Hormone contraceptive use in young women: Altered mood states, neuroendocrine and inflammatory biomarkers.

Oral contraceptives are used by millions of women worldwide, yet there are questions regarding the psychological and biological consequences of these medications. Considering that sex steroid hormones can regulate neuroendocrine and behavioral responses to stress, the current study examined mood and stress symptomatologies, as well as circulating levels of cortisol and inflammatory biomarkers among young women (N = 388), of whom, 47.0 % (n = 182) were using a form of hormonal contraception. Women using hormone contraceptives displayed significantly higher depressive and stress scores compared to non-users, whereas no differences were found for anxiety symptoms. Moreover, contraceptive users had markedly elevated plasma cortisol and C-reactive protein levels in comparison to non-users. Upon assessing women at different phases of their menstrual cycle, hormone contraceptive users displayed higher levels of cortisol compared to women in the follicular and luteal phases, in addition to higher levels of CRP levels compared to women in the luteal phase. Together, these findings suggest that hormone contraceptive use is linked to exaggerated basal neuroendocrine and inflammatory profiles, which could potentially increase sensitivity to the impacts of stressors and mood disturbances.

Modeling the transcriptional regulatory network that controls the early hypoxic response in Candida albicans.

We determined the changes in transcriptional profiles that occur in the first hour following the transfer of Candida albicans to hypoxic growth conditions. The impressive speed of this response is not compatible with current models of fungal adaptation to hypoxia that depend on the depletion of sterol and heme. Functional analysis using Gene Set Enrichment Analysis (GSEA) identified the Sit4 phosphatase, Ccr4 mRNA deacetylase, and Sko1 transcription factor (TF) as potential regulators of the early hypoxic response. Cells mutated in these and other regulators exhibit a delay in their transcriptional responses to hypoxia. Promoter occupancy data for 29 TFs were combined with the transcriptional profiles of 3,111 in vivo target genes in a Network Component Analysis (NCA) to produce a model of the dynamic and highly interconnected TF network that controls this process. With data from the TF network obtained from a variety of sources, we generated an edge and node model that was capable of separating many of the hypoxia-upregulated and -downregulated genes. Upregulated genes are centered on Tye7, Upc2, and Mrr1, which are associated with many of the gene promoters that exhibit the strongest activations. The connectivity of the model illustrates the high redundancy of this response system and the challenges that lie in determining the individual contributions of specific TFs. Finally, treating cells with an inhibitor of the oxidative phosphorylation chain mimics most of the early hypoxic profile, which suggests that this response may be initiated by a drop in ATP production.

The functions of Mediator in Candida albicans support a role in shaping species-specific gene expression.

The Mediator complex is an essential co-regulator of RNA polymerase II that is conserved throughout eukaryotes. Here we present the first study of Mediator in the pathogenic fungus Candida albicans. We focused on the Middle domain subunit Med31, the Head domain subunit Med20, and Srb9/Med13 from the Kinase domain. The C. albicans Mediator shares some roles with model yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe, such as functions in the response to certain stresses and the role of Med31 in the expression of genes regulated by the activator Ace2. The C. albicans Mediator also has additional roles in the transcription of genes associated with virulence, for example genes related to morphogenesis and gene families enriched in pathogens, such as the ALS adhesins. Consistently, Med31, Med20, and Srb9/Med13 contribute to key virulence attributes of C. albicans, filamentation, and biofilm formation; and ALS1 is a biologically relevant target of Med31 for development of biofilms. Furthermore, Med31 affects virulence of C. albicans in the worm infection model. We present evidence that the roles of Med31 and Srb9/Med13 in the expression of the genes encoding cell wall adhesins are different between S. cerevisiae and C. albicans: they are repressors of the FLO genes in S. cerevisiae and are activators of the ALS genes in C. albicans. This suggests that Mediator subunits regulate adhesion in a distinct manner between these two distantly related fungal species.

Cell wall integrity is linked to mitochondria and phospholipid homeostasis in Candida albicans through the activity of the post-transcriptional regulator Ccr4-Pop2.

The cell wall is essential for viability of fungi and is an effective drug target in pathogens such as Candida albicans. The contribution of post-transcriptional gene regulators to cell wall integrity in C. albicans is unknown. We show that the C. albicans Ccr4-Pop2 mRNA deadenylase, a regulator of mRNA stability and translation, is required for cell wall integrity. The ccr4/pop2 mutants display reduced wall ß-glucans and sensitivity to the echinocandin caspofungin. Moreover, the deadenylase mutants are compromised for filamentation and virulence. We demonstrate that defective cell walls in the ccr4/pop2 mutants are linked to dysfunctional mitochondria and phospholipid imbalance. To further understand mitochondrial function in cell wall integrity, we screened a Saccharomyces cerevisiae collection of mitochondrial mutants. We identify several mitochondrial proteins required for caspofungin tolerance and find a connection between mitochondrial phospholipid homeostasis and caspofungin sensitivity. We focus on the mitochondrial outer membrane SAM complex subunit Sam37, demonstrating that it is required for both trafficking of phospholipids between the ER and mitochondria and cell wall integrity. Moreover, in C. albicans also Sam37 is essential for caspofungin tolerance. Our study provides the basis for an integrative view of mitochondrial function in fungal cell wall biogenesis and resistance to echinocandin antifungal drugs.

Developing symptom clusters: linking inflammatory biomarkers to depressive symptom profiles.

Considering the burden of depression and the lack of efficacy of available treatments, there is a need for biomarkers to predict tailored or personalized treatments. However, identifying reliable biomarkers for depression has been challenging, likely owing to the vast symptom heterogeneity and high rates of comorbidity that exists. Examining biomarkers that map onto dimensions of depression as well as shared symptoms/constructs that cut across disorders could be most effective for informing personalized treatment approaches. With a sample of 539 young adults, we conducted a principal component analysis (PCA) followed by hierarchical cluster analysis to develop transdiagnostic clusters of depression and anxiety symptoms. We collected blood to assess whether neuroendocrine (cortisol) and inflammatory profiles (C-reactive protein (CRP), Interleukin (IL)-6, and tumor necrosis factor (TNF) - α) could be used to differentiate symptom clusters. Six distinct clusters were identified that differed significantly on symptom dimensions including somatic anxiety, general anxiety, anhedonia, and neurovegetative depression. Moreover, the neurovegetative depression cluster displayed significantly elevated CRP levels compared to other clusters. In fact, inflammation was not strongly associated with overall depression scores or severity, but rather related to specific features of depression marked by eating, appetite, and tiredness. This study emphasizes the importance of characterizing the biological underpinnings of symptom dimensions and subtypes to better understand the etiology of complex mental health disorders such as depression.

A Qualitative Study to Inform a More Acceptable Pulmonary Rehabilitation Program after Acute Exacerbation of Chronic Obstructive Pulmonary Disease.

Rationale: Current international guidelines for prevention of acute exacerbation of chronic obstructive pulmonary disease (AECOPD) recommend enrolment and participation in a pulmonary rehabilitation (PR) program within 4 weeks of hospital discharge. However, there is poor uptake of these programs as well as low adherence and completion rates.Objectives: The objectives of this study were to explore the views of patients and healthcare professionals (HCPs) on PR after AECOPD and how participation could be enhanced.Methods: A qualitative study was undertaken, and data were analyzed using deductive thematic analysis. A total of 13 patients who had experienced an AECOPD in the previous 6 months and 11 HCPs experienced in the management of COPD participated in face-to-face, semistructured interviews. Patients and HCPs were recruited from both rehabilitation and acute hospital settings.Results: Four main themes were identified: 1) uncertainty about timing of PR-most HCPs endorsed the professional guidelines that advocate for PR programs to begin within 4 weeks of an AECOPD; patients, however, varied drastically in their view of the ideal timing to start a PR program, anywhere from before an exacerbation (perhaps preventing one) and up to 6-8 weeks after exacerbation; 2) tailored and flexible manner to deliver PR programs with a gradual start-patients and HCPs talked about individually tailored programs with a gradual introduction of exercise and teaching sessions; some HCPs advocated allowing patients to pick and choose which elements would help them the most; 3) education for all-patients would like HCPs to be more informed and informative about the PR programs available in their neighborhoods, and HCPs focused on how they could educate patients about their disease and how better to manage it; and 4) logistical, disease-related, and psychological barriers-barriers to PR were discussed by both HCPs and patients; these fell into two categories: a) delivery issues (i.e., transportation and location of PR); and b) patient-specific issues (too sick or too well, high levels of anxiety).Conclusions: Our findings lay the foundation for the development of a flexible, stepped-care approach to delivering PR after AECOPD, which should be tailored according to the needs and preferences of the individual.

Rate of, and barriers and enablers to, pulmonary rehabilitation referral in COPD: A systematic scoping review.

BACKGROUND: Despite pulmonary rehabilitation (PR) being an evidence-based treatment recommended for people with chronic obstructive pulmonary disease (COPD), referral to PR seems to be low. Collating the evidence on referral rates and perceived barriers and enablers may help provide knowledge needed to increase referral. OBJECTIVES: The purpose of this scoping review was to determine 1) the rate of referral of patients with COPD to PR by healthcare professionals (HCPs) and 2) the barriers and enablers to referral perceived by HCPs. METHODS: The JBI scoping review manual was used. Ovid Medline, CINAHL, EMBASE, and ProQuest Dissertations and Theses were searched from inception until July 28, 2017. 245 non-duplicate articles were screened. Included articles reported referral rates and/or HCP perceived barriers and/or enablers to PR referral. RESULTS: Forty-two studies were included. Twenty-eight observational studies reported referral rates ranging from 0% to 85%. Seven studies looking at the effects of different interventions to improve COPD care reported a range of 2%-56% referral pre-intervention, and 8%-71% post-intervention. Nine studies reported HCP perceived barriers to referral. The two most common barriers were low knowledge of what PR is and its benefits and low knowledge of the referral process. Six studies reported HCP perceived enablers to referral. The most common enabler was training/experience in PR. CONCLUSION: The rate of referral to PR is suboptimal, but there are commonly reported barriers and enablers that may help with the creation of actionable changes. HCPs need more knowledge of PR and the benefits it provides.

Outcomes in randomized controlled trials of exercise interventions in solid organ transplant.

AIM: To identify the outcome measures that have been used in randomized controlled trials (RCTs) of exercise training in solid organ transplant (SOT) recipients and to link these outcomes to the International Classification of Functioning, Disability and Health (ICF) framework. METHODS: Electronic literature searches of MEDLINE, EMBASE, CINAHL, Cochrane, Scopus, and Web of Science were performed. We sought RCTs that investigated the effect of exercise training in SOT recipients. Reference lists of all eligible publications were searched for other appropriate studies not identified by the electronic search. A complete list of outcome measures used in the RCTs was generated and each of these was linked to an ICF category. RESULTS: Four hundred and thirteen articles were retrieved, of which 35 met our inclusion criteria. The studies included were designed to compare the effects of exercise training programs to usual care or to another exercise training program and reported on recipients of heart (n = 21), kidney (n = 9), lung (n = 3) or liver (n = 2) transplant. Of the 126 outcome measures identified, 62 were used as primary outcome measures. The most commonly occurring primary outcomes were aerobic capacity using the peak VO2 (n = 13), quality of life using the short-form-36 (n = 8), and muscle strength (n = 7). These outcome measures were linked to 113 ICF categories and the majority of outcomes fall into the body function domain (n = 93). CONCLUSION: There is little standardization in outcome measures used in RCTs of exercise interventions in SOT recipients. The ICF framework can be used to select a core set of outcomes that cross all domains of ICF and that would be appropriate to all SOT recipients.

Hypoglycemic and insulin-sensitizing effects of berberine in high-fat diet- and streptozotocin-induced diabetic rats.

Hypoglycemic effects of berberine (BBR) have been reported in several studies in cell and animal models. However, the mechanisms of action are not fully understood. The present study was therefore aimed at determining the effect and underlying mechanisms of action of BBR on diabetes in a high-fat diet- and streptozotocin-induced diabetic rat model. Ninety male Sprague-Dawley rats, 150 to 170 g, were housed individually in cages. Two groups (n = 12 each) were fed the AIN-93G diet (normal control) and the same diet modified to contain 33% fat and 2% cholesterol (high-fat control), respectively. The third group (n = 66) was fed the high-fat diet and injected intraperitoneally 2 weeks later with 35 mg/kg body weight of streptozotocin in citrate buffer (pH 4.5). The rats in both control groups were injected with the vehicle. After 12 days, rats with semifasting (5 hours) blood glucose levels between 14 and 25 mmol/L were divided into 4 groups (n = 12 each) and treated with 0 (diabetic control), 50, 100, and 150 mg/kg/d of BBR for 6 weeks while continuing on the high-fat diet. Hypoglycemic effects of BBR were consistently demonstrated by semifasting and fasting blood glucose levels, and insulin-sensitizing effects were seen during oral glucose tolerance testing. Berberine also reduced food intake while having no effect on body weight in diabetic rats. No effect of BBR was observed on plasma levels of insulin, adipokines (leptin and adiponectin), or inflammatory cytokines (tumor necrosis factor-α and C-reactive protein). Berberine did not affect the state of oxidative stress as assessed by the activity of superoxide dismutase and the concentrations of malondialdehyde and reduced and oxidized glutathione in the liver. These findings demonstrated the hypoglycemic and insulin-sensitizing capabilities of BBR, with the underlying mechanisms awaiting further investigation.

Berberine and plant stanols synergistically inhibit cholesterol absorption in hamsters.

The present study was conducted to determine the efficacy and underlying mechanism of berberine (BBR), plant stanols (PS) and their combination on plasma lipids. Male Golden Syrian hamsters were randomly divided into 4 groups (n=15/group) and fed a cornstarch-casein-sucrose-based diet containing 0.15% cholesterol and 5% fat. Three treatment groups were supplemented with 0.17% (equivalent to 100mgkg(-1)d(-1)) BBR, 1% PS, or a combination of both (BBRPS) for 4wk. At the end of the study, plasma lipids were analyzed with enzymatic methods, cholesterol absorption and synthesis using stable isotope tracer methodology, and gene and protein expressions in the liver and small intestine using real-time PCR and Western blot, respectively. BBR and PS significantly lowered plasma total- and nonHDL-cholesterol levels, and BBRPS markedly improved cholesterol-lowering efficacy compared to BBR or PS alone. Further examinations revealed that BBR and PS both inhibited cholesterol absorption and by contrast, increased cholesterol synthesis, and exerted a synergistic action when they were combined. Plasma total or nonHDL-cholesterol levels were significantly correlated with cholesterol absorption rates. BBR upregulated sterol 27-hydroxlase gene expression and BBRPS increased both cholesterol-7alpha-hydroxylase and sterol 27-hydroxlase gene expressions. BBR and PS also synergistically decreased plasma triacylglycerides. These findings suggest that the cholesterol-lowering action of BBR might involve a combination of inhibition of cholesterol absorption and stimulation of bile acid synthesis. The combination of BBR and PS improves cholesterol-lowering efficacy through a synergistic action on cholesterol absorption, in addition to synergistically reducing plasma triacylglycerols in hamsters.

Interferon-gamma regulation of intestinal epithelial permeability.

The maintenance and regulation of the barrier function of the epithelial lining of the intestine are important homeostatic events, serving to allow selective absorption from the gut lumen while simultaneously limiting the access of bacteria into the mucosa. Interferon-gamma is a pleiotrophic cytokine produced predominantly by natural kill cells and CD4+ T cells that under normal circumstances, and particularly during infection or inflammation, will be a component of the intestinal milieu. Use of colon-derived epithelial cell lines and, to a less extent, murine in vivo analyses, have revealed that interferon-gamma (IFN-gamma) can increase epithelial permeability as gauged by markers of paracellular permeability and bacterial transcytosis, with at least a portion of the bacteria using the transcellular permeation pathway. In this review, we describe the main characteristics of epithelial permeability and then focus on the ability of IFN-gamma to increase epithelial permeability, and the mechanism(s) thereof.

Topology-dependent transcription in linear and circular plasmids of the segmented genome of Borrelia burgdorferi.

The segmented genome of Borrelia burgdorferi, a causative agent of Lyme disease, contains a mixture of over 20 linear and circular plasmids. Genes encoding five paralogous families of plasmid replication proteins are located on both circular and linear DNA molecules. The effect of DNA topology on the transcription of replication proteins from two B. burgdorferi plasmids, cp9 and lp17, was examined using quantitative reverse transcription polymerase chain reaction. Circular to linear conversion of a cp9-derived plasmid resulted in a 160-fold decrease in transcript levels of bbc01, believed to encode the replication initiator. A 14.9-fold reduction in plasmid copy number was also observed, resulting in a net 10.7-fold lower transcription level per gene copy on a linear versus a circular plasmid. In contrast, expression of the bbd14 replication initiator for the linear plasmid lp17 was 7.2 times higher per gene copy on a linear versus a circular plasmid. Topology-dependent transcription of these genes may help to block topological interconversions during genome evolution, offers a new avenue for global gene regulation and also has important implications for the design of genetic complementation experiments in B. burgdorferi.

Mapping of essential replication functions of the linear plasmid lp17 of B. burgdorferi by targeted deletion walking.

The genome of the Lyme disease pathogen Borrelia burgdorferi strain B31 MI includes one linear chromosome, 10 circular and 12 linear plasmids. Members of four paralogous gene families, revealed by genome sequencing, have been suggested as replication/partition functions for both the linear and circular plasmids. Some of these genes have been experimentally shown to be essential for the replication of the B. burgdorferi replicons that encode them. In this study, we located the region essential for replication of lp17, the second smallest linear plasmid in B. burgdorferi. We used a novel in vivo method, targeted deletion walking, to systematically delete DNA from either the left or right end of lp17. We report that the region essential for replication of lp17 is 1.8 kb (bp 7946-9766) and contains only one intact open reading frame (BBD14). Expression of BBD14 is required for the replication, suggesting that it is the replication initiator for lp17. The BBD14 protein is a member of paralogous family (PF) 62 and we present the first experimental evidence for the role of a PF 62 member. Adjacent non-coding sequences are also required, suggesting that the origin lies at least partially outside the coding region. Surprisingly, deletion of BBD21, the ParA orthologue (PF 32), had little effect upon plasmid stability or incompatibility. Finally, data are presented suggesting that lp17 replication occurs preferentially on a linear rather than a circular DNA molecule.