Evaluation of a novel triplex immunochromatographic test for rapid simultaneous detection of norovirus, rotavirus, and adenovirus on a single strip test.

BACKGROUND: Acute gastroenteritis is one of the major causes of morbidity and mortality in young children worldwide. Among these, rotavirus, norovirus, and adenovirus have been reported as the primary viral pathogens associated with the disease. Rapid diagnosis of viral pathogens is crucial when diarrhea outbreaks occur to ensure the timely administration of appropriate treatment and control measures. METHODS: We evaluated three immunochromatographic test kits designed for the detection of norovirus, rotavirus, and adenovirus in 71 stool specimens collected from children with diarrhea who visited clinics in Japan. The first kit is a triplex immunochromatographic test kit designed for simultaneous detections of norovirus, rotavirus, and adenovirus on a single strip (this kit was referred to as IC-A). The other two immunochromatographic test kits are a dual detection kit for rotavirus and adenovirus, and a single detection kit for norovirus (IC-B). The RT-PCR/PCR was used as the gold standard method. RESULTS: The results revealed that both IC-A and IC-B kits exhibited the same level of sensitivity of detection for rotavirus (72.7%) and adenovirus (22.7%), although the detection rate was lower than that of the RT-PCR/PCR method. However, there was a slight difference in the sensitivity of detection for norovirus between IC-A and IC-B, at 86.7% and 93.3%, respectively. The sensitivity of detection for adenovirus of both kits was relatively lower than those of RT-PCR method. This could be due to low viral load of adenovirus in clinical specimens below the detection limit of IC-A and IC-B kits. However, both immunochromatographic test kits (IC-A and IC-B) exhibited 100% specificity for norovirus, rotavirus, and adenovirus. CONCLUSIONS: The triplex immunochromatographic test kit (IC-A) designed for simultaneous detection of norovirus, rotavirus, and adenovirus has been proved to be more practical and convenient than the use of single or dual detection kits with more or less the same sensitivity and specificity of detections.

Superoxide Dismutase-like Activity of Zeolitic Imidazolate Framework Nanoparticles Comprising Biomimetic Imidazolato-bridged CuZn Units.

Nanoparticles exhibiting enzymatic functions have garnered considerable attention due to their structural robustness and the profusion of active sites that can be introduced to a single nanosized particle. Here we report that nanosized mixed-metal zeolitic imidazolate frameworks (ZIFs) show a superoxide dismutase (SOD)-like catalytic activity. We chose a ZIF composed of copper and zinc ions and 2-methylimidazole, CuZn-ZIF-8, in which the Cu and Zn ions are bridged by an imidazolato ligand. This coordination geometry closely mimics the active site of CuZn superoxide dismutase (CuZnSOD). The CuZn-ZIF-8 nanoparticles exhibit potent SOD-like activity, attributed to their porous nature and numerous copper active sites, and also possess exceptional recyclability.

Effect of sacubitril/valsartan on natriuretic peptide in patients with compensated heart failure.

The time-dependent changes in the natriuretic peptide families during sacubitril/valsartan (S/V) treatment remain obscure in the Asian heart failure (HF) cohort. Eighty-one outpatients with compensated HF were analyzed. The patients were divided into two groups based on the administration of S/V (n = 42) or angiotensin converting enzyme inhibitor (ACE-I; n = 39). Changes to the natriuretic peptide families and the daily dose of loop diuretics were evaluated 3 and 6 months after the intervention. The atrial natriuretic peptide (ANP) level was significantly increased (102 [63-160] pg/mL to 283 [171-614] pg/mL [3 months]; 409 [210-726] pg/mL [6 months]) in the S/V group but not in the ACE-I group. The dose of furosemide was significantly decreased during the six-month follow-up period in the S/V group (40 [20-40] mg to 20 [10-20] mg) but not in the ACE-I group. A multivariate logistic regression model showed that the presence of persistent atrial fibrillation (AF) and HF with a preserved left ventricular ejection fraction (HFpEF) was independently associated with a high delta-ANP ratio (≥ 4.5 ANP value on the start date/ANP value at 6 months; the mean value was used as the cutoff value) (odds ratio [OR]: 4.649, 95% CI 1.032-20.952 and OR: 7.558, 95% CI 1.427-40.042). The plasma level of ANP was increased, and the loop diuretic dose was decreased by the addition of neprilysin inhibitor therapy in patients with compensated HF. In patients with HFpEF and complicated persistent AF, neprilysin inhibitor therapy was associated with an increase in ANP.

The prognostic impact of the serum heart-type fatty acid-binding protein level in patients with sepsis who were admitted to the non-surgical intensive-care unit.

Ongoing myocardial damage at the acme of the sepsis status has not been sufficiently evaluated. The clinical data of 160 sepsis patients who require intensive care and 127 outpatients with chronic heart failure (HF) were compared as a retrospective cohort study. Thereafter, the sepsis patients were divided into 3 groups according to the serum heart-type fatty acid-binding protein (H-FABP) quartiles [low H-FABP = Q1 (n = 39), middle H-FABP = Q2/Q3 (n = 81), and high H-FABP = Q4 group (n = 40)]. The H-FABP level was measured within 15 min of admission. The serum H-FABP levels in the sepsis patients [26.6 (9.3-79.0) ng/ml] were significantly higher than in the choric HF patients [6.6 (4.6-9.7) ng/ml]. A Kaplan-Meier curve showed that the survival rate of the high-H-FABP group was significantly lower than that of the middle- and low-H-FABP groups. The multivariate Cox regression analysis for the 365-day mortality showed that the high-H-FABP group (hazard ratio: 6.544, 95% confidence interval [CI] 2.026-21.140; p = 0.002) was an independent predictor of the 365-day mortality. The same trend in the prognostic impact was significantly (p = 0.015) observed in the cohort that had not been suffering from the cardiac disease before admission. The serum H-FABP level was an independent predictor of the 365-day mortality in the patients who were emergently hospitalized in the intensive-care unit due to sepsis. Ongoing myocardial damage was detected in the majority of patients with sepsis, suggesting that ongoing myocardial damage might be a candidate predictor of adverse outcomes in sepsis patients.

Empagliflozin Administration Can Decrease the Dose of Loop Diuretics and Prevent the Exacerbation of Renal Tubular Injury in Patients With Compensated Heart Failure Complicated by Diabetes.

Background: Whether the dose of loop diuretics can be decreased by administration of a sodium-glucose cotransporter 2 (SGLT2) inhibitor in diabetic outpatients with compensated heart failure (HF) is unclear. Methods and Results: This study prospectively enrolled 60 diabetic outpatients with compensated HF. Patients were randomly divided into 2 groups: those administered the SGLT2 inhibitor empagliflozin (n=28) and those not (n=30). Changes in the daily dose of loop diuretics, blood sampling data, and urinary renal tubular biomarkers were evaluated 6 months after the intervention. The median (interquartile range) furosemide dose decreased significantly over the 6-month follow-up period in the empagliflozin group (from 40 [20-40] to 20 [10-20] mg), but not in the non-empagliflozin group (from 23 [20-40] to 40 [20-40] mg). Hemoglobin levels increased significantly in the empagliflozin group (from 13.2 [11.9-14.6] to 14.0 [12.7-15.0] g/dL). In addition, excretion of acetyl-ß-D-glucosaminidase decreased significantly over the 6-month follow-up in the empagliflozin group (from 4.8 [2.6-11.7] to 3.3 [2.1-5.4] IU/L), especially in the group in which the dose of loop diuretics decreased (from 4.7 [2.5-14.8] to 3.3 [2.1-4.5] IU/L). Conclusions: Empagliflozin administration decreased the dose of loop diuretics and increased the production of erythropoietin, which may help prevent renal tubular injury in diabetic outpatients with HF.

Alarming Situation of Spreading Enteric Viruses Through Sewage Water in Dhaka City: Molecular Epidemiological Evidences.

Global burden of acute viral gastroenteritis remains high, particularly in developing countries including Bangladesh. Sewage water (SW) is an important node to monitor enteric pathogens both in the environment and among the population. Analysis of SW in Dhaka city deems crucially important because a large number of urban-city dwellers live in Dhaka city, the capital of Bangladesh, under a constant threat of precarious sewerage system. In this study, we collected raw SW from five locations of Dhaka city every month from June 2016 to May 2017. It was concentrated with polyethylene glycol (PEG) and investigated for three major enteric viruses, rotavirus A (RVA), norovirus GII (NoV GII) and adenovirus (AdV) using polymerase chain reaction (PCR). Most of these SW samples collected from both hospitals and non-hospital areas yielded enteric viruses: 76% samples were positive for AdV, followed by 53% NoV GII and 38% RVA. Viral load was determined as much as 1 × 107 copies/ml for RVA and 3.5 × 103 copies/ml for NoV GII. Importantly, NoV GII and AdV that can affect people of all ages were predominated during monsoon also when SW overflows and spreads over a wide and crowded area. Genotypes G1, G2, G3, G8, and G9 for RVA, GII.4 for NoV, and type 41 for AdV were detected representing the current profile of circulating genotypes in the population. This study provides the first evidence of distribution of major diarrheal viruses in SW in Dhaka city which is alarming showing grave risk of impending outbreaks through exposure.

Effects of low-energy electron beam irradiation on flexural properties of self-curing acrylic resin.

PURPOSE: The purpose of this study was to confirm the effectiveness of LEB irradiation onto the polymer powder for improving the mechanical properties of self-curing acrylic resin. METHODS: The polymer powder of self-curing acrylic resin was irradiated with total LEB doses of 25, 50, 75 or 100kGy. Non-irradiated powder was used as a control. After LEB irradiation, ESR measurement, weight-average molecular weight measurement and three-point bending test were performed. RESULTS: ESR spectrum of control had no peaks. After LEB irradiation, nine peaks were observed in each ESR spectrum, which indicates the presence of free radicals from main polymer chain. The quantity of free radicals increased linearly up to 100kGy. Calibrated weight-average molecular weights were as follows: control, 960,000; 25kGy, 500,000; 50kGy, 440,000; 75kGy, 410,000; and 100kGy, 390,000. Molecular weight decreased with increasing LEB irradiation dose. The mean values of flexural strength (MPa) were as follows: control, 61.5±3.0; 25kGy, 68.1±4.0; 50kGy, 73.0±1.9; 75kGy, 70.4±3.6; and 100kGy, 67.7±2.3. The flexural strength of the specimens cured with the LEB-irradiated powder was significantly higher than that of control (p<0.01). These results indicate that flexural strength of polymer materials cured with the LEB-irradiated powder increases because of increase in cross-linking structure. CONCLUSION: It is confirmed that LEB irradiation onto the polymer powder of self-curing acrylic resin improves the flexural strength.

Evaluation of newly developed devices for denture placement and removal in the dependent elderly.

OBJECTIVE: The purpose of this study was to subjectively evaluate the utility of newly developed denture placement and removal devices. Objective observations were also made to support the evidence. MATERIALS AND METHOD: Twenty-one subjects were instructed to place and remove their dentures with and without the devices. We evaluated the device based on a questionnaire. Objective observations were based on a 2-D image analysis. We analysed three factors: the time, the area and the circumference required to insert and remove the dentures. RESULTS: Image analysis showed that the effectiveness and ease with which the subject used the device significantly improved with practice. The questionnaire data showed that a majority of the subjects appreciated the device after the first and second time. While there was no significant decrease in time required to place and remove dentures even with the device, the area and circumference of the movement on 2-D images were significantly reduced. CONCLUSION: In this study, the utility of denture placement and removal devices was evaluated both subjectively and objectively. Our data reveal that the device is effective in the elderly. Further minor improvement in the device might be required to increase its effectiveness.

Discrimination between 5-hydroxymethylcytosine and 5-methylcytosine by a chemically designed peptide.

An artificial phosphopeptide recognized the difference between methylated and hydroxymethylated cytosines in DNA. The Sp1 zinc finger peptide substituted by phosphotyrosine effectively discriminated between 5-methylcytosine, 5-hydroxymethylcytosine ((hm)C) and unmethylated cytosine. The DNA recognition properties of the peptide differ from those of other chemicals that detect (hm)C.

Phosphopeptides designed for 5-methylcytosine recognition.

An artificial phosphopeptide has been developed through rational design of the interaction with 5-methylcytosine in duplex DNA. The peptide consists of two tandem zinc finger motifs, in one of which the glutamate was replaced with a phosphotyrosine, the phosphotyrosine in the peptide being effective for methylcytosine selectivity of DNA binding. The flexible modulation of the target methylated sequence by rearrangement of zinc finger peptides is possible, and the phosphopeptide provided us an important hint for expansion of the codes for the interactions of zinc fingers with DNA to methylated DNA sequences. The fluorescence-labeled phosphopeptide provided information on the methylation status of genomic DNA through fluorescence anisotropy after a 10 min incubation.

Effect of Zn doping on improving crystal quality and thermoelectric properties of borosilicides.

Transition-metal (Mo, Mn, Fe, Rh, Ti, Cu, Zn) doping was carried out on the borosilicide compound REB(44)Si(2) (RE = rare earth). REB(44)Si(2) compounds exhibit Seebeck coefficients greater than 200 microV K(-1) at high temperatures and unlike most compounds, the figure of merit shows a steep increase at T > 1000 K making them promising high-temperature thermoelectric materials. Although zinc itself does not remain in the final product, zinc doping was found to improve the crystal quality, which has been a long-standing problem for the borosilicides. As a result, a significant increase of the thermoelectric power factor by more than 30% was achieved.

Osmium complex binding to mismatched methylcytosine: effect of adjacent bases.

We investigated the efficiency of osmium complex formation at 5-methylcytosine in mismatched DNA duplexes. Osmium complexation was not observed in fully matched duplexes, whereas the complexation site and efficiency in mismatched duplexes depended on the 5'-neighboring base of the 5-methylcytosine. In particular, when the base adjacent to the 5' side of the mismatched base pair was thymine, a unique side reaction was observed. However, the mismatched base pairs did not influence the selectivity of osmium complexation with methylated DNA.

Photoresponsive tandem zinc finger peptide.

A photoresponsive tandem zinc finger peptide has been synthesized by attaching an azobenzene unit at its N-terminus resulting in a conjugate that retained a high binding ability toward the target DNA sequence, whereas it showed a significant decrease in DNA-binding ability after photoisomerization by UV irradiation to switch off the inhibition function from the DNA-binding protein.

Osmium complexation of mismatched DNA: effect of the bases adjacent to mismatched 5-methylcytosine.

The efficiency of osmium complex formation at 5-methylcytosine in mismatched DNA duplexes is a key point for the design of sequence-specific detection of DNA methylation. Osmium complexation was not observed in fully matched duplexes, whereas the complexation site and efficiency in mismatched duplexes changed depending on the type of 5'-neighboring base of the 5-methylcytosine forming a mismatched base pair. In particular, when the base adjacent to the 5' side of the mismatched base pair was thymine, a unique "side reaction" was observed. However, the nature of the mismatched base pairs in the reaction site did not influence the selectivity of osmium complex formation with methylated DNA.

Heterogeneity of osmium oxidation efficiency at consecutive thymines.

Consecutive thymine sequences were oxidized more efficiently with a mixture including potassium osmate, potassium hexacyanoferrate(III), and bipyridine than an isolated thymine, and, in particular, osmium oxidation at the 5'T of TT progressed much more rapidly, compared with those of other thymines.

Reactivity of thymine doublet in single strand DNA with osmium reagent.

Osmium oxidation has been frequently utilized for thymine-targeting DNA sequencing because of its high selectivity. In this study, we investigated the reactivity of various DNAs containing multiple thymine bases. A thymine doublet was more easily oxidized than a single thymine in single strand DNA.

Antiviral, antibacterial and vitamin C-synergized radical-scavenging activity of Sasa senanensis Rehder extract.

Sasa senanensis Rehder extract (SE) showed slightly higher cytotoxicity against human squamous cell carcinoma cell lines and human glioblastoma cell lines, as compared with human oral normal cells (gingival fibroblast, pulp cell, periodontal ligament fibroblast), and was more cytotoxic to human myelogenous and T-cell leukemia cell lines. SE showed a bacteriostatic effect on Fusobacterium nucleatum and Prevotella intermedia, but almost completely eliminated hydrogen sulfide (H2S) and methyl mercaptan (CH3SH) produced by these bacteria. SE protected human T-cell leukemia MT-4 cells from the cytopathic effect of human immunodeficiency virus (HIV) infection, and its anti-HIV activity was much higher than that of tannins and flavonoids, comparable with that of natural and synthetic lignins. SE also protected the MDCK cells from the cytopathic effect of influenza virus infection. SE synergistically enhanced the superoxide anion and hydroxyl radical-scavenging activity of vitamin C. The present study suggests the functionality of SE as a complementary alternative medicine.

Anti-HIV and vitamin C-synergized radical scavenging activity of cacao husk lignin fractions.

Cacao husk lignin fractions, prepared by acid precipitation and 50% ethanol precipitation showed unexpectedly higher anti-human immunodeficiency virus (HIV) activity, as compared with the corresponding fractions from the cacao mass, amounting to the level comparable with that of popular anti-HIV compounds. The cacao husk lignin fractions also showed anti-influenza virus activity, but did not show antibacterial activity. The cacao husk lignin fractions synergistically enhanced the superoxide anion and hydroxyl radical scavenging activity of vitamin C. The cacao husk lignin fractions stimulated nitric oxide generation by mouse macrophage-like cells, to a level higher than that attained by lipopolysaccharide (LPS). The present study suggests the functionality of cacao husk lignin fractions as complementary alternative medicine.

DNA methylation analysis using metal complex formation.

Analysis of the cytosine methylation status of a gene is very important for understanding the expression mechanism of genetic information. However, to distinguish 5-methylcytosine (M) from C, i.e., to detect the existence of only one methyl group in a long DNA strand, is not easy. A rapid and selective chemical reaction capable of distinguishing between M and C would become a useful method for efficiently analyzing the status of cytosine methylation at a specific site in a gene. We herein report M-selective oxidation. M was oxidized efficiently by exposure to a reaction mixture containing an osmium complex, making possible a clear distinction from very weak oxidation of C. We readily obtained information on the methylation status at a specific site by means of M-selective oxidation using functional bipyridine ligands.