Identification of suitable mRNAs and microRNAs as reference genes for expression analyses in skin cells under sex hormone exposure.

Quantitative RT-PCR is the most accurate technique for the study of gene expression profiles, however, to ensure the accuracy of qPCR results, suitable reference genes are necessary for data normalization. Hormones influence the development and function of skin cells, regulating the expression of genes and miRNAs. Nevertheless, the stability of reference genes after sex hormone treatment has not been thoroughly investigated. In this study, we evaluated the expression of a set of candidate mRNAs and microRNsA (miRNA) as reference genes in keratinocytes (HaCaT cells), primary human fibroblasts and a melanoma cell line (LM-36 cells) under testosterone or 17ß-estradiol treatment. Two algorithms, namely geNorm, Best-Keeper, and the comparative ΔCt method were used to evaluate the expression stability of the candidate reference genes. The comprehensive ranking showed that TBP and miR-191-5p are the most stable expressed genes across all cultured cells under hormone treatment. Furthermore, we observed that GAPDH, HPRT1 and U6 snRNA expression may be altered by hormone exposure, thus, these genes are not recommended as reference genes. In conclusion, the present study provides, to the best of our knowledge, the first evaluation of expressed mRNA(s) and miRNA(s) as reference genes in three different types of skin cells under the stimulation of sex hormones.

Efficacy of oregano and rosemary essential oils to affect morphology and membrane functions of noncultivable sessile cells of Salmonella Enteritidis 86 in biofilms formed on stainless steel.

AIMS: This study evaluated the efficacy of essential oil from Origanum vulgare L. (oregano; OVEO) and Rosmarinus officinalis L. (rosemary; ROEO) to inactivate sessile cells of Salmonella enterica serovar Enteritidis 86 (SE86) in young and mature biofilms formed on stainless steel. METHODS AND RESULTS: Ultrastructural alterations and damage in different physiological functions caused by OVEO and ROEO in noncultivable sessile cells of SE86 were investigated using scanning electron microscopy and flow cytometry. OVEO (2·5 µl ml-1 ) and ROEO (40 µl ml-1 ) were effective to eradicate young and mature biofilms formed by SE86 sessile cells on stainless steel surfaces; however, the efficacy varied with exposure time. OVEO and ROEO caused alterations in morphology of SE86 sessile cells, inducing the occurrence of bubbles or spots on cell surface. OVEO and ROEO compromised membrane polarization, permeability and efflux activity in noncultivable SE86 sessile cells. These findings show that OVEO and ROEO act by a multitarget mechanism on SE86 membrane functions. CONCLUSIONS: ROEO and OVEO showed efficacy to eradicate SE86 sessile cells in preformed biofilms on stainless steel, displaying a time-dependent effect and multitarget action mode on bacterial cell membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides for the first time the effects of OVEO and ROEO on morphology and physiological functions of noncultivable sessile cells of S. Enteritidis biofilms preformed on stainless steel surfaces.

Detection of Approximal Caries Lesions in Adults: A Cross-sectional Study.

Detection and management of posterior approximal caries lesions are still problematic. Inspection of approximal surfaces is challenging, and bitewing radiographs are used when direct vision is not possible. Unfortunately, there is no definite radiographic appearance to identify lesion cavitation with absolute certainty. Many lesions detected radiographically within the outer half of dentin are not cavitated, often resulting in unnecessary restorative treatment. Our study compared radiographic depth of approximal caries lesions with presence of cavitation in adults using visual inspection following temporary tooth separation (TTS). We conducted this observational descriptive cross-sectional study at two dental schools in two cities in Chile. Clinicians were unaware of radiographic depths of lesions and examined 147 participants (57.3% female and 42.7% male) following TTS. Using the common classification system that consists of E0 (no lesion), E1 (lesion within the outer half of enamel), E2 (lesion within the inner half of enamel), D1 (lesion within the outer third of dentin), D2 (lesion within the middle third of dentin), and D3 (lesion within the inner third of dentin), a trained dentist evaluated all the processed films. Cavitation was detected in only three sites (0.22%) within the E0 category, seven sites (3.41%) in E1, five sites (14.8%) in E2, four sites (14.8%) in D1, six sites (50%) in D2, and eight sites (61.5%) in D3. Considering that restorative treatment should be indicated strictly for cavitated lesions, our findings support indication for restorative treatment for D3 lesions and the rationale for TTS for D1-D2 caries lesions to allow direct visual inspection to determine whether there is surface cavitation.

Expression of TLR4-PTGE2 signaling genes in atherosclerotic carotid plaques and peripheral blood.

Toll-like receptor 4 (TLR4)/prostaglandine synthetase 2 (PTGS2) signaling plays a relevant role in atherosclerotic plaque vulnerability. The purpose of this study was to check the gene expression of 6 genes participating to TLR4/PTGS2 signaling (TLR4, PTGS2, ACSL4, PTGER3, PTGER4, and EPRAP) in carotid plaques and blood samples from the same individual and to evaluate these genes as biomarker of plaque progression. We investigated differential gene expression by qRT-PCR in 62 atherosclerotic patients' carotid plaques and corresponding blood sample. A very weak or no correlation was observed in the overall population or analyzing asymptomatic patients. These analyzed genes are most likely not suitable for inclusion in the clinical routine as biomarkers of plaque instability.

Tuberculosis diseminada de médula ósea en pacientes VIH etapa SIDA. Reporte de 2 casos / Disseminated bone marrow tuberculosis in aids stage hiv (+) patients. 2 case reports

La tuberculosis extrapulmonar se define como tuberculosis que involucra a un órgano diferente al pulmón. Ocurre secundario a una diseminación de un foco pulmonar, la cual se favorece en estados de inmunosupresión, como es en el caso de pacientes portadores de VIH. Dentro de las localizaciones menos frecuentes, se encuentra la médula ósea. En la actualidad no existen reportes de casos de esta forma de tuberculosis extrapulmonar en pacientes VIH. En este artículo reportamos dos casos de pacientes con VIH etapa SIDA con diseminación en médula ósea. Este reporte destaca la importancia de tener presente esta complicación y realizar un estudio amplio, que incluya cultivo de Koch y tinción de Ziehl Nielsen de las muestras obtenidas, especialmente en este tipo de pacientes que pueden presentar clínica pobre e inespecífica.

Extra-pulmonary tuberculosis is defined to be tuberculosis involving any organ other than the lung. Immune suppressed states such as being HIV (+) favor dissemination from pulmonary foci. Bone marrow is one of the least frequent locations. At present there are no recorded cases of this form of extra-pulmonary tuberculosis in HIV (+) patients. We present two cases of HIV (+) patients in the AIDS stage with bone marrow dissemination. This reports underlines the importance of considering this complication and doing a comprehensive study, including testing samples using Koch culture and Ziehl Nielsen stains, especially in patients with few and non-specific clinical signs.

Properties of Particle Size Distribution from Milled White Nixtamalized Corn Kernels as a Function of Steeping Time.

This paper focuses on the particle size distribution (PSD) changes during nixtamalized corn kernels (NCK) as a function of the steeping time (ST). The process to obtain powder or corn flour from NCK was as follows: (i) the NCK with different STs were wet-milled in a stone mill, (ii) dehydrated by a Flash type dryer, and (iii) pulverized with a hammer mill and sieved with a 20 mesh. The powder was characterized by measuring the PSD percentage, calcium percentage (CP), peak viscosity at 90°C (PV), and crystallinity percentage (CP). The PSD of the powder as a function of ST was determined by sieving in Ro-TAP equipment. By sieving, five fractions of powder were obtained employing meshes 30, 40, 60, 80, and 100. The final weight of the PSD obtained from the sieving process follows a Gaussian profile with the maximum corresponding to the average particle obtained with mesh 60. The calcium percentage as a function of ST follows a behavior similar to the weight of the PSD. The study of crystallinity versus the mesh number shows that it decreases for smaller mesh number. A similar behavior is observed as steeping time increases, except around ST = 8 h where the gelatinization of starch is observed. The trend of increasing viscosity values of the powder samples occurs when increasing ST and decreasing particle size. The ST significantly changes the crystallinity and viscosity values of the powder and, in both cases, a minimum value is observed in the region 7-9 h. The experimental results show that the viscosity increases (decreases) if the particle size decreases (increases).

Development of secondary mutations in wild-type and mutant EZH2 alleles cooperates to confer resistance to EZH2 inhibitors.

The histone methyltransferase Enhancer of Zeste Homolog 2 (EZH2) is frequently dysregulated in cancers, and gain-of-function (GOF) EZH2 mutations have been identified in non-Hodgkin lymphomas. Small-molecule inhibitors against EZH2 demonstrated anti-tumor activity in EZH2-mutated lymphomas and entered clinical trials. Here, we developed models of acquired resistance to EZH2 inhibitor EI1 with EZH2-mutated lymphoma cells. Resistance was generated by secondary mutations in both wild-type (WT) and GOF Y641N EZH2 alleles. These EZH2 mutants retained the substrate specificity of their predecessor complexes but became refractory to biochemical inhibition by EZH2 inhibitors. Resistant cells were able to maintain a high level of H3K27Me3 in the presence of inhibitors. Interestingly, mutation of EZH2 WT alone generated an intermediate resistance phenotype, which is consistent with a previously proposed model of cooperation between EZH2 WT and Y641N mutants to promote tumorigenesis. In addition, the findings presented here have implications for the clinical translation of EZH2 inhibitors and underscore the need to develop novel EZH2 inhibitors to target potential resistance emerging in clinical settings.

Factors associated with study attrition among HIV-infected risky drinkers in St. Petersburg, Russia.

BACKGROUND: Participant attrition in HIV longitudinal studies may introduce bias and diminish research quality. The identification of participant characteristics that are predictive of attrition might inform retention strategies. OBJECTIVE: The study aimed to identify factors associated with attrition among HIV-infected Russian risky drinkers from the secondary HIV prevention HERMITAGE trial. We examined whether current injection drug use (IDU), binge drinking, depressive symptoms, HIV status nondisclosure, stigma, and lifetime history of incarceration were predictors of study attrition. We also explored effect modification due to gender. METHODS: Complete loss to follow-up (LTFU), defined as no follow-up visits after baseline, was the primary outcome, and time to first missed visit was the secondary outcome. We used multiple logistic regression models for the primary analysis, and Cox proportional hazards models for the secondary analysis. RESULTS: Of 660 participants, 101 (15.3%) did not return after baseline. No significant associations between independent variables and complete LTFU were observed. Current IDU and HIV status nondisclosure were significantly associated with time to first missed visit (adjusted hazard ratio [AHR], 1.39; 95% CI, 1.03-1.87; AHR, 1.38; 95% CI, 1.03-1.86, respectively). Gender stratified analyses suggested a larger impact of binge drinking among men and history of incarceration among women with time to first missed visit. CONCLUSIONS: Although no factors were significantly associated with complete LTFU, current IDU and HIV status nondisclosure were significantly associated with time to first missed visit in HIV-infected Russian risky drinkers. An understanding of these predictors may inform retention efforts in longitudinal studies.

Facilitadores y barreras que enfrentan las personas al tomarse el test de ELISA para el diagnóstico del VIH: revisión de la literatura / Facilitators and barriersto HIV testing: A literature review

Background: Late diagnosis of HIV is a problem of international and national relevance. Despite the availability of HIV testing in primary health care, it is often performed too late. Aim: To identify facilitators and barriers to early HIV testing in primary health care. Methods: Four databases of nursing, psychological, biomedical, and health related professions areas were examined with a review protocol. Results were grouped into two main subjects: facilitators and barriers occurring among the population, among health care workers, and within primary health care centers. Results: Perception of risk behaviors, self-care, social support, trust, confidentiality of the examination, the offer of the examination, and the knowledge of early treatment have been recognized as facilitators for taking the exam. The lack of information about the test and the disease are recognized as the main barrier to access the test. This information is a cornerstone to design and implement strategies to increase the number of people taking voluntarily HIV testing.

Introducción: El diagnóstico tardío de la infección por VIH es un problema universal. A pesar de la disponibilidad del test de ELISA para el diagnóstico de esta infección en la atención primaria de salud, las personas continúan tomándoselo tardíamente. Objetivo: Conocer los factores que facilitan o dificultan que las personas accedan oportunamente al examen en la atención primaria de salud. Métodos: Cuatro bases de datos del área de la enfermería, psicológica, salud biomédica y profesiones afines (años 2001-2012) fueron examinadas con un protocolo de revisión. Resultados: De 195 artículos detectados, 15 cumplieron con los criterios de inclusión y fueron agrupados en dos grandes temas: elementos facilitadores y elementos obstaculizadores de la persona, de los profesionales y de los centros de atención primaria de salud. Percepción de conductas de riesgo, autocuidado, apoyo social, la confianza, confidencialidad del examen, el ofrecimiento del examen y el conocimiento de un tratamiento oportuno han sido reconocidos como uno de los elementos facilitadores para la toma del examen. La falta de información sobre el test y la enfermedad son reconocidas como los principales obstaculizadores para acceder al test. Discusión: La información obtenida es un pilar fundamental para diseñar e implementar estrategias destinadas a aumentar el número de personas que solicitan voluntariamente al examen.

Identificación de virus respiratorios en pacientes pediátricos con infecciones respiratorias en el Hospital Carlos Van Buren, Valparaíso: época estival 2011-2012 / Identification of respiratory viruses in pediatric patients with respiratory tract infections from Carlos Van Buren Hospital, Valparaíso: 2011-2012 summer season

INTRODUCCIÓN: En pediatría, las infecciones respiratorias agudas (IRA) constituyen globalmente una importante causa de morbimortalidad. Los principales agentes etiológicos son los virus respiratorio sincicial, parainfluenza (PIV), influenza y adenovirus. Actualmente existen pruebas de diagnóstico molecular que permiten identificar agentes no convencionales. En este estudio se propuso estudiar la presencia de: PIV, coronavirus (COV), rinovirus (RV) y enterovirus humano (EV), utilizando reacción en cadena de polimerasa previa transcriptasa reversa (RCP-TR) en muestras de niños hospitalizados en el Hospital Carlos Van Buren, negativas a inmunofluorescencia (IF) paravirus respiratorios. MATERIALES Y MÉTODOS: Se analizaron un total de 82 muestras de aspirado nasofaríngeo de pacientes menores de 5 años hospitalizados por IRA en el Hospital Carlos Van Buren, negativos a IF, recolectadas entre Diciembre del 2011y Marzo del 2012. Se utilizó una RCP-TR anidada de tipo múltiplex específica para virus PIV (1, 2, 3, 4AB), COV-OC43 y 229E, y genérica para RV y EV. RESULTADOS: Los virus más frecuentes fueron los PIV, siendo identificados en 9 (10 por ciento) muestras, siendo: un 2 por ciento (n=2) PIV-2, un 7 por ciento (n=6) PIV-3 y un 1 por ciento (n=1) PIV-4AB. Además se detectó COV-OC43 en un 2 por ciento (n=2), RV y EV en un 6 por ciento (n=5). No se detectaron co-infecciones con dos o más virus. No hubo asociación entre edad de los niños y tipo de infección viral. DISCUSIÓN: Se describió la presencia de estos virus respiratorios en niños con IRA. Se logró detectar falsos negativos para PIV y se demostró la existencia de COV, RV y EV en la región.

INTRODUCTION: Acute Respiratory infection (ARI) is a major cause of morbidity and mortality worldwide in pediatrics. The main etiological agents are respiratory syncytial virus, parainfluenzavirus (PIV), influenza virus and adenovirus. Currently there are molecular diagnostic tests that have helped to identify unconventional agents. Our aim was to study the presence of: PIV, coronavirus (hCoV), rhinovirus (hRV) and human enterovirus (hEV), by using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) in samples of hospitalized children on Carlos Van Buren Hospital negative to immunofluorescence (IF) for respiratory viruses. MATERIALS AND METHODS: A total of 82nasopharyngeal aspirate samples, negative to IF, were collected from children under five years old hospitalized due to ARI at Carlos Van Buren Hospital, between December 2011 and March2012. A Multiplex nested RT-PCR designed for the specifically detection of PIV (1, 2, 3, 4AB), hCoV (OC43 and 229E) and generic detection of RV and EV was used. RESULTS: Parainfluenza was the most frequently identified virus (n=9, 10 percent) specifically PIV-2 (n=2, 2 percent), PIV-3 (n=6, 7 percent) and PIV-4AB (n=1, 1 percent). Followed by hRV and hEV (n=5, 6 percent) and hCoV-OC43 (n=2, 2 percent). There were no co-infections with two or more viruses. There was no correlation between age and type of infection. DISCUSSION: This study described the prevalence of these respiratory viruses in pediatric patients with ARI. It was possible to detect false negative results for parainfluenza virus and also to demonstrate the existence of hCoV, hRV and hEV in the region.

Encapsulation of trans-dehydrocrotonin in liposomes: an enhancement of the antitumor activity.

The aim of this study was the encapsulation of trans-dehydrocrotonin (t-DCTN) and its inclusion complexes with hydropropyl-beta-cyclodextrin (HP-beta-CD) in liposomes to improve t-DCTN antitumor activity. The in vitro kinetic profiles of t-DCTN-loaded liposomes (LD) and t-DCTN:HP-beta-CD-loaded liposomes (LC) were evaluated using the dialysis technique. The antitumor activity of LD and LC were investigated against Sarcoma 180 in Swiss mice. Histopathological and hematological analyses were carried out. The amounts of t-DCTN and t-DCTN:HP-beta-CD inclusion complex encapsulated in liposomes were equivalent to 1 mg of t-DCTN. The encapsulation efficiencies of LD and LC were 95.0 +/- 3.8% and 91.1 +/- 5.6%, respectively. In relation to kinetics, the drug release profiles of t-DCTN are in substantial agreement with the Fickian model. The treatment of animals with LD and LC produced tumor inhibitions of 79.4 +/- 9.6% and 63.5 +/- 5.5%, respectively. The liposomal encapsulation of t-DCTN by entrapment in the phospholipid bilayer increased at twice the antitumor activity. Moreover, the liposomal formulations reduced the hepatotoxicity effect of the drug and no significant hematological toxicity was observed in the treated animals. However, the counting of platelets was slightly decreased. Thus, the results show that the development of liposomal formulations containing t-DCTN or t-DCTN:HP-beta-CD is an important advance for enabling this drug to be use in therapy.

Prevalence of multiple sclerosis in Verona, Italy: an epidemiological and genetic study.

BACKGROUND AND PURPOSE: Recent multiple sclerosis (MS) prevalence studies classify Italy as a high-risk area without intra-regional latitude effect. OBJECTIVES: To determine MS prevalence in Verona, Italy, and frequency of myelin oligodendrocyte glycoprotein (MOG) gene G511C polymorphism and HLA-DRB1*15 locus in a sample of cases and healthy controls. METHODS: The study area population on the prevalence date (31 December 2001) was 253208 (133508 women, 119700 men). Multiple case sources were examined. Patients fulfilling McDonald's criteria (2001) were included. Crude, age- and sex-specific prevalence rates were computed. MOG G511C polymorphism and HLA-DRB1*15 were determined by standard methods. RESULTS: We identified 270 cases of MS yielding a crude prevalence rate of 106.6/100000 (95% CI: 94-120). Prevalence was higher in women (140.8/100000) than in men (68.5/100000). The age-adjusted prevalence rate standardized to the European population was 96.0/100000. MOG G511C polymorphism did not differ between cases and controls. HLA-DRB1*15 frequency was 58/155 (37%) in cases and 24/157 (15%) in controls (P<0.001). There was no HLA-DRB1*15 influence on susceptibility to other autoimmune disorders. CONCLUSIONS: The high MS prevalence in Verona confirms Italy as a high-risk area with a homogenous distribution across the country. HLA-DRB1*15 is a relevant MS susceptibility locus in the Italian population, possibly with little influence on the occurrence of concomitant autoimmune disorders.

[Facilitators and barriers to HIV testing: a literature review]. / Facilitadores y barreras que enfrentan las personas al tomarse el test de ELISA para el diagnóstico del VIH: revisión de la literatura.

BACKGROUND: Late diagnosis of HIV is a problem of international and national relevance. Despite the availability of HIV testing in primary health care, it is often performed too late. AIM: To identify facilitators and barriers to early HIV testing in primary health care. METHODS: Four databases of nursing, psychological, biomedical, and health related professions areas were examined with a review protocol. Results were grouped into two main subjects: facilitators and barriers occurring among the population, among health care workers, and within primary health care centers. RESULTS: Perception of risk behaviors, self-care, social support, trust, confidentiality of the examination, the offer of the examination, and the knowledge of early treatment have been recognized as facilitators for taking the exam. The lack of information about the test and the disease are recognized as the main barrier to access the test. This information is a cornerstone to design and implement strategies to increase the number of people taking voluntarily HIV testing.

Unusual presentation of oral squamous cell carcinoma in a young woman.

Oral squamous cell carcinoma (OSCC) is the most common oral malignant neoplasm, mainly affecting individuals over 50 years old with a history of tobacco and alcohol use. The occurrence of this oral cancer in individuals under 40 years old is unusual and, when it does occur, shows a weaker relation to those risk factors and a more aggressive clinical course. Due to the paucity of reports in this population, it is difficult to prove its increasing trend. A case of oral squamous cell carcinoma in a 39-year-old woman with no history of tobacco or alcohol use is reported. Clinical and histopathological findings, aetiology, and treatment are discussed. The increasing trend of oral squamous cell carcinoma in young women without known risk factors highlights the need for clinicians to be prepared to diagnose this lesion quickly and precisely, providing a better prognosis, chance of survival, and quality of life for the patient.

Cytotoxicity and cellular uptake of newly synthesized fucoidan-coated nanoparticles.

The aim was to synthesize and characterize fucoidan-coated poly(isobutylcyanoacrylate) nanoparticles. The nanoparticles were prepared by anionic emulsion polymerization (AEP) and by redox radical emulsion polymerization (RREP) of isobutylcyanoacrylate using fucoidan as a new coating material. The nanoparticles were characterized, and their cytotoxicity was evaluated in vitro on J774 macrophage and NIH-3T3 fibroblast cell lines. Cellular uptake of labeled nanoparticles was investigated by confocal fluorescence microscopy. Results showed that both methods were suitable to prepare stable formulations of fucoidan-coated PIBCA nanoparticles. Stable dispersions of nanoparticles were obtained by AEP with up to 100% fucoidan as coating material. By the RREP method, stable suspensions of nanoparticles were obtained with only up to 25% fucoidan in a blend of polysaccharide composed of dextran and fucoidan. The zeta potential of fucoidan-coated nanoparticles was decreased depending on the percentage of fucoidan. It reached the value of -44 mV for nanoparticles prepared by AEP with 100% of fucoidan. Nanoparticles made by AEP appeared more than four times more cytotoxic (IC(50) below 2 µg/mL) on macrophages J774 than nanoparticles made by RREP (IC(50) above 9 µg/mL). In contrast, no significant difference in cytotoxicity was highlighted by incubation of the nanoparticles with a fibroblast cell line. On fibroblasts, both types of nanoparticles showed similar cytotoxicity. Confocal fluorescence microscopy observations revealed that all types of nanoparticles were taken up by both cell lines. The distribution of the fluorescence in the cells varied greatly with the type of nanoparticles.

Upregulated expression of Toll-like receptor 4 in peripheral blood of ischaemic stroke patients correlates with cyclooxygenase 2 expression.

OBJECTIVES: An inflammatory process following stroke in human brains and systemic inflammatory responses after stroke in humans have been reported by numerous investigators. The aim of the study was to investigate if genes involved in the cyclooxygenase 2 (COX-2) pathway are upregulated at peripheral level in patients after transient ischaemic attack (TIA) and stroke. DESIGN OF STUDY: Blood samples were obtained from two groups of patients undergoing carotid endarterectomy. The first group included 25 patients who presented TIA or ischaemic stroke. The second group included 35 patients who had an asymptomatic internal carotid artery stenosis. Total RNA was isolated and the expression of Toll-like Receptor 4 (TLR4), COX-2, membrane-associated Prostaglandin E synthase (mPGES-1), Prostaglandin E2 receptors (EP3 and EP4) was analysed by real time RT-PCR. RESULTS: Expression of COX-2 and TLR4 were significantly increased in symptomatic patients (p < 0.001). Correlation analysis showed that TLR4 expression significantly correlated with COX-2 expression (R = 0.65; p < 0.01) in ischaemic stroke patients. This correlation was not observed in TIA and asymptomatic patients. CONCLUSIONS: Our results suggest that the peripheral mechanism of inflammatory injury after stroke may be mediated by TLR4 through a COX-2-dependent pathway.

PTCH1 gene haplotype association with basal cell carcinoma after transplantation.

BACKGROUND: Basal cell carcinoma (BCC) is 10 times more frequent in organ transplant recipients (OTRs) than in the general population. Factors in OTRs conferring increased susceptibility to BCC include ultraviolet radiation exposure, immunosuppression, viral infections such as human papillomavirus, phototype and genetic predisposition. The PTCH1 gene is a negative regulator of the hedgehog pathway, that provides mitogenic signals to basal cells in skin. PTCH1 gene mutations cause naevoid BCC syndrome, and contribute to the development of sporadic BCC and other types of cancers. Associations have been reported between PTCH1 polymorphisms and BCC susceptibility in nontransplanted individuals. OBJECTIVES: To search for novel common polymorphisms in the proximal 5' regulatory region upstream of PTCH1 gene exon 1B, and to investigate the possible association of PTCH1 polymorphisms and haplotypes with BCC risk after organ transplantation. METHODS: Three PTCH1 single nucleotide polymorphisms (rs2297086, rs2066836 and rs357564) were analysed by restriction fragment length polymorphism analysis in 161 northern Italian OTRs (56 BCC cases and 105 controls). Two regions of the PTCH1 gene promoter were screened by heteroduplex analysis in 30 cases and 30 controls. RESULTS: Single locus analysis showed no significant association. Haplotype T(1686)-T(3944) appeared to confer a significantly higher risk for BCC development (odds ratio 2.98, 95% confidence interval 2.55-3.48; P = 0.001). Two novel rare polymorphisms were identified at positions 176 and 179 of the 5'UTR. Two novel alleles of the -4 (CGG)(n) microsatellite were identified. No association of this microsatellite with BCC was observed. CONCLUSIONS: Haplotypes containing T(1686)-T(3944) alleles were shown to be associated with an increased BCC risk in our study population. These data appear to be of great interest for further investigations in a larger group of transplant individuals. Our results do not support the hypothesis that common polymorphisms in the proximal 5' regulatory region of the PTCH1 gene could represent an important risk factor for BCC after organ transplantation.

Método para la evaluación de la confusión en la unidad de cuidados intensivos para el diagnóstico de delírium: adaptación cultural y validación de la versión en idioma español / Confusion assessment method for diagnosing delirium in ICU patients (CAM-ICU): Cultural adaptation and validation of the Spanish version

Objetivo: Realizar la adaptación cultural y la validación del CAM-ICU (Confusion Assessment Method for the Intensive Care Unit «método para la evaluación de la confusión en la unidad de cuidados intensivos») al idioma español para el diagnóstico de delírium. Diseño: Población de validación. Ámbito: La unidad de cuidados intensivos de un hospital universitario de tercer nivel con 600 camas. Pacientes: Veintinueve pacientes críticos que reciben ventilación mecánica. Edad: 70 años (58-77), APACHE II: 16 (13-21) y SOFA: 7 (4-8). Intervención: Aplicación del CAM-ICU en idioma español por 2 operadores independientes y del DSM IV TR (Diagnostic and Statistical Manual of Mental Disorders Fourth Edition-Revised) por un psiquiatra. Variables de interés principales: Concordancia (índice κ), consistencia interna (alfa de Cronbach) y validez (sensibilidad y especificidad) del CAM-ICU en idioma español en relación con el estándar de referencia (DSM IV TR). Resultados: El proceso de traducción y adaptación cultural se desarrolló de acuerdo con las recomendaciones internacionales actualmente vigentes. Se realizó un total de 65 evaluaciones en los 29 pacientes. La concordancia entre los observadores alcanzó un índice K de 0,91 (IC del 95%: 0,86 a 0,96). La consistencia interna del instrumento fue adecuada y alcanzó un alfa de Cronbach de 0,84 (IC del 95% unilateral: 0,77). La sensibilidad del CAM-ICU versión española para el observador A (médico) fue del 80%, con una especificidad del 96%. En tanto que para el observador B (enfermera), la sensibilidad fue del 83%, con una especificidad del 96%. Conclusiones: El CAM-ICU en idioma español es un instrumento válido, confiable y reproducible, que puede aplicarse satisfactoriamente para el diagnóstico de delírium en pacientes de habla hispana (AU)

Objective: To adapt the Confusion Assessment Method for the Intensive Care Unit (CAM-ICU) for the diagnosis of delirium to the language and culture of Spain and to validate the adapted version. Design: Population validation. Setting: Intensive care units in a 600-bed university hospital. Patients: We studied 29 critical patients undergoing mechanical ventilation. Mean age was 70 years (range 58-77 years), mean APACHE II score 16 (range 13-21), and mean SOFA score 7 (range 4-8). Intervention: Two independent operators applied the Spanish version of the CAM-ICU and a psychiatrist applied the Diagnostic and Statistical Manual of Mental Disorders Fourth Edition-Revised (DSM IV TR). Main outcome variables: Concordance (kappa index), internal consistency (Cronbach's alpha), and validity (sensitivity and specificity) of the Spanish version of the CAM-ICU were compared to the DSM IV TR, which is considered the current gold standard. Results: The translation and cultural adaptation was carried out in accordance with current international guidelines. A total of 65 assessments were performed in 29 patients. The interobserver concordance was high: kappa statistic 0.91 (95% CI: 0.86-0.96). The internal consistence was adequate: Cronbach's alpha=0.84 (unilateral 95% CI: 0.77). For observer A (a physician), the sensitivity of the Spanish version of the CAM-ICU was 80% and the specificity was 96%. For observer B, (a nurse) the sensitivity was 83% and the specificity was 96%. Conclusions: The Spanish version of the CAM-ICU is a valid, reliable, and reproducible instrument that can be satisfactorily applied to diagnose delirium in Spanish-speaking ICU patients (AU)

[Confusion Assessment Method for diagnosing delirium in ICU patients (CAM-ICU): cultural adaptation and validation of the Spanish version]. / Método para la evaluación de la confusión en la unidad de cuidados intensivos para el diagnóstico de delírium: adaptación cultural y validación de la versión en idioma español.

OBJECTIVE: To adapt the Confusion Assessment Method for the Intensive Care Unit (CAM-ICU) for the diagnosis of delirium to the language and culture of Spain and to validate the adapted version. DESIGN: Population validation. SETTING: Intensive care units in a 600-bed university hospital. PATIENTS: We studied 29 critical patients undergoing mechanical ventilation. Mean age was 70 years (range 58-77 years), mean APACHE II score 16 (range 13-21), and mean SOFA score 7 (range 4-8). INTERVENTION: Two independent operators applied the Spanish version of the CAM-ICU and a psychiatrist applied the Diagnostic and Statistical Manual of Mental Disorders Fourth Edition-Revised (DSM IV TR). MAIN OUTCOME VARIABLES: Concordance (kappa index), internal consistency (Cronbach's alpha), and validity (sensitivity and specificity) of the Spanish version of the CAM-ICU were compared to the DSM IV TR, which is considered the current gold standard. RESULTS: The translation and cultural adaptation was carried out in accordance with current international guidelines. A total of 65 assessments were performed in 29 patients. The interobserver concordance was high: kappa statistic 0.91 (95% CI: 0.86-0.96). The internal consistence was adequate: Cronbach's alpha=0.84 (unilateral 95% CI: 0.77). For observer A (a physician), the sensitivity of the Spanish version of the CAM-ICU was 80% and the specificity was 96%. For observer B, (a nurse) the sensitivity was 83% and the specificity was 96%. CONCLUSIONS: The Spanish version of the CAM-ICU is a valid, reliable, and reproducible instrument that can be satisfactorily applied to diagnose delirium in Spanish-speaking ICU patients.