Modest antiviral activity of Toll-like receptor 3 (TLR3) against

Human respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory infections in infants and Toll-like receptors (TLRs) are the first line of host defense against such infections. In this study, we aimed to elucidate the antiviral effect of TLR3 against RSV infection. The human TLR3 gene was either transiently or stably overexpressed in A549 cells and they were infected with the Long strain of RSV. In both cases, RSV production determined by plaque assay was modestly but significantly decreased in the TLR3-overexpressed cells compared with control cells. Less interferon (IFN)-β, measured by ELISA, was produced in the supernatant of the TLR3- overexpressed cells. Neutralization of IFN-β in the supernatant of the TLR3-overexpressed cells failed to increase RSV production to the same level as controls. These results indicate that TLR3 has modest anti-RSV activity and IFN-β seems not to be a significant mediator of this activity

LINE-1 hypomethylation status of circulating cell-free DNA in plasma as a biomarker for colorectal cancer.

Colorectal cancer (CRC) is a serious public health problem and non-invasive biomarkers improving diagnosis or therapy are strongly required. Circulating cell-free DNA (cfDNA) has been a promising target for this purpose. In this study, we evaluated the potential of long interspersed nuclear element-1 (LINE-1) hypomethylation as a blood biomarker for CRC. LINE-1 hypomethylation level in plasma cfDNA in 114 CRC patients was retrospectively examined by absolute quantitative analysis of methylated alleles real-time PCR, and was expressed using LINE-1 hypomethylation index (LHI) [unmethylated copy number/ (methylated copy number + unmethylated copy number)]. Greater LHI values indicated enhanced hypomethylation. In our clinicopathological analysis, CRC patients with large tumors (≥6.0 cm), advanced N stage (≥2), and distant metastasis (M1) had statistically significantly higher cfDNA LHI than other CRC patients, suggesting cfDNA LHI as a disease progression biomarker for CRC. Furthermore, early stage I/II (n = 57) as well as advanced stage III/IV (n =57) CRC patients had significantly higher cfDNA LHI than healthy donors (n=53) [stage I/II: median 0.369 (95% confidence interval, 0.360-0.380) vs. 0.332 (0.325-0.339), P \u3c 0.0001; stage III/IV: 0.372 (0.365-0.388) vs. 0.332 (0.325-0.339), P \u3c 0.0001]. The receiver operating characteristic analysis showed that cfDNA LHI had the detection capacity of CRC with area under the curve(AUC) of 0.79 and 0.83 in stage I/II and stage III/IV CRC patients, respectively. The present study demonstrated for the first time the potential of plasma cfDNA LHI as a novel biomarker for CRC, particularly for early stage detection

Direct Renin Inhibitor is Better than Angiotensin II Receptor Blocker for Intrarenal Arterioles

Background/Aims: We have reported that the long-term administration of angiotensin II receptor blockers (ARBs) induced unusual proliferative changes of renal afferent arteriolar smooth muscle cells (SMCs) in rats, associated with the overproduction of renin. In this study, we examined that a direct renin inhibitor (DRI: Aliskilen; Novartis Pharma Co, USA) might induce different changes on afferent arteriolar walls compared to ARBs. Method: Twenty one 6-weeks-old male spontaneous hypertensive rats (SHRs) were divided into the following three groups: high-dose DRI group (n=7), low-dose DRI group (n=5) and control group (n=9). The rats were fed a standard diet (0.4%NaCl) containing high-dose (150mg/kg/day), low-dose (30mg/kg/day) DRI and without DRI for 12 weeks. The kidneys were examined by histological and immunohistochemical studies. Systolic blood pressure, 24-h urine samples and blood samples were also examined. Results: The afferent arteriolar SMC walls in the two DRI groups showed no proliferative changes. The positive renin expression area was the largest in the high-dose DRI group among the three groups (14.3±4.0µm2, 6.7±2.0µm2, 2.6±0.9µm2/glomerlus, p=0.020, p=0.008, p=0.017, respectively). Conclusion: The long-term DRI administration increases tissue and circulatory renin; however, afferent arteriolar proliferative changes as shown in ARBs were not induced

Genetic Variability and Molecular Epidemiology of Respiratory Syncytial Virus Subgroup A Strains in Japan Determined by Heteroduplex Mobility Assay

We used heteroduplex mobility assay (HMA) to determine the genetic variability of 118 respiratory syncytial virus (RSV) field isolates from 19 epidemics occurring in a Japanese urban area between 1980 and 2000. Nucleotides 1 to 584 of the attachment G glycoprotein gene were amplified by reverse transcription-PCR, and the PCR amplicons were analyzed by HMA by using the earliest isolate from 1980 as the reference throughout. We also performed PCR-restriction fragment length polymorphism (RFLP) analysis and phylogenetic analysis on the same nucleotide sequence. PCR-RFLP revealed 9 patterns, whereas HMA produced 31 distinct patterns. The RFLP patterns were divided into two to seven distinct HMA genotypes. Field strains with similar degrees of G gene nucleotide differences from the reference strain often showed distinct HMA types. The RSV genetic heterogeneity detected by direct sequencing of the PCR amplicon was usually identical to HMA analysis. Analysis of the molecular epidemiology of RSV subgroup A isolates obtained by HMA showed that new RSV variants emerged with each epidemic and that previously dominant variants seldom recurred in subsequent epidemics. HMA is useful in detecting genetic variants of RSV subgroup A and has some advantages over other conventional methods

Comparison of an Immunochromatography Test with Multiplex Reverse Transcription-PCR for Rapid Diagnosis of Respiratory Syncytial Virus Infections

A new commercial rapid 10-min one-step immunochromatography (IC) test, SAS RSV test, was compared to another IC test, Directigen EZ RSV, employing RT-PCR as the “gold standard” for detecting respiratory syncytial virus. Of 102 clinical samples, 79 were positive by RT-PCR, 66 (82.5%) were positive with the SAS RSV test, and 55 (69.6%) were positive with Directigen EZ RSV. The specificity of the new test was 91.3% (21 of 23), similar to that of Directigen EZ RSV (100% [23 of 23]). This test performs well enough to be used for patient care