Search

Selective breeding and genetic modification have been the cornerstone of animal agriculture. However, the current strategy of breeding animals over multiple generations to introgress novel alleles is not practical in addressing global challenges such as climate change, pandemics, and the predicted need to feed a population of 9 billion by 2050. Consequently, genome editing in zygotes to allow for seamless introgression of novel alleles is required, especially in cattle with long generation intervals. We report for the first time the use of CRISPR-Cas genome editors to introduce novel PRNP allelic variants that have been shown to provide resilience towards human prion pandemics. From one round of embryo injections, we have established six pregnancies and birth of seven edited offspring, with two founders showing >90% targeted homology-directed repair modifications. This study lays out the framework for in vitro optimization, unbiased deep-sequencing to identify editing outcomes, and generation of high frequency homology-directed repair–edited calves.

The multiplex real‐time PCR assay can simultaneously detect four bacterial species associated with Acute Oak Decline (AOD) from stem bleed swabs, eliminating the need for destructive sampling of high‐value trees to isolate bacteria.

High income jurisdictions are banning menthol/flavored cigarettes and other tobacco products because they attract young people and create dependence. This study explores the importance of menthol and other flavored tobacco products for tobacco markets in low- and middle-income countries (LMICs), identifies countries where the menthol/flavor share is particularly high or rapidly growing, and identifies strategies tobacco companies are using to develop menthol/flavor markets.Research involved analysis of menthol/flavor market data from 2005 to 2019, a systematic review of academic literature, and a scoping exercise with our advocate contacts in LMICs.The median menthol/capsule market share of the cigarette market grew significantly (p0.05) between 2005 and 2019, both in lower and upper middle-income countries [lower: 2.5% (IQR: 0.5-4.0) to 6.5% (IQR: 3.6-15.9); and upper: 4.0% (IQR: 0.8-9.8) to 12.3% (IQR: 3.5-24.3)]. Countries with both high market share and high market share growth were Russia, Guatemala, Peru and Nigeria. No market data were available on low-income countries, but the academic literature suggested high prevalence of menthol use in Zambia. Tobacco industry strategies underpinning growth of menthol/flavored tobacco use in LMICs included in-store marketing and display, colorful packs and non-conventional flavor names.Menthol/flavor tobacco products are a growing problem in LMICs. In addition to menthol/flavor bans, we recommend marketing bans, point of sale display bans and standardized packaging.

Western (WEEV), eastern (EEEV), and Venezuelan (VEEV) equine encephalitis viruses are mosquito-borne pathogens classified as potential biological warfare agents for which there are currently no approved human vaccines or therapies. We aimed to evaluate the safety, tolerability, and immunogenicity of an investigational trivalent virus-like particle (VLP) vaccine, western, eastern, and Venezuelan equine encephalitis (WEVEE) VLP, composed of WEEV, EEEV, and VEEV VLPs.The WEVEE VLP vaccine was evaluated in a phase 1, randomised, open-label, dose-escalation trial at the Hope Clinic of the Emory Vaccine Center at Emory University, Atlanta, GA, USA. Eligible participants were healthy adults aged 18-50 years with no previous vaccination history with an investigational alphavirus vaccine. Participants were assigned to a dose group of 6 μg, 30 μg, or 60 μg vaccine product and were randomly assigned (1:1) to receive the WEVEE VLP vaccine with or without aluminium hydroxide suspension (alum) adjuvant by intramuscular injection at study day 0 and at week 8. The primary outcomes were the safety and tolerability of the vaccine (assessed in all participants who received at least one administration of study product) and the secondary outcome was immune response measured as neutralising titres by plaque reduction neutralisation test (PRNT) 4 weeks after the second vaccination. This trial is registered at ClinicalTrials.gov, NCT03879603.Between April 2, 2019, and June 13, 2019, 30 trial participants were enrolled (mean age 32 years, range 21-48; 16 [53%] female participants and 14 [47%] male participants). Six groups of five participants each received 6 μg, 30 μg, or 60 μg vaccine doses with or without adjuvant, and all 30 participants completed study follow-up. Vaccinations were safe and well tolerated. The most frequently reported symptoms were mild injection-site pain and tenderness (22 [73%] of 30) and malaise (15 [50%] of 30). Dose-dependent differences in the frequency of pain and tenderness were found between the 6 μg, 30 μg, and 60 μg groups (p=0·0217). No significant differences were observed between dosing groups for any other reactogenicity symptom. Two adverse events (mild elevated blood pressure and moderate asymptomatic neutropenia) were assessed as possibly related to the study product in one trial participant (60 μg dose with alum); both resolved without clinical sequelae. 4 weeks after second vaccine administration, neutralising antibodies were induced in all study groups with the highest response seen against all three vaccine antigens in the 30 μg plus alum group (PRNTThe favourable safety profile and neutralising antibody responses, along with pressing public health need, support further evaluation of the WEVEE VLP vaccine in advanced-phase clinical trials.The Vaccine Research Center of the National Institute of Allergy and Infectious Diseases, National Institutes of Health funded the clinical trial. The US Department of Defense contributed funding for manufacturing of the study product.

Background: Accurate and efficient handoff is a critical part of hospital-based patient care. In the current era of smartphones and rapidly advancing technology, multi-platform based applications are an integral part of professional life. This quality improvement project introduced a free, HIPAA compliant, multi-platform handoff application (Listrunner app, Listrunner, Inc. San Francisco, CA) to improve the accuracy and efficiency of patient sign-out in a single academic center within the pediatric cardiology fellowship program. Methods: Listrunner app was implemented to improve …

Objectives Abnormal early angiogenesis appears to impact both placental disorders and fetal congenital heart defects (CHD). We sought to assess the association of placental perfusion defects (PPD) and fetal (CHD). Methods Singleton pregnancies with isolated severe fetal CHD were compared to controls without congenital anomalies or maternal malperfusion (MVM) risk factors. CHD was categorized into group 1: single left ventricle morphology and transposition of the great vessels (TGA) and group 2: single right ventricle and two ventricle morphology. Malperfusion was defined as fetal vascular malperfusion (FVM), MVM, and both FVM and MVM. Results PPD was increased for all CHD (n = 47), CHD with or without risk factors, and CHD groups compared to controls (n = 92). Overall CHD cases and CHD with risk factors had an increased risk of FVM (30% and 80% vs 14%), and MVM (43% and 50% vs 21%), respectively. MVM rates were similar in CHD with and without maternal risk factors. FVM (38% vs 14%) and MVM (44% vs 21%) were increased in Group 1. MVM (42% vs 21%) and both FVM and MVM (16% vs 3%) were increased in Group 2. Conclusions PPD risk is increased in severe isolated fetal CHD. The highest risk is seen in fetal CHD with maternal risk factors.

The genetic defect Jersey Neuropathy with Splayed Forelimbs (JNS) was recently identified by the American Jersey Cattle Association ((AJCA), AJCA 2020). Affected calves are born alive but show several symptoms including being unable to stand on splayed forelimbs and neurologic symptoms including spasticity of head and neck and convulsive behavior. While a missense variant in UCLH1 chromosome 6 60,158,901G->A is the likely causative allele no commercial diagnostic assay currently is offered. Therefore, an animal's JNS status is currently based on a diagnostic SNP haplotype, which is not 100% accurate. To provide a more accurate carrier status information, we developed an in-house diagnostic test for the JNS causative allele. The assay information is provided below to allow diagnostic labs across the globe to quickly implement a causative allele diagnostic test and offer it to their local Jersey producers.

Objective. To investigate the number of authors and unique institutions per paper published in the American Journal of Pharmaceutical Education (AJPE) in 2015 through 2019, and to examine the number of authors and unique institutions for papers that were nominated for the Rufus A. Lyman Award in the same period. Methods. Articles published in AJPE from 2015 through 2019 were reviewed. Data collected for each article included article type, number of authors, and number of institutions. Results. Of the 811 articles published in AJPE during this period, the number of authors increased significantly from a mean (SD) of 3.5 (1.8) to 4.5 (2.2). The number of unique institutions also increased significantly from 1.7 (1.1) to 2.4 (1.8). Conclusion. There is a trend toward a greater number of authors and unique institutions for the publications in one pharmacy education journal. Explanations for this trend may include pressure to publish, increased research complexity, and expanded interprofessional collaboration.

Hexavalent chromium [Cr(VI)] in drinking water is pending regulation in California and is being considered for regulation in other locations. While strong-base anion exchange (SBA-IX) can efficiently remove Cr(VI) to low-levels that may be required to comply with future MCLs, operational and maintenance (O&M) costs can be considerable if the spent brine is disposed of as hazardous waste. Through bench- and pilot-scale experiments and full-scale demonstrations, this study examined the ability of emerging and established brine treatment and reuse techniques as well as recently developed resins to decrease O&M costs. When profiling anion elution during regeneration with nanofiltration treated and untreated spent brine, it appeared that at least 1 and 3 reuse cycles were feasible, respectively. Stannous- and ferrous-based reductants were more efficient than sulfur-based reductants when treating spent brine. Bed volumes to 8 μg/L chromium breakthrough with 7 resins varied by as much as a factor of 2 and correlated (R2 = 0.84) with resin total exchange capacities. Spent brine reuse, segmented regeneration (an optimized brine reuse method), ferrous reduction, and nanofiltration of spent brine were estimated to decrease O&M costs by 30, 70, 63, and 61%, respectively. Selection of high performing resins was the most simple way to decrease O&M costs (up to 70% savings). The sum of nitrate and sulfate raw water equivalent concentrations was found to be the principal water quality parameter that influenced the performance of 4 resins in 7 different groundwaters because nitrate and sulfate concentrations were orders of magnitude greater than chromium concentrations. Resins with higher chromium capacities eluted more co-contaminants including arsenic, selenium, uranium, and vanadium because they likely had higher co-contaminant capacities. Co-contaminant elution was found to be complex because associations can form between regenerant and co-eluting anions. Sodium chloride was the most efficient regenerant, though other regenerants provided benefits such as enhanced uranium elution most likely by complexing with uranium to inhibit its precipitation. Nitrate peaking was found to be limited even when reusing untreated and nanofiltration treated spent brine.

SPECTRA editors Sarah Plummer and Shaun Respess interview a group of rogue historians called History is Illuminating about their work installing unofficial interpretive signs along Monument Avenue in Richmond, Virginia following public protests and discourse surrounding Confederate monuments in Summer 2020.

Twenty-two cream-coloured bacterial strains were isolated from oak trees affected by acute oak decline (AOD) in Southern England. Isolates were Gram-negative, motile, slightly curved rods, aerobic, non-spore-forming, catalase positive and oxidase positive. 16S rRNA gene sequence analysis placed the strains in two separate phylogenetic clusters in the Pseudomonas straminea group, with Pseudomonas flavescens as the closest phylogenetic relative. Multilocus sequence analyses of the gyrB, rpoD and rpoB genes supported the delineation of the strains into two separate taxa, which could be differentiated phenotypically and chemotaxonomically from each other, and their closest relatives. Average nucleotide identity and in silico DNA-DNA hybridization values revealed percentages of genome similarity below the species threshold (95 and 70 %, respectively) between the two taxa and the closest relatives, confirming their novel species status. Therefore, on the basis of this polyphasic approach we propose two novel Pseudomonas species, Pseudomonasdaroniae sp. nov. (type strain FRB 228T=LMG 31087T=NCPPB 4672T) and Pseudomonasdryadis sp. nov. (type strain FRB 230T=LMG 31087T=NCPPB 4673T).

Acute Oak Decline is a syndrome within the Oak Decline complex in Britain. Profuse stem bleeding and larval galleries of the native buprestid, Agrilus biguttatus characterize the disease. A systematic study comparing healthy with diseased trees was undertaken. This work reports the result of isolations from healthy trees, diseased and non-symptomatic tissue within AOD affected trees, at five sites in England. Bacteria and fungi were identified using the DNA gyrase B gene, or ITS 1 sequencing. A significantly higher proportion of diseased tissues (82%) yielded more bacteria than either healthy (18%) or non-symptomatic tissue in diseased trees (33%). Overall bacterial community compositions varied at each site, but significant similarities were evident in diseased tissues at all sites. Enterobacteriaceae dominated in diseased trees whereas Pseudomonadaceae dominated healthy trees. Significant associations between diseased tissues and certain bacterial species occurred, implying that the cause of tissue necrosis was not due to random microbiota. Brenneria goodwinii and Gibbsiella quercinecans were key species consistently isolated from diseased tissue; Rahnella victoriana and an un-named Pseudomonas taxon were also frequently isolated from both healthy and diseased trees. Most fungi isolated were from the outer bark and had no significant association with tree health status. It was concluded that there was a shift in the cultivatable bacterial microbiome of diseased trees, with Enterobacteriaceae strongly represented in symptomatic but not healthy tissues. No single species dominated the isolations from diseased tissues and the tissue degradation in AOD is therefore likely to have a polymicrobial cause.

Background and Purpose: Military and veteran suicide rates exceed those found in the general population. Veterans often reject patient identities, creating barriers to care for mental health within the clinical sector and a mandate for prevention programs. The purpose of this study was to offer a postintervention process evaluation of one peer-led resilience program offered to military veterans of Iraq and Afghanistan at three sites in 2013. Methods: Secondary analysis of survey data collected involved mixed-methods analysis of open and closed-ended questions. In total, the research team reviewed 52 electronic survey responses; participant response rate was 48.1%. Results: Descriptive data analysis found that all participants rated Just Roll With It Bootcamp content as “somewhat useful” (17.9%) or “very useful” (82.1%). Qualitative analysis of open-ended questions found that content was perceived as valuable by participants. Emergent themes included: health practices, social support, and participant quality of life or satisfaction. Comments also informed four subthemes which included: meditation/mindfulness, nutrition, physical practice, and the seminars’ physical environment. Conclusion: Culturally-informed prevention programs that emphasize social support, physical movement, and peer-leadership have a vital role to play in working to prevent suicide by promoting quality of life for veterans.

Summary Background The Zika virus epidemic and associated congenital infections have prompted rapid vaccine development. We assessed two new DNA vaccines expressing premembrane and envelope Zika virus structural proteins. Methods We did two phase 1, randomised, open-label trials involving healthy adult volunteers. The VRC 319 trial, done in three centres, assessed plasmid VRC5288 (Zika virus and Japanese encephalitis virus chimera), and the VRC 320, done in one centre, assessed plasmid VRC5283 (wild-type Zika virus). Eligible participants were aged 18–35 years in VRC19 and 18–50 years in VRC 320. Participants were randomly assigned 1:1 by a computer-generated randomisation schedule prepared by the study statistician. All participants received intramuscular injection of 4 mg vaccine. In VRC 319 participants were assigned to receive vaccinations via needle and syringe at 0 and 8 weeks, 0 and 12 weeks, 0, 4, and 8 weeks, or 0, 4, and 20 weeks. In VRC 320 participants were assigned to receive vaccinations at 0, 4, and 8 weeks via single-dose needle and syringe injection in one deltoid or split-dose needle and syringe or needle-free injection with the Stratis device (Pharmajet, Golden, CO, USA) in each deltoid. Both trials followed up volunteers for 24 months for the primary endpoint of safety, assessed as local and systemic reactogenicity in the 7 days after each vaccination and all adverse events in the 28 days after each vaccination. The secondary endpoint in both trials was immunogenicity 4 weeks after last vaccination. These trials are registered with ClinicalTrials.gov, numbers NCT02840487 and NCT02996461. Findings VRC 319 enrolled 80 participants (20 in each group), and VRC 320 enrolled 45 participants (15 in each group). One participant in VRC 319 and two in VRC 320 withdrew after one dose of vaccine, but were included in the safety analyses. Both vaccines were safe and well tolerated. All local and systemic symptoms were mild to moderate. In both studies, pain and tenderness at the injection site was the most frequent local symptoms (37 [46%] of 80 participants in VRC 319 and 36 [80%] of 45 in VRC 320) and malaise and headache were the most frequent systemic symptoms (22 [27%] and 18 [22%], respectively, in VRC 319 and 17 [38%] and 15 [33%], respectively, in VRC 320). For VRC5283, 14 of 14 (100%) participants who received split-dose vaccinations by needle-free injection had detectable positive antibody responses, and the geometric mean titre of 304 was the highest across all groups in both trials. Interpretation VRC5283 was well tolerated and has advanced to phase 2 efficacy testing. Funding Intramural Research Program of the Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health.

Decline-diseases are complex and becoming increasingly problematic to tree health globally. Acute Oak Decline (AOD) is characterized by necrotic stem lesions and galleries of the bark-boring beetle, Agrilus biguttatus, and represents a serious threat to oak. Although multiple novel bacterial species and Agrilus galleries are associated with AOD lesions, the causative agent(s) are unknown. The AOD pathosystem therefore provides an ideal model for a systems-based research approach to address our hypothesis that AOD lesions are caused by a polymicrobial complex. Here we show that three bacterial species, Brenneria goodwinii, Gibbsiella quercinecans and Rahnella victoriana, are consistently abundant in the lesion microbiome and possess virulence genes used by canonical phytopathogens that are expressed in AOD lesions. Individual and polyspecies inoculations on oak logs and trees demonstrated that B. goodwinii and G. quercinecans cause tissue necrosis and, in combination with A. biguttatus, produce the diagnostic symptoms of AOD. We have proved a polybacterial cause of AOD lesions, providing new insights into polymicrobial interactions and tree disease. This work presents a novel conceptual and methodological template for adapting Koch’s postulates to address the role of microbial communities in disease.

While the impact of service-learning on students and to some extent on communities is well documented, little research addresses the direct impact on instructors. To fulfill the historic civic mission of education and contribute to civic health, a holistic understanding of impacts on all stakeholders will be necessary. This chapter presents the findings of interviews conducted with diverse service-learning practitioner types (higher education faculty members, K-12 teachers, and nonprofit instructors), institutions and years of experience. The findings demonstrate three categories of benefits that accrue to practitioners: pedagogical, personal and relational. Additionally, indirect impacts also contribute to practitioners' satisfaction. Many of these impacts map nicely onto the existing literature that describes the motivations that lead practitioners to undertake service-learning. The link then between motivations and actual benefits received leads to several salient recommendations that can support instructors and administrators wishing to advance engaged work at their institutions.

Beyond Trauma Treatment:Mindfulness Instruction in the Training Environment to Prevent Depression, Lower Suicide Rates and Improve Resilience in the Military and Veteran Communities The suicide numbers among active duty military personnel eclipsed the number of combat deaths in 2011. Before the wars in Iraq and Afghanistan, the incidence of suicide in active duty US service members was consistently 25% lower than in the civilian population. While clinical health services exist for soldiers and Marines with existing mental health conditions like posttraumatic stress, they are not stemming the rising tide of service suicides. A new approach is required, one that speaks to the participatory, hardworking ethos of military culture. Social work and health promotion professionals working to prevent and treat mental health problems like depression and stress illness must understand the confluence of warrior culture and mental health issues in the veteran community. While the research literature does not yet address this confluence issue directly, it contains ample evidence to support the development of a culturally-informed mindfulness training protocol. This protocol would be best implemented in participatory fashion in the training environment, rather than in treatment settings. Veterans often reject patient identities, which creates a major barrier to care for mental health in this population. To combat suicide rates and promote military and veteran mental health, a new approach is required, one that embraces peer-education and speaks to a competitive individualist military culture. A research basis exists for mindfulnessbased training protocols grounded in resiliency theory, and focusing on pre-incident training rather than posttraumatic treatment can reduce stigma and save lives.

The generation of sequence-tagged sites (STSs) has been proposed as a unifying approach to correlating the disparate results generated by genetic and various physical techniques being used to map the human genome. We have developed an STS map to complement the existing physical and genetic maps of 4p16.3, the region containing the Huntington disease gene. A total of 18 STSs span over 4 Mb of 4p16.3, with an average spacing of about 250 kb. Eleven of the STSs are located within the primary candidate HD region of 2.5 Mb between D4S126 and D4S168. The availability of STSs makes the corresponding loci accessibility to the general community without the need for distribution of cloned DNA. These STSs should also provide the means to isolate yeast artificial chromosome clones spanning the HD candidate region.

The RNASEL gene, a strong candidate for the hereditary prostate cancer 1 allele (HPC1), encodes a single-stranded specific endoribonuclease involved in the antiviral actions of IFNs. RNase L is activated enzymatically after binding to unusual 5'-phosphorylated, 2',5'-linked oligoadenylates (2-5A). Biostable phosphorothioate analogues of 2-5A were synthesized chemically and used to study the effects of naturally occurring mutations and polymorphisms in RNASEL. The 2-5A analogues induced RNase L activity and caused apoptosis in cultures of late-stage, metastatic human prostate cancer cell lines DU145, PC3, and LNCaP. However, DU145 and PC3 cells were more sensitive to 2-5A than LNCaP cells, which are heterozygous for an inactivating deletion mutation in RNase L. The RNase activities of missense variants of human RNase L were compared after expression in a mouse RNase L(-/-) cell line. Several variants (G59S, I97L, I220V, G296V, S322F, Y529C, and D541E) produced similar levels of RNase L activity as wild-type enzyme. In contrast, the R462Q variant, previously implicated in up to 13% of unselected prostate cancer cases, bound 2-5A at wild-type levels but had a 3-fold decrease in RNase activity. The deficiency in RNase L(R462Q) activity was correlated with a reduction in its ability to dimerize into a catalytically active form. Furthermore, RNase L(R462Q) was deficient in causing apoptosis in response to 2-5A consistent with its possible role in prostate cancer development. Our findings support the notion that RNASEL mutations and some variants allow tumor cells to escape a potent apoptotic pathway.