Binomial Distribution Test Research Articles

Statins have profound effects on the epicardial fat transcriptome

Abstract Background Epicardial adipose tissue (EAT) has direct contact with the myocardium, which enables a common microcirculation allowing bidirectional signaling between the tissues. It is possible that a change in the balance of protective and damaging mediators produced by EAT is a major driver for myocardial dysfunction. Notably, EAT volume and thickness associate with increased pro-inflammatory secretome and with coronary artery disease (CAD). Purpose This aim of this study was to analyze the total transcriptome in EAT and relate putative differences between patient groups to metabolic parameters as well as ongoing medications. Methods Between 2011 and 2016, 44 patients (mean age 68.0 years, 18.2% women) had a biopsy taken from the epicardial fat during coronoray by-pass surgery (CABG, n=36), or during elective aortic valvular replacement due to aortic stenosis (AVR, n=8). RNA from epicardial fat biopsis was extracted and sequenced. Differentially expressed genes were obtained by custom scripts that performed a negative binomial distribution test on effective counts. Multivariate regression analysis with orthogonal partial least-squares-discriminant analysis (OPLS-DA) was applied to extract and interpret the systematic variation in the resolved RNA sequencing profiles. To identify enrichment of biologically relevant functional gene sets, our data were entered in evidence-based network enrichment analysis (EviNet). Results The CABG surgery was performed approximately 7 days after an acute coronary event (75% acute MI), and 44% of them had previously known CAD. None of the AVR cases had significant CAD. At surgery, 97% of the CABG cases were prescribed statins, and 36% of them had statins alredy before the actual admission (long-term statin treatment). 63% of the AVR cases were on statins at admission. The OPLS analysis revealed only one medication with significant outcome, i.e., patients with long-term treatment with statins (n=18) versus patients without (n=26). Long-term statin treatment associated with down-regulated gene expression involved in beta-oxidation and triglyceride biosynthesis whereas gene expression involved in extracellular matrix organization were up-regulated. Gene expression involved in MHC class II presentation and interferon signalling were also reduced in patients with long-term treatment with statisn. A downregulation of genes in the interleukin- and complement cascade genes among CABG patient (versus AVR patients) without long-term statin treatment showed normal levels in statin pretreated patients. Conclusion Long-term statin treatment has a major influence on transcriptional regulation of genes involved in beta-oxidation, triglyceride biosynthesis, extracellular matrix composition, MHC class II presentation and interferon signalling in epicardial adipose tissue. These findings support important metabolic and immune effects of statins beyond the lipid-lowering function.

Satisfaction survey on graduates of standardised training for resident doctors in Guizhou Province, China

ObjectivesThis study aimed to investigate the satisfaction levels of graduate residents in Guizhou Province and analyse the factors influencing their satisfaction.MethodsA cross-sectional survey was conducted using a simple random sampling...

MRI trigeminal nerve digitalization for trigeminal neuralgia diagnosis

ObjectiveThis study aimed to determine the affected side in trigeminal neuralgia by examining the digitized angle ratio value between the bilateral trigeminal nerve root axis and the pontine tangent line. MethodsTrigeminal nerve-related MRI from 81 patients with trigeminal neuralgia (TNG) and 20 healthy persons were divided into normal and diseased groups. All the patient's films were scanned to form the images stored in compute. The following skull base MRI parameters were measured by Image J software: 1. bilateral trigeminal nerve root-pontine tangent line angle (T-P angle) values were measured. 2. Clinical symptoms were used to record the affected side of diseased patients, and 3. Nerve vascular confliction (NVC) and the side with NVC were subdivided (none/left/right/both sides). The normal T-test proved that P Value between the normal group and the diseased group. The T-P angle Ratio of the normal group was utilized to estimate the normal patient Ratio to establish the 95% confidence interval (CI). In diseased group, if the ratio was ≥ the reference value, the afflicted side was determined by comparing the max T-P angle with the actual affected side for accuracy. A binomial distribution test verified its statistical significance. ResultsThe T-P angle was significantly different between patients with TNG and normal subjects (p < 0.001). The T-P angle ratio (R) ≥ 1.238 was used to diagnose the affected side, achieving an overall accuracy rate of 82% (95% CI: 75%–89%). For patients without NVC, the accuracy was 78% (95% CI: 60%–95%), while for those with unilateral NVC, it was 91% (95% CI: 82%–100%), and for those with bilateral NVC, it was 69% (95% CI: 52%–86%). For 1 ≤ R < 1.238, the accuracy for diagnosing the affected side using NVC was 80% (95% CI: 67%–93%). ConclusionThe T-P angle is significantly higher in TNG patients than in normal patients (p < 0.001)., making it a valuable diagnostic tool. This measurement aids in guiding microvascular decompression surgery, particularly in non-vascular compression cases, and enhances postoperative evaluation. Future integration into standard protocols could improve surgical outcomes and patient care.

CONSTRUÇÃO E VALIDAÇÃO DE TECNOLOGIA EDUCACIONAL SOBRE O PÓS-TRANSPLANTE HEPÁTICO PARA O FAMILIAR CUIDADOR

ABSTRACT Objective: to construct and validate an educational booklet on post-liver transplant for family caregivers. Method: a methodological study carried out in two stages from August 2021 to July 2023: 1) Construction of the educational booklet aimed at liver transplant patient's family caregivers; and 2) Content and face validation by expert judges and semantic validity with the target audience. For the experts to analyze the validity data, the Content Validity Index was used (≥0.78 per item and ≥0.80 in total), as well as the Exact Binomial Distribution test (p&gt;0.05 and 0.80) and the Intraclass Correlation Coefficient (95%) for greater reliability. In the semantic evaluation, the IRaMuTeQ software was used to guide the analysis of the interviews, using Descending Hierarchical Classification and Word Cloud. Results: after the bibliographic survey, four axes guided the construction of the booklet. The total Content Validity Index was 0.99 (p&gt;0.05), with excellent reliability (ICC=0.946). The total Face Validity Index was 0.93 (p&gt;0.05), with good reliability (ICC=0.766). In terms of semantics, the analytical categories pointed to a satisfactory level of perception and understanding of the booklet by the target audience. Conclusion: the technology is innovative, including caregivers in the education process, and is relevant to post-transplant care. The validation process shows the feasibility of understanding the booklet and the need to use it to improve home care.

Short-term Results of the Masking Effect of an Infiltrant Resin on Mild Molar Incisor Hypomineralization Lesions in Anterior Teeth.

This non-controlled clinical study evaluated the masking effect of an infiltrant resin on mild molar incisor hypomineralization (MIH) lesions. Thirty MIH-affected anterior teeth with creamy/ white opacities from 12 children aged 6-15 years received the application of an infiltrant resin (Icon- DMG). Standard photographs were taken before (T1), immediately after (T2), and 1 week after (T3) resin infiltration. Two calibrated examiners qualitatively analyzed the color match using the Fédération Dentaire Internationale (FDI) scale. The binomial distribution test analyzed the scores of the color match at T1 with T2 and T3, and McNemar's test analyzed the scores of the color match between T2 and T3 (α=0.05). There was a significant increase in color match between T1 and T2 (p=0.0005), between T1 and T3 (p=0.0005), and between T2 and T3 (p=0.0019). It was concluded that infiltrant resin was effective in improving the esthetic appearance of creamy/white opacities on MIH-affected anterior teeth.

Uji Distribusi Binomial Pada Data Lama Masa Studi Mahasiswa

The length of study time for students at a higher education institution varies quite a lot. Many factors cause students to finish college on time or not on time. The length of study period is one indicator of the success of a tertiary institution. The faster students complete their education, the better the quality of service from an educational institution. This study aims to test the binomial distribution on data on length of study of students of the Tribuana University Mathematics Study Program (Untrib) Kalabahi. The binomial distribution is a distribution of discrete random variables with n trials and two outcomes. The data was obtained from the mathematics study program (prodi) document, in the form of the length of study period of 105 graduate students in 2017-2021. Then a binomial distribution test was carried out with successful data categories, namely students who completed college ≤ 8 semesters and the failed category, namely students who completed college &gt; 8 semesters. The results of the study show that the average success rate for mathematics students is not equal to 50% of the total students in each batch

Range restriction, climate variability and human‐related risks imperil lizards world‐wide

AbstractAimsIdentifying major reasons for species imperilment is a necessary step for conservation, but the degree to which we can generalize is hard for species‐rich yet less‐studied taxa, such as lizards. Here, we aim to bridge the gap by providing comprehensive analyses of the correlates and processes of species extinction and threats for global lizards.LocationGlobal.Time periodCurrent.Major taxa studiedLizards.MethodsWe compiled a dataset comprising extinction risk status, six intrinsic traits and seven extrinsic factors for 5256 lizard species. We carried out binomial distribution tests for 43 families and seven realms to check the non‐randomness in species extinction risk and used phylogenetic linear regressions to identify the key factors that relate to the extinction proneness of lizards and species subgroups. Based on the International Union for Conservation of Nature (IUCN) threat assessment, we identified major threats for global lizards and for major families and regions.ResultsWe found strong evidence of taxonomic and geographical non‐randomness in the extinction risk of lizards. Geographical range size, human footprint and density, insular endemism, temperature and precipitation seasonality, and body size were key predictors of extinction risk, and the first three factors were also important across families and realms. Moreover, newly described species were more likely to have a restricted range size and a higher extinction risk. Globally, the most detrimental threat was habitat destruction, while overexploitation, species invasion and climate change varied widely in importance among species groups.Main conclusionsOverall, we highlight the detrimental influences of range restriction, climate variability and anthropogenic threats to species persistence. We suggest that lizards are potentially at high risk of extinction owing to widespread human disturbance and that species with extinction‐prone traits require conservation prioritization. Moreover, lizards of different families and regions require different management strategies because of variation in extinction‐risk correlates and threats.

Services personalization in digital academic libraries: a Delphi study

PurposeThis paper aims to explore the realm of literature about personalization of digital library services. This paper focuses on users’ unique needs and will identify different types of personalized services. Therefore, this study has identified different types of services personalization in the context of digital academic libraries.Design/methodology/approachIn this research, the systematic review method has been used to obtain the relevant indicators of different types of personalization in the context of libraries. To explain basic indicators, a Delphi method has been used. The Delphi panel’s members consisted of 15 experts (faculty members, researchers, professional users and software designers). A purposeful sampling and the Delphi fulfillment process were performed in three rounds. After collecting data, descriptive statistics (mean and standard deviation), inferential statistics (binomial distribution test) and the Kendall coordination coefficient were used to determine the consensus rate among experts.FindingsA total of 103 indicators were extracted for different types of personalization through a systematic literature review. Of these, 90 indicators were considered significant in the experts’ view. Generally, content personalization, interactive personalization, collaborative personalization and information retrieval personalization are the main components of personalization types, each of which has its own indicators.Originality/valueThis study has dealt with the issue of what is personalized in the context of digital academic library. The findings should be helpful and effective in the development of a holistic view on personalization of services in digital libraries.

To tether or fuse? Significant equipoise remains in treatment recommendations for idiopathic scoliosis.

Vertebral body tethering (VBT) continues to grow in interest from both a patient and surgeon perspective for the treatment of scoliosis. However, the data are limited when it comes to surgeon selection of both procedure type and instrumented levels. This study sought to assess surgeon variability in treatment recommendation and level selection for VBT versus posterior spinal fusion (PSF) for the management of scoliosis. Surgeon members of the Pediatric Spine Study Group and Harms Study Group were queried for treatment recommendations and proposed upper instrumented vertebra (UIV) and lower instrumented vertebra (LIV) selection for PSF and VBT based on 17 detailed clinical vignettes. Responses were subdivided in each clinical vignette according to surgeon experience and treatment recommendations with assessment of intra-rater reliability. Binomial distribution tests were used to establish equipoise, selecting p < 0.10 to indicate the presence of a treatment choice with consensus set > 70% agreement. For treatment choice, responses were assessed first for consensus on the decision to proceed with PSF or VBT. Thirty-five surgeons with varied experience completed the survey with 26 surgeons (74%) completing the second follow-up survey. Overall, VBT was the recommended treatment by 47% of surgeons, ranging by clinical vignette. Consensus in treatment recommendation was present for 6 clinical vignettes including 3 for VBT and 3 for PSF, with equipoise present for the remaining 11. Of the 17 vignettes, 12 demonstrated moderate intra-observer reliability including the 3 consensus vignettes for VBT. Sanders stage ≤ 3 and smaller curve magnitude were related with VBT recommendation but neither age nor curve flexibility significantly influenced the decision to recommend VBT. Surgeons with high VBT volume, ≥ 11 VBT cases/year, were more likely to recommend VBT than those with low volumes (0-10 cases per year (p < 0.0001)). High VBT volume surgeons demonstrated consensus in VBT recommendation for Lenke 5/6 curves (75% mean recommendation). High VBT volume surgeons had a significantly higher VBT recommendation rate for Lenke 1A, 2A curves (71.8% vs 48.0%, p = 0.012), and Lenke 3 curves (62% vs 26.9%, p = 0.023). Equipoise was present for all vignettes in low volume surgeons. In addition, high VBT volume surgeons trended toward including more instrumented levels than low VBT volume surgeons (7.17 vs 6.69 levels). Significant equipoise is present among pediatric spine surgeons for treatment recommendations regarding VBT and PSF. Surgeon-, patient-, and curve-specific variables were identified to influence treatment recommendations, including surgeon experience, curve subtype, deformity magnitude, and skeletal maturity. This study highlights the need for continued research in identifying the optimal indications for VBT and PSF in the treatment of pediatric spinal deformity.

Development and validation of educational hypermedia for family members and caregivers of people with epidermolysis bullosa.

to develop educational hypermedia to support the care of people with epidermolysis bullosa; and validate its content, functionality, usability, and efficiency. methodological study, based on five phases: analysis and planning; modeling; implementation; evaluation; and distribution. Hypermedia was evaluated by expert judges, using the content validity index, index of agreement, and exact binomial distribution test, considering p > 0.05 and 0.80 agreement ratio to estimate the statistical reliability of CVI and IOA. in the validation with the judges, hypermedia presented excellent indices of total content validity (CVIt=0.99) and agreement for functionality, usability, and efficiency (IOA=100%), with p > 0.05 in all evaluated items. the developed hypermedia was considered adequate, with good screens, descriptions, and animations, presenting itself clearly and objectively to be used by patients with epidermolysis bullosa and their family members/caregivers.

Benchmarking of mobile apps on heart failure.

to analyze the mobile apps on heart failure available in the main operating systems and their usability. benchmarking of mobile applications, systematic research, comprising 38 mobile applications for analysis of general information, functionalities and usability. Usability was assessed using System Usability Scale and Smartphone Usability Questionnaire, followed by the calculation of the agreement index and the exact binomial distribution test, with a significance level of p> 0.05 and a proportion of 0.90. mobile applications had English as the predominant language (73.7%), were directed to patients (71.1%) and the predominant theme was disease knowledge (34.2%). Functionalities ranged from general features to the need for an internet connection. In assessing usability, heart failure was shown to be 92.1% -94.7% and p <0.05. the mobile apps on heart failure have varied content and adequate usability. However, there is a need to develop more comprehensive mobile applications.

Change in amount smoked and readiness to quit among patients undergoing lung cancer screening.

BackgroundThere is mixed evidence regarding whether undergoing computed tomography lung cancer screening (LCS) can serve as a “teachable moment” that impacts smoking behavior and attitudes. The study aim was to assess whether the standard procedures of undergoing LCS and receiving free and low-cost evidence-based cessation resources impacted short-term smoking-related outcomes.MethodsParticipants were smokers (N=87) who were registered to undergo lung screening and were enrolled in a cessation intervention trial. We conducted two phone interviews, both preceding trial randomization: the first interview was conducted prior to lung screening, and the second interview followed lung screening (median =12.5 days post-screening) and participants’ receipt of their screening results. The interviews assessed demographic characteristics, interest in evidence-based cessation intervention methods, and tobacco-related characteristics, including cigarettes per day and readiness to quit. Participants received minimal evidence-based cessation resources following the pre-lung screening interview.ResultsParticipants were 60.3 years old, 56.3% female, and reported a median of 40 pack-years. Participants were interested in using several evidence-based strategies, including counseling from a healthcare provider (76.7%) and receiving nicotine replacement therapy (69.8%). Pre-lung screening, 25.3% smoked ≤10 cigarettes per day, and 29.9% were ready to quit in the next 30 days. We conducted two McNemar binomial distribution tests to assess change from pre- to post-screening. At the post-lung screening assessment, approximately three-quarters reported no change on these variables. However, 23.3% reported smoking fewer cigarettes per day, whereas 4.7% reported smoking more cigarettes per day (McNemar P=0.002), and 17.2% reported increased readiness to quit, whereas 6.9% reported decreased readiness to quit (McNemar P=0.078).ConclusionsFollowing receipt of cessation resources and completion of lung screening, most participants reported no change in smoking outcomes. However, there was a significant reduction in cigarettes per day, and there was a trend for increased readiness to quit. This setting may provide a potential “teachable moment” and an opportunity to assist smokers with quitting. However, more proactive and intensive interventions will be necessary to capitalize on these changes and to support abstinence in the long-term.

Guild burials or family graves? Intrapopulation relationship analysis of 18th century La Concepción church cemetery in Santa Cruz de Tenerife, Canary Islands

La Concepción church is the oldest church in Santa Cruz de Tenerife and it was used as a cemetery for over 200 years, during the 16th century and up to the 18th century. Those who were buried there were from multiple ethnic backgrounds and with different lifestyles. During an emergency archaeological excavation, comingled, poorly preserved bone remains of about 300 individuals were excavated. We here analyze a total of 198 tali, looking for the presence of non-metric traits. The results show that the frequencies of non-metric traits were greater than the majority of previous studies, indicating that a large proportion of the sample had an increased activity in the ankle region. Some traits appeared in the same bone and, more importantly, several tali with uncommon combinations belonged to individuals buried in the same tomb. These results were tested for randomness with binomial distribution tests that resulted in a P < 0.0001. Moreover, tombs containing tali with uncommon trait combinations showed a tendency to cluster in certain areas of the church. The results of this study therefore show that either people who performed similar activities that led to the development of certain patterns of trait combinations were interred together or in neighboring tombs, or these combinations of traits indicate a genetic relationship that explains why the individuals were buried in the same tombs. This study highlights the importance of analysing trait combinations in talar bones, since, together with other bioanthropological research tools – including genetics, and/or historical data – they may provide useful information for the reconstruction of important aspects of the daily lives of past populations. The study of trait combinations in talar bones may also aid in the detection and understanding of relationships within a cemetery, where kinship analysis is unable to be applied.

Clonal Somatic Mutations Are a Biomarker for Inferior Prognosis in Diffuse Large B-Cell Lymphoma

Background Diffuse large B-cell lymphoma (DLBCL) is the most common aggressive non-Hodgkin lymphoma subtype with a 5-year survival of ~64%. While DLBCL is treated using immunochemotherapy (IC) with curative intent, 20%-40% of patients do not reach remission or relapse post IC. Clonal somatic mutations have been associated with aging, hematologic malignancies (predominately myeloid), and reduced OS in the general population. The objective of this study was to evaluate the association of clonal somatic mutations with event-free survival (EFS) and overall survival (OS). Methods We studied newly diagnosed DLBCL cases treated with IC who were enrolled in the Molecular Epidemiology Resource (MER) of the University of Iowa/Mayo Clinic Lymphoma Specialized Program of Research Excellence (SPORE). Clinical and treatment data were abstracted from medical records and all patients were systematically followed for disease outcomes. Pre-treatment DNA was extracted from matched peripheral blood and paraffin-embedded tumor tissue, and whole exome sequencing was conducted at 100x coverage. From the peripheral blood, allele counts from GVCF files produced by HaplotypeCaller in GATK were extracted for 42 genes commonly associated with clonal hematopoiesis (e.g. DNMT3A, TET2, and ASXL1). Mutations were deemed clonal somatic if the population minor allele frequency was &amp;lt;0.5% in ExAC and had a variant allele fraction (VAF) that deviated from 0.5 using a binomial distribution test at p&amp;lt;0.001. Furthermore, mutations had to have a minimum alternate read count of at least 5 reads, a VAF greater than or equal to 10%, and met at least one of the following criteria: CAVA impact of "HIGH"; pathogenic/likely pathogenic per ClinVar; or reported in COSMIC at least 10 times. EFS was defined as time from diagnosis to disease progression, relapse, retreatment, or death due to any cause. The association of clonal somatic mutations with prognosis was estimated using Kaplan-Meier curves, and hazard ratios (HR) and 95% confidence intervals (CI) from Cox regression. Associations were adjusted for age at diagnosis, sex, and the International Prognostic Index (IPI). Results The study consisted of 261 DLBCL patients treated with IC. The median age at diagnosis was 65 years (range 20-90) and 56% were male. With a median follow-up time of 5.1 years (range 0.1-15.0), there were 100 events and 80 deaths. A total of 17 (6.5%) patients had clonal somatic mutations, and16 patients were over the age of 60. A total of 8 (of 42) genes had clonal somatic mutations, with SF3B1, ASXL1, and TET2, being the most frequent (4 individuals per gene). VAFs ranged from 0.10 to 0.28 and none of the patients had multiple mutations. Additionally, the clonal somatic variants found in the peripheral blood were abscent in the tumor sample. Of the 17 patients with clonal somatic mutations, 12 had an event while 88 patients without a mutation had an event. In a univariate analysis, clonal somatic mutations were associated with inferior EFS (HR=2.55, 95% CI 1.39-4.68, p=0.002; Figure 1A). After adjusting for age, sex, and IPI, clonal somatic mutations remained associated with inferior EFS (HR=2.02, 95% CI 1.09-3.74, p=0.026). Clonal somatic mutations were also associated with inferior OS in the univariate analysis (HR=2.06, 95% CI 0.99-4.29, p=0.053), which attenuated after multivariate adjustment (HR=1.59, 95% CI: 0.76-3.34, p=0.22, Figure 1B). Although based on small numbers, mutations in SF3B1 were associated with inferior EFS (HR=3.25, 95% CI 1.16-9.12, p=0.025), but did not reach significance for OS (HR=2.56, 95% CI 0.78-8.38, p=0.120). Conclusions In this novel study of newly diagnosed DLBCL patients, clonal somatic mutations were identified in 6.5% of patients and were associated with inferior outcomes. Additional research is required at deeper sequencing to validate these findings and integration with tumor genomics is required to understand the prognosis of DLBCL patients with smaller clonal populations. Disclosures Shah: Dren Bio: Consultancy. Maurer:Celgene / BMS: Research Funding; Pfizer: Membership on an entity's Board of Directors or advisory committees; Morphosys: Membership on an entity's Board of Directors or advisory committees; Kite: Membership on an entity's Board of Directors or advisory committees; Nanostring: Research Funding. Martin:Celgene: Consultancy; Karyopharm: Consultancy, Research Funding; Morphosys: Consultancy; Regeneron: Consultancy; Incyte: Consultancy; Kite: Consultancy; Cellectar: Consultancy; Bayer: Consultancy; Beigene: Consultancy; Sandoz: Consultancy; I-MAB: Consultancy; Janssen: Consultancy; Teneobio: Consultancy. Witzig:AbbVie: Consultancy; Incyte: Consultancy; Acerta: Research Funding; Karyopharm Therapeutics: Research Funding; Immune Design: Research Funding; Spectrum: Consultancy; Celgene: Consultancy, Research Funding; MorphSys: Consultancy. Nowakowski:NanoString: Research Funding; Seattle Genetics: Consultancy; Curis: Consultancy; Kymera: Consultancy; Kite: Consultancy; Ryvu: Consultancy, Membership on an entity's Board of Directors or advisory committees; MorphoSys: Consultancy, Research Funding; Celgene/BMS: Consultancy, Research Funding. Novak:Celgene/BMS: Research Funding. Cerhan:NanoString: Research Funding; BMS/Celgene: Research Funding.

The Endothelial Protein C Receptor (EPCR) Regulates Endogenous Factor VII Levels in Mice

The endothelial protein C receptor (EPCR) has been demonstrated to bind activated FVII (FVIIa) through the Gla domain with equal affinity to Protein C (PC). Mouse studies suggest that EPCR is involved in the extravasation of infused human FVIIa, leading to an extended extravascular tissue persistence, longer than expected based on its circulating half-life. This provides a plausible explanation for the long-term benefits of hemophilic patients on human FVIIa prophylaxis. Collectively, these data suggest that EPCR sequesters administered FVIIa in tissues where it may have a hemostatic effect. However, the role of the endogenous FVII-EPCR interaction in normal conditions is largely unknown. For this, we have developed a mouse model to better understand this interaction in vivo. Endogenous mouse FVII and FVIIa (mFVII/FVIIa) do not bind mouse EPCR. However, our laboratory has demonstrated that L4F, L8M, and T9R substitutions in the Gla domain of mFVIIa enable its interaction with mouse EPCR while retaining full enzymatic activity in vitro. Based on that data, we utilized CRISPR/Cas9 technology to knock-in L4F, L8M, and T9R into the mFVII Gla domain in the mouse F7 locus (F7FMR), thereby developing mice with a chimeric endogenous FVII capable of binding EPCR. Founder animals were generated and capable of producing offspring, indicating that the gain-of-function in mFVII was compatible with life. Animals were subsequently backcrossed to wildtype C57BL/6 mice in order to remove potential off-target effects of the CRISPR/Cas9. Resultant heterozygous animals (F7FMR/WT) from the final cross were bred to generate F7FMR/FMR, F7FMR/WT, and F7WT/WTlittermates. We generated 59 male and 52 female animals and a binomial distribution test demonstrated that sex is equally distributed in the population. Moreover, the genotypes expected from the heterozygous crosses were inherited in a 1:2:1 ratio, further indicating that the gain-of-function in FVII is not lethal during development. As additional metrics of health, we measured weight longitudinally during weeks 1-10 of life and found no differences between the three genotypes for either gender. Complete blood counts (CBCs) revealed no differences between the F7FMR/FMR, F7FMR/WT, and F7WT/WTgenotypes, with the exception of a mild elevation in F7FMR/WTanimals compared to animals with wildtype FVII. Collectively, we found that the gain-of-function in EPCR binding by endogenous FVII is not detrimental to the overall health of the mice. Subsequently, we determined the mFVII levels in the F7FMR/FMR, F7FMR/WT, and F7WT/WTanimals using an in-house ELISA. We observed that plasmatic mFVII levels were dependent on the EPCR-binding capacity of the endogenous mFVII. Specifically, F7WT/WTmice, whose mFVII does not bind EPCR, had a plasmatic mFVII concentration of ~690 ng/ml. In contrast, F7FMR/FMRhomozygote mice had ~350 ng/ml of mouse FVII, approximately half the plasma levels of the F7WT/WT. Heterozygote animals F7FMR/WThad an intermediate plasmatic mFVII level (~550 ng/ml), suggesting that EPCR may regulate plasmatic FVII levels in vivo. Lastly, we determined the hemostatic response to injury in the F7FMR/FMR, F7FMR/WT, and F7WT/WTanimals. We did this in two ways, by measuring blood loss following tail clip assay and by determining time to vessel occlusion following ferric chloride injury of the carotid artery. We observed no differences between the three genotypes in response to either injury model. In conclusion, we have generated and characterized a novel mouse model in which endogenous FVII is capable of binding EPCR. Using this model, we demonstrated that EPCR can modulate plasmatic FVII levels in vivo but does not appear to affect hemostasis. Since this model mimics the FVII-EPCR interaction in humans, it can now be used to further investigate how this interaction participates in other normal or pathologic states that depend on FVII and/or EPCR. Disclosures Margaritis: Bayer Hemophilia Awards: Research Funding; Bristol-Myers Squibb: Other: Salary (spouse); CSL Behring: Other: Salary (spouse); NovoNordisk A/S: Research Funding.

Multi-omics signature of brain amyloid deposition in asymptomatic individuals at-risk for Alzheimer's disease: The INSIGHT-preAD study.

BackgroundOne of the biggest challenge in Alzheimer's disease (AD) is to identify pathways and markers of disease prediction easily accessible, for prevention and treatment. Here we analysed blood samples from the INveStIGation of AlzHeimer's predicTors (INSIGHT-preAD) cohort of elderly asymptomatic individuals with and without brain amyloid load.MethodsWe performed blood RNAseq, and plasma metabolomics and lipidomics using liquid chromatography-mass spectrometry on 48 individuals amyloid positive and 48 amyloid negative (SUVr cut-off of 0·7918). The three data sets were analysed separately using differential gene expression based on negative binomial distribution, non-parametric (Wilcoxon) and parametric (correlation-adjusted Student't) tests. Data integration was conducted using sparse partial least squares-discriminant and principal component analyses. Bootstrap-selected top-ten features from the three data sets were tested for their discriminant power using Receiver Operating Characteristic curve. Longitudinal metabolomic analysis was carried out on a subset of 22 subjects.FindingsUnivariate analyses identified three medium chain fatty acids, 4-nitrophenol and a set of 64 transcripts enriched for inflammation and fatty acid metabolism differentially quantified in amyloid positive and negative subjects. Importantly, the amounts of the three medium chain fatty acids were correlated over time in a subset of 22 subjects (p < 0·05). Multi-omics integrative analyses showed that metabolites efficiently discriminated between subjects according to their amyloid status while lipids did not and transcripts showed trends. Finally, the ten top metabolites and transcripts represented the most discriminant omics features with 99·4% chance prediction for amyloid positivity.InterpretationThis study suggests a potential blood omics signature for prediction of amyloid positivity in asymptomatic at-risk subjects, allowing for a less invasive, more accessible, and less expensive risk assessment of AD as compared to PET studies or lumbar puncture.FundInstitut Hospitalo-Universitaire and Institut du Cerveau et de la Moelle Epiniere (IHU-A-ICM), French Ministry of Research, Fondation Alzheimer, Pfizer, and Avid.

Abstract 524: Impact of age, sex, smoking, body mass and physical activity on circulating small non-coding RNA expression profiles

Abstract Non-coding RNAs (ncRNA) are regulators of cell functions and circulating ncRNAs from the majority of the RNA classes, such as miRNA, tRNA, piRNAs, lncRNA, snoRNA, snRNA and miscRNAs, are potential non-invasive cancer biomarkers. Understanding how non-disease traits influence ncRNA expression is essential for assessing their biomarker potential. The aim of the study was to investigate associations between sex, age, smoking, body mass, physical activity, technical factors such as sample storage and processing, and serum ncRNA expression profiles. Serum samples from 526 healthy individuals in the Janus Serum Bank Cohort were included in the study. The samples were collected in the time-period 1972-2004 with varying collection procedures, and stored at - 25º C. Information on smoking habits, body mass and physical activity was linked from health examination survey data. RNA was extracted from 400 µl serum using phenol-chloroform separation and the miRNA Neasy Serum/Plasma kit (Qiagen). Small RNAseq was performed using NEBNext Small RNA Library Prep Set for Illumina with an average sequencing depth of 18 million reads per sample. The RNAseq reads were initially trimmed for adapters using Adapter Removal (v2.1.7). We then mapped the collapsed reads (generated by FASTX v0.14) to the human genome (hg38) using Bowtie2 (10 alignments per read were allowed). We compiled a comprehensive annotation set from miRBase (v21) for miRNAs, pirBAse (v1.0) for piRNAs, GENCODE (v26) for other RNAs and tRNAs. We used SeqBuster (v3.1) to get isomiR and miRNA profiles. To count the mapped reads, HTSeq (v0.7.2) was used. Differential gene expression analyses based on the negative binomial distribution and Wald significance tests were performed for each trait using the R package DESeq2 version 1.14.1. We identified associations between all RNA classes and traits. Ageing showed the strongest association with ncRNA expression, both in terms of statistical significance and number of RNAs, regardless of RNA class. Serum processing modifications and storage times significantly altered expression levels of a number of ncRNAs. Smoking cessation generally restored RNA expression to non-smoking levels, although for some isomiRs, mRNA fragments and tRNAs smoking-related expression levels persisted. sncRNA expression levels in serum are considerably age-dependent and age should be adjusted for in studies of circulating sncRNA expression. Certain biomarkers are also influenced by body mass, smoking, physical activity, serum processing and storage conditions. Citation Format: Trine Ballestad Rounge, Sinan Ugur Umu, Andreas Keller, Eckart Meese, Giske Ursin, Steinar Tretli, Robert Lyle, Hilde Langseth. Impact of age, sex, smoking, body mass and physical activity on circulating small non-coding RNA expression profiles [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 524.

Validation of assistive technology on psychoactive substances for visually impaired people

Purpose: To validate the assistive technology “Drugs: reflection for prevention” to be used with visually impaired people.Method: Quantitative and quasi-experimental study, contrasting knowledge before and after the use of the assistive technology with 140 visually impaired people in institutes and associations for people with visual impairment. A questionnaire with identification data, a pre-test, a post-test and a questionnaire to assess the assistive technology were applied. Data were described through means and standard deviations, and analyses included the McNemar test, the exact binomial distribution test, and the intraclass correlation coefficient.Results: Participants were male (65.7%), 84.3% were blind, aged 37.1 years on average and with schooling of 10.1 years on average. There were more correct answers in the post-test (p <.001). The attributes of the assistive technology were considered adequate: objectives, access, clarity, structure and presentation, relevance and efficacy, and interactivity (p <.001).Conclusions: The assistive technology “Drugs: reflection for prevention” was considered valid and reliable to inform visually impaired people about psychoactive substance abuse.Implications for rehabilitationCreated new tool for prevention substance abuse that can be accessed easily.Improved information about substance psychoactive for users of the assistive technology.Improved quality of life for its users.

Percepción de los Estudiantes de Medicina Veterinaria acerca de los Perros vagabundos y sus Estrategias de Control en Lima, Perú

El objetivo del estudio fue determinar la percepción de los estudiantes de la carrera de medicina veterinaria (MV) acerca de la presencia de perros vagabundos y sus estrategias de control en Lima, Perú. Se elaboró un instrumento de auto-aplicación con preguntas de alternativas múltiples, de una o más opciones de respuesta y se aplicó en forma anónima a los estudiantes de MV de dos universidades de Lima. Se hizo una validación previa del contenido por jueces expertos (prueba de distribución binomial) y prueba piloto en estudiantes de MV de una tercera universidad, para evaluar la consistencia interna (prueba estadística alfa de Cronbach). Se analizaron 457 encuestas. El 36.5% de los encuestados perciben que los perros vagabundos en las calles tenían dueño. Los encuestados consideraron que los principales problemas que sufren estos animales eran desnutrición (86.7%), parasitosis (62.8%) y accidentes (54.9%), y los principales problemas que ocasionaban eran la reproducción no deseada (76.1%), zoonosis (47.9%) y ruptura de bolsas de basura (40.7%). Los estudiantes le atribuyen la responsabilidad de enfrentar el problema a la municipalidad del distrito (86.4%) y el Ministerio de Salud (60.6%). Para controlar el problema consideran las esterilizaciones (79.0%), adopciones (70.9%) y castraciones (60.8%) como las opciones más viables.

Negative Assortative Mating Based on Body Coloration in the Freshwater Platyfish (Poecillidae: Xiphophorus maculatus)

The ability of individuals within a population to survive and thrive is highly dependent upon the maintenance of genetic variation and phenotypic diversity, thereby ensuring adaptation to dynamic environments. A fundamental method of maintaining such variation is through a negative assortative mating strategy, in which individuals would be expected to reproductively select members of the opposite sex that exhibit dissimilar phenotypes. Employing three uniform body color morphs, red, yellow and blue, of the platyfish (Xiphophorus maculatus), this study was designed to investigate whether X. maculatus females would preferentially be attracted to males exhibiting an alternative color, thereby enabling an examination of the effect of male body coloration on mate choice by adult females. Mate choice was determined based on the initial preference of each female, as well as the amount of time females spent associating with each male. Initial preferences were analyzed using a binomial distribution test, and overall preference data using Wilcoxon signed rank tests. Red females initially selected for dissimilar colored males, and spent a significantly larger amount of time associating with blue and yellow males, as did yellow females with red and blue males. Blue females initially selected and spent a significantly larger amount of time associating with red males but, interestingly, showed no selective preference between blue and yellow males. In these experimental trials, the overall strong mate selection exhibited by female platyfish for males of dissimilar coloration is suggestive of a negative assortative mating strategy and provides evidence for the maintenance of color polymorphism in nature populations.