Separation of Nucleic Acids by Ion Pair Reversed Phase High Performance Liquid Chromatography (IP RP HPLC)

Ion-pair reversed-phase high performance liquid chromatography (IP RP HPLC) is shown to be an effective method to reliably analyze DNA. IP RP HPLC is a much faster and safer alternative to conventional methods of DNA separation and quantification. The method described here utilized a two-buffer effluent system consisting of triethylammonium acetate (TEAA) and acetonitrile (ACN). The method reliably separated and quantified DNA samples of 54 and 58 nt. This method will be used and optimized to separate similarly sized RNA samples. The ultimate goal is to separate mixtures of nucleotides generated from in-vitro transcription reactions

Inventing Eden: primitivism, millennialism, and the making of New England

Seventeenth-century exegetes described Eden as a three-fold paradise because they believed that Adam and Eve lived in an external garden of delight, possessed incorrupt physiologies, and enjoyed intellectual, spiritual, and social perfections before the Fall. Accordingly, the dissertation is organized thematically, treating the ways in which New England colonists sought to mold their lands, bodies, minds, language, souls, and social spheres after the pattern provided in Eden. Chapter one traces the transition of terms used to describe the New England landscape from the present paradise of John Smith to the hideous and desolate wilderness of William Bradford and the prospective Paradise of Cotton Mather. Chapter two outlines programs of physiological reform, as colonists like Anne Bradstreet disciplined their physical bodies and ministers like Edward Taylor regulated the ecclesiastical body's consumption of communion in order to achieve humoral temperance--the somatic and spiritual state of Adam and Eve in Eden. Chapters three and four document Francis Bacon's influence on educational and linguistic aspirations in New England. I argue that because the encyclopedic knowledge and divinely denotative language of Adam were believed to be inseparably linked, Leonard Hoar's plans to turn Harvard into the world's first experimental laboratory in chemistry situated at a university and John Cotton's attempt to model the language of the Bay Psalm Book after the lingua humana of Eden should be understood as related endeavors, companion contributions from New England to the Baconian project for the instauration of prelapsarian intellectual perfections. Chapter five examines the ways in which ministers of the Great Awakening presented Adam and Eve to their congregants as types of Christian conversion, and chapter six details the process by which theories of natural law distilled from Genesis became the basis for colonial rebellion and republican government through the influence of Oceana, James Harrington's vision of an idealized, edenic republic. Spanning two centuries and surveying the works of major British and American authors from George Herbert and John Milton to Jonathan Edwards and Benjamin Franklin, Inventing Eden is the history of an idea that irrevocably altered the theology, literature, and culture of early modern New England

A Multifactorial Approach to Sport-Related Concussion Prevention and Education: Application of the Socioecological Framework

To offer an overview of sport-related concussion (SRC) prevention and education strategies in the context of the socioecological framework (SEF). Athletic trainers (ATs) will understand the many factors that interact to influence SRC prevention and the implications of these interactions for effective SRC education

Comprehensive Coach Education Reduces Head Impact Exposure in American Youth Football

BACKGROUND: Despite little evidence that defines a threshold of head impact exposure or that participation in youth sports leads to long-term cognitive impairments, it is prudent to identify methods of reducing the frequency of head impacts. PURPOSE: To compare the mean number of head impacts between youth football players in practice and games between leagues that implemented the Heads Up Football (HUF) educational program and those that did not (NHUF). STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: During the 2014 season, head impact exposure was measured using xPatch accelerometers from 70 youth football players aged 8 to 15 years from 5 leagues. Data were collected during both games and practices. The NHUF group comprised 32 players from 8 teams within 3 leagues. The HUF group comprised 38 players from 7 teams within 2 leagues. Independent-sample t tests evaluated differences in head impact exposure across groups (ie, HUF and NHUF). RESULTS: Players (mean ± SD: age, 11.7 ± 1.4 years; height, 152.2 ± 10.5 cm; weight, 51.6 ± 9.6 kg) experienced a total of 7478 impacts over 10g, of which 4250 (56.8%) and 3228 (43.2%) occurred in practices and games, respectively. The majority of impacts occurred within the NHUF group (62.0%), followed by the HUF group (38.0%). With a 10g impact threshold, the mean number of impacts during practice per individual event was lower in the HUF group (mean ± SD, 5.6 ± 2.9) than in the NHUF group (mean ± SD, 8.9 ± 3.1; difference, 3.4; 95% CI, 2.9-3.9). This difference was attenuated when the threshold was changed to 20g but remained significant (difference, 1.0; 95% CI, 0.7-1.3). At both the 10g and 20g impact thresholds, no differences were found in games. CONCLUSION: Players who participated in HUF leagues accumulated fewer head impacts per practice at both the 10g and 20g thresholds. Youth football leagues should consider the HUF educational program, while exploring additional interventions, to help reduce the number of head impacts in players

Injury Rates in Age-Only Versus Age-and-Weight Playing Standard Conditions in American Youth Football

BACKGROUND: American youth football leagues are typically structured using either age-only (AO) or age-and-weight (AW) playing standard conditions. These playing standard conditions group players by age in the former condition and by a combination of age and weight in the latter condition. However, no study has systematically compared injury risk between these 2 playing standards. PURPOSE: To compare injury rates between youth tackle football players in the AO and AW playing standard conditions. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Athletic trainers evaluated and recorded injuries at each practice and game during the 2012 and 2013 football seasons. Players (age, 5-14 years) were drawn from 13 recreational leagues across 6 states. The sample included 4092 athlete-seasons (AW, 2065; AO, 2027) from 210 teams (AW, 106; O, 104). Injury rate ratios (RRs) with 95% CIs were used to compare the playing standard conditions. Multivariate Poisson regression was used to estimate RRs adjusted for residual effects of age and clustering by team and league. There were 4 endpoints of interest: (1) any injury, (2) non-time loss (NTL) injuries only, (3) time loss (TL) injuries only, and (4) concussions only. RESULTS: Over 2 seasons, the cohort accumulated 1475 injuries and 142,536 athlete-exposures (AEs). The most common injuries were contusions (34.4%), ligament sprains (16.3%), concussions (9.6%), and muscle strains (7.8%). The overall injury rate for both playing standard conditions combined was 10.3 per 1000 AEs (95% CI, 9.8-10.9). The TL injury, NTL injury, and concussion rates in both playing standard conditions combined were 3.1, 7.2, and 1.0 per 1000 AEs, respectively. In multivariate Poisson regression models controlling for age, team, and league, no differences were found between playing standard conditions in the overall injury rate (RRoverall, 1.1; 95% CI, 0.4-2.6). Rates for the other 3 endpoints were also similar (RRNTL, 1.1 [95% CI, 0.4-3.0]; RRTL, 0.9 [95% CI, 0.4-1.9]; RRconcussion, 0.6 [95% CI, 0.3-1.4]). CONCLUSION: For the injury endpoints examined in this study, the injury rates were similar in the AO and AW playing standards. Future research should examine other policies, rules, and behavioral factors that may affect injury risk within youth football

Targeting DNA Damage Response and Replication Stress in Pancreatic Cancer

Background and aims: Continuing recalcitrance to therapy cements pancreatic cancer (PC) as the most lethal malignancy, which is set to become the second leading cause of cancer death in our society. The study aim was to investigate the association between DNA damage response (DDR), replication stress and novel therapeutic response in PC to develop a biomarker driven therapeutic strategy targeting DDR and replication stress in PC. Methods: We interrogated the transcriptome, genome, proteome and functional characteristics of 61 novel PC patient-derived cell lines to define novel therapeutic strategies targeting DDR and replication stress. Validation was done in patient derived xenografts and human PC organoids. Results: Patient-derived cell lines faithfully recapitulate the epithelial component of pancreatic tumors including previously described molecular subtypes. Biomarkers of DDR deficiency, including a novel signature of homologous recombination deficiency, co-segregates with response to platinum (P < 0.001) and PARP inhibitor therapy (P < 0.001) in vitro and in vivo. We generated a novel signature of replication stress with which predicts response to ATR (P < 0.018) and WEE1 inhibitor (P < 0.029) treatment in both cell lines and human PC organoids. Replication stress was enriched in the squamous subtype of PC (P < 0.001) but not associated with DDR deficiency. Conclusions: Replication stress and DDR deficiency are independent of each other, creating opportunities for therapy in DDR proficient PC, and post-platinum therapy

Genome-Wide Association Study in BRCA1 Mutation Carriers Identifies Novel Loci Associated with Breast and Ovarian Cancer Risk

BRCA1-associated breast and ovarian cancer risks can be modified by common genetic variants. To identify further cancer risk-modifying loci, we performed a multi-stage GWAS of 11,705 BRCA1 carriers (of whom 5,920 were diagnosed with breast and 1,839 were diagnosed with ovarian cancer), with a further replication in an additional sample of 2,646 BRCA1 carriers. We identified a novel breast cancer risk modifier locus at 1q32 for BRCA1 carriers (rs2290854, P = 2.7×10-8, HR = 1.14, 95% CI: 1.09-1.20). In addition, we identified two novel ovarian cancer risk modifier loci: 17q21.31 (rs17631303, P = 1.4×10-8, HR = 1.27, 95% CI: 1.17-1.38) and 4q32.3 (rs4691139, P = 3.4×10-8, HR = 1.20, 95% CI: 1.17-1.38). The 4q32.3 locus was not associated with ovarian cancer risk in the general population or BRCA2 carriers, suggesting a BRCA1-specific associat

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P < 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead