Determination of antipituitary antibody in patients with endocrine disorders by enzyme-linked immunosorbent assay and Western blot analysis

1998 ◽  
Vol 132 (1) ◽  
pp. 25-31 ◽  
Author(s):  
Shigeki Yabe ◽  
Tsugiyasu Kanda ◽  
Mina Hirokawa ◽  
Shizue Hasumi ◽  
Mizuho Osada ◽  
...  
Keyword(s):  
Western Blot ◽  
Immunosorbent Assay ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Endocrine Disorders

scholarly journals Kinetics and Diagnostic and Prognostic Potential of Quantitative Western Blot Analysis and Antigen-Specific Enzyme-Linked Immunosorbent Assay in Experimental Canine Leishmaniasis

2006 ◽  
Vol 13 (2) ◽  
pp. 271-276 ◽  
Author(s):  
D. Talmi-Frank ◽  
D. Strauss-Ayali ◽  
C. L. Jaffe ◽  
G. Baneth
Keyword(s):  
Western Blot ◽  
Immunosorbent Assay ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Relative Increase ◽  
Early Infection ◽  
Canine Leishmaniasis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Time Points ◽  
Experimental Canine

ABSTRACT Quantitative computerized Western blot analysis of antibody responses during experimental canine Leishmania infantum infection distinguished between immunodominant and nonimmunodominant protein bands. Six infected beagles, positive by both PCR and parasite culture, were monitored over 75 weeks postinfection and during a 12-week allopurinol treatment course. All dogs were symptomatic at the time of treatment. Of 12 antigenic bands examined, the immunodominant bands (12, 14, 24, 29, 48, and 68 kDa) showed significantly increased intensities (P < 0.01) and higher frequencies of recognition than the nonimmunodominant bands at all time points. Detection of the former bands at 6 weeks postinfection preceded seroconversion by enzyme-linked immunosorbent assay (ELISA) both on crude Leishmania antigen or the recombinant proteins rK39 and HSP70. Reactivity with the 14-, 48-, and 68-kDa bands signified early infection, whereas increased reactivity with the 14-, 24-, and 29-kDa bands was associated with posttreatment parasite persistence and potential unfavorable prognosis. Total lane intensity (TLI) emerged as a sensitive marker for early infection and increased as early as 4 weeks postinfection. TLI had a significantly higher (P < 0.01) relative increase rate than crude Leishmania antigen or HSP70 or rK39 ELISA at all time points. These immunodominant antigens and TLI, as determined by quantitative Western blotting, will be valuable for early detection and treatment evaluation of canine leishmaniasis.

scholarly journals Investigations on ornithobacterium rhinotracheale in broiler flocks in elazig province located in the east of turkey

2012 ◽  
Vol 49 (No. 8) ◽  
pp. 305-311 ◽  
Author(s):  
G. Ozbey ◽  
H. Ongor ◽  
D. T Balik ◽  
V. Celik ◽  
A. Kilic ◽  
...  
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Sodium Dodecyl ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rrna Gene ◽  
Major Band ◽  
Cell Extracts ◽  
Sds Page ◽  
Ornithobacterium Rhinotracheale

In the present study, lung, trachea and serum samples from broiler flocks slaughtered at an abattoir in Elazig province located in the East of Turkey were examined for the presence of Ornithobacterium rhinotracheale using culture and enzyme-linked immunosorbent assay (ELISA). The identity was latter proved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot analysis, and polymerase chain reaction (PCR) assays. A total of 324 serum and 250 lung and trachea samples were collected from 10 commercially reared chicken flocks showing respiratory manifestations. The samples were obtained from different flocks. The causative agent (ORT) was isolated from trachea (1.5%) of five chickens and from both lung and trachea (0.4%) of only one chicken in the bacteriological examination of tissues. The presence of antibodies against ORT was detected in 33 (10.2%) of the 324 sera by ELISA. A 784 bp fragment of the 16S rRNA gene was amplified using specific primers in the PCR. All ORT isolates that were positive by culture were also detected to be positive by the PCR. SDS-PAGE protein profiles of whole cell extracts showed a high similarity for all the isolates with a major band of the molecular weight of 33&nbsp;kDa (kiloDalton). Results of Western blot analysis indicate four antigenic fractions predominantly with molecular weights of 33, 42, 52 and 66 kDa.

scholarly journals The diurnal pattern of salivary IL-1β in healthy young adults

2017 ◽  
Vol 31 (5) ◽  
Author(s):  
Nur Basirah Ghazali ◽  
Michael Steele ◽  
David Koh ◽  
Adi Idris
Keyword(s):  
Young Adults ◽  
Circadian Rhythm ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Inflammatory Cytokine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diurnal Pattern ◽  
Healthy Individuals

Abstract Disruption in circadian rhythm affects the production of inflammatory cytokines. Understanding how it behaves in diseased conditions is essential. Despite the role of the interleukin-1β (IL-1β), a potent inflammatory cytokine, in human diseases, little is known about the steady-state circadian rhythm of IL-1β in healthy individuals. This short study investigates the diurnal pattern of salivary IL-1β throughout the day in healthy young adults. Twelve participants provided saliva samples at various times throughout the day. Salivary IL-1β were assessed using enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Salivary IL-1β levels were highest at 0430 h and lowest at 0000 h and shared a similar diurnal pattern to that of salivary IL-6. Western blot analysis showed that these levels correspond to the mature form of IL-1β. Our findings are important as it established the diurnal pattern of salivary IL-1β is fluctuating normally throughout the day. The findings also open an incredible opportunity for developing research conducted in the field with saliva as the diagnostic tool.

Em18 and Em16, new serologic marker epitopes for alveolar echinococcosis in Western blot analysis, are the only two epitopes recognized by commercially available weak positive (cut off) sera for Em2plus-ELISA

1995 ◽  
Vol 69 (4) ◽  
pp. 369-371 ◽  
Author(s):  
A. Ito ◽  
Y. Osawa ◽  
M. Nakao ◽  
T. Horii ◽  
M. Okamoto ◽  
...  
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Alveolar Echinococcosis ◽  
Specific Antigen ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Western Blot Assay ◽  
Molecular Weights ◽  
Serologic Marker

AbstractThe assay system for antibody responses against Em2, the most specific antigen for serodiagnosis of alveolar echinococcosis (AE), has been established by enzyme-linked immunosorbent assay (ELISA) but not by Western blot assay, since Em2 antigen is not protein but carbohydrate in nature. Recently we reported that previously undescribed protein epitopes, designated Em18 and Em16 due to their molecular weights, were good serologic markers for AE by Western blot analysis. It has been shown that Em18 and Em16 are the only two epitopes recognized by commercially available weak positive (cut off) sera for the Em2plus-ELISA.

scholarly journals Quercetin Protects RIMVECs From Inflammatory Damage, Pyroptosis and Cell Migration by Inhibiting the TLR4/NF-κB/NLRP3 Pathway

2021 ◽  
Author(s):  
Huixin Zhang ◽  
Yeye Li ◽  
Zhongjie Liu
Keyword(s):  
Cell Migration ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Quantitative Polymerase Chain Reaction ◽  
Inflammatory Factors ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mrna Levels ◽  
Caspase 1 ◽  
Inflammatory Damage

Abstract Background: Intestinal mucosal microvascular endothelial cells (MEC) have multiple functions and play an important role in intestinal bowel diseases (IBD). Quercetin is a flavonoid found in many plants and fruits. It was reported that quercetin can treat several gastrointestinal cancers, but its effect on bacterial enteritis and pyroptosis-related diseases has been rarely studied. This article aims to explore the effect and mechanism of quercetin on inflammatory injury and pyroptosis of RIMVECs.Methods: The inflammatory damage and pyroptosis in RIMVECs were induced by LPS and ATP. Real-time quantitative polymerase chain reaction (RT-qPCR), western blot analysis, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence methods were used to detect TLR4/NF-κB/NLRP3 pathways, inflammatory factors (IL-1β and IL-18) and pyroptosis-related proteins (Caspase-1 and GSDMD). The expression and distribution of ZO-1 were detected by western blot analysis and immunofluorescence method. The late apoptosis and necrosis of cells were measured by cell flow cytometry. Results: The results showed that different concentrations (5, 10, 20μM) of quercetin not only significantly reduced the protein and mRNA levels of TLR4, NLRP3, Caspase-1 and GSDMD, but also down-regulated the protein expression, mRNA and secretion of IL-1β and IL-18. Quercetin also inhibited the phosphorylation of NF-κB p65 and the degradation of IκB. At the same time, quercetin increased the cell migration rate and the expression level of ZO-1, and reduced the number of late apoptotic cells (P<0.05). Conclusions: Our data indicated that Quercetin reduced the inflammatory response and pyroptosis induced by LPS/ATP through the TLR4/NF-κB/NLRP3 pathway, and protected the migration and tight junctions of RIMVECs.

scholarly journals Effects of Electroacupuncture on Ovarian Expression of the Androgen Receptor and Connexin 43 in Rats with Letrozole-Induced Polycystic Ovaries

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Ge Xu ◽  
Andong Zhang ◽  
Jiandang Liu ◽  
Xi Wang ◽  
Jiwei Feng ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Western Blot ◽  
Real Time ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Connexin 43 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mrna Levels ◽  
Endocrine Dysfunction ◽  
Rt Pcr

Background. Polycystic ovarian syndrome (PCOS) occurs in women of reproductive age and is often characterized by reproductive and endocrine dysfunction. Androgens play a major role in PCOS, and previous studies reported abnormal expression of Connexin 43 (Cx43) in animal models of PCOS, suggesting an association of Cx43 with PCOS pathogenesis. Experimental and clinical evidence indicated that acupuncture may be a safe and effective approach for treating reproductive and endocrine disorders in women with PCOS. This study aimed to determine the effects of electroacupuncture (EA) on PCOS and its relationship with the expression of the androgen receptor (AR) and Cx43. Methods. In total, 30 female Sprague Dawley rats (6 weeks old) were randomly divided into three groups: control group, letrozole (LE) group, and LE + EA group. Rats were administered LE solution (1.0 mg/kg) for 21 consecutive days to induce PCOS. For the LE + EA group, additional EA treatment was conducted (2 Hz, 20 min/d) with “Guanyuan” (CV3) for 14 consecutive days. After hematoxylin-eosin staining, the ovarian structure was observed with an optical microscope, and serum levels of the following hormones were examined via enzyme-linked immunosorbent assay (ELISA): testosterone (T), estradiol (E2), sex hormone-binding globulin (SHBG), follicle-stimulating hormone (FSH); luteinizing hormone (LH), insulin (INS), anti-Müllerian hormone (AMH), and inhibin B (INHB). Fasting blood glucose (FBG) levels were evaluated using glucose oxidase-peroxidase. Ovarian mRNA and protein expressions of AR and Cx43 were determined by real-time RT-PCR and Western blot analysis. Results. EA was found to restore the cyclicity and ovarian morphology in the PCOS rat model. Serum derived from the LE + EA group showed significant decreases in the levels of T, free androgen index (FAI), LH, LH/FSH ratio, AMH, INHB, and fasting serum insulin (FINS), and significant increases in the levels of E2, FSH, and SHBG. Western blot analysis showed a decreased protein expression of ovarian AR and Cx43; real-time RT-PCR showed reduced expression of ovarian mRNA levels of AR and Cx43. Conclusions. In conclusion, our results showed that EA can ease hyperandrogenism and polycystic ovary morphology in PCOS rats. Furthermore, EA counteracted the letrozole-induced upregulation of AR and Cx43. These results suggested that acupuncture can break the vicious cycle initiated by excessive androgen secretion and may be an effective treatment method for improving the reproductive and endocrine dysfunction caused by PCOS.

scholarly journals Helicobacter pylori Infection and Strain Types in Adult Dyspeptic Patients Undergoing Endoscopy in a Specialized Hospital of Dhaka City

2009 ◽  
Vol 3 (1) ◽  
pp. 4-9
Author(s):  
Sufi HZ Rahman ◽  
M Anisur Rahman ◽  
MS Arfin ◽  
M Mahbub Alam ◽  
TM Bhuiyan ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Helicobacter Pylori Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rapid Urease Test ◽  
Type I ◽  
Urease Test ◽  
H Pylori

Helicobacter pylori infection occurs worldwide with a high prevalence in developing countries. Virulence of H. pylori strains varies in different geographic regions. The aim of this study was to see H. pylori infection and its strain types in adult dyspeptic patients in Bangladesh and to analyze association of H. pylori strain types with clinical disease and severity of histological gastritis. Ninety consecutive adult dyspeptic patients undergoing diagnostic endoscopy were tested for H. pylori infection by culture, rapid urease test (RUT), histology and anti H. pylori IgG ELISA (Enzyme linked immunosorbent assay). H. pylori strain types were determined by Western Blot analysis. Association of strain types with clinical gastro-duodenal diseases and grades of histological gastritis were analyzed by χ2 test. Among the selected patients, 53 (58.9%) were culture positive, 48 (53.3%) were RUT positive, 31 (34.4%) were histology positive and 82 (91.1%) were anti-H. pylori IgG ELISA positive. By Western Blot analysis of the 90 sera samples, 48 (53.3%) showed antibodies to Type I strain of H. pylori, 21 (23.3%) Intermediate strain and 3 (3.3%) Type II strain. Endoscopically, 20 (22.2%) patients were found normal, 27 (30.0%) had gastritis, 9 (10.0%) had duodenitis, 28 (31.1%) had peptic ulcer disease, 4 (4.4%) had gastric carcinoma, and 2 (2.2%) had reflux esophagitis. Histologically, 34.4% had H. pylori, 44.4% had polymorhonuclear neutrophil (PMN), 100% had mononuclear cell (MNC) infiltration of different grades, 1.1% had atrophic gastritis and 2.2% had intestinal metaplasia of moderate grade. H. pylori strain types was not associated with clinical gastro-duodenal diseases or grades of PMN or MNC infiltration (p > 0.05) in these patients. Key words: Helicobacter pylori infection, H. pylori strain types, gastro-duodenal diseases, grades of gastritis   doi: 10.3329/bjmm.v3i1.2963 Bangladesh J Med Microbiol 2009; 03 (01): 4-9

scholarly journals Identification of Bartonella-Specific Immunodominant Antigens Recognized by the Feline Humoral Immune System

1999 ◽  
Vol 6 (4) ◽  
pp. 558-566 ◽  
Author(s):  
R. L. Freeland ◽  
D. T. Scholl ◽  
K. R. Rohde ◽  
L. J. Shelton ◽  
K. L. O’Reilly
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Bartonella Henselae ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Whole Cells ◽  
Humoral Immune ◽  
Antibody Titers ◽  
Specific Antigens

ABSTRACT The seroreactivities of both naturally and experimentally infected cats to Bartonella henselae was examined. Serum samples collected weekly from nine cats experimentally infected with B. henselae LSU16 were tested by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. The magnitude and isotype of the antibody response were investigated by ELISA. Western blot analysis allowed the identification of at least 24Bartonella-specific antigens recognized by the cats during infection. Antibody titers to specific antigens, as determined by Western blot analysis, ranged from 10 to 640 and varied among the different antibody-antigen interactions. Absorption of sera from an experimentally infected cat, using whole cells and cell lysates of various Bartonella species and other bacteria that commonly colonize cats, supported the identification of thoseBartonella-specific antigens recognized by the experimentally infected cats. Furthermore, a number of possible species- and type-specific antigens were identified. Finally, sera obtained from cats at local animal shelters were screened for the presence of antibodies directed against theBartonella-specific bands identified in the experimentally infected cats. A number of Bartonella-specific antigens have been identified to which strong antibody responses are generated in both experimentally and naturally infected cats, some of which may be useful in diagnosing species- and/or type-specific infections. In addition, the results from these experiments will lead to the development of monoclonal antibodies targeted against those genus-, species-, and type-specific antigens.

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