Convergent total synthesis of (+)-calcipotriol: A scalable, modular approach to vitamin D analogs.

A convergent approach for the total synthesis of calcipotriol (brand name: Dovonex), a proven vitamin D analog used for the treatment of psoriasis, and medicinally relevant synthetic analogs is described. A complete approach, not wedded to semisynthesis, toward both the A-ring and CD-ring is reported. From a retrosynthetic standpoint, hidden symmetry within the decorated A-ring is disclosed, which allowed for scalable quantities of this advanced intermediate. In addition, a radical retrosynthetic approach is described, which highlights an electrochemical reductive coupling as well as an intramolecular hydrogen atom transfer Giese addition to establish the 6,5-transcarbon skeleton found in the vitamin D family. Finally, a late-stage decarboxylative cross-coupling approach allowed for the facile preparation of various C20-arylated derivatives that show promising biological activity in an initial bioassay.

Dimensional profiling of psychopathology in children and adolescents based on the K-SADS-PL and an analysis of the construct validity of two ADHD symptom dimensions.

OBJECTIVES: The traditional view on psychiatric disorders as categorical and distinct is being challenged by perspectives emphasizing the relevance of dimensional and transdiagnostic assessment. However, most diagnostic instruments are based on a categorical view with a threshold-approach to disease classification. METHODS: We here describe algorithms for dimensionalizing the psychopathological ratings of the widely used diagnostic interview for children and adolescents, the Kiddie-Schedule for Affective Disorders and Schizophrenia - Present and Lifetime Version (K-SADS-PL). We further evaluate the criterion-related construct validity of the dimensionalized attention-deficit/hyperactivity disorder (ADHD) scales using Rasch models in a sample of 590 children (mean age 10.29 (.36), 49% girls). RESULTS: The algorithms generate scores of current symptom load, i.e., the sum of clinician-rated symptoms within each disorder assessed with the interview. We found support for counting symptoms of inattention and hyperactivity/impulsivity, respectively, but not for a single combined ADHD scale. CONCLUSIONS: The algorithms constitute an initial step in creating a framework for clinician-rated dimensional analyses of symptoms derived from the K-SADS-PL, but future studies are needed to further evaluate the construct validity of the remaining scales and the reliability and clinical utility of the method. We believe that our proposed algorithms offer a novel method of dimensional psychopathological assessment, which can be applied in multiple branches of child and adolescent psychiatry.

Real-world outcomes upon second-line treatment in patients with chronic lymphocytic leukaemia.

For chronic lymphocytic leukaemia (CLL), targeted drugs have become the standard of care, in particular for second-line treatment. In this study, overall survival (OS), treatment-free survival (TFS) and adverse events (AE) were registered retrospectively in a Danish population-based cohort upon second-line treatment for CLL. Data were collected from medical records and the Danish National CLL register. For 286 patients receiving second-line treatment, three-year TFS was higher upon targeted treatment (ibrutinib/venetoclax/idelalisib) [63%, 95% confidence interval (CI) 50%-76%] compared with fludarabine, cyclophosphamide and rituximab or bendamustine and rituximab (FCR/BR) (37%, CI: 26%-48%) and chlorambucil+/-CD20-antibody (CD20Clb/Clb) (22%, CI: 10%-33%). Upon targeted treatment, three-year OS estimates were higher for targeted treatment (79%, CI: 68%-91%) compared with FCR/BR (70%, CI: 60%-81%) or CD20Clb/Clb (60%, CI: 47%-74%). The most common AEs were infections and haematological AEs; 92% of patients treated with targeted drugs had AEs, 53% of which were severe. Upon FCR/BR and CD20Clb/Clb, AEs were present for 75% and 53% respectively, of which 63% and 31% were severe. These real-world data demonstrate higher TFS and a tendency towards higher OS following targeted second-line treatment for CLL compared to chemoimmunotherapy, also for patients who may be frailer and more comorbid.

Pharmacology of orismilast, a potent and selective PDE4 inhibitor.

BACKGROUND: There remains an unmet need for oral medications that are safe and efficacious for long-term management of chronic inflammatory skin diseases (CISD). Inhibition of phosphodiesterase 4 (PDE4) can modulate a broad range of pro-inflammatory cytokines that play a major role in CISD pathogenesis. Orismilast is a second generation PDE4 inhibitor in clinical development for CISD treatment. OBJECTIVES: The objective of this study was to examine the PDE4 enzymatic activity and anti-inflammatory effects of orismilast in vitro, ex vivo, and in vivo. METHODS: The PDE1-11 enzymatic activity of orismilast was tested in vitro using a single concentration of 308 nM orismilast. The PDE4 selectivity and inhibitory potency was further examined in a radiometric assay. Orismilast was tested on human whole blood and human peripheral blood mononuclear cells (PBMC) to determine effects on its cytokine secretion and inhibition profile ex vivo. Orismilast was orally administered in a murine model of chronic oxazolone-induced ear skin inflammation. Ear thickness, a marker of inflammation, and inflammatory cytokines were analysed. RESULTS: Orismilast selectively inhibited PDE4 and demonstrated potent inhibition of PDE4B and PDE4D subtype splice variants in vitro. Orismilast inhibited whole blood and PBMC production of tumour necrosis factor α (TNFα), and the secretion of T-helper (Th)1 (TNFα and IFNγ), Th17 (IL-22 and IL-23), and Th2 (IL-4, IL-5, and IL-13) related cytokines in PBMC. In vivo, 10 and 30 mg/kg doses of orismilast significantly reduced ear thickness and inflammation markers (p < 0.0001, respectively). CONCLUSION: Orismilast displayed selective and potent PDE4 inhibition and broad-spectrum anti-inflammatory activity in several pre-clinical models. The results of the study support clinical development of oral orismilast as a novel treatment option for CISD including psoriasis, atopic dermatitis, and hidradenitis suppurativa.

Substitution of unprocessed and processed red meat with poultry or fish and total and cause-specific mortality.

Recent studies found positive associations between intake of red meat and processed meat and total mortality; however, substitution of red meat with poultry and fish has been poorly investigated. We aimed to investigate associations for substitutions of red meat (unprocessed/processed) and total mortality and deaths due to cancer or CVD. We used data from the Danish Diet, Cancer and Health cohort, including 57 053 participants aged 50-64 years at baseline. Information on diet was collected through a validated 192-item FFQ. Information regarding total mortality, deaths due to cancer and deaths due to CVD was obtained by record linkage. Cox proportional hazards models were used to estimate the hazard ratio (HR) of 150 g/week substitutions of red meat with poultry or fish. During a follow-up (mean 16·1 years), 8840 deaths occurred (4567 were due to cancer; 1816 due to CVD). The adjusted HR for total death when substituting 150 g/week total red meat with poultry was 0·96 (95 % CI 0·95, 1·00) and with fish 0·99 (95 % CI 0·97, 1·01). Corresponding HR for cancer death or CVD death were similar. Substitution of processed red meat with fish or poultry was more consistently associated with a lower mortality than substitution of unprocessed red meat. For example, the adjusted HR for total death when substituting 150 g/week processed red meat with poultry was 0·95 (95 % CI 0·92, 0·98). We found that replacing processed red meat with poultry or fish was associated with a lower risk of total mortality and deaths due to cancer, but not deaths due to CVD.

Outcomes of prophylactic cranial irradiation in patients with small cell lung cancer in the modern era of baseline magnetic resonance imaging of the brain.

BACKGROUND: For decades many patients with small cell lung cancer (SCLC) have been offered prophylactic cranial irradiation (PCI) to prevent brain metastases (BM). However, the role of PCI is debated in the modern era of increased brain magnetic resonance imaging (MRI) availability. BM in SCLC patients may respond to chemotherapy, and if a negative MRI is used in the decision to use of PCI in the treatment strategy, the timing of brain MRI may be crucial when evaluating the effect of PCI. This retrospective study investigates the impact of PCI outcomes in patients with SCLC staged with brain MRI prior to chemotherapy. MATERIALS AND METHODS: This study included 245 patients diagnosed SCLC/mixed NSCLC-SCLC treated between 2012 and 2019. The population was analyzed separately for limited disease (LS-SCLC) and extensive disease (ES-SCLC). Patients were divided into groups based on baseline brain MRI prior to chemotherapy and PCI. The primary endpoint was time to symptomatic BM. Secondary endpoints were overall survival (OS), and progression-free survival (PFS). RESULTS: In patients with LS-SCLC staged with brain MRI the probability of developing symptomatic BM at one year was 4% vs. 22% (p < 0.05), median OS was 55 vs. 24 months (p < 0.05), and median PFS was 30 vs. 10 months (p < 0.05) with and without PCI, respectively. No differences in probability of symptomatic BM and survival outcomes were observed in ES-SCLC. In a multivariate regression analysis, no variables were statistically significant associated with the risk of developing symptomatic BM in patients with LS-SCLC and ES-SCLC. For patients with ES-SCLC staged with brain MRI, PS (HR = 3.33, CI; 1.41-7.89, p < 0.05) was associated with poor survival. CONCLUSION: This study found that PCI in LS-SCLC patients staged with brain MRI had lower incidence of symptomatic BM and improved survival outcomes suggesting PCI as standard of care. Similar benefit of PCI in patients with ES-SCLC was not found.

Tumour motion analysis from planning to end of treatment course for a large cohort of peripheral lung SBRT targets.

BACKGROUND: The aim is to quantify and analyse tumour motion during a course of treatment for lung SBRT patients. MATERIAL AND METHODS: Peak-to-peak motion of 483 tumours in 441 patients treated with peripheral lung SBRT at a single institution over a two year period was measured on planning CT and at all treatment fractions. Planning 4D-CT scans were analysed using our clinical workflow involving deformable propagation of the delineated target to all phases. Similarly, acquisition of the 4D-CBCT data followed the clinical workflow based on XVI 5.0 available on Elekta linacs. Differences and correlations of the peak-to-peak motion on the planning CT and at treatment were analysed. RESULTS: On the planning CT, a total of 81.4% of the tumours had a peak-to-peak motion <10 mm, and 96.1% had <20 mm. The largest motion was observed in the CC direction, with largest amplitude for tumours located in the caudal posterior part of the lung. The difference in amplitude in CC between planning CT and first fraction had a mean and standard deviation of 0.3 mm and 3.5 mm, respectively, and the largest differences were observed in the caudal posterior part of the lung. Patients with a difference in tumour motion amplitude exceeding two standard deviations (>7 mm) at the first fraction were evaluated individually, and they all had poor 4DCT image quality. The difference between the first and second/third fractions had a mean and standard deviation of 0.4 mm/0.5 mm and 2.0 mm/1.9 mm. CONCLUSION: Tumour motion at first treatment was similar to motion at planning, and motion at subsequent treatments was very similar to motion at first treatment. Large tumour motions are located towards the caudal posterior tumour locations. Patients with poor 4D-CT image quality should be closely followed at the first treatment to verify the motion.

Measuring Social Support among Soldiers with the Experienced Post-Deployment Social Support Scale (EPSSS): A Rasch-Based Construct Validity Study.

Perceived social support following deployment is a known buffer against post-deployment adverse mental health outcomes. Given contextual sensitivity of social support measures, scales that address specific social support needs of soldiers in the first months after home coming should be developed and validated. In a sample of 553 soldiers deployed to Afghanistan at two different time points (2009 and 2013, respectively), we selected items for and tested the construct validity of an 8-item measure of experienced post-deployment social support (experienced post-deployment social support scale; EPSSS). Within the item response theory framework, we used Rasch models (RM) to conduct item analysis with an emphasis on testing for differential item functioning (DIF) across background variables such as previous deployments and cohort. In short, we found that the scale did not fit the Rasch model, but with exclusion of two items, a 6-item version of the scale did fit an extended graphical loglinear Rasch model (GLLRM) with only one instance of DIF, for which the score can be adjusted. We also demonstrated that when applied as a scale, the DIF will not affect the results substantially. Hence, we conclude that the constructed 6-item EPSSS can be validly applied without score correction to assess the level of social support in Danish soldiers after home coming.

Classification of loneliness using the T-ILS: Is it as simple as it seems?

Student well-being is a growing issue in higher education, and assessment of the prevalence of conditions as loneliness is therefore important. In higher education and population surveys the Three-Item Loneliness Scale (T-ILS) is used increasingly. The T-ILS is attractive for large multi-subject surveys, as it consists of only three items (derived from the UCLA Loneliness Scale). Several ways of classifying persons as lonely based on T-ILS scores exist: dichotomous and trichotomous classification schemes and use of sum scores with rising levels indicating more loneliness. The question remains whether T-ILS scores are comparable across the different population groups where they are used or across groups of students in the higher education system. The aim was to investigate whether the T-ILS suffers from differential item functioning (DIF) that might change the loneliness classification among higher education students, using a large sample just admitted to 22 different academy profession degree programs in Denmark (N = 3,757). DIF was tested relative to degree program, age groups and gender. The framework of graphical loglinear Rasch models was applied, as this allows for adjustment of sum scores for uniform DIF, and thus for assessment of whether DIF might change the classification. Two items showed DIF relative to degree program and gender, and adjusting for this DIF changed the classification for some subgroups. The consequences were negligible when using a dichotomous classification and larger when using a trichotomous classification. Therefore, trichotomous classification should be used with caution unless suitable adjustments for DIF are done prior to classification.

Critical thinking of psychology students: A within- and cross-cultural study using Rasch models.

The primary aim of this study was to conduct a cross-cultural comparison of the level of critical thinking of Danish and Spanish psychology students (N = 788) attending innately different bachelor-level courses; personality psychology and introductory statistics/research methods. Several instruments are available for the assessment of critical thinking (CTh), but most are, however, unsuitable for use in large surveys with many constructs, due to length or single-person administration formats. One brief and much-used scale is the CTh scale in the Motivated Strategies for Learning Questionnaire (MSLQ), used in this study. To ensure unbiased comparisons across countries and courses, our secondary aim was to investigate the content validity, as well as the psychometric properties, of the CTh scale to ensure the most accurate and unbiased comparison. For this purpose, Rasch and graphical loglinear Rasch models were used. After reducing the number of items due to content validity issues, the CTh scale fitted Rasch models within each national sample, and was measurement invariant relative to age group, gender, course and university. In the cross-cultural item analysis, two items functioned differentially relative to nationality, and the scale fitted a graphical loglinear Rasch model. CTh scores related to introductory statistics/research methods courses differed significantly from scores related to personality psychology, for Danish students only.

Real-world outcomes for 205 patients with chronic lymphocytic leukemia treated with ibrutinib.

Ibrutinib has now been approved for treatment of chronic lymphocytic leukemia (CLL) in both front-line setting and as later-line treatment. However, knowledge about the outcomes and adverse events (AE) among patients at a population-based level is still limited. OBJECTIVES: To report outcomes and AEs in a population-based cohort treated with ibrutinib outside clinical trials. METHODS: We conducted a multicenter, retrospective cohort study including all patients with CLL treated with ibrutinib. RESULTS: In total, 205 patients were included of whom 39 (19%) were treatment-naïve. The median follow-up was 21.4 months (interquartile range (IQR), 11.9,32.8), the estimated overall survival at 12 months was 88.8% (95% confidence interval (CI); 84.3%, 93.3%), and the estimated progression-free survival at 12 months was 86.3% (95% CI; 81.3%, 91.2%). During follow-up, 200 (97.6%) patients had at least one AE and 100 (48.8%) patients had at least one grade ≥3 AE. Eighty-six patients (42.0%) discontinued ibrutinib, hereof 47 (54.7%) due to AEs and 19 (22.1%) had progression of CLL or Richter transformation. CONCLUSIONS: In our study, we find comparable, though slightly inferior, overall, and progression-free survival, and discontinuation due to toxicity was higher compared with clinical trials. Patient training and information may improve treatment adherence outside clinical trials.

The Parental Stress Scale revisited: Rasch-based construct validity for Danish parents of children 2-18 years old with and without behavioral problems.

BACKGROUND: Experiencing parental stress is common among parents of children of all ages and is elevated in families characterized by stressors such as poverty, mental health problems, and developmental problems. The Parental Stress Scale (PSS) is a short measure for the assessment of perceived stress resulting from being a parent. METHODS: This study examines the construct validity and psychometric properties of the Danish PSS using Rasch and graphical loglinear Rasch models in a sample of parents of 2-18-year-old children with and without known behavior problems. We emphasized analyses of differential item functioning, to ascertain whether the scale yields unbiased scores for subgroups of parents. RESULTS: The 18-item PSS did not fit the Rasch model or a graphical loglinear Rasch model. After dichotomizing item responses and eliminating items 2 and 11, we found the PSS to consist of two distinct subscales measuring parental stress and lack of parental satisfaction. For the total sample, the Parental Stress subscale fit a very complex graphical loglinear Rasch model with differential item functioning relative to parental education and whether children had behavior problems or not. The Lack of Parental Satisfaction subscale fit a simple graphical loglinear Rasch model with differential item functioning only relative to subsample. When dividing into subsamples of parents of children with and without behavior problems, the Parental Stress subscale fit a simple graphical loglinear Rasch model, though still with differential item functioning, while the Lack of Parental Satisfaction subscale fit the Rasch model in each subsample of parents. Both subscales performed best for parents of children with behavior problems. CONCLUSIONS: The PSS should be used in a 16-item version and scored as two subscales. The PSS appears better suited for use among parents of children with behavior problems than within a sample without any known difficulties.

Measuring stress in Australia: validation of the perceived stress scale (PSS-14) in a national sample.

BACKGROUND: In Australia, the stress levels have increased over the years, impacting on the physical and mental health of the general population. The aim of the present study was to evaluate the validity and reliability of the PSS-14 in an Australian population. METHODS: The PSS-14 was applied to a large national sample comprising 3857 Australians in the population-based cross-sectional study Australia's National Survey of Adult Oral Health 2004-2006. The psychometric properties analyzed with the Rasch model and Graphical Log-linear Rasch models were: model fit, item fit, local dependence, differential item functioning, unidimensionality, reliability, targeting and criterion validity. RESULTS: The PSS-14 did not fit the pure RM (χ2 (55) = 3828.3, p = < 0.001) and the unidimensionality of the whole scale was rejected (p = < 0.001). The Perceived Stress (χ2 (27) = 1409.7, p = < 0.001) and Perceived Control (χ2 (27) = 713.4, p = < 0.001) subscales did not fit the pure RM. After the deletion of two items, the Perceived Stress subscale (χ2 (96) = 94.4, p = 0.440) fitted a GLLRM, while the Perceived Control scale (χ2 (55) = 62.50, p = 0.224) fitted a GLLRM after the exclusion of four misfitting items. CONCLUSIONS: The Perceived Stress subscale displayed adequate psychometric properties after the deletion of two items; however, the majority of problems centered around the Perceived Control subscale. The presence of differential item functioning among four items indicates that adjustment of total scores is required to avoid measurement bias. Recommendations for future applications in Australia are provided.

Assessment of cognitive ability at conscription for the Danish army: Is a single total score sufficient?

Børge Priens Prøve (BPP) was developed for the Danish Army by psychologist Børge Prien in the 1950s, as a test of general cognitive ability for use at conscription for the Danish armed forces. The final BPP (1957) had four subtests; a Raven-like matrix subtest, and three subtests measuring verbal, numerical, and visuospatial ability. The BPP is a speeded test counting the number of correct responses within 45 minutes. Thus, we consider the BPP as a measure of "cognitive efficiency" rather than a pure measure of cognitive ability. The BPP is still in use.Using techniques available in 1960, Rasch concluded that the matrices and numerical tests appeared to satisfy the requirements of the Rasch (Probabilistic models for some intelligence and attainment tests, Danish Institute for Educational Research, Copenhagen; 1960) model, while the verbal and visuospatial tests did not. Since then, there have been, to our knowledge, no published studies of the psychometric scaling properties of the BPP, partly because the practice of the Danish draft board has been to record only the total score. We examine these properties by analysis of data from two cohorts (n = 9,491), using the Leunbach (A probabilistic measurement model for assessing whether two tests measure the same personal factor. The Danish Institute of Educational, Copenhagen, Denmark; 1976) model to assess whether the sum of the four subtests provides a statistically sufficient measure of a common latent trait. Since we found only weak evidence against fit to the Leunbach model, we claim that this warrants the use of a summarized total BPP score. We examined whether BPP subscales suffered from differential test functioning (DTF) relative to samples. Weak, and for practical purposes too weak, DTF was suggested for one subscale.

Structural outline of the detailed mechanism for elongation factor Ts-mediated guanine nucleotide exchange on elongation factor Tu.

Translation elongation factor EF-Tu belongs to the superfamily of guanine-nucleotide binding proteins, which play key cellular roles as regulatory switches. All G-proteins require activation via exchange of GDP for GTP to carry out their respective tasks. Often, guanine-nucleotide exchange factors are essential to this process. During translation, EF-Tu:GTP transports aminoacylated tRNA to the ribosome. GTP is hydrolyzed during this process, and subsequent reactivation of EF-Tu is catalyzed by EF-Ts. The reaction path of guanine-nucleotide exchange is structurally poorly defined for EF-Tu and EF-Ts. We have determined the crystal structures of the following reaction intermediates: two structures of EF-Tu:GDP:EF-Ts (2.2 and 1.8Å resolution), EF-Tu:PO4:EF-Ts (1.9Å resolution), EF-Tu:GDPNP:EF-Ts (2.2Å resolution) and EF-Tu:GDPNP:pulvomycin:Mg(2+):EF-Ts (3.5Å resolution). These structures provide snapshots throughout the entire exchange reaction and suggest a mechanism for the release of EF-Tu in its GTP conformation. An inferred sequence of events during the exchange reaction is presented.

Protein interaction screening for the ankyrin repeats and suppressor of cytokine signaling (SOCS) box (ASB) family identify Asb11 as a novel endoplasmic reticulum resident ubiquitin ligase.

The ankyrin and SOCS (suppressor of cytokine signaling) box (ASB) family of proteins function as the substrate recognition subunit in a subset of Elongin-Cullin-SOCS (ECS) E3 ubiquitin ligases. Despite counting 18 members in humans, the identity of the physiological targets of the Asb proteins remains largely unexplored. To increase our understanding of the function of ASB proteins, we conducted a family-wide SILAC (stable isotope labeling by amino acids in cell culture)-based protein/protein interaction analysis. This investigation led to the identification of novel as well as known ASB-associated proteins like Cullin 5 and Elongins B/C. We observed that several proteins can be bound by more than one Asb protein. The additional exploration of this phenomenon demonstrated that ASB-Cullin 5 complexes can oligomerize and provides evidence that Cullin 5 forms heterodimeric complexes with the Cullin 4a-DDB1 complex. We also demonstrated that ASB11 is a novel endoplasmic reticulum-associated ubiquitin ligase with the ability to interact and promote the ubiquitination of Ribophorin 1, an integral protein of the oligosaccharyltransferase (OST) glycosylation complex. Moreover, expression of ASB11 can increase Ribophorin 1 protein turnover in vivo. In summary, we provide a comprehensive protein/protein interaction data resource that can aid the biological and functional characterization of ASB ubiquitin ligases.

Direct binding between BubR1 and B56-PP2A phosphatase complexes regulate mitotic progression.

BubR1 is a central component of the spindle assembly checkpoint that inhibits progression into anaphase in response to improper kinetochore-microtubule interactions. In addition, BubR1 also helps stabilize kinetochore-microtubule interactions by counteracting the Aurora B kinase but the mechanism behind this is not clear. Here we show that BubR1 directly binds to the B56 family of protein phosphatase 2A (PP2A) regulatory subunits through a conserved motif that is phosphorylated by cyclin-dependent kinase 1 (Cdk1) and polo-like kinase 1 (Plk1). Two highly conserved hydrophobic residues surrounding the serine 670 Cdk1 phosphorylation site are required for B56 binding. Mutation of these residues prevents the establishment of a proper metaphase plate and delays cells in mitosis. Furthermore, we show that phosphorylation of serines 670 and 676 stimulates the binding of B56 to BubR1 and that BubR1 targets a pool of B56 to kinetochores. Our data suggest that BubR1 counteracts Aurora B kinase activity at improperly attached kinetochores by recruiting B56-PP2A phosphatase complexes.

Post-myocardial infarction anxiety or depressive symptoms and risk of new cardiovascular events or death: a population-based longitudinal study.

OBJECTIVE: To examine the association between anxiety symptoms 3 months after myocardial infarction (MI) and/or new cardiovascular events and death, taking into account established risk factors, and to compare the results with those of the impact of depressive symptoms. Post-MI anxiety symptoms have been associated with a composite outcome of new cardiovascular events or death, but previous studies have not fully adjusted for potential confounders. It remains unclear whether anxiety symptoms are independently associated with both new cardiovascular events and death. METHODS: A population-based cohort study of 896 persons (70% of eligible) with first-time MI between 1 January 2009 and 31 December 2009, completing the Hospital Anxiety and Depression Scale, were followed up until 31 July 2012. RESULTS: A total of 239 new cardiovascular events and 94 deaths occurred during 1975 person-years of follow-up. Cox proportional hazards models showed that anxiety symptoms were associated with both new cardiovascular events and death in analysis adjusted for age only. The estimates decreased when adjusted for dyspnea score, physical activity, and depressive symptoms, and anxiety symptoms were no longer associated with new cardiovascular events (hazard ratio [HR] = 1.02, 95% confidence interval [CI] = 0.98-1.07) or with death (HR = 0.94, 95% CI = 0.88-1.01). In fully adjusted models, depressive symptoms remained associated with death (HR = 1.13, 95% CI = 1.05-1.21), but not with new cardiovascular events (HR = 1.02, 95% CI = 0.99-1.06). CONCLUSIONS: Post-MI anxiety symptoms were not an independent prognostic risk factor for new cardiovascular events or for death, whereas depressive symptoms were associated with an increased risk of mortality.

In silico prediction of Gallibacterium anatis pan-immunogens.

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.