An effective live-attenuated Zika vaccine candidate with a modified 5' untranslated region.

Zika virus (ZIKV) is a mosquito-transmitted flavivirus that has caused devastating congenital Zika syndrome (CZS), including microcephaly, congenital malformation, and fetal demise in human newborns in recent epidemics. ZIKV infection can also cause Guillain-Barré syndrome (GBS) and meningoencephalitis in adults. Despite intensive research in recent years, there are no approved vaccines or antiviral therapeutics against CZS and adult Zika diseases. In this report, we developed a novel live-attenuated ZIKV strain (named Z7) by inserting 50 RNA nucleotides (nt) into the 5' untranslated region (UTR) of a pre-epidemic ZIKV Cambodian strain, FSS13025. We used this particular ZIKV strain as it is attenuated in neurovirulence, immune antagonism, and mosquito infectivity compared with the American epidemic isolates. Our data demonstrate that Z7 replicates efficiently and produces high titers without causing apparent cytopathic effects (CPE) in Vero cells or losing the insert sequence, even after ten passages. Significantly, Z7 induces robust humoral and cellular immune responses that completely prevent viremia after a challenge with a high dose of an American epidemic ZIKV strain PRVABC59 infection in type I interferon (IFN) receptor A deficient (Ifnar1-/-) mice. Moreover, adoptive transfer of plasma collected from Z7 immunized mice protects Ifnar1-/- mice from ZIKV (strain PRVABC59) infection. These results suggest that modifying the ZIKV 5' UTR is a novel strategy to develop live-attenuated vaccine candidates for ZIKV and potentially for other flaviviruses.

Draft Genome Sequences of Six Strains Isolated from the Rhizosphere of Wheat Grown in Cadmium-Contaminated Soil.

This study presents high-quality draft genome assemblies of six bacterial strains isolated from the roots of wheat grown in soil contaminated with cadmium. The results of this study will help to elucidate at the molecular level how heavy metals affect interactions between beneficial rhizobacteria and crop plants.

Optimal bedside urinalysis for the detection of proteinuria in hypertensive pregnancy: a study of diagnostic accuracy.

OBJECTIVE: To compare semi-quantitative visual and automated methods of urine testing with fully quantitative point of care urinalysis for the detection of significant proteinuria (0.3 g/24 hours) in pregnancy complicated by hypertension. DESIGN: A prospective comparative study. SETTING: A large teaching maternity hospital. SAMPLE: One hundred and seventy-one pregnant women referred to the obstetric day-care unit for assessment of newly arisen hypertension. METHODS: Early morning urine specimens were tested with four dipstick techniques (Multistix 8SG visual and automated and microalbumin/creatinine ratio visual and automated; Bayer, Elkhart, USA) as well as a fully quantitative measure of the microalbumin creatinine ratio with the DCA 2000 (a point of care assay for albumin; Bayer). These results were compared to a 24-hour urine protein measurement and measures of diagnostic accuracy/prediction are reported. MAIN OUTCOME MEASURES: Significant proteinuria (> or =0.3 g/24 hours) measured by laboratory assay. RESULTS: Automated dipstick urinalysis using the Clinitek 50 has significantly better predictive values for significant proteinuria (LR(+) 4.27, 95% CI 2.78 to 6.56; LR(-) 0.225, 95% CI 0.14 to 0.37) than conventional visual dipstick urinalysis (LR(+) 2.27, 95% CI 1.47 to 3.51; LR(-) 0.635, 95% CI 0.49 to 0.82). Dipstick microalbumin/creatinine ratio testing did not improve overall detection rates with automated or visual testing. Fully quantitative point of care measurement of albumin/creatinine ratio (ACR) was significantly better than any dipstick technique (LR(+) 14.6, 95% CI 6.74 to 31.8; LR(-) 0.069, 95% CI 0.030 to 0.16). CONCLUSIONS: This study confirms that in pregnancy automated dipstick urinalysis is a more accurate screening test for the detection of proteinuria than visual testing. ACR testing can offer a significant improvement over conventional urinalysis if a fully quantitative method of detection is employed that uses pregnancy-specific thresholds. Dipstick assessment of ACR does not improve the detection rate of significant proteinuria.

Validation of the DCA 2000 microalbumin:creatinine ratio urinanalyzer for its use in pregnancy and preeclampsia.

OBJECTIVE: To determine the accuracy of the DCA 2000 albumin/creatinine ratio urinanalyzer (Bayer Corp., Elkhart, IN) in uncomplicated pregnancy and preeclampsia. METHODS: This was a prospective observational study in a large teaching maternity hospital. Ninety one uncomplicated pregnant women and 100 women referred for assessment of de novo hypertension in pregnancy had albumin concentrations, creatinine concentrations, and albumin/creatinine ratios (ACR) compared between the DCA 2000 and the laboratory gold standard assays (Dade Dimension clinical chemistry autoanalyzer), for both early morning urines (EMU) and 24-hr urine collections. RESULTS: The interassay and intra-assay variability for the DCA 2000 were less than 5.1%. In uncomplicated pregnancy the mean difference in ACR between the DCA 2000 and the laboratory assay was 0.08 mg/mmol (SD 0.28; 95% limits of agreement, -0.47, 0.63) for EMU and 0.06 (SD 0.23; 95% limits of agreement, -0.39, 0.51) for 24-hr samples. In the hypertensive cohort the ACR mean differences were -0.82 (SD 7.13; 95% limits of agreement, -14.79, 13.15) for EMU and -0.76 (SD 4.14; 95% limits of agreement, -8.87, 7.35) for 24-hr samples. The mean differences between assays in the hypertensive group had broader 95% limits of agreement due to greater variability in the samples with high albumin concentrations (>40 mg/L). CONCLUSIONS: The DCA 2000 is accurate for the measurement of albumin creatinine ratios in the uncomplicated pregnant population. In the hypertensive population the DCA 2000 remains accurate though when the albumin concentration is greater than 40 mg/L the 95% limits of agreement are broader. We would recommend that all other automated urinalysis devices be validated by similar protocols to allow meaningful comparisons of accuracy.

Urinary microalbumin/creatinine ratios: reference range in uncomplicated pregnancy.

During uncomplicated pregnancy, the development of proteinuria is accepted as a poor prognostic sign and is associated with increasing maternal and perinatal mortality and morbidity. Physiological proteinuria increases with increasing gestation and one of its largest constituents is albumin. The reference range for the (micro)albumin/creatinine ratio (ACR) has not been described for normal pregnancy. This prospective cross-sectional study describes the gestation-specific 95% reference ranges for urinary microalbumin concentration, creatinine concentration and ACR in uncomplicated pregnancy. There is a significant increase ( P =0.016) in the ACR in the third trimester. The mean difference is 0.091 mg of albumin/mmol of creatinine (95% confidence interval, 0.014-0.168). Our results describe the first well-defined gestation-specific 95% reference range for a point-of-care measurement of the ACR. These data are essential if such testing is to be employed in antenatal care.