Point-of-care electroencephalography enables rapid evaluation and management of non-convulsive seizures and status epilepticus in the emergency department.

Objectives: To describe our institutional experience with point-of-care electroencephalography (pocEEG) and its impact on the evaluation/management of suspected non-convulsive seizures in the emergency department (ED). Methods: We retrospectively identified 157 adults who underwent pocEEG monitoring in our community hospital ED in 1 year. We calculated the time to obtain pocEEG in the ED (door-to-EEG time) and examined the impact of pocEEG findings (categorized as seizure, highly epileptiform patterns, slowing, or normal activity) on antiseizure medication treatment. Results: PocEEG revealed seizures (14%, n = 22), highly epileptiform patterns (22%, n = 34), slowing (44%, n = 69), and normal activity (20%, n = 32). The median door-to-EEG time (from initial ED evaluation to pocEEG monitoring) was only 1.2 hours (interquartile range 0.1-2.1) even though 55% of studies were performed after-hours (5 pm-9 am). Most patients were admitted (54% to the intensive care unit, 41% to floor). Antiseizure medication treatment occurred pre-pocEEG in 93 patients (59%) and post-pocEEG in 88 patients (56%). By reviewing the relationship between pocEEG monitoring and antiseizure medication management, we found a significant association between pocEEG findings and changes in management (P < 0.001). Treatment escalation occurred more frequently in patients with epileptiform activity (seizures or highly epileptiform patterns, 52%) than patients with non-epileptiform activity (normal or slow, 25%, P < 0.001), and avoidance of treatment escalation occurred more frequently in patients with normal or slow activity (27%) than patients with seizures or highly epileptiform patterns (2%, P < 0.001). Conclusion: Our study, the largest to date describing the real-world use of pocEEG in emergency medicine, found that rapid EEG acquisition in the ED was feasible in a community hospital and significantly affected the management of suspected non-convulsive seizures.

Recommendations for Medical Discharge Documentation and Academic Supports for University Students Recovering From Concussion.

OBJECTIVE: This study sought to (1) collate the experiences of university students with concussion history and academic stakeholders through interviews and (2) develop concussion management recommendations for institutions of higher learning using a multidisciplinary Delphi procedure. SETTING: Remote semistructured interviews and online surveys. PARTICIPANTS: The first aim of this study included undergraduate university students with concussion history who did not participate in varsity athletics ( n = 21; 57.1% female), as well as academic faculty/staff with experience assisting university students with their postconcussion academic needs ( n = 7; 71.4% female). The second aim enrolled 22 participants (54.5% female) to serve on the Delphi panel including 9 clinicians, 8 researchers, and 5 academic faculty/staff. DESIGN: An exploratory-sequential mixed-methods approach. MAIN MEASURES: Semistructured interviews were conducted to unveil barriers regarding the return-to-learn (RTL) process after concussion, with emergent themes serving as a general framework for the Delphi procedure. Panelists participated in 3 stages of a modified Delphi process beginning with a series of open-ended questions regarding postconcussion management in higher education. The second stage included anonymous ratings of the recommendations, followed by an opportunity to review and/or modify responses based on the group's consensus. RESULTS: The results from the semistructured interviews indicated students felt supported by their instructors; however, academic faculty/staff lacked information on appropriate academic supports and/or pathways to facilitate the RTL process. Of the original 67 statements, 39 achieved consensus (58.2%) upon cessation of the Delphi procedure across 3 main categories: recommendations for discharge documentation (21 statements), guidelines to facilitate a multidisciplinary RTL approach (10 statements), and processes to obtain academic supports for students who require them after concussion (8 statements). CONCLUSIONS: These findings serve as a basis for future policy in higher education to standardize RTL processes for students who may need academic supports following concussion.

Achieving Consensus Through a Modified Delphi Technique to Create the Post-concussion Collegiate Return-to-Learn Protocol.

BACKGROUND: Sport-related concussions (SRCs) affect millions of adolescents and young adults annually in the USA; however, current SRC consensus statements provide limited guidance on academic support for students within higher education. OBJECTIVE: To generate consensus on appropriate academic recommendations for clinicians, students, and academic stakeholders to support university students during their recovery. METHODS: Panelists participated in three stages of a modified Delphi procedure: the first stage included a series of open-ended questions after reviewing a literature review on post-SRC return-to-learn (RTL) in higher education; the second stage asked panelists to anonymously rate the recommendations developed through the first Delphi stage using a 9-point scale; and the final stage offered panelists the opportunity to change their responses and/or provide feedback based on the group's overall ratings. RESULTS: Twenty-two panelists including clinicians, concussion researchers, and academic stakeholders (54.5% female) from 15 institutions and/or healthcare systems participated in a modified Delphi procedure. A total of 42 statements were developed after round one. Following the next two rounds, 27 statements achieved consensus amongst the panel resulting in the four-stage Post-Concussion Collegiate RTL Protocol. CONCLUSION: There are several unique challenges when assisting university students back to the classroom after SRC. Explicit guidelines on when to seek additional medical care (e.g., if they are experiencing worsening or persistent symptoms) and how to approach their instructor(s) regarding academic support may help the student self-advocate. Findings from the present study address barriers and provide a framework for universities to facilitate a multidisciplinary approach amongst medical and academic stakeholders.

Atypical Posterior Nutcracker Syndrome in a 17-Year-Old Male Without Hematuria.

Retro-aortic left renal vein (RLRV) is an anatomical variation, where the left renal vein (LRV) courses posterior to the aorta and anterior to the vertebrae before it eventually drains into the inferior vena cava (IVC). RLRV is a rare finding, with a prevalence of around 1%-2%, and only a small minority of RLRVs cause symptoms. RLRV symptoms occur secondary to compression of the LRV between the abdominal aorta and vertebrae, otherwise known as posterior nutcracker syndrome (PNCS). The most common symptoms of PNCS are hematuria and flank pain. We present a 17-year-old male, who came in with recurring left flank pain without hematuria, initially thought to be renal colic secondary to nephrolithiasis. On further investigations, an aberrant posterior renal vein was seen on CT suggestive of PNCS. The patient was treated successfully with RLRV vascular stent placement by interventional radiology. This case report adds to the limited number of PNCS cases observed in children and to the even rarer cases of PNCS without hematuria. This case also acts as a reminder for pediatricians to keep a wide scope of differentials in patients presenting with flank pain and provides an outline of both diagnostic and treatment modalities available for these patients.

Orthogonal SARS-CoV-2 Serological Assays Enable Surveillance of Low-Prevalence Communities and Reveal Durable Humoral Immunity.

We conducted a serological study to define correlates of immunity against SARS-CoV-2. Compared to those with mild coronavirus disease 2019 (COVID-19) cases, individuals with severe disease exhibited elevated virus-neutralizing titers and antibodies against the nucleocapsid (N) and the receptor binding domain (RBD) of the spike protein. Age and sex played lesser roles. All cases, including asymptomatic individuals, seroconverted by 2 weeks after PCR confirmation. Spike RBD and S2 and neutralizing antibodies remained detectable through 5-7 months after onset, whereas α-N titers diminished. Testing 5,882 members of the local community revealed only 1 sample with seroreactivity to both RBD and S2 that lacked neutralizing antibodies. This fidelity could not be achieved with either RBD or S2 alone. Thus, inclusion of multiple independent assays improved the accuracy of antibody tests in low-seroprevalence communities and revealed differences in antibody kinetics depending on the antigen. We conclude that neutralizing antibodies are stably produced for at least 5-7 months after SARS-CoV-2 infection.

Detection, prevalence, and duration of humoral responses to SARS-CoV-2 under conditions of limited population exposure.

We conducted an extensive serological study to quantify population-level exposure and define correlates of immunity against SARS-CoV-2. We found that relative to mild COVID-19 cases, individuals with severe disease exhibited elevated authentic virus-neutralizing titers and antibody levels against nucleocapsid (N) and the receptor binding domain (RBD) and the S2 region of spike protein. Unlike disease severity, age and sex played lesser roles in serological responses. All cases, including asymptomatic individuals, seroconverted by 2 weeks post-PCR confirmation. RBD- and S2-specific and neutralizing antibody titers remained elevated and stable for at least 2-3 months post-onset, whereas those against N were more variable with rapid declines in many samples. Testing of 5882 self-recruited members of the local community demonstrated that 1.24% of individuals showed antibody reactivity to RBD. However, 18% (13/73) of these putative seropositive samples failed to neutralize authentic SARS-CoV-2 virus. Each of the neutralizing, but only 1 of the non-neutralizing samples, also displayed potent reactivity to S2. Thus, inclusion of multiple independent assays markedly improved the accuracy of antibody tests in low seroprevalence communities and revealed differences in antibody kinetics depending on the viral antigen. In contrast to other reports, we conclude that immunity is durable for at least several months after SARS-CoV-2 infection.

Climate change, extinction, and Sky Island biogeography in a montane lizard.

Around the world, many species are confined to "Sky Islands," with different populations in isolated patches of montane habitat. How does this pattern arise? One scenario is that montane species were widespread in lowlands when climates were cooler, and were isolated by local extinction caused by warming conditions. This scenario implies that many montane species may be highly susceptible to anthropogenic warming. Here, we test this scenario in a montane lizard (Sceloporus jarrovii) from the Madrean Sky Islands of southeastern Arizona. We combined data from field surveys, climate, population genomics, and physiology. Overall, our results support the hypothesis that this species' current distribution is explained by local extinction caused by past climate change. However, our results for this species differ from simple expectations in several ways: (a) their absence at lower elevations is related to warm winter temperatures, not hot summer temperatures; (b) they appear to exclude a low-elevation congener from higher elevations, not the converse; (c) they are apparently absent from many climatically suitable but low mountain ranges, seemingly "pushed off the top" by climates even warmer than those today; (d) despite the potential for dispersal among ranges during recent glacial periods (~18,000 years ago), populations in different ranges diverged ~4.5-0.5 million years ago and remained largely distinct; and (e) body temperatures are inversely related to climatic temperatures among sites. These results may have implications for many other Sky Island systems. More broadly, we suggest that Sky Island species may be relevant for predicting responses to future warming.

High-throughput transformation of Saccharomyces cerevisiae using liquid handling robots.

Saccharomyces cerevisiae (budding yeast) is a powerful eukaryotic model organism ideally suited to high-throughput genetic analyses, which time and again has yielded insights that further our understanding of cell biology processes conserved in humans. Lithium Acetate (LiAc) transformation of yeast with DNA for the purposes of exogenous protein expression (e.g., plasmids) or genome mutation (e.g., gene mutation, deletion, epitope tagging) is a useful and long established method. However, a reliable and optimized high throughput transformation protocol that runs almost no risk of human error has not been described in the literature. Here, we describe such a method that is broadly transferable to most liquid handling high-throughput robotic platforms, which are now commonplace in academic and industry settings. Using our optimized method, we are able to comfortably transform approximately 1200 individual strains per day, allowing complete transformation of typical genomic yeast libraries within 6 days. In addition, use of our protocol for gene knockout purposes also provides a potentially quicker, easier and more cost-effective approach to generating collections of double mutants than the popular and elegant synthetic genetic array methodology. In summary, our methodology will be of significant use to anyone interested in high throughput molecular and/or genetic analysis of yeast.

Promoting healthy eating and active playtime by connecting to nature families with preschool children: evaluation of pilot study "Play&Grow".

BACKGROUND: This pilot project aimed to evaluate the "Play&Grow" program which promotes age-appropriate dietary habits and playtime healthy routines through "connectedness to nature" experiences in Hong Kong families with young children. METHOD: Thirty-eight preschoolers (aged 33.97 ± 9.38 mo), mothers, and their domestic workers were recruited. The families attended one workshop/week for a 4-mo period, consisting of: (i) health topic; (ii) food games; (iii) nature-related outdoor activities. RESULTS: Feeding practices, particularly Promoting and Encouragement to eat (PE) and Instrumental Feeding (IF) improved after the intervention (P = 0.008 and P = 0.016, respectively). Mother's BMI, responsibility for child's meal, child's birth weight had a bearing on the improvement of PE, r2 = 0.243, F(3,33) = 3.54, P = 0.025. Domestic helper's responsibility for child's cooking and her IF practices could predict child's picky eating (r2 = 0.203, F(2,34) = 4.322, P = 0.021). Mother's responsibility for child and helper's responsibility for cooking could predict child's consumption of salty foods (r2 = 0.252, F(2,34) = 5.737, P = 0.007). Physical activity of caregivers improved after the intervention. CONCLUSION: The pilot confirmed the design, protocols, evaluation instruments, and logistics of the study. Modified "Play&Grow" intervention will be conducted in a more rigorous randomized controlled trial to determine the long-term impact on obesity prevention in Hong Kong.

A randomized controlled trial for families with preschool children - promoting healthy eating and active playtime by connecting to nature.

BACKGROUND: Promotion of healthy lifestyles in children focuses predominantly on proper nutrition and physical activity, elements now widely recognised as essential for a healthy life. Systematic reviews have shown that nature-related activities also enhance general well-being as reflected in increased physical activity, a healthier diet, reduced stress and better sleep. Recent research suggests that many young children in Hong Kong between the ages of two and four in Hong Kong are more sedentary than recommended and seldom participate in active play, placing them at risk of becoming overweight or obese. The proposed project aims to investigate whether connecting families to nature positively influences physical activity (i.e., active playtime) and healthy eating routines in children aged 2 to 4. METHODS: We recently conducted a pilot study in Hong Kong to develop a programme, Play & Grow, based on the most successful evidence-based international preschool interventions. In addition to adopting the healthy eating and physical activity elements of these interventions, this project will additionally include a third novel element of Connectedness to nature: discovering nature through games and awareness of sounds, touch, smells, and temperature. To test the effectiveness of this modified intervention, a randomised controlled trial (RCT) involving 240 families with children aged 2 to 4 will be conducted. Families and children will take part in weekly one-hour activity sessions for 10-weeks. Lifestyle-related habits will be assessed before and immediately after the 10-week intervention, with follow up testing at 6 and 12 months' post intervention. DISCUSSION: A novel measuring tool created specifically for assessing Connectedness to nature, Nature Relatedness Scale (NRS), will be validated and tested for reliability prior to the RCT. The results of the RCT are intended to be used to understand which components of the intervention are most effective. The objectives of this project will be achieved over a 30-month period and will contribute to the research that examines key components of successful healthy lifestyle promotion programmes during early childhood. We predict that the inclusion of Connectedness to nature will significantly improve recognised preschool interventions. Finally, the aim of targeting family involvement will hopefully increase the sustainability of longer-term lifestyle modifications in children. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02715544 . Registered 22 March 2016.

Genome-Wide Analysis of the TORC1 and Osmotic Stress Signaling Network in Saccharomyces cerevisiae.

The Target of Rapamycin kinase Complex I (TORC1) is a master regulator of cell growth and metabolism in eukaryotes. Studies in yeast and human cells have shown that nitrogen/amino acid starvation signals act through Npr2/Npr3 and the small GTPases Gtr1/Gtr2 (Rags in humans) to inhibit TORC1. However, it is unclear how other stress and starvation stimuli inhibit TORC1, and/or act in parallel with the TORC1 pathway, to control cell growth. To help answer these questions, we developed a novel automated pipeline and used it to measure the expression of a TORC1-dependent ribosome biogenesis gene (NSR1) during osmotic stress in 4700 Saccharomyces cerevisiae strains from the yeast knock-out collection. This led to the identification of 440 strains with significant and reproducible defects in NSR1 repression. The cell growth control and stress response proteins deleted in these strains form a highly connected network, including 56 proteins involved in vesicle trafficking and vacuolar function; 53 proteins that act downstream of TORC1 according to a rapamycin assay--including components of the HDAC Rpd3L, Elongator, and the INO80, CAF-1 and SWI/SNF chromatin remodeling complexes; over 100 proteins involved in signaling and metabolism; and 17 proteins that directly interact with TORC1. These data provide an important resource for labs studying cell growth control and stress signaling, and demonstrate the utility of our new, and easily adaptable, method for mapping gene regulatory networks.

Influence of cadmium and mycorrhizal fungi on the fatty acid profile of flax (Linum usitatissimum) seeds.

BACKGROUND: The soil environment can affect not only the quantity of crops produced but also their nutritional quality. We examined the combined effects of below-ground cadmium (0, 5, and 15 ppm) and mycorrhizal fungi (presence and absence) on the concentration of five major fatty acids within flax seeds (Linum usitatissimum). RESULTS: Plants grown with mycorrhizal fungi produced seeds that contained higher concentrations of unsaturated (18:1, 18:2 and 18:3), but not saturated (16:0 and 18:0) fatty acids. The effects of mycorrhizal fungi on the concentration of unsaturated fatty acids in seeds were most pronounced when plant roots were exposed to 15 ppm Cd (i.e. the concentrations of 18:1, 18:2 and 18:3 increased by 169%, 370% and 150%, respectively). CONCLUSIONS: The pronounced effects of mycorrhizal fungi on the concentration of unsaturated fatty acids at 15 ppm Cd may have been due to the presence of elevated levels of Cd within seeds. Our results suggest that, once the concentration of cadmium within seeds reaches a certain threshold, this heavy metal may improve the efficiency of enzymes that convert saturated fatty acids to unsaturated fatty acids.

From cheek swabs to consensus sequences: an A to Z protocol for high-throughput DNA sequencing of complete human mitochondrial genomes.

BACKGROUND: Next-generation DNA sequencing (NGS) technologies have made huge impacts in many fields of biological research, but especially in evolutionary biology. One area where NGS has shown potential is for high-throughput sequencing of complete mtDNA genomes (of humans and other animals). Despite the increasing use of NGS technologies and a better appreciation of their importance in answering biological questions, there remain significant obstacles to the successful implementation of NGS-based projects, especially for new users. RESULTS: Here we present an 'A to Z' protocol for obtaining complete human mitochondrial (mtDNA) genomes - from DNA extraction to consensus sequence. Although designed for use on humans, this protocol could also be used to sequence small, organellar genomes from other species, and also nuclear loci. This protocol includes DNA extraction, PCR amplification, fragmentation of PCR products, barcoding of fragments, sequencing using the 454 GS FLX platform, and a complete bioinformatics pipeline (primer removal, reference-based mapping, output of coverage plots and SNP calling). CONCLUSIONS: All steps in this protocol are designed to be straightforward to implement, especially for researchers who are undertaking next-generation sequencing for the first time. The molecular steps are scalable to large numbers (hundreds) of individuals and all steps post-DNA extraction can be carried out in 96-well plate format. Also, the protocol has been assembled so that individual 'modules' can be swapped out to suit available resources.

Benefits of a unit-based skin care group.

The medical patient population presents with various types of wounds when admitted to the hospital. A clinical nurse specialist (CNS) is often consulted to help nurses to assess wounds and dress them appropriately. Because CNS consultation is available only during business hours, staff identified a need for a core group of nurses to be knowledgeable about wound care. To address this need, a skin group was developed and given additional education. The members of this group were able to provide several interventions geared toward the nursing staff. In conjunction with the unit-based CNS, the skin group members have helped to improve the skin and wound care that patients receive and have consistently delivered this care in a timely manner.

Chiral calcium VAPOL phosphate mediated asymmetric chlorination and Michael reactions of 3-substituted oxindoles.

We disclose a novel high yielding and highly enantioselective chiral calcium VAPOL phosphate-catalyzed chlorination of 3-substituted oxindoles with N-chlorosuccinimide (NCS). The reaction conditions are also shown to be effective for the catalytic enantioselective Michael addition of 3-aryloxindoles to methyl vinyl ketone.

Chiral phosphoric acid catalyzed addition of dihydropyrans to N-acyl imines: stereocontrolled access to enantioenriched spirocyclic oxazoletetrahydropyrans with three contiguous stereocenters.

Dihydropyran derivatives readily undergo addition to N-acyl imines in the presence of chiral phosphoric acids. This addition process yields an attractive product that is capable of a tandem oxidative-cyclization via an epoxide intermediate.

Machine-learning approaches for classifying haplogroup from Y chromosome STR data.

Genetic variation on the non-recombining portion of the Y chromosome contains information about the ancestry of male lineages. Because of their low rate of mutation, single nucleotide polymorphisms (SNPs) are the markers of choice for unambiguously classifying Y chromosomes into related sets of lineages known as haplogroups, which tend to show geographic structure in many parts of the world. However, performing the large number of SNP genotyping tests needed to properly infer haplogroup status is expensive and time consuming. A novel alternative for assigning a sampled Y chromosome to a haplogroup is presented here. We show that by applying modern machine-learning algorithms we can infer with high accuracy the proper Y chromosome haplogroup of a sample by scoring a relatively small number of Y-linked short tandem repeats (STRs). Learning is based on a diverse ground-truth data set comprising pairs of SNP test results (haplogroup) and corresponding STR scores. We apply several independent machine-learning methods in tandem to learn formal classification functions. The result is an integrated high-throughput analysis system that automatically classifies large numbers of samples into haplogroups in a cost-effective and accurate manner.

Intergenerational family conversations and decision making about eating healthfully.

OBJECTIVE: To explore how youth, parents, and grandparents discuss issues related to eating healthfully and unhealthfully and to identify intergenerational strategies for educators to improve this communication. DESIGN: In three intergenerational focus groups, each with 4-8 families, a trained moderator asked questions about family practices and conversations for eating healthfully and unhealthfully. SETTING: Three focus group sites, each with Pennsylvania Nutrition Education Program sites (PANEP) programs serving low-income populations and multigenerational clientele, based in geographically and culturally diverse communities in Pennsylvania. PARTICIPANTS: Forty-four individuals (21 pre-teens, 16 parents, and 7 grandparents) from 17 families. PHENOMENON OF INTEREST: How youth, parents, and grandparents discuss and influence each other's healthful and unhealthful eating practices. ANALYSIS: "Strength" of evidence determined by repetition of ideas across focus groups and from the respondents' quotes providing in-depth information. RESULTS: Families demonstrated a wide range of ways that family communication is associated with the adoption of healthful and unhealthful patterns of eating. Parents and grandparents expressed anguish over their struggle and inability to help their children eat more healthfully. All three generations enumerated strategies for dealing with disagreement. CONCLUSIONS AND IMPLICATIONS: Grandparents, parents and children indicate that they need opportunities to learn together and communicate about ways to improve nutrition behaviors.

MtDNA evidence for a genetic bottleneck in the early history of the Ashkenazi Jewish population.

The relative roles of natural selection and accentuated genetic drift as explanations for the high frequency of more than 20 Ashkenazi Jewish disease alleles remain controversial. To test for the effects of a maternal bottleneck on the Ashkenazi Jewish population, we performed an extensive analysis of mitochondrial DNA (mtDNA) hypervariable segment 1 (HVS-1) sequence and restriction site polymorphisms in 565 Ashkenazi Jews from different parts of Europe. These patterns of variation were compared with those of five Near Eastern (n=327) and 10 host European (n=849) non-Jewish populations. Only four mtDNA haplogroups (Hgs) (defined on the basis of diagnostic coding region RFLPs and HVS-1 sequence variants) account for approximately 70% of Ashkenazi mtDNA variation. While several Ashkenazi Jewish mtDNA Hgs appear to derive from the Near East, there is also evidence for a low level of introgression from host European non-Jewish populations. HVS-1 sequence analysis revealed increased frequencies of Ashkenazi Jewish haplotypes that are rare or absent in other populations, and a reduced number of singletons in the Ashkenazi Jewish sample. These diversity patterns provide evidence for a prolonged period of low effective size in the history of the Ashkenazi population. The data best fit a model of an early bottleneck (approximately 100 generations ago), perhaps corresponding to initial migrations of ancestral Ashkenazim in the Near East or to Europe. A genetic bottleneck followed by the recent phenomenon of rapid population growth are likely to have produced the conditions that led to the high frequency of many genetic disease alleles in the Ashkenazi population.