Time-regulated transcripts with the potential to modulate human pluripotent stem cell-derived cardiomyocyte differentiation.

BACKGROUND: Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are a promising disease model, even though hiPSC-CMs cultured for extended periods display an undifferentiated transcriptional landscape. MiRNA-target gene interactions contribute to fine-tuning the genetic program governing cardiac maturation and may uncover critical pathways to be targeted. METHODS: We analyzed a hiPSC-CM public dataset to identify time-regulated miRNA-target gene interactions based on three logical steps of filtering. We validated this process in silico using 14 human and mouse public datasets, and further confirmed the findings by sampling seven time points over a 30-day protocol with a hiPSC-CM clone developed in our laboratory. We then added miRNA mimics from the top eight miRNAs candidates in three cell clones in two different moments of cardiac specification and maturation to assess their impact on differentiation characteristics including proliferation, sarcomere structure, contractility, and calcium handling. RESULTS: We uncovered 324 interactions among 29 differentially expressed genes and 51 miRNAs from 20,543 transcripts through 120 days of hiPSC-CM differentiation and selected 16 genes and 25 miRNAs based on the inverse pattern of expression (Pearson R-values < - 0.5) and consistency in different datasets. We validated 16 inverse interactions among eight genes and 12 miRNAs (Person R-values < - 0.5) during hiPSC-CMs differentiation and used miRNAs mimics to verify proliferation, structural and functional features related to maturation. We also demonstrated that miR-124 affects Ca2+ handling altering features associated with hiPSC-CMs maturation. CONCLUSION: We uncovered time-regulated transcripts influencing pathways affecting cardiac differentiation/maturation axis and showed that the top-scoring miRNAs indeed affect primarily structural features highlighting their role in the hiPSC-CM maturation.

Ppia is the most stable housekeeping gene for qRT-PCR normalization in kidneys of three Pkd1-deficient mouse models.

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most common inherited renal disorder, characterized by renal cyst development leading to end-stage renal disease. Although the appropriate choice of suitable reference is critical for quantitative RNA analysis, no comparison of frequently used "housekeeping" genes is available. Here, we determined the validity of 7 candidate housekeeping genes (Actb, Actg1, B2m, Gapdh, Hprt, Pgam1 and Ppia) in kidney tissues from mouse models orthologous to ADPKD, including a cystic mice (CY) 10-12 weeks old (Pkd1flox/flox:Nestincre/Pkd1flox/-:Nestincre, n = 10) and non-cystic (NC) controls (Pkd1flox/flox/Pkd1flox/-, n = 10), Pkd1-haploinsufficient (HT) mice (Pkd1+/-, n = 6) and wild-type (WT) controls (Pkd1+/+, n = 6) and a severely cystic (SC) mice 15 days old (Pkd1V/V, n = 7) and their controls (CO, n = 5). Gene expression data were analyzed using six distinct statistical softwares. The estimation of the ideal number of genes suggested the use of Ppia alone as sufficient, although not ideal, to analyze groups altogether. Actb, Hprt and Ppia expression profiles were correlated in all samples. Ppia was identified as the most stable housekeeping gene, while Gapdh was the least stable for all kidney samples. Stat3 expression level was consistent with upregulation in SC compared to CO when normalized by Ppia expression. In conclusion, present findings identified Ppia as the best housekeeping gene for CY + NC and SC + CO groups, while Hprt was the best for the HT + WT group.

Wording Effect in the Measurement of Attitudes towards Dating Violence / La eficacia de una intervención breve basada en la Teoría Incremental de la Personalidad para la prevención de la violencia en las relaciones de noviazgo adolescentes: un ensayo clínico aleatorizado

Positive and negative-worded items affect respondents' answering style in attitude surveys and have psychometric implications. This research was conducted to assess the wording effect on the validation of the Spanish adaptation of the Intimate Partner Violence Attitude Scale-Revised (IPVAS-R). Five competing models were tested with confirmatory factor analyses in two samples of young people between 15 and 25 years old (sample A, n = 402, Mage = 17.5, SDage = 2.21, 60.4% girls; sample B, n = 451, Mage = 19.55, SDage = 3.11, 57.6% girls). An outstanding method effect related to the writing of items in a positive (acceptance) versus negative (rejection) sense affecting the original three-factor structure of the IPVAS-R was supported by this research. The wording effect produced inconsistent factorial loadings and flawed internal consistency. Most of the relationships with other criterion measures, after controlling for the method effect, were moderate, in line with previous studies. Our results imply that the wording effect should be considered both in the building and the validation of instruments on attitudes towards dating violence

La redacción de los ítems en sentido directo e inverso afecta al estilo de respuesta de los encuestados en las escalas de actitud y tienen implicaciones psicométricas. Esta investigación se llevó a cabo para evaluar el efecto de la redacción en sentido directo o inverso en la validación de la adaptación española de la Escala de Actitud hacia la Violencia en la Pareja-revisada (IPVAS-R). Se probaron cinco modelos competitivos con análisis factorial confirmatorio en dos muestras de jóvenes entre 15 y 25 años de edad (muestra A, n = 402, Medad = 17.5, DTedad = 2.21, 60.4% chicas; muestra B, n = 451, Medad = 19.55, DTedad = 3.11, 57.6% chicas). Un notable efecto del método relacionado con la redacción de los ítems en un sentido positivo (aceptación) frente a negativo (rechazo) que afectaba a la estructura original de tres factores del IPVAS-R fue avalado por esta investigación. El efecto de la redacción produjo cargas factoriales incongruentes y una consistencia interna defectuosa. La mayoría de las relaciones con otras medidas criterio, tras controlar el efecto del método, fueron moderadas, en línea con estudios anteriores. Nuestros resultados implican que el efecto de la redacción debe ser considerado tanto en la construcción como en la validación de los instrumentos que miden actitudes hacia la violencia en el noviazgo

FoxN1 mediates thymic cortex-medulla differentiation through modifying a developmental pattern based on epithelial tubulogenesis.

The mechanisms that determine the commitment of thymic epithelial precursors to the two major thymic epithelial cell lineages, cTECs and mTECs, remain unknown. Here we show that FoxN1 nu mutation, which abolishes thymic epithelium differentiation, results in the formation of a tubular branched structure according to a typical branching morphogenesis and tubulogenesis developmental pattern. In the presence of FoxN1, in alymphoid NSG and fetal Ikaros-/- thymi, there is no lumen formation and only partial apical differentiation. This initiates cortex-medulla differentiation inducing expression of medullary genes in the apically differentiating cells and of cortical genes in the non-apically differentiating cells, which will definitely differentiate in wt and postnatal Ikaros-/- mice. Therefore, the thymus development is based on a branching morphogenesis and tubulogenesis developmental pattern: FoxN1 expression in the thymic primordium inhibits tubulogenesis and induces the expression of genes involved in TEC differentiation, which culminates with the expression of functional cell markers, i.e., MHCII, CD80, Aire in both postnatal Ikaros-/- and WT thymi after arrival of lymphoid progenitor cells.

Older Workers and Affective Job Satisfaction: Gender Invariance in Spain.

Older employees' affective job satisfaction is an aspect that arouses growing interest among researchers. Among the affective measures of job satisfaction, the Brief Index of Affective Job Satisfaction (BIAJS) is one of the most used in the last decade. This study is intended to the test the gender invariance of the BIAJS in two samples of workers over age 40 in Spain. The first sample, of 300 participants and the second sample, of 399 participants, have been used to test gender invariance of the BIAJS. In comparison with the original English version, the Spanish version of the BIAJS has adequate psychometric properties. The findings allow us to consider it a valid and reliable tool to assess older people's affective expressions about their work. In addition, this study provides evidence of its factorial invariance as a function of gender.

Variables associated with sleep quality in chronic tension-type headache: A cross-sectional and longitudinal design.

OBJECTIVE: To investigate variables associated at baseline (cross-sectional design) and at one year (longitudinal design) with the quality of sleep in chronic tension-type headache (CTTH). METHODS: One hundred and eighty (n = 180) and 135 individuals with CTTH participated in the cross-sectional and longitudinal design respectively. Clinical features were collected with a 4-weeks headache diary at baseline and one-year follow-up. Sleep quality was assessed at baseline and 1-year follow-up with the Pittsburgh Sleep Quality Index. Anxiety and depression (Hospital Anxiety and Depression Scale-HADS), burden of headache (Headache Disability Inventory-HDI), quality of life (SF-36 questionnaire), and pressure pain thresholds (PPTs) at trigeminal, extra-trigeminal and widespread area were assessed at baseline. Hierarchical regression analyses were conducted to determine the associations between variables at baseline and 1-year follow-up with sleep quality. RESULTS: At baseline positive correlations between sleep quality and headache intensity, headache frequency, headache duration, emotional and physical burden of headache and depression were observed. The regression analyses found that depression and emotional burden of headache explained 27.5% of the variance in sleep quality at baseline (r2 = .262; F = 23.72 P < .001). At one-year, sleep quality was significantly associated with baseline burden of headache, depression, widespread PPTs, vitality and mental health domains. Regression analyses revealed that vitality, PPT over the second metacarpal and PPT over the neck explained 30.0% of the variance of sleep quality at one-year (r2 = .269, F = 9.71, P < .001). CONCLUSIONS: It seems that sleep quality exhibits a complex interaction in individuals with CTTH since depression and the emotional burden were associated with sleep quality at baseline, but vitality and PPTs over extra-trigeminal areas were associated with the quality of sleep at one-year.

Sleep disturbances in tension-type headache and migraine.

Current research into the pathogenesis of tension-type headache (TTH) and migraine is focused on altered nociceptive pain processing. Among the potential factors that influence sensitization mechanisms, emotional stress, depression, or sleep disorders all have an essential role: they increase the excitability of nociceptive firing and trigger hyperalgesic responses. Sleep disturbances and headache disorders share common brain structures and pathogenic mechanisms and TTH, migraine, and sleep disturbances often occur together; for example, 50% of individuals who have either TTH or migraine have insomnia. Moreover, insomnia and poor sleep quality have been associated with a higher frequency and intensity of headache attacks, supporting the notion that severity and prevalence of sleep problems correlate with headache burden. It should be noted that the association between headaches and sleep problems is bidirectional: headache can promote sleep disturbances, and sleep disturbances can also precede or trigger a headache attack. Therefore, a better understanding of the factors that affect sleep quality in TTH and migraine can assist clinicians in determining better and adequate therapeutic programs. In this review, the role of sleep disturbances in headaches, and the association with depression, emotional stress, and pain sensitivity in individuals with TTH or migraine are discussed.

Is the association between health-related quality of life and fatigue mediated by depression in patients with multiple sclerosis? A Spanish cross-sectional study.

OBJECTIVES: To determine the mediating effects of depression on health-related quality of life and fatigue in individuals with multiple sclerosis (MS). DESIGN: A cross-sectional study. SETTING: Tertiary urban hospital. PARTICIPANTS: One hundred and eight patients (54% women) with MS participated in this study. OUTCOME MEASURES: Demographic and clinical data (weight, height, medication and neurological impairment), fatigue (Fatigue Impact Scale), depression (Beck Depression Inventory-II) and health-related quality of life (Short-Form Health Survey 36) were collected. RESULTS: Fatigue was significantly associated with bodily pain, physical function, mental health and depression. Depression was associated with bodily pain and mental health. The path analysis found direct effects from physical function, bodily pain and depression to fatigue (all, P<0.01). The path model analysis revealed that depression exerted a mediator effect from bodily pain to fatigue (B=-0.04, P<0.01), and from mental health to fatigue (B=-0.16, P<0.01). The amount of fatigue explained by all predictors in the path model was 37%. CONCLUSIONS: This study found that depression mediates the relationship between some health-related quality of life domains and fatigue in people with MS. Future longitudinal studies focusing on proper management of depressive symptoms in individuals with MS will help determine the clinical implications of these findings.

Gender differences in variables associated with sleep quality in chronic tension type headache.

We aimed to evaluate gender differences in the relationships between headache features, sleep quality, anxiety, depressive symptoms, and burden of headache in 193 patients (73 percent women) with chronic tension type headache (CTTH). Sleep quality was assessed with the Pittsburgh Sleep Quality Index. Headache features were collected with a four-week diary. The Hospital Anxiety and Depression Scale was used to assess anxiety/depressive symptoms. Headache Disability Inventory was used to evaluate the burden of headache. In men with CTTH, sleep quality was positive correlated with headache frequency (r = 0.310; p = .018), emotional (r = 0.518; p < .001) and physical (r = 0.468; p < .001) burden of headache, and depressive symptoms (r = 0.564; p < .001). In women, positive correlations were observed between sleep quality and headache intensity (r = 0.282; p < .001), headache frequency (r = 0.195; p = .021), emotional burden (r = 0.249; p = .004), and depressive symptoms (r = 0.382; p < .001). The results of stepwise regression analyses revealed that depressive symptoms and emotional burden of headache explained 37.2 percent of the variance in sleep quality in men (p < .001), whereas depressive symptoms and headache intensity explained 17.4 percent of the variance in sleep quality in women (p < .001) with CTTH. Gender differences associated with poor sleep should be considered for proper management of individuals with CTTH.

The burden of headache is associated to pain interference, depression and headache duration in chronic tension type headache: a 1-year longitudinal study.

BACKGROUND: To investigate variables associated at one year (longitudinal design) with the physical or emotional component of burden in chronic tension type headache (CTTH). METHODS: One hundred and thirty (n = 130) individuals with CTTH participated in this longitudinal study. Clinical features were collected with a 4-weeks headache diary at baseline and 1-year follow-up. The burden of headache was assessed at baseline and one -year follow-up with the Headache Disability Inventory (HDI), physical (HDI-P) or emotional (HDI-E) component. Sleep quality (Pittsburgh Sleep Quality Index), anxiety and depression (Hospital Anxiety and Depression Scale-HADS), and quality of life (SF-36) were also assessed at baseline. Hierarchical regression analyses were conducted to determine the associations between the baseline variables and the headache burden at 1-year. Simple mediation models were also applied to determine the potential mediation effect of any intermediary variable. RESULTS: Regression analyses revealed that baseline pain interference and depression explained 32% of the variance in the emotional burden of headache, whereas baseline emotional burden of the headache, pain interference, and headache duration explained 51% of the variance in the physical burden of headache (P < .01) at 1-year. The mediation models observed that the effect of baseline pain interference on emotional burden of headache at 1-year was mediated through baseline depression, whereas the effect of baseline pain interference on the physical burden of headache at 1-year was mediated through baseline emotional burden of headache (both P < .05). CONCLUSIONS: The current study found a longitudinal interaction between pain interference and depression with the burden of headache in individuals with CTTH.

Valor del «test de bipedestación» en el diagnóstico y la evaluación de la respuesta al tratamiento con bloqueadores beta en el síndrome de QT largo / Value of the 'standing test' in the diagnosis and evaluation of beta-blocker therapy response in long QT syndrome

Introducción y objetivos: Los pacientes con síndrome de QT largo (SQTL) tienen una adaptación anormal del QT a los cambios bruscos de la frecuencia cardiaca producidos con la bipedestación. Este trabajo estudia la utilidad del test de bipedestación en una cohorte de pacientes con SQTL y evalúa si el fenómeno de «mala adaptación» del QT se normaliza con el tratamiento con bloqueadores beta. Métodos: Se realizó un electrocardiograma basal y otro inmediatamente tras la bipedestación a 36 pacientes con SQTL (6 [17%] con QTL1, 20 [56%] con QTL2, 3 [8%] con QTL7 y 7 [19%] con genotipo no identificado) y 41 controles. Se midió el intervalo QT corregido (QTc) basal (QTcdecúbito) y tras la bipedestación (QTcbipedestación) y el incremento del QTc (ΔQTc = QTcbipedestación - QTcdecúbito). Se repitió el test en 26 de los pacientes bajo tratamiento con bloqueadores beta. Resultados: El QTcbipedestación y el ΔQTc fueron mayores en el grupo de SQTL que en el grupo control (QTcbipedestación, 528 ± 46 frente a 420 ± 15 ms; p < 0,0001; ΔQTc, 78 ± 40 frente a 8 ± 13 ms; p < 0,0001). No hubo diferencias significativas entre los pacientes con QTL1 y QTL2. Los pacientes con SQTL presentaron patrones típicos del segmento ST-onda T tras la bipedestación. Las curvas receiver operating characteristic del QTcbipedestación y ΔQTc mostraron un incremento significativo del valor diagnóstico comparadas con la del QTcdecúbito(área bajo la curva de ambos, 0,99 frente a 0,85; p < 0,001). El tratamiento con bloqueadores beta atenuó la respuesta a la bipedestación de los pacientes con SQTL (en tratamiento, QTcbipedestación, 440 ± 32 ms [p < 0,0001] y ΔQTc, 14 ± 16 ms [p < 0,0001]). Conclusiones: La evaluación del intervalo QTc tras la bipedestación proporciona un alto rendimiento diagnóstico y podría ser de gran utilidad en la monitorización del tratamiento con bloqueadores beta en los pacientes con SQTL (AU)

Introduction and objectives: Patients with congenital long QT syndrome (LQTS) have an abnormal QT adaptation to sudden changes in heart rate provoked by standing. The present study sought to evaluate the standing test in a cohort of LQTS patients and to assess if this QT maladaptation phenomenon is ameliorated by beta-blocker therapy. Methods: Electrographic assessments were performed at baseline and immediately after standing in 36 LQTS patients (6 LQT1 [17%], 20 LQT2 [56%], 3 LQT7 [8%], 7 unidentified-genotype patients [19%]) and 41 controls. The corrected QT interval (QTc) was measured at baseline (QTcsupine) and immediately after standing (QTcstanding); the QTc change from baseline (ΔQTc) was calculated as QTcstanding - QTcsupine. The test was repeated in 26 patients receiving beta-blocker therapy. Results: Both QTcstanding and ΔQTc were significantly higher in the LQTS group than in controls (QTcstanding, 528 ± 46 ms vs 420 ± 15 ms, P < .0001; ΔQTc, 78 ± 40 ms vs 8 ± 13 ms, P < .0001). No significant differences were noted between LQT1 and LQT2 patients. Typical ST-T wave patterns appeared after standing in LQTS patients. Receiver operating characteristic curves of QTcstanding and ΔQTc showed a significant increase in diagnostic value compared with the QTcsupine (area under the curve for both, 0.99 vs 0.85; P < .001). Beta-blockers attenuated the response to standing in LQTS patients (QTcstanding, 440 ± 32 ms, P < .0001; ΔQTc, 14 ± 16 ms, P < .0001). Conclusions: Evaluation of the QTc after the simple maneuver of standing shows a high diagnostic performance and could be important for monitoring the effects of beta-blocker therapy in LQTS patients (AU)

The association of headache frequency with pain interference and the burden of disease is mediated by depression and sleep quality, but not anxiety, in chronic tension type headache.

BACKGROUND: A better understanding of potential relationship between mood disorders, sleep quality, pain, and headache frequency may assist clinicians in determining optimal therapeutic programs. The aim of the current study was to analyze the effects of sleep quality, anxiety, depression on potential relationships between headache intensity, burden of headache, and headache frequency in chronic tension type headache (CTTH). METHODS: One hundred and ninety-three individuals with CTTH participated. Headache features were collected with a 4-weeks headache diary. The Hospital Anxiety and Depression Scale was used for assessing anxiety and depression. Headache Disability Inventory evaluated the burden of headache. Pain interference was determined with the bodily pain domain (SF-36 questionnaire). Sleep quality was assessed with Pittsburgh Sleep Quality Index. Path analyses with maximum likelihood estimations were conducted to determine the direct and indirect effects of depression, anxiety, and sleep quality on the frequency of headaches. RESULT: Two paths were observed: the first with depression and the second with sleep quality as mediators. Direct effects were noted from sleep quality, emotional burden of disease and pain interference on depression, and from depression to headache frequency. The first path showed indirect effects of depression from emotional burden and from sleep quality to headache frequency (first model R 2 = 0.12). Direct effects from the second path were from depression and pain interference on sleep quality and from sleep quality on headache frequency. Sleep quality indirectly mediated the effects of depression, emotional burden and pain interference on headache frequency (second model R 2 = 0.18). CONCLUSIONS: Depression and sleep quality, but not anxiety, mediated the relationship between headache frequency and the emotional burden of disease and pain interference in CTTH.

Value of the "Standing Test" in the Diagnosis and Evaluation of Beta-blocker Therapy Response in Long QT Syndrome.

INTRODUCTION AND OBJECTIVES: Patients with congenital long QT syndrome (LQTS) have an abnormal QT adaptation to sudden changes in heart rate provoked by standing. The present study sought to evaluate the standing test in a cohort of LQTS patients and to assess if this QT maladaptation phenomenon is ameliorated by beta-blocker therapy. METHODS: Electrographic assessments were performed at baseline and immediately after standing in 36 LQTS patients (6 LQT1 [17%], 20 LQT2 [56%], 3 LQT7 [8%], 7 unidentified-genotype patients [19%]) and 41 controls. The corrected QT interval (QTc) was measured at baseline (QTcsupine) and immediately after standing (QTcstanding); the QTc change from baseline (ΔQTc) was calculated as QTcstanding - QTcsupine. The test was repeated in 26 patients receiving beta-blocker therapy. RESULTS: Both QTcstanding and ΔQTc were significantly higher in the LQTS group than in controls (QTcstanding, 528 ± 46ms vs 420 ± 15ms, P < .0001; ΔQTc, 78 ± 40ms vs 8 ± 13ms, P < .0001). No significant differences were noted between LQT1 and LQT2 patients. Typical ST-T wave patterns appeared after standing in LQTS patients. Receiver operating characteristic curves of QTcstanding and ΔQTc showed a significant increase in diagnostic value compared with the QTcsupine (area under the curve for both, 0.99 vs 0.85; P < .001). Beta-blockers attenuated the response to standing in LQTS patients (QTcstanding, 440 ± 32ms, P < .0001; ΔQTc, 14 ± 16ms, P < .0001). CONCLUSIONS: Evaluation of the QTc after the simple maneuver of standing shows a high diagnostic performance and could be important for monitoring the effects of beta-blocker therapy in LQTS patients.

Integrative miRNA and mRNA analysis in penile carcinomas reveals markers and pathways with potential clinical impact.

Penile carcinoma (PeCa) is an important public health issue in poor and developing countries, and has only recently been explored in terms of genetic and epigenetic studies. Integrative data analysis is a powerful method for the identification of molecular drivers involved in cancer development and progression. miRNA and mRNA expression profiles followed by integrative analysis were investigated in 23 PeCa and 12 non-neoplastic penile tissues (NPT). Expression levels of eight miRNAs and 10 mRNAs were evaluated in the same set of samples used for microarray and in a validation set of cases (PeCa = 36; NPT = 27). Eighty-one miRNAs and 2,697 mRNAs were identified as differentially expressed in PeCa. Integrative data analysis revealed 255 mRNAs potentially regulated by 68 miRNAs. Using RT-qPCR, eight miRNAs and nine transcripts were confirmed as altered in PeCa. We identified that MMP1, MMP12 and PPARG and hsa-miR-31-5p, hsa-miR-224-5p, and hsa-miR-223-3p were able to distinguish tumors from NPT with high sensitivity and specificity. Higher MMP1 expression was detected as a better predictor of lymph node metastasis than the clinical-pathological data. In addition, PPARG and EGFR were highlighted as potential pathways for targeted therapy in PeCa. The analysis based on HPV positivity (7 of 23 cases) revealed five miRNA and 13 mRNA differentially expressed. Although in a limited number of cases, HPV positive PeCa presented less aggressive phenotype in comparison with negative cases. Overall, an integrative analysis using mRNA and miRNA profiles revealed markers related with tumor development and progression. Furthermore, MMP1 expression level was a predictive marker for lymph node metastasis in patients with PeCa.

Identification of Subgroups of Women with Carpal Tunnel Syndrome with Central Sensitization.

OBJECTIVE: Identification of subjects with different sensitization mechanisms can help to identify better therapeutic strategies for carpal tunnel syndrome (CTS). The aim of the current study was to identify subgroups of women with CTS with different levels of sensitization. METHODS: A total of 223 women with CTS were recruited. Self-reported variables included pain intensity, function, disability, and depression. Pressure pain thresholds (PPT) were assessed bilaterally over median, ulnar, and radial nerves, C5-C6 joint, carpal tunnel, and tibialis anterior to assess widespread pressure pain hyperalgesia. Heat (HPT) and cold (CPT) pain thresholds were also bilaterally assessed over the carpal tunnel and the thenar eminence to determine thermal pain hyperalgesia. Pinch grip force between the thumb and the remaining fingers was calculated to determine motor assessment. Subgroups were determined according to the status on a previous clinical prediction rule: PPT over the affected C5-C6 joint < 137 kPa, HPT on affected carpal tunnel <39.6ºC, and general health >66 points. RESULTS: The ANOVA showed that women within group 1 (positive rule, n = 60) exhibited bilateral widespread pressure hyperalgesia (P < 0.001) and bilateral thermal thresholds (P < 0.001) than those within group 2 (negative rule, n = 162). Women in group 1 also exhibited higher depression than those in group 2 (P = 0.023). No differences in self-reported variables were observed. CONCLUSION: This study showed that a clinical prediction rule originally developed for identifying women with CTS who are likely to respond favorably to manual physical therapy was able to identify women exhibiting higher widespread pressure hyper-sensitivity and thermal hyperalgesia. This subgroup of women with CTS exhibiting higher sensitization may need specific therapeutic programs.

Safety of targeting tumor endothelial cell antigens.

The mechanisms underlying discrimination between "self" and "non-self", a central immunological principle, require careful consideration in immune oncology therapeutics where eliciting anti-cancer immunity must be weighed against the risk of autoimmunity due to the self origin of tumors. Whole cell vaccines are one promising immunotherapeutic avenue whereby a myriad of tumor antigens are introduced in an immunogenic context with the aim of eliciting tumor rejection. Despite the possibility collateral damage to healthy tissues, cancer immunotherapy can be designed such that off target autoimmunity remains limited in scope and severity or completely non-existent. Here we provide an immunological basis for reconciling the safety of cancer vaccines, focusing on tumor endothelial cell vaccines, by discussing the following topics: (a) Antigenic differences between neoplastic and healthy tissues that can be leveraged in cancer vaccine design; (b) The layers of tolerance that control T cell responses directed against antigens expressed in healthy tissues and tumors; and, (c) The hierarchy of antigenic epitope selection and display in response to whole cell vaccines, and how antigen processing and presentation can afford a degree of selectivity against tumors. We conclude with an example of early clinical data utilizing ValloVax™, an immunogenic placental endothelial cell vaccine that is being advanced to target the tumor endothelium of diverse cancers, and we report on the safety and efficacy of ValloVax™ for inducing immunity against tumor endothelial antigens.

A comprehensive characterization of cell cultures and xenografts derived from a human verrucous penile carcinoma.

This study aimed to establish and characterize primary cell cultures and xenografts derived from penile carcinoma (PeCa) in order to provide experimental models for cellular processes and efficacy of new treatments. A verrucous squamous cell carcinoma (VSCC) was macrodissected, dissociated, and cultivated in KSFM/DF12 medium. Cell cultures were evaluated at passage 5 (P5) using migration and invasion assays and were serially propagated, in vivo, in BALB/c nude mice until passage 3 (X1-X3). Immunophenotypic characterization of cultures and xenografts was performed. Genomic (CytoScan HD, Affymetrix) and transcriptomic profiles (HTA 2.0 platform, Affymetrix) for VSCC, cell cultures, and xenografts were assessed. P5 cells were able to migrate, invade the Matrigel, and produce tumors in immunodeficient mice, demonstrating their malignant potential. The xenografts unexpectedly presented a sarcomatoid-like carcinoma phenotype. Genomic analysis revealed a high similarity between the VSCC and tumor-derived xenograft, confirming its xenograft origin. Interestingly, a subpopulation of P5 cells presented stem cell-related markers (CD44(+)CD24(-) and ALDH1(high)) and sphere-forming capacity, suggesting their potential xenograft origin. Cell cultures and xenografts retained the genomic alterations present in the parental tumor. Compared to VSCC, differentially expressed transcripts detected in all experimental conditions were associated with cellular morphology, movement, and metabolism and organization pathways. Malignant cell cultures and xenografts derived from a verrucous penile carcinoma were established and fully characterized. Nevertheless, xenograft PeCa models must be used with caution, taking into consideration the selection of specific cell populations and anatomical sites for cell/tumor implantation.

Pain is Associated to Clinical, Psychological, Physical, and Neurophysiological Variables in Women With Carpal Tunnel Syndrome.

OBJECTIVES: To investigate potential relationships of clinical (age, function, side of pain, years with pain), physical (cervical range of motion, pinch grip force), psychological (depression), and neurophysiological (pressure and thermal pain thresholds) outcomes and hand pain intensity in carpal tunnel syndrome (CTS). METHODS: Two hundred and forty-four (n=224) women with CTS were recruited. Demographic data, duration of the symptoms, function and severity of the disease, pain intensity, depression, cervical range of motion, pinch tip grip force, heat/cold pain thresholds (HPT/CPT), and pressure pain thresholds (PPT) were collected. Correlation and regression analysis were performed to determine the association among those variables and to determine the proportions of explained variance in hand pain intensity. RESULTS: Significant negative correlations existed between the intensity of pain and PPTs over the radial nerve, C5/C6 zygapophyseal joint, carpal tunnel and tibialis anterior muscle, HPT over the carpal tunnel, cervical extension and lateral-flexion, and thumb-middle, fourth, and little finger pinch tip forces. Significant positive correlations between the intensity of hand pain with function and depression were also observed. Stepwise regression analyses revealed that function, thumb-middle finger pinch, thumb-little finger pinch, depression, PPT radial nerve, PPT carpal tunnel, and HPT carpal tunnel were significant predictors of intensity of hand pain (R²=0.364; R² adjusted=0.343; F=16.87; P<0.001). CONCLUSION: This study showed that 36.5% of the variance of pain intensity was associated to clinical (function), neurophysiological (localized PPT and HPT), psychological (depression), and physical (finger pinch tip force) outcomes in women with chronic CTS.

Fenotipo heterogéneo del síndrome de QT largo causado por la mutación KCNH2-H562R: importancia del estudio genético familiar / Heterogeneous phenotype of long QT Syndrome caused by the KCNH2-H562R Mutation: Importance of familial genetic testing

Introducción y objetivos: El síndrome de QT largo es una canalopatía hereditaria que se asocia a síncope y muerte súbita. La heterogeneidad fenotípica de esta enfermedad hace que el estudio genético sea fundamental para detectar a los sujetos con síndrome de QT largo oculto. En este trabajo se exponen las características de una familia con 13 portadores de la mutación missense KCNH2-H562R que afecta a la región del poro del canal HERG. Métodos: Se describió la mutación KCNH2-H562R en un varón de 65 años con intervalo QTc prolongado que presentó un episodio de torsade de pointes. Posteriormente, se identificaron 13 portadores de la mutación en la familia. Se realizó evaluación clínica, electrocardiograma y ecocardiograma a los portadores (edad, 48 ± 26 años; el 46% varones). Resultados: El QTc medio en los portadores fue de 493 ± 42 ms (3 [23%] mostraron QTc normal); 6 (46%) tuvieron síntomas (4, síncope; 1, muerte súbita; 1, muerte súbita resucitada [probando]). Durante el tratamiento con bloqueadores beta, 11 (92%) de los 12 portadores permanecieron asintomáticos a los 5 años de seguimiento (1 paciente requirió simpatectomía cardiaca izquierda). El acortamiento del QTc con bloqueadores beta fue de 50 ± 37 ms. Hubo 1 muerte súbita en un paciente que rechazó tratamiento con bloqueadores beta. Conclusiones: El estudio familiar es fundamental en la interpretación de los resultados de los tests genéticos en la actualidad. Este artículo describe el fenotipo variable y heterogéneo de una amplia familia portadora de la mutación KCNH2-H562R y destaca el papel del estudio genético en la identificación de los individuos en riesgo que se beneficiarían del tratamiento con bloqueadores beta (AU)

Introduction and objectives: Long QT syndrome is an inherited ion channelopathy that leads to syncope and sudden death. Because of the heterogeneous phenotype of this disease, genetic testing is fundamental to detect individuals with concealed long QT syndrome. In this study, we determined the features of a family with 13 carriers of the KCNH2-H562R missense mutation, which affects the pore region of the HERG channel. Methods: We identified the KCNH2-H562R mutation in a 65-year-old man with a prolonged QTc interval who had experienced an episode of torsade de pointes. Subsequently, a total of 13 mutation carriers were identified in the family. Carriers (age 48 [26] years; 46% males) underwent clinical evaluation, electrocardiography and echocardiography. Results: The mean (standard deviation) QTc in carriers was 493 (42) ms (3 [23%] showed normal QTc); 6 (46%) had symptoms (4, syncope; 1, sudden death; 1, aborted sudden death [proband]). While under treatment with beta-blockers, 11 of 12 carriers (92%) remained asymptomatic at 5 years of follow-up (1 patient required left cardiac sympathectomy). The QTc shortening with beta-blockers was 50 (37) ms. There was 1 sudden death in a patient who refused treatment. Conclusions: Family study is essential in the interpretation of a genetic testing result. This article describes the heterogeneous and variable phenotype of a large family with the KCNH2-H562R mutation and highlights the role of genetic study for the appropriate identification of at-risk individuals who would benefit from treatment (AU)

Direct and Indirect Effects of Function in Associated Variables Such as Depression and Severity on Pain Intensity in Women with Carpal Tunnel Syndrome.

OBJECTIVE: To determine the direct and indirect effects of function on clinical variables such as age, pain intensity, years of the disease, severity of symptoms, and depression in women with electrodiagnostic and clinical diagnosis of carpal tunnel syndrome (CTS). DESIGN: A cross-sectional study. SETTING: Patients from an urban hospital referred to a university clinic. METHODS: Two hundred and forty-four (n = 224) women with CTS were included. Demographic and clinical data, duration of symptoms, function, symptom's severity of the symptoms, pain intensity, and depression were self-reported collected. Correlation and path analysis with maximum likelihood estimation were conducted to assess the direct and indirect effect of hand function on pain, age, years with the disease, symptoms severity, and depression. RESULTS: Significant positive correlations between function and pain intensity, years with pain and symptoms severity were observed. The path analysis found direct effects from depression, symptoms severity, and years with pain to function (all, P < 0.01). Paths between function and depression on pain intensity (both, P < 0.01) were also observed. The amount of function explained by all predictors was 22%. The indirect effects in the path analysis revealed that function exerted an indirect effect from depression to pain intensity (B = 0.18; P < 0.01), and from symptoms severity to the intensity of pain (B = 0.10; P < 0.01). Overall, the amount of current pain intensity explained by all predictors in the model was R(2) = 0.22. CONCLUSIONS: Our study demonstrated that function mediates the relationship between depression and symptoms severity with pain intensity in women with CTS. Future longitudinal studies will help to determine the clinical implications of these findings.