Vortex fluidics-mediated DNA rescue from formalin-fixed museum specimens.

DNA from formalin-preserved tissue could unlock a vast repository of genetic information stored in museums worldwide. However, formaldehyde crosslinks proteins and DNA, and prevents ready amplification and DNA sequencing. Formaldehyde acylation also fragments the DNA. Treatment with proteinase K proteolyzes crosslinked proteins to rescue the DNA, though the process is quite slow. To reduce processing time and improve rescue efficiency, we applied the mechanical energy of a vortex fluidic device (VFD) to drive the catalytic activity of proteinase K and recover DNA from American lobster tissue (Homarus americanus) fixed in 3.7% formalin for >1-year. A scan of VFD rotational speeds identified the optimal rotational speed for recovery of PCR-amplifiable DNA and while 500+ base pairs were sequenced, shorter read lengths were more consistently obtained. This VFD-based method also effectively recovered DNA from formalin-preserved samples. The results provide a roadmap for exploring DNA from millions of historical and even extinct species

Substituted Anthraquinones Represent a Potential Scaffold for DNA Methyltransferase 1-Specific Inhibitors

In humans, the most common epigenetic DNA modification is methylation of the 5-carbon of cytosines, predominantly in CpG dinucleotides. DNA methylation is an important epigenetic mark associated with gene repression. Disruption of the normal DNA methylation pattern is known to play a role in the initiation and progression of many cancers. DNA methyltransferase 1 (DNMT1), the most abundant DNA methyltransferase in humans, is primarily responsible for maintenance of the DNA methylation pattern and is considered an important cancer drug target. Recently, laccaic acid A (LCA), a highly substituted anthraquinone natural product, was identified as a direct, DNA-competitive inhibitor of DNMT1. Here, we have successfully screened a small library of simplified anthraquinone compounds for DNMT1 inhibition. Using an endonuclease-coupled DNA methylation assay, we identified two anthraquinone compounds, each containing an aromatic substituent, that act as direct DNMT1 inhibitors. These simplified anthraquinone compounds retain the DNA-competitive mechanism of action of LCA and exhibit some selectivity for DNMT1 over DNMT3a. The newly identified compounds are at least 40-fold less potent than LCA, but have significantly less complex structures. Collectively, this data indicates that substituted anthraquinone compounds could serve as a novel scaffold for developing DNMT1-specific inhibitors

Solid State Proton Spin Relaxation and Methyl and \u3ci\u3et\u3c/i\u3e-Butyl Reorientation

We have measured the temperature T and Larmor frequency ω/2π dependence of the proton spin‐lattice relaxation rate R in solid 1‐hydroxy‐2,4,6‐tri‐butylbenzene. The data is interpreted in terms of the rotational motion of the t‐butyl groups and their constituent methyl groups. Our data is much more extensive than a previous report [J. Yamauchi and C. A. McDowell, J. Chem. Phys. 75, 1051 (1981)] resulting in a revised dynamical model and considerably larger rotational barriers. Interesting thermal history effects are discussed

Epitope-Specific Anti-C1q Autoantibodies in Systemic Lupus Erythematosus

OBJECTIVE: In patients with systemic lupus erythematosus (SLE) complement C1q is frequently targeted by autoantibodies (anti-C1q), that correlate best with active renal disease. Anti-C1q bind to largely unknown epitopes on the collagen-like region (CLR) of this highly functional molecule. Here we aimed at exploring the role of epitope-specific anti-C1q in SLE patients. METHODS: First, 22 sera of SLE patients, healthy controls and anti-C1q positive patients without SLE were screened for anti-C1q epitopes by a PEPperMAP(®) microarray, expressing CLR of C1q derived peptides with one amino acid (AA) shift in different lengths and conformations. Afterwards, samples of 378 SLE patients and 100 healthy blood donors were analyzed for antibodies against the identified epitopes by peptide-based ELISA. Relationships between peptide-specific autoantibodies and SLE disease manifestations were explored by logistic regression models. RESULTS: The epitope mapping showed increased IgG binding to three peptides of the C1q A- and three of the C1q B-chain. In subsequent peptide-based ELISAs, SLE sera showed significantly higher binding to two N-terminally located C1q A-chain peptides than controls (p < 0.0001), but not to the other peptides. While anti-C1q were associated with a broad spectrum of disease manifestations, some of the peptide-antibodies were associated with selected disease manifestations, and antibodies against the N-terminal C1q A-chain showed a stronger discrimination between SLE and controls than conventional anti-C1q. CONCLUSION: In this large explorative study anti-C1q correlate with SLE overall disease activity. In contrast, peptide-antibodies are associated with specific aspects of the disease suggesting epitope-specific effects of anti-C1q in patients with SLE

Primary Care Provider Perceptions of Colorectal Cancer Screening Barriers: Implications for Designing Quality Improvement Interventions

Aims. Colorectal cancer (CRC) screening is underutilized. Increasing CRC screening rates requires interventions targeting multiple barriers at each level of the healthcare organization (patient, provider, and system). We examined groups of primary care providers (PCPs) based on perceptions of screening barriers and the relationship to CRC screening rates to inform approaches for conducting barrier assessments prior to designing and implementing quality improvement interventions. Methods. We conducted a retrospective cohort study linking EHR and survey data. PCPs with complete survey responses for questions addressing CRC screening barriers were included (N=166 PCPs; 39,430 patients eligible for CRC screening). Cluster analysis identified groups of PCPs. Multivariate logistic regression estimated odds ratios and 95% confidence intervals for predictors of membership in one of the PCP groups. Results. We found two distinct groups: (1) PCPs identifying multiple barriers to CRC screening at patient, provider, and system levels (N=75) and (2) PCPs identifying no major barriers to screening (N=91). PCPs in the top half of CRC screening performance were more likely to identify multiple barriers than the bottom performers (OR, 4.14; 95% CI, 2.43–7.08). Conclusions. High-performing PCPs can more effectively identify CRC screening barriers. Targeting high-performers when conducting a barrier assessment is a novel approach to assist in designing quality improvement interventions for CRC screening

Menstrual health and school absenteeism among adolescent girls in Uganda (MENISCUS): a feasibility study.

BACKGROUND: Management of menstruation can present substantial challenges to girls in low-income settings. In preparation for a menstrual hygiene intervention to reduce school absenteeism in Uganda, this study aimed to investigate menstruation management practices, barriers and facilitators, and the influence of menstruation on school absenteeism among secondary school students in a peri-urban district of Uganda. METHODS: Qualitative and quantitative studies were conducted among consenting girls and boys aged 14-17 years in four secondary schools in Entebbe sub-District, Uganda. Methods included group and in-depth interviews with students, a quantitative cross-sectional questionnaire, a prospectively self-completed menstrual diary, key informant interviews with policy makers, and observations of school water, sanitation and hygiene facilities. Multiple logistic regression was used to assess factors associated with school absenteeism during the most recent menstrual period. RESULTS: Girls reported substantial embarrassment and fear of teasing related to menstruation in the qualitative interviews, and said that this, together with menstrual pain and lack of effective materials for menstrual hygiene management, led to school absenteeism. All policy makers interviewed reported poverty and menstruation as the key factors associated with school attendance. The 352 girls with questionnaire data had a median age of 16 (inter-quartile range (IQR) = 15,16) years, with median age at menarche of 13 (IQR = 13,14) years. Of these, 64 girls (18.7%) reported having stained their clothes and 69 (19.7%) reported missing at least 1 day of school, during their most recent period. Missing school during the most recent period was associated with physical symptoms (headache (odds ratio (OR) = 2.15, 95%CI:1.20, 3.86), stomach pain (OR = 1.89, 95%CI:0.89, 4.04), back pain (OR = 1.75, 95%CI:0.97, 3.14), and with changing protection 4 or more times per 24 h period (OR = 2.08, 95%CI:1.06, 4.10). In the diary sub-study among 40 girls, school absence was reported on 28% of period-days, compared with 7% of non-period days (adjusted odds ratio = 5.99, 95%CI:4.4, 8.2; p < 0.001). CONCLUSION: In this peri-urban Ugandan population, menstruation was strongly associated with school attendance. Evaluation of a menstrual management intervention that address both psychosocial (e.g. self-confidence, attitudes) and physical (e.g. management of pain, use of adequate menstrual hygiene materials, improved water and sanitation facilities) aspects of menstruation are needed

Chesterfield County and the Cities of Colonial Heights, Petersburg, and Richmond, Virginia - Shoreline Inventory Report: Methods and Guidelines

The 2017Inventory for Chesterfield County and the Cities of Colonial Heights, Petersburg, and Richmond was generated using on-screen, digitizing techniques in ArcGIS® -ArcMap v10.4.1while viewing conditions observed in Bing high resolution oblique imagery, Google Earth, and2013imagery from the Virginia Base Mapping Program (VBMP).Four GIS shapefiles are developed. The first describes land use and bank conditions (Chesterfield_lubc_2017). The second portrays the presence of beaches (Chesterfield_beaches_2017). The third reports shoreline structures that are described as arcs or lines(e.g. riprap)(Chesterfield_sstru_2017). The final shapefile includes all structures that are represented as points (e.g. piers)(Chesterfield_astru_2017).The metadata file accompanies the shapefiles and defines attribute accuracy, data development, and any use restrictions that pertain to data

Acute posterior cingulum integrity post-trauma prospectively predicts depression but not PTSD symptoms

Background: Little is known about what distinguishes those who are resilient after trauma from those at risk for developing posttraumatic stress disorder (PTSD). Previous work indicates white matter integrity may be a useful biomarker in predicting PTSD. Research has shown changes in the integrity of three white matter tracts—the cingulum bundle, corpus callosum (CC), and uncinate fasciculus (UNC)—in the aftermath of trauma relate to PTSD symptoms. However, few have examined the predictive utility of white matter integrity in the acute aftermath of trauma to predict prospective PTSD symptom severity in a mixed traumatic injury sample. Method: Thus, the current study investigated acute brain structural integrity in 148 individuals being treated for traumatic injuries in the Emergency Department of a Level 1 trauma center. Participants underwent diffusion-weighted magnetic resonance imaging 2 weeks post-trauma and completed several self-report measures at 2-weeks (T1) and 6 months (T2), including the Clinician Administered PTSD Scale for DSM-V (CAPS-5), post-injury. Results: Consistent with previous work, T1 lesser anterior cingulum fractional anisotropy (FA) was marginally related to greater T2 total PTSD symptoms. No other white matter tracts were related to PTSD symptoms. Conclusions: Results demonstrate that in a traumatically injured sample with predominantly subclinical PTSD symptoms at T2, acute white matter integrity after trauma is not robustly related to the development of chronic PTSD symptoms. These findings suggest the timing of evaluating white matter integrity and PTSD is important as white matter differences may not be apparent in the acute period after injury