scholarly journals Western blot method as a necessary step of serodiagnostics of Lyme disease

2012 ◽  
Vol 5 ◽  
pp. 397-402
Author(s):  
Jacek Noworyta ◽  
Maja Machcińska ◽  
Maria Brasse-Rumin ◽  
Jakub Ząbek ◽  
Jolanta Gago
Keyword(s):  
Lyme Disease ◽  
Western Blot ◽  
Western Blot Method

scholarly journals Quality Control Assessment of Canadian Laboratories Testing for Lyme Disease

1991 ◽  
Vol 2 (1) ◽  
pp. 41-45
Author(s):  
Harvey Artsob ◽  
Maxwell Garvie
Keyword(s):  
Public Health ◽  
Quality Control ◽  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Western Blot ◽  
Etiological Agent ◽  
Immunofluorescent Assay ◽  
Satisfactory Performance ◽  
Endemic Areas ◽  
Negative Controls

In June 1990 a quality control assessment was undertaken of Canadian public health laboratories testing for antibodies toBorrelia burgdorferi, the etiological agent of Lyme disease. Twenty sera were distributed to nine laboratories, including 12 obtained from patients in Lyme endemic areas and presumed to be serological positives, and eight prescreened negative controls. Seventeen serological reports were submitted, comprising nine enzyme-linked immunosorbent assays (elisa), six immunofluorescent assays and two Western blot assessments. Antibodies were detected in 11 of the 12 sera which had been presumed to be positive. Assuming 11 positive sera had been submitted, the test sensitivities varied from 88.9 to 100% byelisa, and 54.5 to 90.1% by immunofluorescent assay. Specificities were 100% for all but oneelisaand one immunofluorescent assay assessment. The results indicate a satisfactory performance byelisabut a need for upgrading or replacement of some immunofluorescent assay tests.

scholarly journals Erythropoietin production by the kidney and the liver in response to severe hypoxia evaluated by Western blotting with deglycosylation

2020 ◽  
Author(s):  
Yukiko Yasuoka ◽  
Takashi Fukuyama ◽  
Yuichiro Izumi ◽  
Yushi Nakayama ◽  
Hideki Inoue ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mass Spectrometry ◽  
Western Blot ◽  
Western Blotting ◽  
Fold Increase ◽  
Interstitial Cells ◽  
Severe Hypoxia ◽  
Liquid Chromatography Mass Spectrometry ◽  
Western Blot Method ◽  
Erythropoietin Production

AbstractThe detection of erythropoietin (Epo) protein by Western blotting has required pre-purification of the sample. We developed a new Western blot method to detect plasma and urinary Epo using deglycosylation. Epo in urine and tissue and erythropoiesis-stimulating agents (ESAs) in urine were directly detected by our Western blotting. Plasma Epo and ESAs were detected by our Western blotting after deglycosylation. The broad bands of Epo and ESAs were shifted to 22 kDa by deglycosylation except PEG-bound epoetin β pegol. The 22 kDa band from anemic patient urine was confirmed by Liquid Chromatography/Mass Spectrometry (LC/MS) to contain human Epo.Sever hypoxia (7% O2, 4 hr) caused a 400-fold increase in deglycosylated Epo expression in rat kidneys, which is consistent with the increases in both Epo gene expression and plasma Epo concentration. Immunohistochemistry showed Epo expression in nephrons but not in interstitial cells under control conditions, and hypoxia increased Epo expression in interstitial cells but not in tubules.These data show that intrinsic Epo and all ESAs can be detected by Western blot either directly in urine or after deglycosylation in blood, and that the kidney is the main and sole site of Epo production in control and severe hypoxia. Our method will completely change Epo doping and detection.

scholarly journals Borrelia burgdorferi and Borrelia miyamotoi seroprevalence in California blood donors

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243950
Author(s):  
Sharon I. Brummitt ◽  
Anne M. Kjemtrup ◽  
Danielle J. Harvey ◽  
Jeannine M. Petersen ◽  
Christopher Sexton ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Western Blot ◽  
Sierra Nevada ◽  
Blood Donors ◽  
Test System ◽  
Emerging Diseases ◽  
Borrelia Miyamotoi ◽  
Increased Risk ◽  
Significant Difference

The western blacklegged tick, Ixodes pacificus, an important vector in the western United States of two zoonotic spirochetes: Borrelia burgdorferi (also called Borreliella burgdorferi), causing Lyme disease, and Borrelia miyamotoi, causing a relapsing fever-type illness. Human cases of Lyme disease are well-documented in California, with increased risk in the north coastal areas and western slopes of the Sierra Nevada range. Despite the established presence of B. miyamotoi in the human-biting I. pacificus tick in California, clinical cases with this spirochete have not been well studied. To assess exposure to B. burgdorferi and B. miyamotoi in California, and to address the hypothesis that B. miyamotoi exposure in humans is similar in geographic range to B. burgdorferi, 1,700 blood donor sera from California were tested for antibodies to both pathogens. Sampling was from high endemic and low endemic counties for Lyme disease in California. All sera were screened using the C6 ELISA. All C6 positive and equivocal samples and nine randomly chosen C6 negative samples were further analyzed for B. burgdorferi antibody using IgG western blot and a modified two ELISA test system and for B. miyamotoi antibody using the GlpQ ELISA and B. miyamotoi whole cell sonicate western blot. Of the 1,700 samples tested in series, eight tested positive for antibodies to B. burgdorferi (0.47%, Exact 95% CI: 0.20, 0.93) and two tested positive for antibodies to B. miyamotoi (0.12%, Exact 95% CI: 0.01, 0.42). There was no statistically significant difference in seroprevalence for either pathogen between high and low Lyme disease endemic counties. Our results confirm a low frequency of Lyme disease and an even lower frequency of B. miyamotoi exposure among adult blood donors in California; however, our findings reinforce public health messaging that there is risk of infection by these emerging diseases in the state.

scholarly journals Anti-inflammatory activity of pectic enzyme-treated pectin on lipopolysaccharide-induced RAW 264.7 cells

2019 ◽  
Vol 1 (1) ◽  
pp. 23-30 ◽  
Author(s):  
Cian-Song Huang ◽  
Qiao-Lin Li ◽  
Diana Lo ◽  
Yuh-Tai Wang ◽  
Ming-Chang Wu
Keyword(s):  
Western Blot ◽  
Raw 264.7 Cells ◽  
No Production ◽  
Raw 264.7 ◽  
Pectic Enzyme ◽  
Western Blot Method ◽  
Tnf Α ◽  
Cox 2 ◽  
Necrosis Factor ◽  
Normal Macrophage

The purpose of this study was to investigate the ability and pathway of the pectic enzyme-treated (PET) pectin to inhibit the inflammation of macrophage RAW 264.7 induced by lipopolysaccharide. Results showed that PET-pectin produced from 1% substrate and 48 h reaction time had the highest antioxidative activity, thus these parameters were used to produce PET-pectin used in this study. PET-pectin showed no cell cytotoxicity to normal macrophage RAW 264.7 and reduce the nitrite secretion from LPS-induced RAW 264.7 by 20%. Finally, the expression of cytokines, including NO synthase (iNOS), nitric oxide (NO), cyclooxygenase-2 (COX-2), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and tumor necrosis factor (TNF-α) were analyzed by western blot. In the western blot method, it was found that iNOS, COX-2, NF-κB, TNF-α and other proteins that activated NO production had a downtrend. It was found that PET-pectin possess promising activity to mitigate the inflammatory response.

Potentially misleading Western blot results in Lyme disease diagnosis

2006 ◽  
Vol 63 (3) ◽  
pp. 142-144
Author(s):  
S. Mavin ◽  
R. Evans ◽  
K. Appleyard ◽  
A.W.L. Joss ◽  
D.O. Ho-Yen
Keyword(s):  
Lyme Disease ◽  
Western Blot ◽  
Disease Diagnosis

Effect of PGK1 on radiosensitivity through regulation of CIN and CFL1 in human glioma cells.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e14631-e14631
Author(s):  
Hongyi Liu ◽  
Yuchi Liu ◽  
Hong Xiao ◽  
Mengjie Zhao ◽  
Yong Xiao
Keyword(s):  
Western Blot ◽  
Phosphoglycerate Kinase ◽  
Glioma Cells ◽  
Underlying Mechanism ◽  
Migration And Invasion ◽  
Normal Cells ◽  
Western Blot Method ◽  
Successful Establishment ◽  
U373 Cells

e14631 Background: Phosphoglycerate kinase 1 (PGK1) has been detected overexpressed in many malignancies and usually correlated with increased tumorigenesis and poor survival. Our previous study revealed that PGK1 significantly up-regulated and positively correlated with cofilin-1 (CFL1) expression in radioresistant astrocytomas samples. However, the further mechanism remains unknown. Methods: In the present study, functional interactions among PGK1, chronophin (CIN) and CFL1 were analyzed by co-immunoprecipitation and immunofluorescence experiments. Transfection of shRNA and pcDNA3.1-plasmid were respectively utilized to silence and overexpress PGK1 or CIN in both normal glioma cells (U251 and U373) and estabilshed radioresistant (RR-U251 and RR-U373) cells. The effect of PGK1 on CIN and CFL1 expression levels was investigated with Western-blot method; while the effect of PGK1 or CIN on radiosensitivity of glioma cells was evaluated using cell viability, migration and invasion assays after irradiation. Results: Cell proliferation, migration and invasion abilities significantly increased in RR-U251 and RR-U373 cells compared with those of normal cells, indicating the successful establishment of RR-glioma cell models. Laser confocal experiment in vitro showed the colocalization of PGK1 and CIN. Meanwhile, PGK1 specifically and directly combined with CIN, while did not interplay with CFL1. Elevated PGK1 expression level was observed in radioresistant cells compared with that of normal cells. The western blot analysis showed that CFL1 and CIN levels were positively correlated with the PGK1 expression in both normal and radioresistant cells. The proliferation, migration and invasion capabilities significantly decreased in PGK1-silenced and CIN-silenced glioma cells, indicating the improvement of the radiosensitivity of radioresistant cells. Conclusions: Our research implied the effect of CIN-mediated PGK1 regulation on CFL1 level, thereby altering the migration and invasion abilities in nomal and notably radioresistant glioma cells. These findings clarify a novel underlying mechanism of radioresistance in glioma cells and provide valid basis for further explorations concerning radiotherapy.

Effects of Intermittent Cisplatin Intervention on 6PGD Levels and Tumor Growth and Migration in A549 Lung Cancer Cells

2021 ◽  
Vol 8 ◽  
Author(s):  
WU Qiang ◽  
TANG Lin-dong ◽  
WU Yan-hui ◽  
LI Qiang ◽  
YU Li
Keyword(s):  
Lung Cancer ◽  
Tumor Growth ◽  
Western Blot ◽  
Cancer Cells ◽  
A549 Cells ◽  
Lung Cancer Cells ◽  
Migration Ability ◽  
Western Blot Method ◽  
And Migration ◽  
A549 Lung

Aim: To investigate the changes of A549 protein 6PGD expression, tumor growth and migration, NADPH and ROS levels in lung cancer cells after intermittent intervention with DDP. Methods: The sensitivity of A549 cells to DDP was detected by cck-8 method, A549 cells were treated by DDP intermitten, 6PGD protein expression was detected by Western blot method, and the ROS level of A549 cells was determined by ROS kit. NADPH levels of A549 cells were determined by NADPH kit. Result: After DDP intermittent intervention, the expression of 6PGD protein in lung cancer cells A549 was increased, the growth and migration of A549 cells were significantly inhibited, and ROS and NADPH levels were significantly increased. Conclusion: 6PGD were upregulated in after DDP intermittent intervention and affected the migration ability in lung cancer cells.

The Expression and Significance of Matrix Metalloproteinase, Wnt5a and β-Catenin in Placental Tissue of Preeclampsia

2021 ◽  
Vol 11 (12) ◽  
pp. 2375-2380
Author(s):  
Hui Zhang ◽  
Wei Chen ◽  
Shenmiao Chen ◽  
Junrong Wang ◽  
Shunfen Mao
Keyword(s):  
Matrix Metalloproteinase ◽  
Western Blot ◽  
Pregnant Women ◽  
Placental Tissue ◽  
Normal Pregnancy ◽  
Expression Level ◽  
Control Group ◽  
Cytotrophoblast Cell ◽  
Western Blot Method ◽  
Low Expression

In order to investigate the expression and clinical significance of MMPs (Matrix Metalloproteinase) (MPP-2 and MPP-9), Wnt5a as well as β-catenin in placental tissues of preeclampsia (PE) patients, 36 PE patients (PE group) and 25 pregnant women (control group) with normal pregnancy were selected for cesarean delivery. The expression and localization of MPP-2, MPP-9, Wnt5a as well as β-catenin proteins in placental tissue were detected by Western blot method and immunohistochemistry method. The correlation between the poor prognosis of mother and child and the expression level of MMPs, Wnt5a as well as β-catenin is analyzed. The results showed that Western blot results showed that MPP-2, MPP-9, Wnt5a as well as β-catenin proteins were expressed in placental tissue, and the expression level of them of placental tissue in PE group was significantly lower than that in control group (P < 0.01). Immunohistochemistry showed that MMPs, Wnt5a, as well as β-catenin proteins were expressed in the cytotrophoblast cell (CTB) and syncytiotrophoblast cell (STB) of placental tissue, among which Wnt5a proteins as well as β-catenin proteins were mainly expressed in the cytoplasm of STB and CTB respectively. MMPs proteins were mainly expressed in the cytoplasm of these two types of cells. Low expression of MPPs, Wnt5a, β-catenin proteins may be related to blood pressure as well as 24 h urine protein of pregnant women at admission. In conclusion, Low expression of MPPs, Wnt5a, β-catenin proteins may be involved in the PE pathogenesis.

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