High sensitivity of an ELISA kit for detection of the gamma-isoform of 14-3-3 proteins: usefulness in laboratory diagnosis of human prion disease

<p>Abstract</p> <p>Background</p> <p>The gamma-isoform of the 14-3-3 protein (14-3-3 gamma) is expressed in neurons, and could be a specific marker for neuronal damage. This protein has been reported as a detectable biomarker, especially in the cerebrospinal fluid (CSF) of Creutzfeldt-Jakob disease (CJD) patients by Western blotting (WB) or enzyme-linked immunosorbent assays (ELISAs). Western blotting for 14-3-3 gamma is not sensitive, and the reported data are conflicting among publications. An ELISA specific for 14-3-3 gamma is not available.</p> <p>Methods</p> <p>CJD patients (n = 114 sporadic CJD patients, 7 genetic CJD, and 3 iatrogenic CJD) and 99 patients with other neurodegenerative diseases were examined in this study. The CSF samples obtained were analyzed by Western blotting for 14-3-3 gamma, and by ELISA for total tau protein. We evaluated the sensitivity and specificity of the newly developed sandwich ELISA for 14-3-3 gamma.</p> <p>Results</p> <p>The cut-off value of the 14-3-3 gamma ELISA was > 1, 683 AU/ml; and sensitivity was 95.2%, with 72.7% specificity. This specificity was the same for the total tau protein ELISA. Seven CJD cases were negative by WB but positive using the 14-3-3 gamma ELISA, indicating that the ELISA is more sensitive. All 21 cases of early stage CJD could be diagnosed using a combination of the 14-3-3γ ELISA and diffusion weighted MR imaging (DWI-MRI).</p> <p>Conclusion</p> <p>The 14-3-3 gamma ELISA was more sensitive than conventional WB, and was useful for laboratory diagnosis of CJD, similar to the ELISA for the tau protein. Using DWI-MRI and these ELISA tests on CSF, diagnosis of CJD will be possible even at early stages of the disease.</p

New auroral spectrometer using an acousto-optic tunable filter

This paper reports the performance and capability of a newly developed zenith spectrometer (for measurements of airglow and aurora) that uses an acousto-optic tunable filter (AOTF). The AOTF can scan the pass-band of the spectrometer between 450 and 700 nm with a bandwidth of 2-3 nm by changing the RF driver frequency from 180 to 100 MHz. The absolute sensitivity of the spectrometer is ～0.1-1.5 counts/Rayleigh/s per spectral step. The spectrometer is fully automated. The O I (557.7 nm) airglow line can be clearly identified in test observations of midlatitude airglow performed at Shigaraki Observatory, Japan. Based on an estimate of the signal-to-noise ratio, we conclude that the full auroral spectrum (450-700) nm can be measured by the AOTF spectrometer with a time resolution of ～100 s and a signal-to-noise ratio of ～100 for an auroral emission intensity of 10 kR. An example of the auroral spectra is shown for observations made at Syowa Station in Antarctica

Detection of reverse transcriptase activity by enzyme-linked immunosorbent assay in human immunodeficiency virus type 1.

An enzyme-linked immunosorbent assay (ELISA) using biotin-labelled oligo-dT primer and digoxigenin (Dig)-dUTP was designed to measure the reverse transcriptase (RT) activity of human immunodeficiency virus type 1 (HIV-1). The ELISA system involves the selective detection step of a newly synthesized cDNA by two specific bindings, biotin-streptavidin binding and alkaline phosphatase (AP)-conjugated anti-Dig-Dig binding, and the enzymatic amplification step to increase coloring generated by AP. This method was used to measure the activity of RT in the culture supernatants of peripheral leukocytes obtained from four anti-HIV-1-positive persons cocultivated with those from four anti-HIV-1-negative persons. RT activity was detected in all of four anti-HIV-1-positive culture supernatants but not in those cultivated with anti-HIV-1-negative supernatants alone. Thus, our improved ELISA for detection of HIV-1 appears to be sensitive enough and useful for routine laboratory work. This non-radioactive method will also be useful for detecting other retroviruses and for screening of RT inhibitors.</p

VERA monitoring of the radio jet 3C 84 during 2007--2013: detection of non-linear motion

We present a kinematic study of the subparsec-scale radio jet of the radio galaxy 3C 84/NGC 1275 with the VLBI Exploration of Radio Astrometry (VERA) array at 22 GHz for 80 epochs from 2007 October to 2013 December. The averaged radial velocity of the bright component "C3" with reference to the radio core is found to be $0.27 pm 0.02c$ between 2007 October and 2013 December. This constant velocity of C3 is naturally explained by the advancing motion of the head of the mini-radio lobe. We also find a non-linear component in the motion of C3 with respect to the radio core. We briefly discuss possible origins of this non-linear motion.Comment: 11 pages, 7 figures, 8 tables (table 1 - 5 are supplementaries), accepted for publication on PAS

Effect of Partial Ileal Bypass on Cholesterol and Bile Acid Metabolism in Rats

In order to clarify the effect of ileal bypass on cholesterol and bile acid metabolism, partial (20 cm) ileal bypass rats were fed a 2% cholesterol supplemented diet for a week after 4 weeks of the operation. The serum and liver cholesterol and phospholipid levels, biliary cholesterol, phospholipid and bile acid secretions, and fecal cholesterol, coprostanol and bile acid excretions were examined. The serum cholesterol level in ileal bypass rats was lower than in normal rats and no hypercholesterolemia was brought about in ileal bypass rats by feeding them the cholesterol diet. The liver cholesterol level increased by feeding the cholesterol diet even in ileal bypass rats but the increase was far less than that in normal rats (22% versus 57%). Biliary bile acid secretion decreased and fecal bile acid excretion increased markedly in ileal bypass rats. Deoxycholic acid increased remarkably in both bile and feces and resulted in an increase in the ratio of the sum of bile acids derived from cholic acid over the sum of bile acids derived from chenodeoxycholic acid (CA/CDCA ratio) in the feces. These results suggest that the absorption of bile acids is impaired, the pool size of bile acids decreases and the hepatic synthesis of bile acids, especially that of cholic acid, increases in ileal bypass rats. As a result, cholesterol feeding to ileal bypass rats produces neither hypercholesterolemia nor a further increase in bile acid synthesis