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Comparison of Thymic Stromal Lymphopoietin Concentration in Various Human Biospecimens from Asthma and COPD Patients Measured with Two Different ELISA Kits

Abstract
Thymic stromal lymphopoietin (TSLP) seems a promising asthma biomarker. In earlier studies, mainly the serum concentration of TSLP was investigated. The aim of the present study was to compare the TSLP concentration measured by two different ELISA kits in the serum, induced sputum, and exhaled breath condensate in asthma, COPD, and control subjects. The study included 24 asthmatics, 36 patients with COPD, and 12 controls. TSLP concentration was measured with the use of R&D and EIAab commercial ELISA kits. The results obtained with the EIAab kit were 3 to even 45-fold higher than those measured with the R&D kit. Significant differences between the investigated groups were found only for the TSLP concentration in induced sputum. When the R&D kit was used, the highest TSLP levels in induced sputum were found in asthmatics, while the EIAab kit showed the highest TSLP levels in controls. The distribution of results in the Bland-Altman plot was typical for a proportional constant error. TSP concentration in induced sputum might be a more reliable asthma biomarker than serum TSLP. We conclude that TSLP level is highly dependent on the ELISA kit used for the measurement. Thus, judgement on TSLP results obtained with different assays might be confusing and lead to wrong conclusions.

 

Caveolin-1, Ring finger protein 213, and endothelial function in Moyamoya disease

Posted by Bang O Y, Chung J W, Kim S J, et al. on 2016-12-12 22:48:08

Abstract
Background

Moyamoya disease is a unique cerebrovascular occlusive disease of unknown etiology. Ring finger protein 213 (RNF213) was identified as a susceptibility gene for Moyamoya disease in East Asian countries. However, the pathogenesis of Moyamoya disease remains unclear.
Methods

We prospectively analyzed clinical data for 139 patients with Moyamoya disease (108 bilateral Moyamoya disease, 31 unilateral Moyamoya disease), 61 patients with intracranial atherosclerotic stroke, and 68 healthy subjects. We compared the genetic (RNF213 variant) and protein biomarkers for caveolae (caveolin-1), angiogenesis (vascular endothelial growth factor (VEGF) and receptor (VEGFR2), and antagonizing cytokine (endostatin)) and endothelial dysfunction (asymmetric dimethylarginine (ADMA), and nitric oxide and its metabolites (nitrite and nitrate)) between patients with Moyamoya disease and intracranial atherosclerotic stroke. We then performed path analysis to evaluate whether a certain protein biomarker mediates the association between genes and Moyamoya disease.
Results

Caveolin-1 level was decreased in patients with Moyamoya disease and markedly decreased in RNF213 variant carriers. Circulating factors such as VEGF and VEGFR2 did not differ among the groups. Markers for endothelial dysfunction were significantly higher in patients with intracranial atherosclerotic stroke but normal in those with Moyamoya disease. Path analysis showed that the presence of the RNF213 variant was associated with caveolin-1 levels that could lead to Moyamoya disease. The level of combined marker of Moyamoya disease (caveolin-1) and intracranial atherosclerotic stroke (ADMA, an endothelial dysfunction marker) predicted Moyamoya disease with good sensitivity and specificity.
Conclusion

Our results suggest that Moyamoya disease is a caveolae disorder but is not related to endothelial dysfunction or dysregulation of circulating cytokines.

 

Abstract
Macrophage migration inhibitory factor (MIF) is a key pro-inflammatory cytokine in immuno-inflammatory diseases. For the first time, we examined the expression of MIF in Epinephelus awoara (E. awoara). MIF expressions have been detected in the head kidney, spleen, liver, brain, intestine, gill, heart, stomach, and muscle of E. awoara infected with Vibrio parahaemolyticus. The mRNA levels observed in infected groupers were higher than those in healthy groupers. MIF, tumor necrosis factor-α (TNF-α), and interleukin-1 (IL-1) tissue levels have been measured by ELISA. A significant increase in MIF, TNF-α, and IL-1 tissue levels have been found in the treatment groups compared with those in controls. MIF, TNF-α and IL-1 tissue levels in the spleen, head kidney, intestine, and liver of E. awoara during the challenge trial with V. parahaemolyticus were significantly higher than those in controls. There was evidence of functions of MIF in a positive feedback loop with TNF-α and IL-1 that could perpetuate the inflammatory process in grouper infected with V. parahaemolyticus. In conclusion, these results indicated that MIF was related to pathogen-induced immune response.

 

COMPARATIVE STUDY BETWEEN DKK-1 AND AFP FOR DIAGNOSING OF HEPATOCELLULAR CARCINOMA AMONG EGYPTIAN PATIENTS

Posted by Mohamed F Z, Barakat L A A, Radwan N H, et al. on 2016-12-12 22:45:30

ABSTRACT
Background: Hepatocellular carcinoma is a major health problem. It was ranked 2nd most common cancer site among males and 7th among females. AFP is the main routinely parameter for HCC diagnosis, it can also be elevated in liver cirrhosis. It represents a liver cell- specific, not a tumor-specific marker, for these reasons, our suggestion is to use AFP as a supplementary marker for diagnosis of HCC. So identification of sensitive biomarkers to improve early diagnosis of HCC is needed. Our aim is to evaluate serum Dickkopf-1 (DKK1) as biomarkers for early diagnosis of HCC.

Methods: The study included 40 HCCs patients (stage I (9) cases, stage II (31) cases), 30 liver cirrhosis patients, and 20 healthy subjects were enrolled. Serum DKK1 using ELISA kit was measured in all included subjects.

Results: Serum DKK1 was significantly elevated in HCC group compared to cirrhotic and healthy control groups. Receiver operating characteristic curve showed the best cutoff values for DKK1, and AFP were (1 ng/ml and 400 ng/ml) respectively. Area under the curve of DKK1 was significantly larger than that of AFP (0.990 vs. 0.763). The sensitivity of DKK1 and AFP was (98% vs. 28%) respectively.
Conclusion: Serum DKK1 might be potential diagnostic markers for HCC.
KEY WORDS: HCC patients, DKK1, AFP.

 

Electrochemical immunosensor for the determination of 8-isoprostane aging biomarker using carbon nanohorns-modified disposable electrodes

Posted by Sánchez-Tirado E, González-Cortés A, Yudasaka M, et al. on 2016-12-12 22:44:00

Abstract

The first electrochemical immunosensor for the determination of 8-isoprostane (8-iso prostaglandin F2α, ISO), one of the most reliable biomarkers of lipid peroxidation in the human body and of aging related to Alzheimer’s disease or atherosclerosis is reported in this article. Disposable screen-printed carbon electrodes modified with carboxylated carbon nanohorns (CNHs) were employed as scaffolds for covalent immobilization of a specific anti-ISO antibody). A competitive immunoassay involving ISO and HRP-labeled antigen was designed and the determination of ISO was carried out by amperometry at − 200 mV using the H2O2/hydroquinone (HQ) system. Under the optimized conditions, the immunosensor provides a linear response for ISO (r2 = 0.998) extending up to 700 pg/mL, which is suitable for the determination of the target compound in human serum. The analytical performance of the immunosensor improves that claimed for ELISA kits in terms of linearity of the calibration plot, precision, with RSD values lower than 1% , and assay time (1 h 30 min), and exhibits a low limit of detection, 12 pg/mL, a long storage stability (30 days), and an excellent selectivity against other proteins that may be found in human serum. The analytical utility of the developed immunosensor was demonstrated by determining ISO in two types of human serum samples: lyophilized spiked serum, and real human serum from healthy male and female individuals with good results.

 

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