Mild-to-Moderate Kidney Dysfunction and Cardiovascular Disease: Observational and Mendelian Randomization Analyses

BACKGROUND: End-stage renal disease is associated with a high risk of cardiovascular events. It is unknown, however, whether mild-to-moderate kidney dysfunction is causally related to coronary heart disease (CHD) and stroke. METHODS: Observational analyses were conducted using individual-level data from 4 population data sources (Emerging Risk Factors Collaboration, EPIC-CVD [European Prospective Investigation into Cancer and Nutrition-Cardiovascular Disease Study], Million Veteran Program, and UK Biobank), comprising 648 135 participants with no history of cardiovascular disease or diabetes at baseline, yielding 42 858 and 15 693 incident CHD and stroke events, respectively, during 6.8 million personyears of follow-up. Using a genetic risk score of 218 variants for estimated glomerular filtration rate (eGFR), we conducted Mendelian randomization analyses involving 413 718 participants (25917 CHD and 8622 strokes) in EPIC-CVD, Million Veteran Program, and UK Biobank. RESULTS: There were U-shaped observational associations of creatinine-based eGFR with CHD and stroke, with higher risk in participants with eG FR values 105 mL.min(-1).1.73 m(-2), compared with those with eG FR between 60 and 105 mL.min(-1).1.73 m(-2). Mendelian randomization analyses for CHD showed an association among participants with eGFR 105 mL.min(-1).1.73 m(-2). Results were not materially different after adjustment for factors associated with the eGFR genetic risk score, such as lipoprotein(a), triglycerides, hemoglobin Alc, and blood pressure. Mendelian randomization results for stroke were nonsignificant but broadly similar to those for CHD. CONCLUSIONS: In people without manifest cardiovascular disease or diabetes, mild-to-moderate kidney dysfunction is causally related to risk of CHD, highlighting the potential value of preventive approaches that preserve and modulate kidney function

Generation of ozone foam and its application for disinfection

Generated ozone foam was applied to the disinfection of Pseudomonas fluorescens. The effect of disinfection has been confirmed experimentally and new equipment for the disinfection of hands using this ozone foam has been put on the market for the practical use. The ozone foam was produced in the foam generator after mixing the water including surfactant (30 mL/min) and air including ozone (1000 ppm = 2.14 g/m3 ~ 1600 ppm = 3.4 g/m3, 300 mL/min). The liquid-to-gas ratio is 100 L/m3. The concentration of dissolved ozone in the thin liquid films of the bubbles was about 3 mg/L which was measured by the chemical method of the KI absorption and titration of sodium thiosulfate solution. The disinfection test samples were prepared using the PET disk on which Pseudomonas fluorescens of its number of more than 108 were attached. Test sample was inserted into ozone foam set on the glass plate for one to 6 min. The survival rate log (N/N0 decreased with time and its value of about−2.6 (i.e., ~1/400) was obtained at 6 min (2 min × 3 times repeated). It was also confirmed that the ozone foam was useful for the disinfection of hands. For more effective disinfection (in case of taking a long time for foam melting), the ozone foam was broken by force and changed into ozone water by which the survival rate decreased ×4 (i.e., N/N0 = 1/10 000) at 4 ~ 6 min

Quantitative assay for TALEN activity at endogenous genomic loci

Summary Artificially designed nucleases such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) can induce a targeted DNA double-strand break at the specific target genomic locus, leading to the frameshift-mediated gene disruption. However, the assays for their activity on the endogenous genomic loci remain limited. Herein, we describe a versatile modified lacZ assay to detect frameshifts in the nuclease target site. Short fragments of the genome DNA at the target or putative off-target loci were amplified from the genomic DNA of TALEN-treated or control embryos, and were inserted into the lacZα sequence for the conventional blue–white selection. The frequency of the frameshifts in the fragment can be estimated from the numbers of blue and white colonies. Insertions and/or deletions were easily determined by sequencing the plasmid DNAs recovered from the positive colonies. Our technique should offer broad application to the artificial nucleases for genome editing in various types of model organisms