Search results for "western blot"
showing 10 items of 171 documents
Diagnostic follow-up of blood donors with indeterminate results to Western Blot for HIV infection in Palermo, Sicily
2002
27,343 blood donors, were tested for HIV infection at the Poli- clinico of Palermo in the period 1999-2001 by using a com- mercial ELISA as screening test and a commercial Western Blot (WB) as confirmatory test. HIV infection was detected in two subjects, whereas five individuals reacted faintly at the screening and tested indeterminate at the confirmatory test showing the presence of only one band for gp41 (four cases) or p24 (one case). A one year follow-up was performed in these patients. All donors but one, spontaneously seroreverted during the follow-up; the only one who persisted as WB inde- terminate, tested also negative for HIV RNA by PCR assay at the end of follow-up. In conclusio…
Role of nitric oxide synthases from Klebsormidium nitens: structural characterization and identification of protein partners
2022
Nitric oxide (NO) is an important cellular signalling molecule regulating various physiological processes, in both animals andplants. In animals, NO synthesis is mainly catalysed by NO synthase (NOS) enzymes. During biotic or abiotic stresses, NOSlike activities that are sensitive to mammalian NOS inhibitors have been detected in plants, although no sequences encodingthe well described mammalian NOS are highlighted in land plants. Interestingly, we identified NOS-like sequences in 20 algaespecies. Among them, NOSs are found in Klebsormidium nitens the model alga to study the early transition step from aquaticalgae to land plants.As mechanisms governing NO synthesis and signalling in green l…
Soluble liver antigen (SLA) antibody detection by ELISA and multiplexing technologies
2002
Abstract Objective: Develop and evaluate assays for the detection of antibodies to soluble liver antigen (SLA). SLA, also known as liver/pancreas antibody, was found to be 100% specific for autoimmune hepatitis (AIH) in a recent study of 2000 sera collected from individuals with various disease conditions and healthy individuals (Wies et al., Lancet 2000; 355:1510). Although SLA antibodies occur in only about 30% of patients with autoimmune hepatitis, they are found in some individuals with AIH who are negative for other autoantibodies. Methods: Specimens from patients with autoimmune hepatitis, non-autoimmune liver disease, various autoimmune conditions, as well as specimens from healthy i…
Type V collagen-induced upregulation of capn2 (large subunit of m-calpain) gene expression and DNA fragmentation in 8701-BC breast cancer cells
2011
Abstract Type V collagen is known to be over-deposited in the stroma of ductal infiltrating carcinomas of the breast. When used as a substrate, type V collagen restrains growth and invasion, and affects gene expression of 8701-BC ductal infiltrating carcinomas cells. Here we supplement existing data by demonstrating type V collagen dependent upregulation of capn2 gene expression in 8701-BC cells through differential display-PCR and Western blot assays. Furthermore, we suggest that our data obtained by centrifugal sedimentation and electrophoresis strongly suggest a correlation between calpain overproduction and DNA fragmentation, since the incubation with calpain inhibitor partly reverts th…
Purification and Characterization of Glutamine Synthetase Isoenzymes from Leaves and Roots of Brassica napus (L.)
1995
Summary The glutamine synthetase enzymes from leaves (GS2) and roots (GSR) of Brassica napus L. have been purified to homogeneity by the application of a three-stage isolation procedure comprising anion-exchange chromatography, adsorption by hydroxylapatite and gel filtration on Sephacryl 5-300 HR. The isoforms of the enzyme show a differential distribution in leaf and root tissues. Elution profiles of hydroxylapatite chromatography showed a distinct behaviour for GS proteins found in leaves and roots. Denaturing SDS-PAGE and Western blot experiments revealed molecular masses of approximately 43.5 and 40.5 kD for GS2 and GSR subunits, respectively. Moreover, kinetic properties determined us…
Properties of sea urchin coelomocyte agglutinins
1996
We examined some biological activities of a 200-kDa glycoprotein, referred to as Paracentrotus lividus vitellogenin, contained both in the coelomic fluid and in a subpopulation of coelomocytes called «colourless spherula cells». Cell-free coelomic fluid, coelomocyte lysate and supernatant obtained after coelomocyte washings were assayed for hemagglutinating activity. All samples agglutinated rabbit erythrocytes in a calcium-dependent way. The comparison between the electrophoretic patterns of erythrocyte lysates, before and after incubation with the coelomic fluid, revealed that a 200-kDa band was obtained from membranes of agglutinated erythrocytes. In addition, polyclonal antibodies again…
Identification and characterization of autoantibodies against catalase and α-enolase in patients with primary sclerosing cholangitis
1998
SUMMARY Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease of unknown aetiology. Recent studies have shown that genetic factors and both cellular and humoral immunological abnormalities are important in the pathogenesis of PSC. The most prominent autoantibodies in PSC are anti-neutrophil cytoplasmic antibodies (ANCA). The autoepitopes of ANCA in PSC are not well defined. The aim of this study was to identify corresponding ANCA autoantigens in patients with PSC. A biochemical approach with enrichment and partial purification of soluble neutrophil proteins, detection of autoantibodies by Western blot and partial amino acid sequencing were used. Two new autoantigen/aut…
Influence of nerve growth factor in endometriosis-associated symptoms
2011
To investigate the role of the nerve growth factor (NGF) in the development of dysmenorrhea/pelvic pain in patients with endometriosis, we performed a prospective, clinical, blind study. Peritoneal fluids (PFs) were obtained from patients with histologically proven endometriosis. Patients with endometriosis were divided into 7 different groups depending on their preoperative pain score and symptomatology: patients with no pain, patients with minimal pain (dysmenorrhea, pelvic pain, or both), and patients with severe pain (dysmenorrhea, pelvic pain, or both) and were used for the neuronal growth assay with cultured chicken dorsal root ganglia (DRG) and for Western blot analyses. Dorsal root …
Estradiol deficiency and skeletal muscle apoptosis: Possible contribution of microRNAs.
2020
Background Menopause leads to estradiol (E2) deficiency that is associated with decreases in muscle mass and strength. Here we studied the effect of E2 deficiency on miR-signaling that targets apoptotic pathways. Methods C57BL6 mice were divided into control (normal estrous cycle, n = 8), OVX (E2 deficiency, n = 7) and OVX + E2 groups (E2-pellet, n = 4). Six weeks following the OVX surgery, mice were sacrificed and RNA isolated from gastrocnemius muscles. miR-profiles were studied with Next-generation sequencing (NGS) and candidate miRs verified using qPCR. The target proteins of the miRs were found using in silico analysis and measured at mRNA (qPCR) and protein levels (Western blot). Resu…
Antibodies to alpha B-crystallin, vimentin, and heat shock protein 70 in aqueous humor of patients with normal tension glaucoma and IgG antibody patt…
2007
To show the existence of IgG antibodies against retinal antigens in aqueous humor of normal tension glaucoma patients.Forty-two patients were included in this study. Aqueous humor was collected from control subjects (CO; n = 21) and patients with normal tension glaucoma (NTG; n = 21). Western blot methods against bovine retinal antigens were used to detect the IgG antibody patterns. The complex antibody repertoires were analyzed by multivariate statistical techniques. Mass spectrometry was used to identify the most important antigens.Very complex IgG antibody patterns against retinal antigens were found in all analyzed aqueous humor samples. Our multivariate approach could quantify differen…