Highly active CRISPR-adaptation proteins revealed by a robust enrichment technology.

Natural prokaryotic defense via the CRISPR-Cas system requires spacer integration into the CRISPR array in a process called adaptation. To search for adaptation proteins with enhanced capabilities, we established a robust perpetual DNA packaging and transfer (PeDPaT) system that uses a strain of T7 phage to package plasmids and transfer them without killing the host, and then uses a different strain of T7 phage to repeat the cycle. We used PeDPaT to identify better adaptation proteins-Cas1 and Cas2-by enriching mutants that provide higher adaptation efficiency. We identified two mutant Cas1 proteins that show up to 10-fold enhanced adaptation in vivo. In vitro, one mutant has higher integration and DNA binding activities, and another has a higher disintegration activity compared to the wild-type Cas1. Lastly, we showed that their specificity for selecting a protospacer adjacent motif is decreased. The PeDPaT technology may be used for many robust screens requiring efficient and effortless DNA transduction.

Transaxial gunshot injuries.

OBJECTIVE: To define the clinical characteristics of gunshot injuries across the truncal midline (transaxial injuries). DESIGN: A retrospective case-control study MATERIALS AND METHODS: Analysis of a consecutive series of 223 patients with gunshot injuries to the torso treated at one trauma center. Sixty-three patients with transaxial injuries were compared to 148 patients with unilateral bullet trajectories. MAIN RESULTS: The incidence of cardiac (8%), vascular (17%), and spinal (29%) injuries and the mortality rate (29%) were significantly higher among the 63 patients with transaxial injuries compared with 148 patients with unilateral injuries. Initial operative access to the wrong visceral cavity (10%), missed injuries (14%), and early reoperations (19%) were also more common in the transaxial group. CONCLUSIONS: A transaxial bullet trajectory identifies a distinct injury pattern associated with a high incidence of major visceral damage, intraoperative misadventures, and very high mortality.

Emergency medicine in Israel: state of the art.

Emergency medicine is in its infancy in Israel but is developing rapidly. Medical and government authorities such as the Israeli Medical Association and the Israeli Ministry of Health have already recognized the need for this field in Israel, although it remains for emergency medicine to be recognized as an independent medical specialty. Those involved in the intense process of obtaining formal recognition believe this will occur in the next year.

Hyperinsulinemia, sex, and risk of atherosclerotic cardiovascular disease.

BACKGROUND: The possibility that hyperinsulinemia may be involved in the etiology of atherosclerotic cardiovascular disease (CVD) was first suggested 20 years ago. During the last decade, this possibility has received support from three large prospective studies. METHODS AND RESULTS: In the present study, the association between CVD, glucose intolerance, obesity, and hypertension (the GOH conditions) and hyperinsulinemia was examined cross-sectionally in a representative sample (n = 1,263) of the adult Jewish population aged 40-70 years in Israel. Previously known diabetics were excluded. CVD comprising clinical or ECG evidence of ischemic heart disease, as well as clinical evidence of cerebrovascular or peripheral vascular disease, was identified in 97 men and 39 women. A significant (p less than 0.01) hyperinsulinemia-sex interaction was found for CVD rate, with the adjusted risk ratios (followed by 95% confidence limits), relative to the rate in 298 normoinsulinemic women, being 1.15 (0.68-1.95) in 328 normoinsulinemic men, 0.85 (0.48-1.49) in 277 hyperinsulinemic women, and 2.27 (1.33-3.08) in 360 hyperinsulinemic men. Age-adjusted CVD rates in men versus women were: a) similar and low among all normoinsulinemic normotensives and hyperinsulinemics free of any of the GOH conditions (all rates less than or equal to 6.5%); b) similar and high among normoinsulinemic hypertensives (13.4% versus 10.4%); c) significantly higher in men among hyperinsulinemic normotensives with glucose intolerance and/or obesity (15.2% versus 3.3%; p = 0.02) and all hyperinsulinemic hypertensives (21.5% versus 12.8%; p = 0.04). These trends remained significant after adjusting for age, ethnic group, and blood lipids. CONCLUSIONS: Therefore, hyperinsulinemia was associated with excess CVD risk in men but not in women, and all excess CVD risk in men was confined to hyperinsulinemic individuals in the presence of glucose intolerance, obesity, or hypertension.

Smoking accounts for adverse effect of antihypertensive medications on plasma lipids. A population-based study.

Mean plasma levels of total cholesterol, high density lipoprotein cholesterol, total-to-high density lipoprotein cholesterol ratio, and total triglycerides were determined in a representative sample of the adult Israeli Jewish population, excluding known diabetics and individuals with overt atherosclerotic morbidity (n = 1,153). Levels were compared in normotensive and untreated and treated hypertensive individuals by glucose tolerance category and smoking, after adjustment for sex, age, and body mass index. In the presence of normal glucose tolerance, lipid levels in the nonsmoking normotensive and treated hypertensive groups were similar, whereas in the smoking, treated hypertensive group, lipids were significantly affected, as indicated by the respective adjusted mean levels (mg/dl): total cholesterol, 219, 221, and 240; high density lipoprotein cholesterol, 45.0, 43.6, and 42.0; ratio, 5.2, 5.3, and 6.0; and triglycerides, 114, 107, and 144. In individuals with glucose intolerance, trends were the same with the exception of triglycerides, which were significantly elevated in the nonsmoking, treated hypertensive group also; the respective values were 217, 225, and 257 for total cholesterol; 45.1, 44.9, and 41.4 for high density lipoprotein cholesterol; 5.2, 5.5, and 6.8 for the ratio; and 133, 152, and 187 for triglycerides. Lipid disturbances in treated smokers were not due to heavier smoking or differences in dietary intake. We conclude that disturbance of plasma lipid profile in treated hypertensive individuals may be mainly due to an interaction with smoking, with an additional effect of glucose intolerance.