Cas9-expressing chickens and pigs as resources for genome editing in livestock.

Genetically modified animals continue to provide important insights into the molecular basis of health and disease. Research has focused mostly on genetically modified mice, although other species like pigs resemble the human physiology more closely. In addition, cross-species comparisons with phylogenetically distant species such as chickens provide powerful insights into fundamental biological and biomedical processes. One of the most versatile genetic methods applicable across species is CRISPR-Cas9. Here, we report the generation of transgenic chickens and pigs that constitutively express Cas9 in all organs. These animals are healthy and fertile. Functionality of Cas9 was confirmed in both species for a number of different target genes, for a variety of cell types and in vivo by targeted gene disruption in lymphocytes and the developing brain, and by precise excision of a 12.7-kb DNA fragment in the heart. The Cas9 transgenic animals will provide a powerful resource for in vivo genome editing for both agricultural and translational biomedical research, and will facilitate reverse genetics as well as cross-species comparisons.

Evidence of egg-laying grounds for critically endangered flapper skate (Dipturus intermedius) off Orkney, UK.

Essential fish habitats (EFHs) are critical for fish life-history events, including spawning, breeding, feeding or growth. This study provides evidence of EFHs for the critically endangered flapper skate (Dipturus intermedius) in the waters around the Orkney Isles, Scotland, based on citizen-science observation data. The habitats of potential egg-laying sites were parametrised as >20 m depth, with boulders or exposed bedrock, in moderate current flow (0.3-2.8 knots) with low sedimentation. This information provides a significant contribution to the understanding of EFHs for flapper skate.

Surface-Water Nutrient Conditions and Sources in the United States Pacific Northwest.

The SPAtially Referenced Regressions On Watershed attributes (SPARROW) model was used to perform an assessment of surface-water nutrient conditions and to identify important nutrient sources in watersheds of the Pacific Northwest region of the United States (U.S.) for the year 2002. Our models included variables representing nutrient sources as well as landscape characteristics that affect nutrient delivery to streams. Annual nutrient yields were higher in watersheds on the wetter, west side of the Cascade Range compared to watersheds on the drier, east side. High nutrient enrichment (relative to the U.S. Environmental Protection Agency's recommended nutrient criteria) was estimated in watersheds throughout the region. Forest land was generally the largest source of total nitrogen stream load and geologic material was generally the largest source of total phosphorus stream load generated within the 12,039 modeled watersheds. These results reflected the prevalence of these two natural sources and the low input from other nutrient sources across the region. However, the combined input from agriculture, point sources, and developed land, rather than natural nutrient sources, was responsible for most of the nutrient load discharged from many of the largest watersheds. Our results provided an understanding of the regional patterns in surface-water nutrient conditions and should be useful to environmental managers in future water-quality planning efforts.

A novel automated SARS-CoV-2 saliva PCR test protects a global asymptomatic workforce.

Regular PCR testing of nasopharyngeal swabs from symptomatic individuals for SARS-CoV-2 virus has become the established method by which health services are managing the COVID-19 pandemic. Businesses such as AstraZeneca have also prioritised voluntary asymptomatic testing to keep workplaces safe and maintain supply of essential medicines to patients. We describe the development of an internal automated SARS-CoV-2 testing programme including the transformative introduction of saliva as an alternative sample type.

High-Mobility Multipurpose Wheeled Vehicle Rollover Accidents and Injuries to U.S. Army Soldiers by Reported Occupant Restraint Use, 1992-2013.

OBJECTIVE: The high-mobility multipurpose wheeled vehicle (HMMWV) is a light military tactical vehicle. During Operation Iraqi Freedom and Operation Enduring Freedom, the U.S. Army modified the HMMWV into a combat vehicle by adding vehicle armor, which made the vehicle more difficult to control and more likely to roll over. Consequently, reports of fatal rollover accidents involving up-armored HMMWVs began to accumulate during the up-armoring period (August 2003 to April 2005). Furthermore, the lack of occupant restraint use prevalent in a predominantly young, male, and enlisted military population compounded the injuries resulting from these accidents. In this retrospective case series analysis, we describe the characteristics of U.S. Army HMMWV rollover accidents, occupants, and injuries reported worldwide from fiscal year 1992 to 2013 based on reported occupant restraint use. METHODS: We conducted all analyses using Microsoft Excel 2010 and SAS version 9.1. Because this analysis does not constitute human subjects research, no institutional review board review was required. First, we obtained U.S. Army HMMWV accident records from the U.S. Army Combat Readiness Center, and selected those records indicating a HMMWV rollover had occurred. Next, we successively deduplicated the records at the accident, vehicle, occupant, and injury levels for descriptive analysis of characteristics at each level. For each occupant position, we calculated relative, attributable, and population attributable risks of nonfatal and fatal injury based on reported occupant restraint use. Finally, we analyzed body part injured and nature of injury to characterize the injury patterns that HMMWV occupants in each position sustained based on restraint use. We performed a χ2 test of homogeneity to assess differences in injury patterns between restrained and unrestrained occupants. RESULTS: A total of 819 U.S. Army HMMWV rollover accidents worldwide were reported from October 1991 through May 2013 involving 821 HMMWVs and 1,395 occupants (828 nonfatally injured, 151 fatally injured, and 416 noninjured). Thirty-five percent of more severe (class A and B) accidents involved the M1114 up-armored variant, whereas 32% of less severe (class C and D) accidents involved the M998 nonarmored variant. Unrestrained occupants were 20% more likely to be nonfatally injured and 5.6 times more likely to be fatally injured than were restrained occupants. Among unrestrained occupants, restraint use could have potentially saved 82% of lives lost. Among all occupants involved in a HMMWV rollover, an estimated 56% of fatalities could have been prevented by restraint use. Unrestrained drivers and vehicle commanders had greater than expected torso injuries, while restrained vehicle commanders and passengers had greater than expected upper extremity injuries. Unrestrained drivers had greater than expected fractures, whereas restrained drivers and vehicle commanders had greater than expected sprains/strains. CONCLUSION: While reporting bias may exist, nevertheless these results show that occupant restraint use confers substantial life-saving protection to HMMWV occupants in rollover accidents. Therefore, commanders, safety officers, and peers should continue to promote and enforce restraint use consistently during all Army ground operations and training involving HMMWVs. Doing so will save Soldiers' lives in rollover accidents during the remaining years of the HMMWV program.

Addressing the global cardiovascular risk of hypertension, dyslipidemia, diabetes mellitus, and the metabolic syndrome in the southeastern United States, part II: treatment recommendations for management of the global cardiovascular risk of hypertension, dyslipidemia, diabetes mellitus, and the metabolic syndrome.

An aggressive global approach to screening and to the management of the metabolic syndrome is recommended to slow the growth of the syndrome throughout the United States. Prevention should begin in childhood with healthy nutrition, daily physical activity, and annual measurement of weight, height, and blood pressure beginning at 3 years of age. Such screenings will identify cardiovascular risk factors early, allow the health care provider to define global cardiovascular risk with the COSEHC Cardiovascular Risk Assessment Tool, and allow treatment of each risk factor. Lifelong lifestyle modifications and pharmacologic therapy will be required in most patients. Antihypertensive therapy for these patients should begin with an angiotensin-converting enzyme inhibitor or an angiotensin receptor blocker unless a compelling indication for another drug is present. Metformin should be considered the first drug for glucose control in the patient with type 2 diabetes. A statin should be used initially for hyperlipidemia unless contraindicated. Combinations of antihypertensive, antiglycemic, and lipid-lowering agents will often be required.

Addressing the global cardiovascular risk of hypertension, dyslipidemia, and insulin resistance in the southeastern United States.

An expanded occurrence of the metabolic syndrome in the U.S. population, especially in the Southeastern United States, has raised awareness of a need to revise our approach to the management of global cardiovascular risk factors while underscoring a need for more aggressive interventions and prevention measures. In defining the components of the metabolic syndrome and the interrelationship among obesity, hypertension, dyslipidemia, and insulin resistance, a basic framework for the medical management of this syndrome has been defined. In Part I of the consensus report prepared by the Workgroup on Medical Guidelines of the Consortium for Southeastern Hypertension Control (COSEHC), we analyze the components of the metabolic syndrome, discuss its pathophysiology, and recommend an approach to the quantitative analysis of the risk factors contributing to excess cardiovascular death in the region.

Modulation of Innate Immune Signalling by Lipid-Mediated MAVS Transmembrane Domain Oligomerization.

RIG-I-like receptors detect viral RNA in infected cells and promote oligomerization of the outer mitochondrial membrane protein MAVS to induce innate immunity to viral infection through type I interferon production. Mitochondrial reactive oxygen species (mROS) have been shown to enhance anti-viral MAVS signalling, but the mechanisms have remained obscure. Using a biochemical oligomerization-reporter fused to the transmembrane domain of MAVS, we found that mROS inducers promoted lipid-dependent MAVS transmembrane domain oligomerization in the plane of the outer mitochondrial membrane. These events were mirrored by Sendai virus infection, which similarly induced lipid peroxidation and promoted lipid-dependent MAVS transmembrane domain oligomerization. Our observations point to a role for mROS-induced changes in lipid bilayer properties in modulating antiviral innate signalling by favouring the oligomerization of MAVS transmembrane domain in the outer-mitochondrial membrane.

The role of noninvasive hemodynamic monitoring in the evaluation and treatment of hypertension.

Advances in the understanding of the mechanisms accounting for the elevation of arterial pressure in essential hypertension suggest that there is value in assessing the relative contribution of hemodynamic factors in tailoring specific therapies to control arterial pressure. The non-invasive method of impedance cardiography (ICG) to measure hemodynamic abnormalities in hypertensive patients has emerged as a valuable adjuvant in the decision-making process of selecting antihypertensive agents. The technique is both accurate and reproducible in delineating the hemodynamic mechanisms of hypertension, comparing age-and gender-related changes in hemodynamics, detecting the presence of left ventricular dysfunction, and demonstrating clinically significant improvement in blood pressure control using ICG-guided therapy.

YouthFriends: outcomes from a school-based mentoring program.

This evaluation explores the effectiveness of school-based mentoring as a universal prevention strategy. The impact of mentoring on students displaying risk factors is also addressed. The evaluation of YouthFriends, a school-based mentoring program, employed a pretest-posttest control group design. Students (n = 170) across five school districts provided data on eight dependent variables. At posttest, there was a statistically significant difference favoring YouthFriends over controls on sense of school membership. In addition, for students who had low scores at baseline, results indicated a statistically significant improvement unique to YouthFriends on community connectedness and goal-setting. Analyses of students' academic performance also indicated a positive effect for those YouthFriends who had low grades at baseline. Editors' Strategic Implications: Important lessons are provided for school administrators and mentoring program staff and evaluators. As a universal prevention program, school-based mentoring may produce few (and small) short-term effects. A greater understanding of the effects of dosage and quality of the mentoring is needed as we seek to evaluate the efficacy of school-based mentoring across a variety of student risk levels.