MicroRNA-profiling of miR-371~373- and miR-302/367-clusters in serum and cerebrospinal fluid identify patients with intracranial germ cell tumors.

PURPOSE: Intracranial germ cell tumors (iGCT) comprise germinoma and non-germinoma. Their diagnosis predominantly relies on biopsy as only one-fifth of patients present with elevated biomarkers (AFP/ß-HCG) in serum or cerebrospinal fluid (CSF). MicroRNAs (miR/miRNA) have emerged as non-invasive biomarkers in extracranial GCT and may potentially facilitate non-invasive diagnosis in iGCT. METHODS: We analyzed eight miRNAs in serum and CSF from the miR-371~373- and miR-302/367-clusters and four miRNAs differentially expressed in iGCT tissue (miR-142-5p/miR-146a-5p/miR-335-5p/miR-654-3p) from eight iGCT patients (age 10-33 years) and 12 control subjects by pre-amplified RT-qPCR. MiR-30b-5p (serum) and miR-204-5p (CSF) acted as reference genes. ΔCt-values were expressed as [Formula: see text] after standardization against controls. RESULTS: Between iGCT and control patients' serum ΔCt-values of miR-371a-3p (p = 0.0159), miR-372-3p (p= 0.0095, miR-367 (p = 0.0190), miR-302a (p = 0.0381) and miR-302d-3p (p = 0.0159) differed significantly. Discriminatory pattern in CSF was similar to serum as miR-371a (p = 0.0286), miR-372-3p (p = 0.0028), miR-367-3p (p = 0.0167) and miR-302d-3p (p = 0.0061) distinguished between patients and controls. Abundant [Formula: see text] levels of each of these miRNAs were found across all serum and CSF samples including biomarker-negative patients. CONCLUSION: With the largest data set so far, we underline the suitability of miR-371a, miR-372, miR-367 and miR-302d in serum and CSF for diagnosis of iGCT, particularly in biomarker-negative germinoma. Diagnosis of iGCT by miRNA analysis is a feasible and valid approach, particularly as serum can be readily obtained by a less invasive procedure. MiRNA analysis may discriminate iGCT from other tumors with similar radiological findings and may allow to monitor response to therapy as well as early relapse during follow-up.

Nonsense-mediated mRNA decay does not restrict influenza A virus propagation.

Nonsense-mediated mRNA decay (NMD) was identified as a process to degrade flawed cellular messenger RNA (mRNA). Within the last decades it was also shown that NMD carries virus-restricting capacities and thus could be considered a part of the cellular antiviral system. As this was shown to affect primarily positive-sense single stranded RNA ((+)ssRNA) viruses there is only scarce knowledge if this also applies to negative-sense single stranded RNA ((-)ssRNA) viruses. Influenza A viruses (IAVs) harbour a segmented (-)ssRNA genome. During their replication IAVs produce numerous RNA transcripts and simultaneously impair cellular transcription and translation. The viral mRNAs hold several molecular patterns which can elicit NMD and in turn would lead to their degradation. This, in consequence, may mitigate viral propagation. Thus, we examined if a knockdown or a pharmacological inhibition of NMD key components may influence IAV replication. Additionally, we performed similar experiments with respiratory syncytial virus (RSV), another (-)ssRNA virus, but with a non-segmented genome. Although it seemed that a knockdown of up-frameshift protein 1 (UPF1), the central NMD factor, slightly increased viral mRNA and protein levels, no significant alteration of viral replication could be observed, implying that the NMD machinery may not have restricting capacities against (-)ssRNA viruses.

Creating a Queer Ontology: The Gender, Sex, and Sexual Orientation (GSSO) Ontology.

Cultural attitudes, linguistic variation, and historical pathology have led to a plethora of terms concerning gender, sex, and sexual orientation that have caused confusion and uneasiness among both lay people and relevant professionals. For members of the LGBTQIA+ community, these negative reactions are compounded by identities which have historically and contemporarily been mistreated by medical professionals. In an effort to provide a reliable resource for patients and clinicians, we have created the Gender, Sex, and Sexual Orientation (GSSO) ontology, which currently includes over 4,000 entities from multiple disciplines. The GSSO is a manually curated resource utilizing related glossaries from biology, medicine, psychology, sociology, and gender studies. With links to over 20 other ontology resources such as SNOMED-CT and MedDRA, the GSSO aims for accessibility and interoperability with existing systems. It is also open-source and features an easy-to-use web interface (https://github.com/Superraptor/GSSO). Future work involves multiple language support efforts and empirical evaluation.

Late activation of the Raf/MEK/ERK pathway is required for translocation of the respiratory syncytial virus F protein to the plasma membrane and efficient viral replication.

Activation of the Raf/MEK/ERK cascade is required for efficient propagation of several RNA and DNA viruses, including human respiratory syncytial virus (RSV). In RSV infection, activation of the Raf/MEK/ERK cascade is biphasic. An early induction within minutes after infection is associated with viral attachment. Subsequently, a second activation occurs with, so far, unknown function in the viral life cycle. In this study, we aimed to characterise the role of Raf/MEK/ERK-mediated signalling during ongoing RSV infection. Our data show that inhibition of the kinase MEK after the virus has been internalised results in a reduction of viral titers. Further functional investigations revealed that the late-stage activation of ERK is required for a specific step in RSV replication, namely, the secretory transport of the RSV fusion protein F. Thus, MEK inhibition resulted in impaired surface accumulation of the F protein. F protein surface expression is essential for efficient replication as it is involved in viral filament formation, cell fusion, and viral transmission. In summary, we provide detailed insights of how host cell signalling interferes with RSV replication and identified the Raf/MEK/ERK kinase cascade as potential target for novel anti-RSV strategies.

Hepatitis C Virus Exploits Death Receptor 6-mediated Signaling Pathway to Facilitate Viral Propagation.

The life cycle of hepatitis C virus (HCV) is highly dependent on host proteins for virus propagation. By transcriptome sequencing analysis, we identified host genes that were highly differentially expressed in HCV-infected cells. Of these candidates, we selected Death receptor 6 (DR6) for further characterization. DR6 is an orphan member of the tumor necrosis factor receptor superfamily. In the present study, we demonstrated that both mRNA and protein levels of DR6 were increased in the context of HCV replication. We further showed that promoter activity of DR6 was increased by HCV infection. By employing promoter-linked reporter assay, we showed that HCV upregulated DR6 via ROS-mediated NF-κB pathway. Both mRNA and protein levels of DR6 were increased by NS4B or NS5A. However, NS5A but not NS4B specifically interacted with DR6. We showed that HCV modulated JNK, p38 MAPK, STAT3, and Akt signaling pathways in a DR6-dependent manner. Interestingly, Akt signaling cascade was regulated by protein interplay between DR6 and NS5A. Silencing of DR6 expression resulted in decrease of infectious HCV production without affecting viral entry, replication, and translation. Together, these data indicate that HCV modulates DR6 signaling pathway for viral propagation and may contribute to HCV-mediated pathogenesis.

Ankyrin Repeat Domain 1 is Up-regulated During Hepatitis C Virus Infection and Regulates Hepatitis C Virus Entry.

Hepatitis C virus (HCV) is highly dependent on host proteins for its own propagation. By transcriptome sequencing (RNA-Seq) analysis, we identified 30 host genes that were significantly differentially expressed in cell culture-grown HCV (HCVcc)-infected cells. Of these candidate genes, we selected and characterized ankyrin repeat domain 1 (ANKRD1). Here, we showed that protein expression of ANKRD1 was up-regulated in HCVcc-infected cells. We further showed that protein expression level of ANKRD1 was increased by nonstructural 5A (NS5A) protein. ANKRD1 specifically interacted with NS5A both in vitro and coimmunoprecipitation assays. Protein interaction was mediated through the domain II of NS5A and the C-terminal region of ANKRD1. Promoter activity of ANKRD1 was also increased by NS5A protein. Moreover, up-regulation of ANKRD1 expression was mediated through alteration in intracellular calcium homeostasis and ER stress in HCVcc-infected cells. We showed that silencing of ANKRD1 impaired HCV propagation without affecting HCV replication. By using HCV-like infectious particle (HCV-LP), we demonstrated that HCV single-cycle infection was drastically impaired in ANKRD1 knockdown cells. Finally, we verified that ANKRD1 was required for HCV entry. These data suggest that HCV coopts ANKRD1 for its own propagation and up-regulation of ANKRD1 may contribute to HCV-mediated liver pathogenesis.

Nonstructural 5A Protein of Hepatitis C Virus Regulates Soluble Resistance-Related Calcium-Binding Protein Activity for Viral Propagation.

UNLABELLED: Hepatitis C virus (HCV) is a major cause of chronic liver disease and is highly dependent on cellular proteins for virus propagation. To identify the cellular factors involved in HCV propagation, we recently performed protein microarray assays using the HCV nonstructural 5A (NS5A) protein as a probe. Of 90 cellular protein candidates, we selected the soluble resistance-related calcium-binding protein (sorcin) for further characterization. Sorcin is a calcium-binding protein and is highly expressed in certain cancer cells. We verified that NS5A interacted with sorcin through domain I of NS5A, and phosphorylation of the threonine residue 155 of sorcin played a crucial role in protein interaction. Small interfering RNA (siRNA)-mediated knockdown of sorcin impaired HCV propagation. Silencing of sorcin expression resulted in a decrease of HCV assembly without affecting HCV RNA and protein levels. We further demonstrated that polo-like kinase 1 (PLK1)-mediated phosphorylation of sorcin was increased by NS5A. We showed that both phosphorylation and calcium-binding activity of sorcin were required for HCV propagation. These data indicate that HCV modulates sorcin activity via NS5A protein for its own propagation. IMPORTANCE: Sorcin is a calcium-binding protein and regulates intracellular calcium homeostasis. HCV NS5A interacts with sorcin, and phosphorylation of sorcin is required for protein interaction. Gene silencing of sorcin impaired HCV propagation at the assembly step of the HCV life cycle. Sorcin is phosphorylated by PLK1 via protein interaction. We showed that sorcin interacted with both NS5A and PLK1, and PLK1-mediated phosphorylation of sorcin was increased by NS5A. Moreover, calcium-binding activity of sorcin played a crucial role in HCV propagation. These data provide evidence that HCV regulates host calcium metabolism for virus propagation, and thus manipulation of sorcin activity may represent a novel therapeutic target for HCV.

Pharmaco-economic analysis of direct medical costs of metastatic colorectal cancer therapy with XELOX or modified FOLFOX-6 regimens: implications for health-care utilization in Australia.

AIM: The objective of this economic evaluation, which was based on patients from two randomized controlled clinical trials (NO16966 and NO16967), was to compare direct medical costs to the Australian health-care system of capecitabine plus oxaliplatin (XELOX) and bolus and/or infusional 5-fluorouracil (5-FU) plus folinic acid combined with oxaliplatin (modified [m] FOLFOX-6) in first-line and second-line treatment of advanced or metastatic colorectal cancer (mCRC). METHODS: Direct medical costs were estimated for five treatment settings from a public and private hospital. The costs included in evaluation were for drug acquisition, preparation (oxaliplatin, bolus and infusional 5-FU), administration and wastage. The cost of drug acquisition was calculated based on dosage data and the mean number of treatment cycles from the pivotal studies NO16966 and NO16967. There were no costs associated with preparing capecitabine and leucovorin. An oncology grouping and costing study was performed to determine the relevant administration costs associated with central venous access devices, their placement, maintenance and removal (for oxaliplatin administration) and the continuous infusion of 5-FU via a continuous ambulatory delivery device pump or infuser. RESULTS: This economic evaluation has shown that treating mCRC patients with XELOX in the first and second-line settings results in average cost savings of $9110 and $7113, respectively, compared with mFOLFOX-6. A multi-way sensitivity analysis demonstrated that the use of XELOX remained cost-saving from an Australian government health budget perspective. CONCLUSION: The use of XELOX, compared with mFOLFOX-6, for the treatment of mCRC is cost-saving in the Australian government health budget.

Gingival recession associated with predisposing factors in young vietnamese: a pilot study.

AIMS: Several studies have shown a large diversity in the prevalence, extent and severity of gingival recession as well as controversial conclusions of its associated factors. Therefore, the aim of this pilot study was to evaluate gingival recession with predisposing factors in young Vietnamese. METHODS: A cross-sectional study using clinical examination was performed in 120 dental students. Oral hygiene status, tooth malposition and fraenal attachment were recorded. The width of keratinised gingiva was measured after mucosa staining with Lugol's iodine solution. Measurements of gingival recession were performed on labial tooth surfaces. Chisquare test, t-test and Pearsonâs correlation were used for data analysis. RESULTS: The prevalence of gingival recession was 72.5% of the studied population. The extent of affected teeth was 11.1% of the examined teeth. The proportion of root-surface exposure was statistically higher (P<0.05) in the maxilla (12.5%) than in the mandible (9.6%). Premolars and right canines were the teeth most frequently and most seriously associated with gingival recession, respectively. There was a strong negative correlation between narrow width of keratinised gingiva and gingival recession (P<0.001). The recession was statistically associated with tooth malposition (P<0.001) but it was not related to high fraenal attachment and gender. CONCLUSIONS: A high prevalence of gingival recession was found in Vietnamese dental students. Gingival recession was associated with narrow width of keratinised gingiva, tooth malposition and maxillary teeth. Further studies performed in larger populations with more extended age groups are needed to confirm these findings.

Alcohol expectancies and social self-efficacy as mediators of differential intervention outcomes for college hazardous drinkers with social anxiety.

The current pilot study examined the roles of two cognitive factors - positive alcohol expectancies of social anxiety reduction and drink refusal self-efficacy relevant to social situations - in mediating greater reduction in alcohol behaviors by the Brief Intervention for Socially Anxious Drinkers (BISAD; n=21) compared to an alcohol psychoeducation (n=20) in a sample of college hazardous drinkers with social anxiety. Mediation analysis results indicated that decreased positive alcohol expectancies and increased drink refusal self-efficacy relevant to social situations accounted for an average of 67% of the variance in treatment outcomes as measured by total quantity of alcohol consumption, heavy drinking days and problems related to alcohol use in the past month. Study results may enhance the understanding of cognitive factors' role in alcohol treatment outcomes, which could in turn improve the efficacy of interventions aimed to reduce hazardous drinking and comorbid social anxiety.

Drinking refusal self-efficacy and tension-reduction alcohol expectancies moderating the relationship between generalized anxiety and drinking behaviors in young adult drinkers.

Despite the substantial comorbidity between generalized anxiety disorder (GAD) and alcohol use disorders (AUD), little is known about contributing factors to this relationship. This lack of knowledge has limited the development of theoretical models explicating the interesting yet complex relationship between GAD and AUD. The current study examined the roles of generalized anxiety, tension-reduction alcohol expectancies, and drinking refusal self-efficacy in accounting for the variance of alcohol consumption and alcohol-related consequences in a sample of young adult drinkers (N = 474; 18-25 years of age, median age 19, 66% female) from a large, urban Midwestern university. Results showed that generalized anxiety level interacted with both tension-reduction alcohol expectancies and drinking refusal self-efficacy to predict alcohol consumption and alcohol-related consequences. Findings support the assessment of both alcohol-related consequences and alcohol consumption, and highlight the importance of drinking refusal self-efficacy, which is a currently underexamined variable. Study results also enhance the knowledge about the underlining mechanisms of GAD and AUD comorbidity, which facilitates the development of an empirically based theoretical paradigm for their relationship.

Combining motivational interviewing with compliance enhancement therapy (MI-CET): development and preliminary evaluation of a new, manual-guided psychosocial adjunct to alcohol-dependence pharmacotherapy.

OBJECTIVE: Psychosocial interventions that are practical, transportable, and effective in promoting treatment adherence and efficacy are greatly needed in both research and clinical settings involving alcohol-dependence pharmacotherapy. In this article, we describe the development and preliminary evaluation of an integrative treatment blending motivational interviewing and compliance enhancement therapy (MI-CET) as a means of enhancing adherence and retention in an ongoing clinical trial. METHOD: Medication adherence, session attendance, and study completion rates were examined for 121 treatment-seeking, alcohol-dependent adults participating in a randomized clinical trial of citalopram (n = 81) versus placebo (n = 40). All participants received the manual-guided MI-CET intervention as an adjunct to pharmacotherapy. Preliminary adherence and retention data for this trial were compared with data from prior studies involving treatment for alcohol dependence with a selective serotonin reuptake inhibitor. RESULTS: High rates of medication adherence (79% of citalopram and 91% of placebo completers took > or = 80% of doses), session attendance (average of 90% for citalopram and 93% for placebo groups), and study completion (81% for citalopram and 88% for placebo groups) were obtained in the present study using MI-CET. These rates were at least comparable to or were, in some cases, 20%-30% higher than rates obtained in the comparison trials. CONCLUSIONS: These results suggest that MI-CET is feasible as a psychosocial adjunct to alcohol-dependence pharmacotherapy. Given its strengths as a clinical and research intervention (e.g., practicality, transportability), further evaluation of its efficacy is warranted.

Intimate partner violence experience and expectations among college women in dating relationships: implications for behavioral interventions.

Studies suggest that experience and expectations of intimate partner violence (IPV) among college women may be positively related. We investigated their association using modified versions of two standard measures: the Conflict Tactics Scale and the Multidimensional Emotional Abuse Scale. Five hundred and thirty-four college women completed each measure twice: once based on their partner's actual behaviors and once based on what they expected a male partner would do in a relationship. Results demonstrated that IPV experience and IPV expectations are significantly and positively related. Results also showed that respondents expected more IPV than they reportedly experienced, suggesting that college women may believe that IPV occurs more frequently in others' relationships than in their own. Findings suggest that behavioral interventions for IPV victims should address IPV expectations.

Alcohol expectancies and drinking motives in college drinkers: Mediating effects on the relationship between generalized anxiety and heavy drinking in negative-affect situations.

The current study tested the hypotheses that drinking to cope motives and alcohol expectancies of tension- and worry-reduction mediate the relationship between generalized anxiety (GA) and negative-affect heavy drinking in a cross-sectional sample of 782 college drinkers. As expected, structural equation modeling results indicated that alcohol expectancies mediated the relationship between GA and drinking to cope motives, and drinking to cope motives mediated the relationship between alcohol expectancies and heavy drinking in negative-affect situations. Unexpectedly, drinking to cope motives also mediated the relationship between GA and negative-affect heavy drinking. The model predicting negative-affect heavy drinking was tested in subsamples of 413 hazardous and 366 nonhazardous drinkers and did not differ structurally; however, omnibus measurement of model indirect effects was stronger for hazardous than nonhazardous drinkers. Finally, the results of a similar post-hoc model to predict general problem drinking support the specificity of the interrelationships among GA, cognitive mediators and to negative-affect drinking. These results inform cognitive-behavioral theories and interventions for comorbid GA and alcohol use problems.

Development and initial validation of the Worry-Reduction Alcohol Expectancy Scale.

Recent research on the relation between generalized anxiety and heavy drinking highlighted a need for a measure of worry-reduction alcohol expectancies. The current study aimed to develop and to assess the psychometric properties of the Worry-Reduction Alcohol Expectancy Scale. The development and initial validation of the WRAES occurred across two phases with two separate non-clinical undergraduate samples. Phase I was focused on scale construction and item selection, while Phase II included an assessment of the WRAES' reliability, validity and cross-validation of factor structure. Results from both study phases support a two-factor model. Cronbach's alpha for the total scale was .96 and Pearson r test-retest reliability was .75. Additionally, the mean correlation between the WRAES and convergent measures was significantly higher than the mean correlation between the WRAES and discriminant measures. Overall, the results provide initial support for the WRAES as a measure of worry-reduction alcohol expectancies.

Does negative affect increase change readiness among college hazardous drinkers?

The Transtheoretical Stages of Change Model specifies that when the costs outweigh the benefits of substance abuse, the resulting discomfort can be a catalyst for change. The current study evaluated the roles of depression and anxiety in motivating readiness to change current drinking behaviors. Results from regression analyses of self-report data from 233 undergraduate hazardous drinkers indicated that higher levels of depression and anxiety were associated with elevated readiness to change. Additionally, study findings showed that when considered together, anxiety accounted for more of the individual differences in alcohol change readiness than depressive symptoms. Study results were discussed in the context of existing models of change readiness and implications for further research and clinical practice.

Effects of training intensity on observers' ratings of anxiety, social skills, and alcohol-specific coping skills.

Few studies have addressed the amount of training needed to obtain reliable ratings in behavioral observation data. The current study examined the effects of differing intensities of frame-of-reference (FOR) rater training on observers' ratings of anxiety, social skills, and alcohol-specific coping skills in community volunteers with and without social anxiety and alcohol use disorders. Interrater reliability was assessed by comparing three training conditions (no-training, moderate FOR, and intensive FOR) on discrepancies between raters'scores and the strength of association between raters'scoring patterns. The discrepancies between raters' scores were significantly larger in the control condition than in the intensive and moderate training conditions. Generally, small and nonsignificant differences were found between intensive and moderate training's discrepancy scores. Strength of association results showed significantly lower correlations in the control group compared to the intensive group. However, these correlational results showed less consistent differences between the moderate and other training conditions; differences when found were in the expected directions. Study findings suggest that differing training intensities can affect rating scores and that interrater reliability may be meaningfully assessed through multiple methods.

Correlates for posttraumatic stress disorder in Gulf War veterans: a retrospective study of main and moderating effects.

With a sample of 120 Gulf War veterans, the present study investigated the main effects of childhood and lifetime trauma, combat exposure, and coping strategies on posttraumatic stress disorder (PTSD), as well as combat exposure's moderating effects on the other variables' relationships with PTSD. Logistic regression results indicated correct classification of PTSD diagnosis for 88% of the participants, with combat exposure and avoidant coping making significant contributions to this classification. Multiple regression results indicated that lifetime trauma, combat exposure, and avoidant coping were strongly related to PTSD symptoms. Multiple regression results also revealed that combat exposure moderated the strength and direction of PTSD's relationships with childhood trauma and avoidant coping. Study findings have implications for longitudinal investigation of PTSD development and preventive interventions.

Partner violence impacts the psychosocial and psychiatric status of pregnant, drug-dependent women.

The present study compared pregnant, drug-dependent women reporting current partner abuse (n=26) or no partner abuse (n=76) at treatment enrollment on initial psychosocial and psychiatric functioning. Abused pregnant women presented with more severe alcohol and family/social problems and greater rates of psychiatric problems and comorbidity than nonabused women did. The abused women reported that their partners had greater rates of alcohol and illicit drug use, relative to the partners of the nonabused women. Results suggest that treatment should address the women's relationships with their partners, especially when substance abuse and violence issues are present.

Bulimic symptoms and mood predict food relevant Stroop interference in women with troubled eating patterns.

Cognitive processing differences based on attentional biases of words pertaining to eating disorders were investigated to assess people's pathological thoughts. Participants were 165 undergraduate women (mean age=19.2) at a large Midwestern university. This Stroop task that included color identification of three word groups (food-related words, neutral words, and color words) was administered to measure differential speed in cognitive processing of salient words in individuals with and without troubled eating patterns. As predicted by the moderator hypothesis, a statistically significant interaction effect between bulimic symptoms and negative mood was found on food-related reaction time. Post hoc analysis of the interaction showed that women endorsing more bulimic symptoms responded slower to food-related cues than women with fewer bulimic symptoms among individuals who reported negative mood. The study results indicate that women who have problematic eating patterns and experience negative mood are hyperattentive to food-related cues.