Dental Caries: Early Intervention and the Role of the Pediatrician.

Despite improvements in oral morbidity levels and access to care among the pediatric population, there are still major disparities in the United States. Results of national surveys have documented a decrease in the number of children receiving either a dental examination or a cleaning. This finding is particularly concerning for toddlers and infants, as early preventive dental visits and the establishment of a dental home is cost-effective and leads to enhanced oral health outcomes over the life span. Many infants and toddlers do not visit a dentist, suggesting that the recommendations of the American Academy of Pediatrics and the American Academy of Pediatric Dentistry to establish a dental home are not appropriately adopted.

Reliability of a digital system for models measurements in BBO grading: A pilot study.

INTRODUCTION: Currently, no method is considered effective for the evaluation of digital models in the Certification Examination of the Brazilian Board of Orthodontics (BBO), considering the parameters of the currently used manual method. OBJECTIVE: Thus, the aim of this study is to verify the reliability of an evaluation method for digital models that could be used in the BBO exam, compared to the gold standard. METHODS: Measurements were performed by five previously calibrated examiners. Samples of ten sets of plaster models of the final phase of orthodontic treatment were measured using a manual method (Objective Grading System, OGS). These models were digitized using a 3D scanner and exported to Geomagic Qualify software, in which the measurements were made with the proposed digital method. These measurements were repeated using five models, after fifteen days. The intra-examiner performance with this method was analyzed with a paired t-test, whereas the inter-examiner analysis was carried out with analysis of variance and Tukey's test. To compare the manual and digital methods, a paired t-test and Pearson's correlation analysis were performed. RESULTS: A statistically significant difference was found. The results showed that, when compared to the manual method, the digital method was effective in measuring the OGS in four of the seven variables studied: Marginal Ridge, Overjet, Occlusal Contact, and Interproximal Contact. The variables Alignment, BL inclination, and Occlusal Relationship showed a great amount of dispersion in the findings. CONCLUSION: Further studies are needed to develop an adequate digital methodology that can be used for all OGS variables.

Reliability of a digital system for models measurements in BBO grading: A pilot study

ABSTRACT Introduction: Currently, no method is considered effective for the evaluation of digital models in the Certification Examination of the Brazilian Board of Orthodontics (BBO), considering the parameters of the currently used manual method. Objective: Thus, the aim of this study is to verify the reliability of an evaluation method for digital models that could be used in the BBO exam, compared to the gold standard. Methods: Measurements were performed by five previously calibrated examiners. Samples of ten sets of plaster models of the final phase of orthodontic treatment were measured using a manual method (Objective Grading System, OGS). These models were digitized using a 3D scanner and exported to Geomagic Qualify software, in which the measurements were made with the proposed digital method. These measurements were repeated using five models, after fifteen days. The intra-examiner performance with this method was analyzed with a paired t-test, whereas the inter-examiner analysis was carried out with analysis of variance and Tukey's test. To compare the manual and digital methods, a paired t-test and Pearson's correlation analysis were performed. Results: A statistically significant difference was found. The results showed that, when compared to the manual method, the digital method was effective in measuring the OGS in four of the seven variables studied: Marginal Ridge, Overjet, Occlusal Contact, and Interproximal Contact. The variables Alignment, BL inclination, and Occlusal Relationship showed a great amount of dispersion in the findings. Conclusion: Further studies are needed to develop an adequate digital methodology that can be used for all OGS variables.

RESUMO Introdução: Ainda não há um método considerado eficaz para análise dos modelos digitais no exame do Board Brasileiro de Ortodontia (BBO), considerando-se os parâmetros do método manual atual. Objetivo: Assim, o presente estudo objetiva verificar a confiabilidade de um método de avaliação em modelos digitais para o exame do BBO, comparando com o padrão-ouro. Métodos: As medições foram realizadas por 5 examinadores, previamente calibrados. A amostra de 10 pares de modelos de gesso da fase final do tratamento ortodôntico foi medida no método manual (Sistema Objetivo de Avaliação, SOA). Os modelos foram digitalizados por meio de um scanner 3D e exportados para o software Geomagic Qualify, onde foram feitas as medidas no método digital proposto. As medidas foram refeitas em 5 modelos após 15 dias. A análise intraexaminador desse método foi realizada por meio do teste t pareado; já a interexaminadores, feita com ANOVA e teste de Tukey, sendo encontrada diferença estatisticamente significativa. Para a comparação dos métodos manual e digital, foram utilizados o teste t pareado e a correlação de Pearson. Resultados: Uma diferença estatisticamente significativa foi encontrada. Os resultados mostraram que, comparada ao método manual, a metodologia digital mostrou-se eficaz para medição do SOA em quatro das sete variáveis estudadas: Margem interproximal, Sobressaliência, Contato oclusal e Contato interproximal. As variáveis Alinhamento, Inclinação V-L e Relação oclusal mostraram muita dispersão nos achados. Conclusão: Mais estudos são necessários para o desenvolvimento de uma metodologia digital adequada em todas as variáveis do SOA.

Intrastriatal Administration of AAV5-miHTT in Non-Human Primates and Rats Is Well Tolerated and Results in miHTT Transgene Expression in Key Areas of Huntington Disease Pathology.

Huntington disease (HD) is a fatal, neurodegenerative genetic disorder with aggregation of mutant Huntingtin protein (mutHTT) in the brain as a key pathological mechanism. There are currently no disease modifying therapies for HD; however, HTT-lowering therapies hold promise. Recombinant adeno-associated virus serotype 5 expressing a microRNA that targets HTT mRNA (AAV5-miHTT) is in development for the treatment of HD with promising results in rodent and minipig HD models. To support a clinical trial, toxicity studies were performed in non-human primates (NHP, Macaca fascicularis) and Sprague-Dawley rats to evaluate the safety of AAV5-miHTT, the neurosurgical administration procedure, vector delivery and expression of the miHTT transgene during a 6-month observation period. For accurate delivery of AAV5-miHTT to the striatum, real-time magnetic resonance imaging (MRI) with convection-enhanced delivery (CED) was used in NHP. Catheters were successfully implanted in 24 NHP, without neurological symptoms, and resulted in tracer signal in the target areas. Widespread vector DNA and miHTT transgene distribution in the brain was found, particularly in areas associated with HD pathology. Intrastriatal administration of AAV5-miHTT was well tolerated with no clinically relevant changes in either species. These studies demonstrate the excellent safety profile of AAV5-miHTT, the reproducibility and tolerability of intrastriatal administration, and the delivery of AAV5-miHTT to the brain, which support the transition of AAV5-miHTT into clinical studies.

A Novel Organoid Model of Damage and Repair Identifies HNF4α as a Critical Regulator of Intestinal Epithelial Regeneration.

BACKGROUND & AIMS: Recent evidence has suggested that the intact intestinal epithelial barrier protects our body from a range of immune-mediated diseases. The epithelial layer has an impressive ability to reconstitute and repair upon damage and this process of repair increasingly is seen as a therapeutic target. In vitro models to study this process in primary intestinal cells are lacking. METHODS: We established and characterized an in vitro model of intestinal damage and repair by applying γ-radiation on small-intestinal organoids. We then used this model to identify novel regulators of intestinal regeneration. RESULTS: We identified hepatocyte nuclear factor 4α (HNF4α) as a pivotal upstream regulator of the intestinal regenerative response. Organoids lacking Hnf4a were not able to propagate in vitro. Importantly, intestinal Hnf4a knock-out mice showed impaired regeneration after whole-body irradiation, confirming intestinal organoids as a valuable alternative to in vivo studies. CONCLUSIONS: In conclusion, we established and validated an in vitro damage-repair model and identified HNF4α as a crucial regulator of intestinal regeneration. Transcript profiling: GSE141515 and GSE141518.

[Liver transplantation: development, learning curve and results after the first 300 cases]. / Trasplante hepático: evolución, curva de aprendizaje y resultados después de los primeros 300 casos.

BACKGROUND: Liver transplantation (LT) is an option for people with liver failure who cannot be cured with other therapies and for some people with liver cancer. AIM: To describe, and analyze the first 300 LT clinical results, and to establish our learning curve. MATERIAL AND METHODS: Retrospective cohort study with data obtained from a prospectively collected LT Program database. We included all LT performed at a single center from March 1994 to September 2017. The database gathered demographics, diagnosis, indications for LT, surgical aspects and postoperative courses. We constructed a cumulative summation test for learning curve (LC-CUSUM) using 30-day post-LT mortality. Mortality at 30 days, and actuarial 1-, and 5-year survival rate were analyzed. RESULTS: A total of 281 patients aged 54 (0-71) years (129 women) underwent 300 LT. Ten percent of patients were younger than 18 years old. The first, second and third indications for LT were non-alcoholic steatohepatitis, chronic autoimmune hepatitis and alcoholic liver cirrhosis, respectively. Acute liver failure was the LT indication in 51 cases (17%). The overall complication rate was 71%. Infectious and biliary complications were the most common of them (47 and 31% respectively). The LC-CUSUM curve shows that the first 30 patients corresponded to the learning curve. The peri-operative mortality was 8%. Actuarial 1 and 5-year survival rates were 82 and 71.4%, respectively. CONCLUSIONS: Outcome improvement of a LT program depends on the accumulation of experience after the first 30 transplants and the peri-operative mortality directly impacted long-term survival.

Enhanced Factor IX Activity following Administration of AAV5-R338L "Padua" Factor IX versus AAV5 WT Human Factor IX in NHPs.

Gene therapy for severe hemophilia B is advancing and offers sustained disease amelioration with a single treatment. We have reported the efficacy and safety of AMT-060, an investigational gene therapy comprising an adeno-associated virus serotype 5 capsid encapsidating the codon-optimized wild-type human factor IX (WT hFIX) gene with a liver-specific promoter, in patients with severe hemophilia B. Treatment with 2 × 1013 gc/kg AMT-060 showed sustained and durable FIX activity of 3%-13% and a substantial reduction in spontaneous bleeds without T cell-mediated hepatoxicity. To achieve higher FIX activity, we modified AMT-060 to encode the R338L "Padua" FIX variant that has increased specific activity (AMT-061). We report the safety and increased FIX activity of AMT-061 in non-human primates. Animals (n = 3/group) received intravenous AMT-060 (5 × 1012 gc/kg), AMT-061 (ranging from 5 × 1011 to 9 × 1013 gc/kg), or vehicle. Human FIX protein expression, FIX activity, and coagulation markers including D-dimer and thrombin-antithrombin complexes were measured. At equal doses, AMT-060 and AMT-061 resulted in similar human FIX protein expression, but FIX activity was 6.5-fold enhanced using AMT-061. Both vectors show similar safety and transduction profiles. Thus, AMT-061 holds great promise as a more potent FIX replacement gene therapy with a favorable safety profile.

Trasplante hepático: evolución, curva de aprendizaje y resultados después de los primeros 300 casos / Liver transplantation: development, learning curve and results after the first 300 cases

Background: Liver transplantation (LT) is an option for people with liver failure who cannot be cured with other therapies and for some people with liver cancer. Aim: To describe, and analyze the first 300 LT clinical results, and to establish our learning curve. Material and Methods: Retrospective cohort study with data obtained from a prospectively collected LT Program database. We included all LT performed at a single center from March 1994 to September 2017. The database gathered demographics, diagnosis, indications for LT, surgical aspects and postoperative courses. We constructed a cumulative summation test for learning curve (LC-CUSUM) using 30-day post-LT mortality. Mortality at 30 days, and actuarial 1-, and 5-year survival rate were analyzed. Results: A total of 281 patients aged 54 (0-71) years (129 women) underwent 300 LT. Ten percent of patients were younger than 18 years old. The first, second and third indications for LT were non-alcoholic steatohepatitis, chronic autoimmune hepatitis and alcoholic liver cirrhosis, respectively. Acute liver failure was the LT indication in 51 cases (17%). The overall complication rate was 71%. Infectious and biliary complications were the most common of them (47 and 31% respectively). The LC-CUSUM curve shows that the first 30 patients corresponded to the learning curve. The peri-operative mortality was 8%. Actuarial 1 and 5-year survival rates were 82 and 71.4%, respectively. Conclusions: Outcome improvement of a LT program depends on the accumulation of experience after the first 30 transplants and the peri-operative mortality directly impacted long-term survival.

Determinación de las medidas de bioseguridad en clínicas y consultorios de pequeños animales en la ciudad de pasto, nariño / Determination of biosecurity measures in small animals clinics and veterinary offices in pasto, nariño

La medicina veterinaria es considerada como una de las profesiones más expuestas a sufrir daños a la salud humana como consecuencia de accidentes y enfermedades profesionales relacionadas con los animales, las enfermedades zoonóticas son el principal riesgo laboral que enfrenta el personal veterinario a diario. El presente estudio se realizó en la ciudad de Pasto, en ocho clínicas y consultorios veterinarios, con el objetivo de determinar las medidas de bioseguridad que implementan durante la prestación de sus servicios profesionales. La recolección de los datos se realizó mediante encuestas para determinar las medidas profilácticas que usan los médicos veterinarios en su ejercicio profesional para prevenir el contagio de enfermedades zoonóticas. Los resultados obtenidos a partir de la encuesta personal revelan que el 87,5% de los profesionales desconocen los alcances de la ley 776 de 2002 sobre accidentes de trabajo y enfermedades profesionales. Las enfermedades zoonóticas que conocen son principalmente rabia (100%) y brucelosis (87,5%). El 87,5% de los profesionales se encuentran vacunados profilácticamente contra tétanos y solo el 25% contra rabia. El 37% de los profesionales ha sido afectado por alguna enfermedad zoonótica durante el tiempo de su ejercicio profesional. Las principales medidas profilácticas implementadas son el lavado de manos y depósito de material cortopunzantes en recipientes adecuados. El uso de barreras de protección varía dependiendo del procedimiento y tipo de paciente. En conclusión, los profesionales encuestados dedicados a clínica y cirugía de pequeñas especies, conocen los riesgos a los que están expuestos en su práctica profesional y la necesidad de implementar medidas profilácticas, sin embargo no cuentan con un esquema de bioseguridad que sigan estrictamente.

Veterinary medicine is considered one of the most vulnerable professions to human health damage as a result of occupational accidents and diseases related to animals, being zoonotic diseases the primary occupational hazard faced by the veterinary staff daily. This study was conducted in the city of Pasto, in eight clinics and veterinary offices with the purpose of determining the biosecurity measures implemented during the provision of professional services. The collection of data was done using surveys to determine the protective measures used by veterinarians in their professional practice to prevent the spread of zoonotic diseases. The results obtained from the study show that 87.5% of the professionals are unaware of the scope of law 776 of 2002 on occupational accidents and diseases. Zoonotic diseases mainly known by veterinaries are rabies (100%) and brucellosis (87.5%). The 87.5% of the professionals surveyed are prophylactically vaccinated against tetanus and only 25% against rabies. A 37% of professionals have been affected by any zoonotic disease at the time of their professional practice. The primary preventive measures implemented are handwashing and deposit of sharp pointed material into suitable containers. The use of protective barriers varies depending on the method and type of patient. In conclusion, professionals surveyed dedicated to clinical and surgery of small species in Pasto know the risks they are exposed to in their professional practice and the need to implement prophylactic measures, but they do not have a biosafety scheme to follow strictly.

Perfil dos pacientes e contexto das consultas em que se fez pela primeira vez o diagnóstico de depressão no Centro de Saúde de Eiras, durante o ano de 2011 / Patient's profile and the context of consultations in which the diagnose of depression was made for the first time in Eiras Health Centre in 2011 / Perfil de losusuarios y contexto de las consultas en que se hizoel primer diagnóstico de depresiónenel Centro de Salud de Eiras durante elaño de 2011

Objetivo: Este estudo pretendeu conhecer aspectos dos utentes e consultas em que se fez pela primeira vez o diagnóstico de depressão. Métodos: Estudo observacional, transversal e descritivo. População obtida por aplicação de critérios de exclusão aos utentes que, em consulta durante 2011, tiveram em "Avaliação" a codificação "Perturbações depressivas". Pela análise do registro da consulta, estudaram-se as variáveis: idade, sexo, mês, tipo de consulta, consulta presencial/não presencial, sinais/sintomas depressivos anotados e/ou codificados, prescrição de psicofármacos, prescrição pela primeira vez/renovação de receituário. No caso de prescrição pela primeira vez: psicofármacos segundo grupo farmacológico e Denominação Comum Internacional, prescrição de antidepressivo na dose terapêutica e referência ao tempo de tratamento antidepressivo. Resultados: População de 105 indivíduos. Consultas maioritariamente presenciais (79%). Maior codificação de sinais/sintomas depressivos que anotação apenas ou que anotação e codificação. O sinal/sintoma mais codificado foi "Sensação de depressão" (28%). Houve prescrição de ansiolíticos isoladamente e um caso de prescrição subterapêutica do antidepressivo. Quanto à duração do tratamento antidepressivo, em 13,7% das receitas houve menção de que o tratamento deveria prolongar-se no mínimo por 6 meses. Conclusão: A obtenção de uma população pequena e possíveis vieses de informação foram limitações encontradas. Achamos curioso que o sinal/sintoma depressivo mais codificado fosse "Sensação de depressão". É necessário melhorar os registros clínicos e prescrição na depressão.

Objective: This investigation intended to know about aspects about patients and consultations in which depression was diagnosed for the first time. Methods: Observational, cross-sectional and descriptive study. Population obtained through the application of exclusion criteria in patients that during 2011 were in "Evaluation" with the codification "Depressive disturbance". Through the analysis of the consultation, we studied the variables: age, sex, month, type of consultation, consultation in the presence of the patient/patient not attending, depressive signs/symptoms noted and/or coded, prescription of psychiatric medication, if it was prescribed for the first time/renewed. In case it is prescribed for the first time: group of medication and name of the drug. If antidepressant was prescribed for the first time: therapeutic dosage/not and mention the time of treatment. Results: Population of 105 individuals. The consultations were mainly in the presence of the patient (79%). More encoding of depressive signs/symptoms than annotation only or than annotation and codification. The most coded depressive sign/symptom was "Sensation of depression" (28%). There was prescription of isolated anxiolytics and a case of under-therapeutic prescription of antidepressant. 13.7% of the prescription had the reference that the antidepressive treatment should last at least 6 months. Conclusion: Obtainment of a small population and possible information bias were limitations encountered. It is worthy to mention that the most coded depressive sign/symptom was "Sensation of depression". We must improve our clinical records and the prescription in depression.

Objetivo: Este estudio se destinó a comprender los aspectos de los usuarios y las consultas en que se hizo el primer diagnóstico de depresión. Métodos: Se realizó un estudio observacional, transversal y descriptivo. Población obtenida mediante la aplicación de criterios de exclusión a los usuarios que en consulta durante 2011, tuvieran la codificación "trastornos depresivos". Por el análisis de registros de consulta, se estudiaron las variables: edad, sexo, mes, tipo de consulta, consulta presencial/no, signos/síntomas depresivos anotados y/o codificados, la prescripción psicotrópica, si esa prescripción fue por la primera vez/renovación de la prescripción. En el caso de la prescripción por la primera vez: los psicotrópicos según grupo farmacológico y las denominaciones comunes internacionales, referencia, en la prescripción de terapia antidepresiva, a la dosis y tiempo del tratamiento. Resultados: Población de 105 individuos. Sobre todo consultas presenciales (79%). Más codificación de señales/síntomas depresivos que la anotación sola o que anotación y codificación. El signo/síntoma que se codificó más fue "Sentirse deprimido" (28%). Fueron prescritos ansiolíticos aislados y hubo un caso de prescripción sub-terapéutica de antidepresivo. En cuanto a duración del tratamiento antidepresivo, en 13,7% de las prescripciones se mencionaba que el tratamiento debía extenderse por lo menos 6 meses. Conclusión: La obtención de una pequeña población y los posibles sesgos de información fueron limitaciones encontradas. Fue curioso el hecho de que la señal/síntoma más codificada fue "Sentirse deprimido". Es necesario mejorar las historias clínicas y la prescripción en depresión.

Percepção do ruído ocupacional e perda auditiva em estudantes de Odontologia / Perception of occupational noise and hearing loss in dental students

Este estudo teve como objetivo analisar a percepção do ruído ocupacional e a perda auditiva em estudantes de Odontologia de uma instituição pública. O estudo transversal foi realizado em Parnaíba, Brasil, em uma amostra intencional com alunos a partir do terceiro ano do curso (n= 62). Os dados coletados em questionários autoaplicáveis foram analisados pelos Testes do Quiquadrado de Pearson e Exato de Fisher. Os estudantes consideraram o ruído ocupacional nocivo à saúde (82,3%) e o cirurgião-dentista susceptível à perda auditiva (88,7%). As queixas mais relatadas foram cefaleia constante (35,5%) e irritação/nervosismo (32,3%). Medidas de prevenção não são utilizadas (98,4%), apesar de serem conhecidas (75,8%). Os estudantes já receberam informações sobre a perda auditiva (53,2%), a maioria na própria Instituição de Ensino Superior (81,8%). Audiometria nunca foi realizada (80,6%). A legislação sobre tolerância ao ruído não é conhecida (77,4%) e este conhecimento foi estatisticamente associado com o período em curso (p=0,004). Os estudantes de Odontologia desta instituição conhecem as consequências e medidas de prevenção, embora não as utilizem (AU).

This study aimed to analyze the perception of occupational noise and hearing loss in dental students of a public institution. The crosssectional study was conducted in Parnaiba, Brazil, in intentional sample with of third year students (n = 62). Data collected through questionnaires were analyzed by Pearson's chi-square and Fisher's exact test. Students considered the occupational noise harmful to health (82.3%) and the dentist susceptible to hearing loss (88.7%). The most frequently reported complaints were constant headache (35.5%) and irritation/nervousness (32.3%). Preventative measures aren't used (98.4%), despite being known (75.8%). Students already have received information about hearing loss (53.2%), most of on own institution of higher education (81.8%). Audiometry has never been performed (80.6%). The legislation concerning tolerance to noise isn't known (77.4%) and this knowledge was statistically associated with the course period (p=0,004). Dental students at this institution know the consequences and preventive measures, although don't use it (AU).

A informação na consulta presencial em Medicina Geral e Familiar: classificações segundo a ICPC-2 e anotações livres para a memória futura no SOAP / Information available in general practice/family medicine consultations: coding and annotations for future use / La información en la consulta médica presencial en Medicina General y Familiar: clasificaciones según ICPC-2 y apuntes libres para la memoria futura en el SOAP

Objetivos: assumindo a obrigatoriedade de classificação ICPC-2 em cada consulta, conhecer a informação, em consultas passadas pelo método de Weed-SOAP segundo o gênero e idade de quem consulta o médico (consulente ou paciente), caracterizando o nível de registro pelo método SOAP em Subjetivo (S) - classificação e anotações - em Objetivo (O) anotações sobre o estado do paciente, em Avaliação (A) da classificação e em Plano (P) da classificação e anotações. Métodos: estudo observacional, transversal em outubro de 2012, em amostra aleatorizada das consultas presenciais de dois médicos orientadores de internato de especialidade, em três meses sorteados do 1º semestre de 2012, e em quatro dias sorteados em cada mês, em amostra representativa com intervalo de confiança de 95% e margem de erro de 6%. Utilizou-se estatística descritiva e inferencial. Resultados: amostra de 318 consultas, n=149 (46,9%) no gênero masculino, n=61 (19,2%) no grupo etário <18 anos e n=194 (61,0%) no ≥18 e <65 anos, ns por grupos etários e gênero. Em S, há classificação em 98,7% e anotação em 47,2% das consultas; Em O, verificamos "As anotações demonstram o estado do paciente" em 66,0% e "As anotações são explícitas e entendíveis" em 79,9%; em A, 97,8% das consultas têm classificação; Em P, há classificação em 96,5% e anotações explicando o plano em 23,0% das consultas. Distribuição sem significado por grupo etário para as variáveis estudadas. É mais frequente haver no gênero feminino em S "As anotações são explícitas e entendíveis" e em P "Há classificação de procedimentos". Conclusão: há campo para mais completa coleta da informação na consulta, permitindo, assim, melhor conhecimento de cada consulta e caso para o futuro.

Objectives: assuming the mandatoriness of ICPC2 classification in every consultation, the objective of this study was to ascertain the frequency of this type of classification in past consultations. This analysis was performed using the Weed-SOAP method, where Subjective (S) is measured by classifications and annotations, Objective (O) by annotations, Avaliation (A) by classification, and Plan (P) by classification and annotations. Methods: a cross-sectional observational study was performed in October 2012, with a 95% confidence interval and 6% error margin, on a representative random sample of consultations conducted on 4 random days of 3 random months of the first semester of 2012, and data were analyzed using descriptive and inferential statistics. Results: among the sample of 318 consultations, 149 (46.9%) were with male patients and 61 (19.2%) were aged under 18 years, while 194 (61.0%) were aged 18 or above but under 65 years, after sorting by age groups and gender. In terms of S, 98.7% of consultations had an ICPC-2 classification and 47.2% had an annotation; in terms of O, 66.0% had an annotation demonstrating the state of the patient and 79.9% were explicit; in terms of A, 97.8% had a classification; and in terms of P, 96.5% had a classification and 23.0% had an annotation explaining the plan. There was no statistically significant difference by age group for the studied variables. However, for gender, women had more "Clear and explicit" annotations in S, and "Classification of plan" instances in P. Conclusion: more complete recall of information in consultations is required in order to gain better knowledge about individual consultations and patients for future use.

Objetivos: asumiendo la obligatoriedad de clasificar con la ICPC-2 en cada consulta, conocer la información, en consultas tras-efectuadas pelo método Weed-SOAP según el género y edad de quien consulta al médico (los pacientes), caracterizando el nivel de registro por el método SOAP en Subjetivo (S) - clasificación y anotaciones - en Objetivo (O) apuntes sobre el estado del paciente, en Evaluación (A) de la clasificación y en Pleno (P) de la clasificación y apuntes. Métodos: estudio observacional, transversal en octubre del 2012, en muestra aleatoria de las consultas presenciales de dos médicos tutores de residencia de la especialidad, por tres meses sorteados del 1º semestre del 2012 y en cuatro días sorteados en cada mes, en muestra representativa con intervalo de seguridad del 95% y margen de error del 6%. Se utilizó estadística descriptiva e inferencial. Resultados: muestra de 318 consultas, n=149 (46,9%) en el género masculino, n=61 (19,2%) en el grupo de edad <18 años y n=194 (61,0%) en el ≥18 y <65 años, ns por grupos de edad y género. En S, hay clasificación en el 98,7% y apuntes en el 47,2% de las consultas; En O, verificamos "Los apuntes demuestran el estado del paciente" en 66,0% y "Los apuntes son explícitos y comprensibles" en el 79,9%; en A, el 97,8% de las consultas tienen clasificación; En P, hay clasificación en el 96,5% y apuntes explicando el plan en el 23,0% de las consultas. Distribución sin significado por grupo de edad para las variables estudiadas. Es más frecuente haber en el género femenino en S "Los apuntes son explícitos y comprensibles" y en P "Hay clasificación de procedimientos". Conclusión: hay campo para un recogido de informaciones más completo, permitiendo de esta manera, mejor conocimiento de cada consulta en el futuro.

A protocol for lentiviral transduction and downstream analysis of intestinal organoids.

Intestinal crypt-villus structures termed organoids, can be kept in sustained culture three dimensionally when supplemented with the appropriate growth factors. Since organoids are highly similar to the original tissue in terms of homeostatic stem cell differentiation, cell polarity and presence of all terminally differentiated cell types known to the adult intestinal epithelium, they serve as an essential resource in experimental research on the epithelium. The possibility to express transgenes or interfering RNA using lentiviral or retroviral vectors in organoids has increased opportunities for functional analysis of the intestinal epithelium and intestinal stem cells, surpassing traditional mouse transgenics in speed and cost. In the current video protocol we show how to utilize transduction of small intestinal organoids with lentiviral vectors illustrated by use of doxycylin inducible transgenes, or IPTG inducible short hairpin RNA for overexpression or gene knockdown. Furthermore, considering organoid culture yields minute cell counts that may even be reduced by experimental treatment, we explain how to process organoids for downstream analysis aimed at quantitative RT-PCR, RNA-microarray and immunohistochemistry. Techniques that enable transgene expression and gene knock down in intestinal organoids contribute to the research potential that these intestinal epithelial structures hold, establishing organoid culture as a new standard in cell culture.

Polyinosinic acid blocks adeno-associated virus macrophage endocytosis in vitro and enhances adeno-associated virus liver-directed gene therapy in vivo.

Adeno-associated virus serotype 8 (AAV8) has been demonstrated to be effective for liver-directed gene therapy in humans. Although hepatocytes are the main target cell for AAV8, there is a loss of the viral vector because of uptake by macrophages and Kupffer cells. Reducing this loss would increase the efficacy of viral gene therapy and allow a dose reduction. The receptor mediating this uptake has not been identified; a potential candidate seems the macrophage scavenger receptor A (SR-A) that is involved in the endocytosis of, for instance, adenovirus. In this study we show that SR-A can mediate scAAV8 endocytosis and that blocking it with polyinosinic acid (poly[i]) reduces endocytosis significantly in vitro. Subsequently, we demonstrate that blocking this receptor improves scAAV-mediated liver-directed gene therapy in a model for inherited hyperbilirubinemia, the uridine diphospho-glucuronyl transferase 1A1-deficient Gunn rat. In male rats, preadministration of poly[i] increases the efficacy of a low dose (1×10¹¹ gc/kg) but not of a higher dose (3×10¹¹ gc/kg) scAAV8-LP1-UT1A1. Administration of poly[i] just before the vector significantly increases the correction of serum bilirubin in female rats. In these, the effect of poly[i] is seen by both doses but is more pronounced in the females receiving the low vector, where it also results in a significant increase of bilirubin glucuronides in bile. In conclusion, this study shows that SR-A mediates the endocytosis of AAV8 in vitro and in vivo and that blocking this receptor can improve the efficacy of AAV-mediated liver-directed gene therapy.

Peripheral blood pressure by Dinamap and central blood pressure by applanation tonometry in outpatient general practice.

INTRODUCTION: Central blood pressure (CBP) is the pressure exerted by the blood column at any given moment on the aortic and carotid artery walls, which is a close proxy for the blood pressure inside the brain and the heart, and is thus a better marker of cardiovascular morbidity and mortality than peripheral blood pressure (PBP). OBJECTIVE: To assess how the augmentation index (AI), peripheral pulse pressure (pPP), central pulse pressure (cPP) and subendocardial viability ratio (SEVR) vary in hypertensive patients according to level of control of CBP and PBP. METHODS: We performed an observational, cross-sectional study in a convenience sample from a general practice in Central Portugal over a period of four days in May 2010. Measurements were taken after a four-minute resting period. The following values were considered to reflect controlled pressures: PBP <140/90 mmHg, CBP <130/80 mmHg, pPP <55 mmHg and cPP <45 mmHg. RESULTS: The sample included 92 patients, 38 male (41.3%), mean age 62.3±11.1 years, with no significant difference in gender distribution. PBP was controlled in 55 (59.8%), and CBP in 53 (57.6%). Both PBP and CBP were controlled in 50 patients (54.3%) and neither was controlled in 34 (37.9%). pPP and cPP were significantly lower in those with controlled PBP (p<0.001) and CBP (p<0.001). AI was non-significantly lower in those with controlled PBP (78±9 vs. 80.7) and those with controlled CBP (78±9 vs.81±7) (p=0.02). SEVR was within the desirable range in 92 patients (92.2%). 78.4% of individuals were taking drugs acting on the renin angiotensin aldosterone system (RAAS). CONCLUSIONS: In a convenience sample of 92 patients, PBP and CBP were controlled in 59.8% and 57.6%, respectively. Those with controlled PBP had significantly better peripheral systolic and diastolic blood pressure, CBP, pPP and cPP; the same was true of those with controlled CBP, who also had a significantly better AI. The percentage of the cardiac cycle in diastole had a desirable value for 92,2% of the subjects.

Tendência de classificação no Capítulo Z da CIAP-2 entre 2006 e 2011 em um centro de saúde de Medicina Familiar em Coimbra, Portugal / Classification trends with the ICPC-2 Z Chapter from 2006 to 2011 in a Family Medicine Health Centre in Coimbra, Portugal / Tendencia de la clasificación en el Capitulo Z de la CIAP-2 entre 2006 y 2011 en un Centro de Salud de Medicina Familiar en Coimbra, Portugal

Objetivo: Estudar a tendência evolutiva da classificação com a CIAP-2 no Capítulo Z, no período de 2006 a 2011 quanto a: variação de frequência do número total de componentes de sinais e sintomas; volume de "classificação ajustada à população" e, através da coleta de informação SOAP, quais dos seus componentes foram mais frequentemente registrados nos campos Subjetivo (S) e Avaliação (A). Métodos: Estudo transversal, observacional e descritivo dos registros eletrônicos efetuados por todos os médicos na plataforma Serviço de Apoio ao Médico (SAM) utilizando a ferramenta de coleta de dados (SAM-Estatísticas) em agosto de 2012 em um centro de saúde em Coimbra, região central de Portugal. O volume de codificação foi estudado em código/1.000 hab./dia, tendo como base de cálculo a metade de cada ano estudado. Foram selecionados os seis códigos mais frequentes de cada ano. Resultados: Verificou-se uma dinâmica de crescimento positivo nos componentes registrados nos campos Subjetivo e Avaliação entre 2006 e 2011, tanto no número total de códigos (S:+4,83 e A:+6,44) e volume de codificação ­ código/1.000 hab./dia (S:+4,40 e A:+6,44) como na percentagem de diferentes componentes de sinais e sintomas do Capítulo Z (S:+0,30; A:+0,56). Conclusão: Entre 2006 e 2011 verificou-se uma dinâmica de crescimento positivo na classificação no Capítulo Z da CIAP-2 que foi mais importante no campo Avaliação e nos tipos de componente. O desenvolvimento profissional e a educação médica continuada são necessários para melhorar o desempenho na tarefa de classificar e registrar adequadamente, bem como no criterioso registro das anotações clínicas feitas na consulta, evitando perda de informação clínica.

Objective: To study the classification trend in ICPC-2, Z Chapter, in the period 2006-2011, in terms of frequency in variation of the total number of sign and symptom components; volume of "population-adjusted classification"; and - based on SOAP for collecting information - which of its components were more frequently recorded in the Subjective (S) and Assessment (A) fields. Methods: Cross-sectional, observational, descriptive study of electronic registries in the SAM (Serviço de Apoio ao Médico - medical visit assistant software) using data collection tool (SAM-Statistics) in August 2012 in Coimbra, central Portugal. Classification volume was studied using codes/1000 inhab/day. The population for each year was calculated based on the middle of each study period. The six most frequent codes in each year were selected. Results: A positive increasing trend was found between 2006 and 2011 in the Subjective and Assessment recording fields, in terms of the total of codes (S:+4,83; A:+6,44) and classification volume - codes/1000 inhab/day (S:+4,40; A:+6,44), as well as in the percentage of different components of the Z sign and symptom codes (S: +0,30; A: +0,56). Conclusions: From 2006 to 2011, a positive trend was found in the ICPC-2 Z Chapter classification, which was more important in Assessment (A), and the type of components recorded through the SOAP method. Hence, continuing medical education is still necessary to improve performance in the crucial task of classification, registering and clinical noting to avoid lack of clinical information.

Objetivo: Estudiar la tendencia evolutiva de la clasificación con CIAP-2 en el capítulo Z, en el período 2006-2011, en términos de: variación de la frecuencia del número total de los componentes de los signos y síntomas; el volumen de "clasificación ajustada a la población" y, a través de la recolección de información SOAP, cuáles de sus componentes se registraron con mayor frecuencia en los campos Subjetivo (S) y Evaluación (E). Métodos: Estudio transversal, observacional y descriptivo de los registros electrónicos realizados por todos los médicos en la plataforma Servicio de Apoyo Médico (SAM) en agosto de 2012 en un centro de salud de Coimbra, región central de Portugal, utilizando la herramienta de recolección de datos (SAM-estadística). El volumen de codificación fue estudiado en código/1000 hab./día, teniendo como base de cálculo la mitad de cada año estudiado. Fueron seleccionados los seis códigos más comunes de cada año. Resultados: Se observó una dinámica de crecimiento positivo, entre 2006-2011, de los componentes registrados en los campos Subjetivo y Evaluación, tanto en el número total de códigos (S: +4,83; E: +6,44) y volumen de codificación - código/1000/hab./día (S: +4,40; E: +6,44), como en el porcentaje de los diversos componentes de los signos y síntomas del Capítulo Z (S: +0,30; E: +0,56). Conclusión: Entre los años 2006 y 2011 hubo una dinámica de crecimiento positivo de la clasificación en el capítulo Z de la CIAP-2, más importante en el campo Evaluación y en los tipos de componente. El desarrollo profesional y la educación médica continua son necesarios para mejorar el rendimiento en la tarea de clasificar y registrar correctamente, así como para el registro cuidadoso de las notas clínicas realizadas durante las consultas, evitando, así, la pérdida de información clínica.

Adeno-associated viral vector serotype 5 poorly transduces liver in rat models.

Preclinical studies in mice and non-human primates showed that AAV serotype 5 provides efficient liver transduction and as such seems a promising vector for liver directed gene therapy. An advantage of AAV5 compared to serotype 8 already shown to provide efficient correction in a phase 1 trial in patients suffering from hemophilia B, is its lower seroprevalence in the general population. Our goal is liver directed gene therapy for Crigler-Najjar syndrome type I, inherited severe unconjugated hyperbilirubinemia caused by UGT1A1 deficiency. In a relevant animal model, the Gunn rat, we compared the efficacy of AAV 5 and 8 to that of AAV1 previously shown to be effective. Ferrying a construct driving hepatocyte specific expression of UGT1A1, both AAV8 and AAV1 provided an efficient correction of hyperbilirubinemia. In contrast to these two and to other animal models AAV5 failed to provide any correction. To clarify whether this unexpected finding was due to the rat model used or due to a problem with AAV5, the efficacy of this serotype was compared in a mouse and two additional rat strains. Administration of an AAV5 vector expressing luciferase under the control of a liver specific promoter confirmed that this serotype poorly performed in rat liver, rendering it not suitable for proof of concept studies in this species.