oalib

Publish in OALib Journal

ISSN: 2333-9721

APC: Only $99

Submit

Any time

2019 ( 12 )

2018 ( 68 )

2017 ( 59 )

2016 ( 64 )

Custom range...

Search Results: 1 - 10 of 2340 matches for " western blot. "
All listed articles are free for downloading (OA Articles)
Page 1 /2340
Display every page Item
Commensal E. Coli Strains Uniquely Alter the ECM Topography Independent of Colonic Epithelial Cells  [PDF]
Shruthi S. Bharadwaj, Victor Nekrasov, Ramana Vishnubhotla, Crystal Foster, Sarah C. Glover
Journal of Biomaterials and Nanobiotechnology (JBNB) , 2012, DOI: 10.4236/jbnb.2012.31009
Abstract: The relationship between commensal bacteria and the epithelial cells lining the colon is normally symbiotic. However, in the setting of diseases which lead to a loss of the protective mucosal layer such as inflammatory bowel disease or colon cancer, commensal bacteria gain the ability to alter both the behavior of epithelial cells as well as their surrounding extra cellular matrix (ECM). While much work has been done to understand the effects of bacteria on diseased epithelial cells in the colon, very little has been done to understand their impact on the ECM. In our previous work, we have shown that topographical changes in the ECM on the luminal side of the colon have a profound influence on the behavior of colonic epithelial cells. However, we do not understand all of the mechanisms that lead to changes in the ECM topography. This study aimed to understand the role that commensal E. coli strains play in altering the ECM topography of type-1 collagen scaffolds. To do this, 1.2 mg/ml type 1 collagen scaffolds were infected with various commensal bacterial strains. At 24 hours post-infection collagen fiber dimensions and substrate topography were determined using standard molecular biology techniques and advanced imaging. Intriguingly, all of the commensal E. coli strains showed some form of substrate degradation. Especially in the case of commensal E. coli strain HS4, maximum nano-scaled protrusions were observed. This data suggests, for the first time, that studying the effects of bacteria alone on the ECM may be critical to improving our understanding of how the cellular microenvironment changes in both health and disease.
Voltage profile generation for simultaneous multi-protein detection in western blot analysis  [PDF]
Matthew Blair, Mina Wanis, Gaurav Swarnkar, Hiroki Yokota, Stanley Chien
Journal of Biomedical Science and Engineering (JBiSE) , 2012, DOI: 10.4236/jbise.2012.59067
Abstract: Western blotting is a popular technique for examining expression levels of proteins using gel-based electrophoretic fractionation followed by blotting and antibody reactions. Although this is a mature technique, one of the major limitations is the need to prepare an individual electrophoretic gel for each of the protein species to be analyzed. Since most analyses require the detection of multiple protein species, a procedure that allows utilization of a single gel for detecting multiple protein species should significantly save time and resources. In this paper, we developed a novel multiprotein detection device, which enabled simultaneous detection of several proteins species from a single electrophoretic gel. In this device, a protein transfer unit utilized a multi-anode plate that generated a non-uniform voltage profile. This voltage profile enabled uniform transfer regardless of molecular mass of proteins. In vitro experiments using samples, isolated from boneforming osteoblast cells, showed that the expression levels of 5 - 7 different proteins were detectable in the presence and absence of mechanical stimulation that activated genes necessary for bone formation. The result supports the notion that through simultaneous detection of multiple protein species, the described device contributes to reduction in procedural time and sample amounts, as well as a removal of variations among multiple gels.
HIV Diagnosis in Resource Constrained Setting: How Good Is the Current Algorithm?  [PDF]
Nayana Ingole, Purva Sarkate, Supriya Paranjpe, Rahul Sarode, Sameer Shinde, Preeti Mehta
World Journal of AIDS (WJA) , 2014, DOI: 10.4236/wja.2014.41006
Abstract:

Background: The conventional HIV testing algorithm in most of the developed countries consists of two tests: an HIV enzyme immunoassay capable of identifying HIV-1 and HIV-2 antibodies and a confirmatory HIV-1 Western blot or immunofluorescence assay. However, the current algorithm for HIV diagnosis in India uses three sequential antibody assays. There has always been doubt regarding the benefits of this algorithm. Objective: To determine the utility of the current diagnostic algorithm and to find out the proportion of indeterminate or discrepant results. Methods: Retrospective analysis of HIV antibody testing data was carried out over a period of five years after institutional ethics committee approval. The specimens positive with the screening test and negative with both the supplemental tests were labeled as discrepant. Specimens positive with any of the two tests (screening and one of the supplemental tests) and negative with the remaining supplemental test were labeled as indeterminate. These indeterminate specimens were confirmed by immunoblotting. Results: A total of 141,296 samples were tested. Of these, 71 (0.05%) samples were indeterminate and 292 (0.21%) were discrepant. Western blot was done on 60 indeterminate samples of which 10 (16.67%) were positive for HIV 1 antibodies, 14 (23.33%) were negative for HIV antibodies and 36 (60%) had indeterminate result. Conclusion: In view of the low numbers of indeterminate and discrepant results, the current algorithm appears to be appropriate in our resource constrained setting. However, the algorithm for HIV testing should also include DNA PCR testing facility to resolve western blot indeterminate results.

Immunobiochemical Characteristics of Purified Native Leptin Protein from Indian Major Carp, Rohu (Labeo rohita Ham.)  [PDF]
Seikh Sahanawaz Alam, Siddhartha Narayan Joardar, Ashis Kumar Panigrahi, Thangapalam Jawahar Abraham, Sabyasachi Mukherjee, Anupama Mukherjee
Open Journal of Immunology (OJI) , 2014, DOI: 10.4236/oji.2014.44016
Abstract: Information regarding molecular characteristics of leptin protein in different animal species in-cluding fish is scarce. With the aim of characterizing the native leptin protein of Indian major carprohu (Labeo rohita), at molecular level, the present study was designed to isolate rohu leptin from its hepatocytes (the prime source of leptin in fish) and immunobiochemical characterization of the same, subsequently. In the present study, chemical treatment and ultra-sonication technique was used for isolating leptin from rohu liver tissue. Purification of the protein was attempted using affinity column chromatography. The molecular, biophysical and serological characterization of rohu leptin was carried out by 2D-gel electrophoreis, SDS-PAGE, MALDI-TOF Mass spectroscopy and Western blot. The SDS-PAGE and 2D gel analysis revealed that rohu native leptin possesses molecular mass of 16 kDa. Western blot analysis showed that the fish hepatocytes possessed the sero-reactive leptin protein of 16 kDa. MALDI-TOF mass spectroscopy and peptide analysis showed the molecular mass of rohu leptin as 16283.38 Da. The serodiagnostic potential of native leptin of rohu was revealed for the first time while assessing its serological responses by ELISA using anti-leptin antibodies.
Patterns of serologic response to human immunodeficiency virus type 1 (HIV-1) in brazilians with different clinical forms of HIV infection
Santos, Jairo Ivo dos;Campos, Deise Luci;Castro, Bernardo Galv?o;
Memórias do Instituto Oswaldo Cruz , 1989, DOI: 10.1590/S0074-02761989000100003
Abstract: in order to investigate the igg hiv-1 antibodies rectivity to structural components of the virus, 85 sera from infected brazilians, comprising the total spectrum of hiv infection, were analysed by western blot assay. the sera were confirmed as being positive to hiv with enzyme linked immuno assay (elisa) and indirect immunofluorescence (iif). although the sera from patients reacted less intensively to the gag polypeptide of 55kda, no distinctive antigen reaction patterns were observed between sera patients with different clinical forms. because of the higher frequency of reactivity to the gag p24 in aids patients, the patterns of anti-hiv igg responses are similar to those observed in their african counterparts.
Brote familiar de fascioliasis en Venezuela
Alarcón de Noya,Belkisyolé; Rojas,Elina; Colmenares,Cecilia; Morales,Carmen; Contreras,Rosa; Kay Valero,Sharon; Hernández,Dalila; Brice?o,Sonia; Vicente Scorza,Jose; Noya,Oscar;
Boletín de Malariología y Salud Ambiental , 2007,
Abstract: of the 9 cases of human fascioliasis recorded in venezuela, 4 have been published in the last 10 years, 2 of them originating from andean region. the well known transmission of fascioliasis in cattle and the clinical condition of a 14-year-old girl with abdominal discomfort, vomiting, eosinophilia and previous abundant ingestion of watercress, motivated the field study in the site of occurrence (timotes, edo. mérida in the andean region) to learn the risk of community infection. sixty-five people were evaluated clinically and serologically. coprology was done in 37 and abdominal ultrasound in 33 persons. elisa with adult fasciola hepatica soluble extract (afhes- elisa) was performed with 9 persons being positive. western blot (wb) was carried out on all elisa positive sera, resulting in 5 that recognized 9, 14, 27 and 65 kda specific molecules. the remaining four sera, with weak reactivity by elisa, did not have this recognition pattern. by stool examination, we found entamoeba coli, e. histolytica/dispar, blastocystis hominis, giardia lamblia, endolimax nana and ascaris lumbricoides. three of the 5 positive people by afhes-elisa and afhes-wb had f. hepatica eggs in their feces. the 5 positive cases are girls of a family, sisters and whose parents do not have the infection, although all consume watercress in salads with other vegetables from that region. the present study constitutes the first report of a well established focus of fascioliasis with human repercussion in venezuela.
Humoral response to low molecular weight antigens of Mycobacterium tuberculosis by tuberculosis patients and contacts
Beck, S.T.;Leite, O.M.;Arruda, R.S.;Ferreira, A.W.;
Brazilian Journal of Medical and Biological Research , 2005, DOI: 10.1590/S0100-879X2005000400013
Abstract: much effort has been devoted to the identification of immunologically important antigens of mycobacterium tuberculosis and to the combination of target antigens to which antibodies from serum of tuberculous patients could react specifically. we searched for igg antibodies specific for antigens of 45 to 6 kda obtained after sonication of the well-characterized wild m. tuberculosis strain in order to detect differences in the antibody response to low molecular weight antigens from m. tuberculosis between patients with pulmonary tuberculosis and contacts. specific igg antibodies for these antigens were detected by western blot analysis of 153 serum samples collected from 51 patients with confirmed pulmonary tuberculosis. three samples were collected from each patient: before therapy, and after 2 and 6 months of treatment. we also analyzed 25 samples obtained from contacts, as well as 30 samples from healthy individuals with known tuberculin status, 50 samples from patients with other lung diseases and 200 samples from healthy blood donors. the positive predictive value for associated igg reactivity against the 6-kda and 16-kda antigens, 6 and 38 kda, and 16 and 38 kda was 100% since simultaneous reactivity for these antigens was absent in healthy individuals and individuals with other lung diseases. this association was observed in 67% of the patients, but in only 8% of the contacts. the humoral response against antigens of 16 and 6 kda seems to be important for the detection of latent tuberculosis since the associated reactivity to these antigens is mainly present in individuals with active disease.
Western blot detection of infectious bursal disease virus infection
Pereira, S.R.F.G.;Travassos, C.E.P.F.;Huguenim, A.;Guimar?es, A.C.C.;Silva, A.G.;Guimar?es, M.A.A.M.;
Brazilian Journal of Medical and Biological Research , 1998, DOI: 10.1590/S0100-879X1998000500011
Abstract: in order to evaluate the use of a western blot methodology for the diagnosis of infectious bursal disease virus (ibdv) infection, chickens were experimentally infected with ibdv strains and tested for the presence of viral antigens and antibodies by a blocking western blot test (bwb). the viral proteins obtained from the bursa of fabricius (bf) were transferred to a nitrocellulose membrane after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page), and the chicken sera obtained by heart puncture were used for the detection of these proteins. in order to eliminate nonspecific reactions, we used a rabbit anti-chicken serum (blocking tool). by the use of the bwb test, two distinct viral proteins of 43-kda (vp2) and 32-kda (vp3) were detected. we suggest the use of this methodology for the detection of ibdv infection in animals suspected of having ibdv reinfection and a chronic subclinical form of the disease. with the use of the rabbit anti-chicken sera for blocking, this method is practical, sensitive and less time consuming
Western blot detection of infectious bursal disease virus infection
Pereira S.R.F.G.,Travassos C.E.P.F.,Huguenim A.,Guimar?es A.C.C.
Brazilian Journal of Medical and Biological Research , 1998,
Abstract: In order to evaluate the use of a Western blot methodology for the diagnosis of infectious bursal disease virus (IBDV) infection, chickens were experimentally infected with IBDV strains and tested for the presence of viral antigens and antibodies by a blocking Western blot test (bWB). The viral proteins obtained from the bursa of Fabricius (BF) were transferred to a nitrocellulose membrane after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the chicken sera obtained by heart puncture were used for the detection of these proteins. In order to eliminate nonspecific reactions, we used a rabbit anti-chicken serum (blocking tool). By the use of the bWB test, two distinct viral proteins of 43-kDa (VP2) and 32-kDa (VP3) were detected. We suggest the use of this methodology for the detection of IBDV infection in animals suspected of having IBDV reinfection and a chronic subclinical form of the disease. With the use of the rabbit anti-chicken sera for blocking, this method is practical, sensitive and less time consuming
Humoral response to low molecular weight antigens of Mycobacterium tuberculosis by tuberculosis patients and contacts
Beck S.T.,Leite O.M.,Arruda R.S.,Ferreira A.W.
Brazilian Journal of Medical and Biological Research , 2005,
Abstract: Much effort has been devoted to the identification of immunologically important antigens of Mycobacterium tuberculosis and to the combination of target antigens to which antibodies from serum of tuberculous patients could react specifically. We searched for IgG antibodies specific for antigens of 45 to 6 kDa obtained after sonication of the well-characterized wild M. tuberculosis strain in order to detect differences in the antibody response to low molecular weight antigens from M. tuberculosis between patients with pulmonary tuberculosis and contacts. Specific IgG antibodies for these antigens were detected by Western blot analysis of 153 serum samples collected from 51 patients with confirmed pulmonary tuberculosis. Three samples were collected from each patient: before therapy, and after 2 and 6 months of treatment. We also analyzed 25 samples obtained from contacts, as well as 30 samples from healthy individuals with known tuberculin status, 50 samples from patients with other lung diseases and 200 samples from healthy blood donors. The positive predictive value for associated IgG reactivity against the 6-kDa and 16-kDa antigens, 6 and 38 kDa, and 16 and 38 kDa was 100% since simultaneous reactivity for these antigens was absent in healthy individuals and individuals with other lung diseases. This association was observed in 67% of the patients, but in only 8% of the contacts. The humoral response against antigens of 16 and 6 kDa seems to be important for the detection of latent tuberculosis since the associated reactivity to these antigens is mainly present in individuals with active disease.
Page 1 /2340
Display every page Item


Home
Copyright © 2008-2017 Open Access Library. All rights reserved.