhibit a senescence-like phenotype indicated by SA-β-galactosidase positive staining. Importantly, carboplatin-induced senescence-like phenotype is reversible. In ovarian cancer cells, EGFR positively regulated cells proliferation, decreased carboplatin-induced senescence and upregulated the NF-κB1 protein level. EGFR/NF-κB1 upregulation promoted the recovery of ovarian cancer cells from senescence and chemoresistance to carboplatin. Conclusions: Ovarian cancer cells treated with carboplatin displayed a reversible senescence-like phenotype that could be combined with EGFR or NF-κB1 inhibitors to improve treatment effects.

Transfusion Of RBCs With Low-Density KEL Induces Tolerance To The KEL Antigen

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1160-1160
Author(s):  
Sean R. Stowell ◽  
Connie M Arthur ◽  
Nicole H. Smith ◽  
Kathryn R. Girard-Pierce ◽  
James C. Zimring ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Antibody Formation ◽  
Research Funding ◽  
Flow Cytometric Analysis ◽  
P Value ◽  
The Impact ◽  
Rbc Transfusion

Abstract Background Patients requiring repeat transfusion often develop RBC specific alloantibodies that decrease the therapeutic efficacy of transfused cells and limit the availability of compatible RBCs for future transfusion. However, not all RBC antigens possess equal ability to induce alloantibodies. While many factors likely influence this process, several studies suggest that antigen density may independently influence rates of RBC alloimmunization. To directly examine this, we generated transgenic founders with normal or lower levels of the human KEL antigen specifically on RBCs and examined the impact of RBC antigen levels on the development of anti-KEL antibodies following transfusion. Materials and methods Transgenic C57BL/6 founders expressing the human KEL antigen specifically on RBCs were generated using the β-globin promoter and screened for levels of KEL antigen using monoclonal anti-KEL antibodies by flow cytometric analysis. The number of KEL antigens on RBCs isolated from different founders was estimated using QIFIKIT beads. The molecular weight of KEL on RBCs isolated from each founder was assessed by Western blot analysis. C57BL/6 recipients were transfused with RBCs that expressed normal levels of KEL (KEL RBCs) or reduced levels of KEL (KELlo RBCs), followed by harvesting blood on days 3, 5, 7, 14, 21 and 28 following transfusion and analysis of serum for anti-KEL antibodies by indirect immunofluorescence using flow cytometry with KEL and control C57BL/6 RBCs as targets. In addition, C57BL/6 recipients were transfused with KELlo RBCs followed by KEL RBCs and similar analysis for anti-KEL antibody formation on days 3, 5, 7, 14, 21 and 28 following KEL RBC transfusion. All experiments were completed at least three times with 3–5 recipients per group per experiment. Results While KEL RBCs express approximately 1200 antigens per cell, KELlo RBCs express fewer than 200 KEL antigens. However, each KEL transgenic expressed a KEL protein of the predicted molecular weight (83 kD) as assessed by Western blot analysis. Transfusion of KEL RBCs induced IgM anti-KEL antibodies as early as 3 days post transfusion followed by peak IgG anti-KEL antibody levels 14 days following transfusion. In contrast, transfusion of KELlo RBCs failed to induce detectable IgM or IgG anti-KEL antibody formation following transfusion. Similarly, while antibodies could be detected on the surface of KEL RBCs following the development of detectable anti-KEL antibodies in the serum, no antibodies could be detected on KELlo RBCs following transfusion, although anti-KEL generated following KEL RBC transfusion readily bound KELlo RBCs in vitro. Although subsequent KEL RBC exposure following initial KEL RBC transfusion induced considerable increases in anti-KEL antibody formation, KEL RBC transfusion following initial KELlo RBCs transfusion completely failed to induce detectable IgM or IgG anti-KEL antibody formation. (All the above differences achieved a p value of <0.05). Conclusions These results suggest that RBC alloantigen density may significantly impact the immunological outcome of RBC transfusion. KELlo RBC transfusion not only failed to induce anti-KEL antibodies, but also induced an apparent state of tolerance to KEL RBCs following subsequent KEL RBC transfusion. Thus, antigen density may not only influence whether RBC alloimmunization occurs, but may also alter a recipient’s subsequent response to the same antigen. These results also suggest that manipulation of RBCs to express lower levels of RBC antigens may provide a unique tool to tolerize individuals against RBC alloantigens. Disclosures: Zimring: Immucor Inc.: Research Funding; Terumo: Research Funding; Haemonetics: Consultancy; Cerus: Honoraria.

scholarly journals Autophagy and Mitophagy Promotion in a Rat Model of Endometriosis

2021 ◽  
Vol 22 (10) ◽  
pp. 5074
Author(s):  
Rosalba Siracusa ◽  
Ramona D’Amico ◽  
Daniela Impellizzeri ◽  
Marika Cordaro ◽  
Alessio Filippo Peritore ◽  
...  
Keyword(s):  
Western Blot ◽  
Rat Model ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Mammalian Target Of Rapamycin ◽  
Immunohistochemical Analysis ◽  
Reproductive Age ◽  
Serine Threonine Kinase ◽  
Autophagy Pathway ◽  
The Impact

Endometriosis is a gynecological condition affecting patients in reproductive age. The aim of this paper was to assess the effects of the autophagy and mitophagy induction in a rat model of endometriosis. Endometriosis was induced by the injection of uterine fragments, and rapamycin (0. 5 mg/kg) was administered once per week. One week from the induction, rats were sacrificed, and laparotomy was performed to collect the endometriotic implants and to further process them for molecular analysis. Western blot analysis was conducted on explanted lesions to evaluate the autophagy pathway during the pathology. Elevated phospho-serine/threonine kinase (p-AKT) and mammalian target of rapamycin (mTOR) expressions were detected in vehicle-treated rats, while Beclin and microtubule-associated protein 1A/1B-light chain 3 II (LC3II) expressions were low. Additionally, samples collected from vehicle groups indicated low Bnip3, Ambra1, and Parkin expressions, demonstrating impaired autophagy and mitophagy. Rapamycin administration reduced p-AKT and mTOR expressions and increased Beclin and LC3II, Bnip3, Ambra1, and Parkin expressions, activating both mechanisms. We also evaluated the impact of the impaired autophagy and mitophagy pathways on apoptosis and angiogenesis. Rapamycin was administered by activating autophagy and mitophagy, which increased apoptosis (assessed by Western blot analysis of Bcl-2, Bax, and Cleaved-caspase 3) and reduced angiogenesis (assessed by immunohistochemical analysis of vascular endothelial grow factor (VEGF) and CD34) in the lesions. All of these mechanisms activated by the induction of the autophagy and mitophagy pathways led to the reduction in the lesions’ volume, area and diameter.

scholarly journals Intermittent Stretching and Osteogenic Differentiation of Bone Marrow Derived Mesenchymal Stem Cells via the p38MAPK-Osterix Signaling Pathway

2015 ◽  
Vol 36 (3) ◽  
pp. 1015-1025 ◽  
Author(s):  
Wen-lin Xiao ◽  
Dai-zun Zhang ◽  
Cun-hui Fan ◽  
Bao-jun Yu
Keyword(s):  
Western Blot ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Mechanical Strain ◽  
Control Group ◽  
Rt Pcr ◽  
Rnai Technology ◽  
P38mapk Signaling

Aims: The relationship between the p38MAPK signaling pathway and osterix in osteogenic differentiation of BMMSCs subjected to intermittent stretching was investigated. Methods: BMMSCs derived from C57BL/6J mice were divided into the following groups: 1) control, 2) stretch, and 3) SB203580+stretch (SB203580 is a p38MAPK signal pathway inhibitor). BMMSCs were exposed to an intermittent mechanical strain of 0.8% (8000μ strain) at 0.5 Hz, twice a day for 30 min each application. BMMSCs were harvested on days 1, 3, and 5 post-treatment. The expression of ALP, COL I, OCN, and osterix mRNA was assessed utilizing RT-PCR while the expression of P-p38MAPK and osterix protein was assessed by Western blot analysis. The osterix gene in mouse BMMSCs was knocked down using RNAi technology and its protein expression was also assessed by Western blot. RT-PCR was used to detect ALP, COL I, and OCN mRNA expression. Results: Intermittent stretching was found to promote expression of ALP, COL I, OCN, and osterix mRNA. Silencing the osterix gene was found to reduce levels of ALP, COL I, and OCN mRNA. Western blot analysis demonstrated that the levels of osterix and P-p38MAPK proteins in the stretch group were significantly higher than in the control group (P<0.05). There was less expression of ALP, COL I, OCN, and osterix mRNA in the SB203580+stretch group than in the control and stretch groups. Conclusions: Data demonstrate that intermittent stretching promotes osteogenic differentiation of BMMSCs, and the p38MAPK-osterix pathway has an important role in the control of osteogenesis-related gene expression.

scholarly journals EA Ameliorated Depressive Behaviors in CUMS Rats and Was Related to Its Suppressing Autophagy in the Hippocampus

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Zhinan Zhang ◽  
Xiaowen Cai ◽  
Zengyu Yao ◽  
Feng Wen ◽  
Zhiyi Fu ◽  
...  
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Preference Test ◽  
Control Group ◽  
Mild Stress ◽  
Sprague Dawley ◽  
Transmission Electron ◽  
Nonsignificant Difference ◽  
Sucrose Preference Test

Autophagy is confirmed to be involved in the onset and development of depression, and some antidepressants took effect by influencing the autophagic process. Electroacupuncture (EA), as a common complementary treatment for depression, may share the mechanism of influencing autophagy in the hippocampus like antidepressants. To investigate that, sixty Sprague-Dawley rats firstly went through chronic unpredictable mild stress (CUMS) model establishment, and 15 rats were assigned to a control group. After modeling, 45 successfully CUMS-induced rats were randomly divided to 3 groups: CUMS, selective serotonin reuptake inhibitor (SSRI), and EA groups (15 rats per group), to accept different interventions for 2 weeks. A sucrose preference test (SPT), weighing, and open field test (OFT) were measurement for depressive behaviors of rats. Transmission electron microscope (TEM), immunohistochemistry (IHC), and western blot analysis were used to evaluate the autophagic changes. After that, depression-like behaviors were successfully induced in CUMS models and reversed by SSRI and EA treatments (both p<0.05), but these two therapies had nonsignificant difference between each other (p>0.05). Autolysosomes observed through TEM in the CUMS group were more than that in the control group. Their number and size in the SSRI and EA groups also decreased significantly. From IHC, the CUMS group showed enhanced positive expression of both Beclin1 and LC3 in CA1 after modeling (p<0.05), and the LC3 level declined after EA treatments, which was verified by decreased LC3-II/LC3-I in western blot analysis. We speculated that CUMS-induced depression-like behavior was interacted with an autophagy process in the hippocampus, and EA demonstrated antidepressant effects by partly inhibiting autophagy with a decreased number of autolysosomes and level of LC3 along with LC3-II/LC3-I.

scholarly journals Anti-Toxocara Seropositivity in the Patients with Eye Diseases in Ophthalmology Clinics in Tehran, Iran

2019 ◽  
Author(s):  
Soodabeh EINIPOUR ◽  
Farid TAHVILDAR BIDEROUNI ◽  
Alireza RAMEZANI
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Inflammatory Diseases ◽  
Research Work ◽  
Parasitic Infection ◽  
Toxocara Canis ◽  
Eye Diseases ◽  
Control Group ◽  
Limited Information

Background: Eye diseases, including, Uveitis, inflammation of the retina, and choroid are caused by different agents in humans and has a variety of anterior medial and posterior types. The agent of eye diseases is very different from simple bacterial to acute viral, fungal and parasitic infection. There is limited information regarding the type of eye diseases and Toxocara canis. Methods: Blood samples of 359 individuals (339 patients including endophthalmitis, uveitis, DCR, glaucoma, and cataract 20 individuals as control group) together with their information were collected from ophthalmology hospitals in Tehran, Iran from Feb 2013 to Jan 2015. The patient's serum was evaluated for the presence of anti-Toxocara antibodies by ELISA kits and blood smears for high eosinophilia. The positive samples were confirmed by Western blot analysis for T. canis infection. Results: Overall, 339 patients sera with eye diseases and control group were tested for anti-T. canis antibodies. Nineteen (5.6%) patients had anti-T. canis IgG that 14 (6.1%) were male and 5 (4.5%) were female, and all the patients had negative eosinophilia as well. The results of Western blot analysis for 19 positive patients indicated that 15 were infected by T. canis and 4 were infected by other parasitic infection. The results for control group were negative. Conclusion: All the patients with inflammatory eye diseases such as endophthalmitis, uveitis DCR, glaucoma and cataract were studied for Toxocara infection in this research work were at risk. Therefore, in contrast to the previous idea, all eye inflammatory diseases in ocular patients should be considered for Toxocara antibodies in addition to Uveitis.

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