Quantitative and qualitative evaluation of microorganism profile identified in bioburden analysis in a biopharmaceutical facility in Brazil: Criteria for classification and management of results.

Microbiological contamination may cause microbial proliferation and consequently additional problems for pharmaceutical companies through production stoppage, product contamination, investigations of process deviations, out-of-specification results and product disposal. This is one of the major concerns of the regulatory health agencies. Microbiological load (bioburden) may represent a potential risk for patients if the sterilization process is not effective and/or due to the production of toxins. Although bioburden can be eliminated by terminal sterilization or filtration processes, it is important to monitor the amount and determine the identity and characteristics of the microorganisms present prior to final processing. The application of microorganism identification systems is crucial for identifying the type of contamination, which can be extremely useful for investigating. The aim of this study was to evaluate the profiles of microorganisms identified in bioburden assays from solutions, culture medias, and products (SCP) from a pharmaceutical industry facility. From 2018-2020, a total of 1,078 samples from 857 different lots of SCP were analyzed and isolated microorganisms were identified. A prefiltering step was included after March 2020, in order to reduce the bioburden before sterilizing filtration. Criteria for the definition and management of microorganisms identified were evaluated after an integrative bibliographic review, and three groups were proposed (critical, objectionable, and nonobjectionable microorganisms). For the samples that did not include prefiltering (n=636), 227 (35.7%) presented microbial growth. For those that included prefiltering, before prefiltering (n=221), 60.6% presented microbial growth, and after prefiltering, this value was reduced to 4.1%, which can be attributed to a contamination during the sampling or a wrong filtering. From the samples that presented microbial growth, 678 microorganisms were identified as bacteria and 59 as molds and yeasts. A total of 120 microorganisms (56 and 27 Gram-positive and negative bacteria, respectively, 31 yeasts, and six filamentous molds) could not be identified, and the remaining microorganisms were classified as objectionable (n=507; 82.2%), nonobjectionable (n=103; 16.7%) and critical (n=7; 1.1%). Most of the bioburden species (>80.0%) were considered objectionable microorganisms. A process for classification and management of bioburden analysis results based on a literature review of pathogenic and physiological characteristics of the microorganisms was proposed.

Alignment of multiple metabolomics LC-MS datasets from disparate diseases to reveal fever-associated metabolites.

Acute febrile illnesses are still a major cause of mortality and morbidity globally, particularly in low to middle income countries. The aim of this study was to determine any possible metabolic commonalities of patients infected with disparate pathogens that cause fever. Three liquid chromatography-mass spectrometry (LC-MS) datasets investigating the metabolic effects of malaria, leishmaniasis and Zika virus infection were used. The retention time (RT) drift between the datasets was determined using landmarks obtained from the internal standards generally used in the quality control of the LC-MS experiments. Fitted Gaussian Process models (GPs) were used to perform a high level correction of the RT drift between the experiments, which was followed by standard peakset alignment between the samples with corrected RTs of the three LC-MS datasets. Statistical analysis, annotation and pathway analysis of the integrated peaksets were subsequently performed. Metabolic dysregulation patterns common across the datasets were identified, with kynurenine pathway being the most affected pathway between all three fever-associated datasets.

Evaluation of MALDI-TOF MS, sequencing of D2 LSU rRNA and internal transcribed spacer regions (ITS) for the identification of filamentous fungi isolated from a pharmaceutical facility.

The identification of filamentous fungi through culture characterization may be hampered by phenotypic variability. Information obtained from the identification of microorganisms are important for investigation of sources of contamination of a product or process. The aim of this study was to identify filamentous fungal strains (n = 50) isolated from a pharmaceutical facility by using Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), as well as D2 domain of the large-subunit (LSU) ribosomal RNA gene and internal transcribed spacer regions (ITS) sequencing. MALDI-TOF MS system only identified five strains at the species level, while 45 were not identified. The analysis through GenBank allowed the identification of up to 19 strains at the species level, while MycoBank allowed the identification of up to nine strains at the species level. The databases identified up to 11 genera: Penicillium, Aspergillus, Cladosporium, Chaetomium, Coniochaeta, Curvularia, Diaporthe, Fusarium, Trichoderma, Rhizopus and Microdochium. MALDI-TOF MS showed an insufficient database to identify the species of fungi. DNA sequencing was the best methodology to identify to the genus level but was unable to differentiate between closely related species. Therefore further methods for the identification of filamentous fungi from pharmaceutical areas at species level need to be developed.

Identification of Sutcliffiella horikoshii strains in an immunobiological pharmaceutical industry facility.

The pharmaceutical industry must comply with the requirements for good manufacturing practices to reduce inherent contamination risks in the production process. Bacillus and related genera are among the main bacterial isolated from clean areas, raw material, and products in the pharmaceutical industries, but the correct identification of these species is still a challenge. The aim of this study was to characterize by phenotyping, protein profiling, and 16S rRNA gene sequencing Sutcliffiellahorikoshii strains (n = 6) isolated from an immunobiological pharmaceutical facility, and to propose the reclassification of Bacillus tianshenii to the genus Sutcliffiella, and Sutcliffiella tianshenii sp. nov. The strains were characterized by VITEK®2, matrix-assisted laser desorption ionization-time of flight/mass spectrometry (MALDI-TOF/MS) using VITEK®MS, and 16S rRNA gene sequencing analysis. MALDI-TOF/MS did not identify any strains that were identified by 16S rRNA as S. horikoshii. VITEK®2 showed false-positive results, with misidentification as B. sporothermodurans (reclassified as Heyndrickxia sporothermodurans) and Geobacillus thermoleovorans. After MALDI-TOF/MS database expansion, with the creation of SuperSpectrum, the strains were correctly identified as S. horikoshii. This study is the first report of isolation of S. horikoshii strains from a pharmaceutical industry. More studies are necessary to better understand the ability of S. horikoshii to contaminate the environment and products.

MALDI-TOF MS database expansion for identification of Bacillus and related genera isolated from a pharmaceutical facility.

Bacillus and related genera are among the main bacterial groups isolated from pharmaceutical production areas. The identification of Bacillus species and related genera by classical methods is particularly difficult, due to similarities between closely related species. The Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) is one of the most promising techniques for chemotaxonomic characterization of microorganisms, being an alternative to genotypic methods. This study aimed to identify Bacillus strains and related genera isolated from immunobiological production areas by phylogenetic analysis of housekeeping genes and expand the database associated with MALDI-TOF MS to improve their identification. In a previous study, 97 aerobic endospore-forming bacteria isolated from a pharmaceutical facility were analyzed by MALDI-TOF MS and 16S rRNA gene full-length sequencing. All strains were identified as Bacillus and related genera by the latest methodology. Among the 97 strains, 22 were unidentified and 2 strains were misidentified by MALDI-TOF MS. In the present study, these 24 strains were subjected to 16S rRNA gene phylogenetic analysis. Strains not identified at species level by this methodology were submitted to rpoB gene phylogenetic analysis. After identifying the strains, 19 of the 24 strains were incubated for 24, 48, and 72 h on Tryptic Soy Agar and Sheep Blood Agar and subjected to analysis by MALDI-TOF MS. A SuperSpectrum for each strain was created and entered into the equipment database. Finally, the 24 strains were again submitted to proteomic analysis by MALDI-TOF MS, and, at this time, all were correctly identified. The genotypic identification of in-house isolated strains and the introduction of these spectra in MALDI-TOF MS, in order to obtain a customized database, proved to be an extremely effective tool in the identification of Bacillus and related genera from pharmaceutical industry origin.

Vitamin A status, inflammation adjustment, and immunologic response in the context of acute febrile illness: A pilot cohort study among pediatric patients.

BACKGROUND: Vitamin A is necessary for an adequate immune response to infections. Infection also alters vitamin A biomarkers, which interferes with assessment of vitamin A deficiency and thus impairs clinical management. Here we apply multiple strategies to adjust vitamin A biomarkers for inflammation during acute infection and evaluate associations between adjusted vitamin A status and immunologic response markers. METHODS: We measured biomarkers in pediatric patients presenting with acute febrile illness in Guayaquil, Ecuador at paired acute and convalescent visits. Four adjustment strategies were applied to retinol-binding protein (RBP) concentrations: Thurnham correction factor (TCF), BRINDA regression correction (BRC), CRP-only adjustment factor (CRP), and proof-of-concept for a proposed interleukin 6 regression model (IL-6 RM). Adjusted RBP concentrations were compared between visits using the paired Wilcoxon signed-rank test. Multivariate regression analysis was used to assess associations between adjusted vitamin A status and immunologic response markers. RESULTS: A sample of 57 participants completed the acute visit 1, and 18 of these individuals completed the convalescent visit 2. The IL-6 RM was the only strategy resulting in adjusted RBP concentrations that were not significantly different between paired visits (p = 0.20). Following RBP adjustment, 0.0% of participants were classified as vitamin A deficient (RBP ≤ 0.70 µmol/L) and 14.0% were classified as vitamin A insufficient (RBP ≤ 1.05 µmol/L). Adjusted vitamin A insufficiency was associated with an increase in macrophage inflammatory protein 1-alpha (MIP-1α, p = 0.03) and a pro-inflammatory immune response profile (p = 0.03) during the acute visit. CONCLUSIONS: We introduce a strategy for adjusting vitamin A in the context of clinical illness based on IL-6 concentrations that will need to be validated in larger studies. Assessment of vitamin A during infection allows for further understanding of how vitamin A status modulates immunopathology and enables targeted strategies for vitamin A supplementation in the context of infection among children in settings with high burdens of undernutrition and infectious diseases.

Presence of Ebola virus in breast milk and risk of mother-to-child transmission: synthesis of evidence.

To help inform global guidelines on infant feeding, this systematic review synthesizes evidence related to the presence of the Ebola virus (EBOV) in breast milk and its potential risk of viral transmission to the infant when breastfeeding. We relied on a comprehensive search strategy to identify studies including women with suspected, probable, or confirmed EBOV infection, intending to breastfeed or give breast milk to an infant. Our search identified 10,454 records, and after deduplication and screening, we assessed 148 full texts. We included eight studies reporting on 10 breastfeeding mothers and their children (one mother with twins), who provided breast milk samples for assessment. EBOV was detected via RT-PCR or viral culture in seven out of ten breast milk samples. Four out of the five-breastfed infants with EBOV-positive breast milk were found positive for EBOV infection, and all of these EBOV-positive infants died. Since previous reports have detected EBOV in tears, saliva, sweat, and contaminated surfaces, with the current evidence, it is not possible to conclude with certainty that breast milk was the main route of EBOV transmission.

Dengue associated with severe cutaneous leukocytoclastic vasculitis and pericardial effusion: a case report.

Dengue is a viral disease, caused by an arbovirus of the genus Flavivirus. In Brazil, its incidence rate is high with a broad clinical spectrum. This report discusses a rare case of dengue associated with cutaneous leukocytoclastic vasculitis and pericardial effusion with eminence of cardiac tamponade in a previously healthy patient with no comorbidities. The serology for dengue was positive and the histopathological analysis of the cutaneous lesions confirmed the diagnosis of leukocytoclastic vasculitis. After receiving treatment, the patient's condition greatly improved.

Ocorrência e caracterização da tontura em idosos atendidos em uma Estratégia Saúde da Família (ESF) / Occurrence and characterization of dizziness in the elderly assisted in a Family Health Strategy (FHS) / Ocurrencia y caracterización de mareos en ancianos asistidos en una Estrategia de Salud Familiar (ESF)

Introdução: A tontura é vista como um dos sintomas mais comuns na população em geral, ocorrendo com maior incidência na população idosa que frequentemente apresenta distúrbios do equilíbrio e comprometimento do sistema vestibular. A caracterização de um grupo de idosos da região centro sul do estado do Paraná com relação à ocorrência de tontura tem sua importância a fim de viabilizar a promoção de ações que visem um cuidado ampliado a esta população por parte da Atenção Básica. Objetivo: caracterizar as relações da queixa de tontura, sua autopercepção dos efeitos incapacitantes, o auto relato de uso de medicamentos e ocorrência de quedas em idosos atendidos por uma equipe de Estratégia Saúde da Família. Método: Setenta e cinco idosos de ambos os sexos foram questionados sobre presença de tontura; os que a referiram, foram submetidos ao Dizziness Handicap Inventory (DHI) na versão brasileira e a um questionário para caracterização do sintoma. Resultados: Dos 75 idosos questionados, 42 (56,00%) relataram queixa de tontura, a média etária foi de 67,61±5,68 anos (média ± desvio padrão), sendo 25 (59,52%) do sexo feminino e 17 (40,48%) do masculino. O escore total médio do DHI foi de 12,52±6,80 pontos no domínio físico, de 11,90±7,93 pontos no domínio funcional, 9,33±8,25 pontos no domínio emocional e 33,67±21,05 pontos no domínio total. Conclusão: A tontura, caracterizada por impacto moderado, de tipo não rotatória, de persistência superior a 12 meses e associada com uso de polifármacos, foi um sintoma frequente na população idosa deste estudo, sobretudo no sexo feminino. A auto percepção de qualidade de vida indicou diminuição no domínio físico impactando negativamente nas atividades cotidianas.

Introduction: Dizziness is seen as one of the most common symptoms in the general population, occurring with a greater incidence in the elderly population, which frequently presents balance disorders and impaired vestibular systems. The characterization of a group of elderly people in the south central region of the Brazilian state of Paraná in relation to the occurrence of dizziness has its importance in order to enable a promotion of actions that aim an expanded care for this population by Primary Care. Objective: to characterize the relation between the dizziness complaints, their self-perception of the disabling effects, the self-report of medication use and the occurrence of falls in the elderly assisted by a Family Health Strategy team .Method: Seventy-five elderly people of both genders were asked about the presence of dizziness; those who reported it were submitted to the Dizziness Handicap Inventory (DHI) in the Brazilian version and a questionnaire to characterize the symptom. Results: In 75 elderly people questioned, 42 (56.00%) reported complaints of dizziness, the average age was 67.61 ± 5.68 years (mean ± standard deviation), 25 (59.52%) being female and 17 (40.48%) were male. The mean total DHI score was 12.52 ± 6.80 points in the physical domain, 11.90 ± 7.93 points in the functional domain, 9.33 ± 8.25 points in the emotional domain and 33.67 ± 21, 05 points in the total domain. Conclusion: Dizziness, characterized by moderate impact, of a non-rotating type, persisting for more than 12 months and associated with the use of polypharmaceuticals, was a frequent symptom in the elderly population of this study, especially in females. The self-perception of quality of life indicated a reduction in the physical domain, negatively impacting daily activities.

Introducción: Se considera que los mareos son uno de los síntomas más comunes en la población en general, y se producen con mayor incidencia en la población de edad avanzada, que con frecuencia presenta trastornos del equilibrio y deterioro del sistema vestibular. La caracterización de un grupo de ancianos de la región centro-sur del estado de Paraná en relación con la aparición de mareos, tiene su importancia para permitir la promoción de acciones encaminadas a una atención prolongada de esta población por parte de la Atención Básica. Objetivo: Caracterizar las relaciones de la queja de mareo, su autopercepción de los efectos incapacitantes, la auto información sobre el uso de medicamentos y la ocurrencia de caídas en ancianos atendidos por un equipo de Estrategia de Salud Familiar. Método: Se interrogó a 75 hombres y mujeres de edad avanzada sobre la presencia de mareo; los que lo mencionaron fueron sometidos al Inventario de Discapacidades por Mareos (DHI) en la versión brasileña y a un cuestionario para caracterizar el síntoma. Resultados: De los 75 ancianos interrogados, 42 (56,00%) informaron de mareos, la edad media fue de 67,61 ± 5,68 años (media ± desviación estándar), siendo 25 (59,52%) mujeres y 17 (40,48%) hombres. La media total de la puntuación del DHI fue de 12,52±6,80 puntos en el dominio físico, 11,90±7,93 puntos en el dominio funcional, 9,33±8,25 puntos en el dominio emocional y 33,67±21,05 puntos en el dominio total. Conclusión: El mareo, caracterizado por un impacto moderado, de tipo no rotacional, con persistencia superior a 12 meses y asociado al uso de poli farmacéuticos, fue un síntoma frecuente en la población de edad avanzada de este estudio, especialmente en las mujeres. La autopercepción de la calidad de vida indicaba una disminución del dominio físico, lo que afectaba negativamente a las actividades cotidianas.

Prevalence and risk of potentially adverse drug interactions in the treatment of acute alcohol poisoning

ABSTRACT The aim of this paper is to determine the profile of acute alcohol poisoning and to estimate the risk of potentially adverse drug interactions (ADIs) in patients intoxicated by alcohol when attended in emergency care at hospital. A descriptive serial cross-sectional study was performed with 4,271 individuals intoxicated by alcohol, from January 2009 to July 2011. Possible correlations were measured by Pearson's chi-square test. The data show high consumption in the population, especially in males between 25 and 59 years. The main circumstances for poisoning were alcohol misuse (96.3%). After treatment complete recovery from the signs or symptoms of the poisoning was observed in 96.88% cases; and death in 0.70%. The demonstration of potential risk for ADIs in medical care included 300 medical records which contained a history of acute alcohol poisoning. Possible drug-drug interactions (44.2%) and drug-alcohol interactions (55.8%) were demonstrated in 60.60% of analyzed medical records. Among these cases, 3%, 92.4% and 4.6% were classified as mild, moderate and severe, respectively. The measurement of ADIs aims to prevent clinical complications in medical care for alcohol misuse disorders.

RESUMO O objetivo deste trabalho foi definir o perfil de intoxicação alcoólica aguda e estimar o risco de interações medicamentosas adversas (IMAs) potenciais em pacientes com intoxicação alcoólica atendidos na emergência hospitalar. Um estudo descritivo, serial, de corte transversal foi realizado com 4.271 indivíduos com intoxicação alcoólica, de janeiro 2009 a julho 2011. Correlações foram medidas pelo teste qui-quadrado. Os dados mostram alto consumo na população estudada, especialmente em homens de 25 a 59 anos. A principal circunstância de intoxicação foi o abuso (96,3%). Após tratamento, cura foi observada em 96,88% dos casos e morte em 0,7%. O risco de IMAs potenciais no atendimento médico incluiu 300 prontuários médicos com histórico de intoxicação alcoólica aguda. Possíveis interações medicamentosas (44,2%) e interações fármaco-álcool (55,8%) foram observadas em 60,6% dos prontuários analisados. Entre elas, 3%, 92,4% e 4,6% foram classificadas como leve, moderada e grave, respectivamente. A medição das IMAs visa a prevenir complicações clínicas no atendimento dos agravos devido ao abuso de álcool.

A Educação permanente e a cooperação internacional em saúde: um olhar sobre a experiência de fortalecimento da Rede haitiana de vigilância, pesquisa e educação em saúde, no âmbito do projeto Tripartite Brasil-Haiti-Cuba / Continuing education and international cooperation in health: a look at the experience of strengthening Haitian Network of Surveillance, research and education in Health, under the Tripartite project Brazil-Cuba-Haiti

O tema deste artigo é a cooperação internacional em saúde entre o Brasil-Haiti-Cuba, com foco no fortalecimento da Rede Haitiana de Vigilância, Pesquisa e Educação em Saúde, com vistas a contribuir para o fortalecimento da capacidade institucional do Ministério de Saúde Pública e da População MSPP) em Vigilância em Saúde. Abordaremos o tema por meio da experiência desenvolvida entre a Escola Nacional de Saúde Pública Sérgio Arouca/Fiocruz (ENSP), a Brigada Médica Cubana e a Direção Nacional de Epidemiologia do Ministério da Saúde Pública e População do Haiti. O modelo de cooperação aqui proposto busca romper com tradicionais paradigmas da cooperação norte sul, que se apoiam em transferencias verticais de tecnologias e traduzir na prática das atividades desenvolvidas os conceitos de horizontalidade e sustentabilidade propostos no âmbito da Cooperação Triparte em processos de trabalho se constitui um desafio e objeto de aprendizagem para a equipe condutora do processo. A educação permanente em saúde, como política brasileira e como abordagem metodológica da aprendizagem na saúde, demonstrou-se oportuna para configurar o projeto e as ações desenvolvidas. Neste relato de cooperação esperamos que a presença do Brasil com sua experiência no âmbito da educação permanente em saúde traga inovações ao processo de cooperação e efetivas contribuições ao desenvolvimento de capacidades institucionais no Ministério da Saúde do Haiti, assim como contribua para o desenvolvimento de conhecimentos e tecnologias de cooperação horizontal entre países.

El tema del presente artículo es la cooperación internacional en Salud entre Brasil-Haiti-Cuba, con acento en el fortalecimento de la Red. Haitiana de Vigilância, Investigación y Educación en Salud, con el objetivo de contribuir al fortalecimento de la capacidad institucional en vigilância en salud del Ministerio de Salud Publica y de la Poblacion (MSPP). Abordaremos el tema con base en la experiência desarrollada entre la Escuela Nacional de Salud Publica Sergio Arouca de la Fiocruz , la Brigada Medica Cubana y la Direccion Nacional de Epidemiologia del Ministerio de Salud Publica y de la Poblacion de Haiti. El modelo de cooperación aqui propuesto, busca romper los tradicionales paradigmas de la cooperación norte-sur apoyadosen transferência vertical de tecnologia. El modelo propuesto intenta traducir en la práctica, los conceptos de horizontalidad y sustentabilidad en los procesos de trabajo, en el ámbito de lacooperación tripartita. Estas concepciones se constituyen en un desafio y son sin dudas, objetode aprendizaje para el equipo conductor del proceso. La Educación Permanente en Salud como política Brasileña y como abordaje metodológico del proceso de aprendizaje en salud, demostróse oportuna para configurar el proyecto y orientar las acciones desarrolladas. En este relato sobre cooperación, esperamos que la presencia del Brasil, con su experiência en Educación Per-manente em salud,traiga innovaciones al proceso de cooperación,y efectivas contribuciones al desarrollo de la capacidad institucional del Ministerio de Salud de Haiti, asi como contribuya aldesarrollo de conocimientos y tecnologias de cooperación horizontal entre países.

Adiposidade e perfil metabólico em crianças de escolas da zona urbana de Ouro Preto-MG / Adiposity and metabolic profile of schoolchildren in the urban areas of Ouro Preto, Minas Gerais

A obesidade, doença multifatorial definida como excesso de gordura corporal, apresenta concomitância entre fatores de risco genéticos e ambientais. O diagnóstico precoce e as intervenções no período crítico do desenvolvimento da obesidade - infância e adolescência - têm sido recomendados, buscando-se evitar desfechos desfavoráveis na idade adulta. Este estudo transversal teve como objetivo caracterizar perfil lipídico, glicemia, adiponectina, leptina e grelina de escolares entre seis e nove anos, portadores de sobrepeso e obesidade, do município de Ouro Preto-MG. Os dados foram analisados a partir do teste de normalidade Shapiro Wilk; e nas comparações entre os grupos foi aplicado o teste paramétrico (Teste t) ou não paramétrico (Teste Mann Whitney), adotando-se intervalo de confiança de 95% e nível de significância para valores ≤ 0,05. A idade média da população escolar foi de 7,8 ± 1,1 anos, com prevalência de 8,9% de sobrepeso e 3% de obesidade. Foram identificados hipercolesterolemia em 5,5%, HDL alterado em 98,7%, taxa limítrofe de LDL em 32,4% e glicemia alterada em 46,6% das crianças. Na análise estratificada quanto ao gênero, foram observados valores maisaltos para leptina em meninas (p=0,032) e grelina nos meninos (p=0,033), não havendo diferença para as demais variáveis. Os resultados demonstram ser o excesso de peso entre escolares problema de saúde relevante no município, ressaltando-se a importância de implementação de programa de intervenção precoce por parte dos gestores. Elucidar os precursores da obesidade na infância pode levar a intervenções capazes de atenuar ou impedir suas consequências na juventude e fase adulta.

Obesity, defined as an excess in body fat, is a multifactorial disease involving both genetic and environmental risk factors. Early diagnosis and interventions during critical periods of development of obesity - childhood and adolescence - have been recommended, aiming at preventing unfavorable outcomes at a later age. This cross-sectional study sought tocharacterize the lipid profile, glucose, adiponectin, leptin and ghrelin in schoolchildren between six and nine years of age with overweight and obesity in the city of Ouro Preto (MG). The data was analyzed with the Shapiro Wilk normality test, groups comparisons were made with either a parametric (T test) or a nonparametric (Mann Whitney) test, adopting confidence intervals of 95% and a significance level of ≤ 0.05. The average age of the school population was 7.8 ± 1.1 years, with a prevalence of 8.9% of overweight and 3% of obesity. Hypercholesterolemia was found in 5.5%, HDL was abnormal in 98.7%, LDL levels were borderline in 32.4% and altered glucose levels were present in 46.6% of the children. In stratified analysis by gender, higher values of leptin in girls (p = 0.032) and ghrelin in boys (p = 0.033) were found, with no difference for the other variables. The results show that overweight/obesity among schoolchildren should be considered a significant health problem in this population, highlighting the importanceof implementing early intervention programs. Uncovering the precursors of childhood obesity could lead to interventions so as to prevent or mitigate its consequences in youth and adulthood.

La carga de la enfermedad en países de América Latina / Burden of disease in Latin America

OBJETIVO. Describir los estudios de carga de la enfermedad realizados en la región e identificar las principales prioridades en salud a partir del indicador años de vida saludable perdidos (AVISA). MATERIAL Y MÉTODOS: Mediante el uso de AVISA, identificar la carga de enfermedad en los distintos países. RESULTADOS: Los AVISA destacan la emergencia de los trastornos mentales, la diabetes mellitus en las mujeres y los trastornos por consumo de alcohol y lesiones en los hombres. CONCLUSIONES: América Latina es la región con más estudios nacionales de carga de la enfermedad realizados con una metodología estandarizada, que permiten identificar problemas de salud que están presionando a los servicios de atención; por ello estos resultados constituyen un elemento a tomar en cuenta en el establecimiento de políticas públicas en cada país.

OBJECTIVE: To describe the burden of disease studies made in the region, identify the main priorities in health from the indicator Disability Adjusted Life Years (DALYs). MATERIAL AND METHODS: By the use of DALYs identify the burden of disease in the countries in the network. RESULTS: DALYs emphasize the emergency of mental disorders, diabetes mellitus in women and the disorders associated with alcohol consumption and injuries in men. CONCLUSIONS: Latin America is the region with more national studies of burden of disease, using a standardized methodology, that allows identifying new health priorities which are pressing to the health services; for that reason these results constitute an element to take into account in the establishment of public policies in each country.

Expression of major vault protein gene in osteosarcoma patients.

Osteosarcoma (OS) is a primary malignant tumor of bone. Despite the successful use of multiple chemotherapeutic agents in the treatment of OS, more than 30% of OS tumors remain resistant to treatment. Elucidation of cellular resistance mechanisms may lead to better treatments for cancer patients. In this study, we used the low-density expression cDNA array, GEArray Q Series Human Cancer Drug Resistance and Metabolism Gene Array to screen genes related to drug resistance in 15 OS tumors. Expression patterns of the MPV gene were validated by real time PCR on 45 OS patient tumor samples and correlated with clinical and pathological data. Major vault protein (MVP) expression was present in 24 (53%) tumor samples and absent in 21 (47%). Samples from surgery showed correlation between the expression of MVP, metastatic disease at diagnosis and event free survival (EFS). The MVP gene expression correlates with metastatic disease at diagnosis after neoadjuvant chemotherapy (p=0.048), and is also associated with worse EFS (p=0.036). These findings suggest that MVP expression is involved in one of the mechanisms of drug resistance in OS and is induced by chemotherapy.

Effects of THBS3, SPARC and SPP1 expression on biological behavior and survival in patients with osteosarcoma.

BACKGROUND: Osteosarcoma is a very aggressive tumor with a propensity to metastasize and invade surrounding tissue. Identification of the molecular determinants of invasion and metastatic potential may guide the development of a rational strategy for devising specific therapies that target the pathways leading to osteosarcoma. METHODS: In this study, we used pathway-focused low density expression cDNA arrays to screen for candidate genes related to tumor progression. Expression patterns of the selected genes were validated by real time PCR on osteosarcoma patient tumor samples and correlated with clinical and pathological data. RESULTS: THBS3, SPARC and SPP1 were identified as genes differentially expressed in osteosarcoma. In particular, THBS3 was expressed at significantly high levels (p = 0.0001) in biopsies from patients with metastasis at diagnosis, which is a predictor of worse overall survival, event-free survival and relapse free survival at diagnosis. After chemotherapy, patients with tumors over-expressing THBS3 have worse relapse free survival. High SPARC expression was found in 51/55 (96.3%) osteosarcoma samples derived from 43 patients, and correlated with the worst event-free survival (p = 0.03) and relapse free survival (p = 0.07). Overexpression of SPP1 was found in 47 of 53 (89%) osteosarcomas correlating with better overall survival, event-free survival and relapse free survival at diagnosis. CONCLUSION: In this study three genes were identified with pattern of differential gene expression associated with a phenotypic role in metastasis and invasion. Interestingly all encode for proteins involved in extracellular remodeling suggesting potential roles in osteosarcoma progression. This is the first report on the THBS3 gene working as a stimulator of tumor progression. Higher levels of THBS3 maintain the capacity of angiogenesis. High levels of SPARC are not required for tumor progression but are necessary for tumor growth and maintenance. SPP1 is not necessary for tumor progression in osteosarcoma and may be associated with inflammatory response and bone remodeling, functioning as a good biomarker.

Metaphase and array comparative genomic hybridization: unique copy number changes and gene amplification of medulloblastomas in South America.

Tumors of the central nervous system are the second most frequent malignancy of childhood, accounting for the majority of cancer-related deaths in this age group. Among these tumors, medulloblastomas (MB) remain in need of further genomic characterization toward understanding of pathogenesis and outcome predictors. Eight pediatric embryonal brain tumors were analyzed: five MB (one being desmoplastic), one PNET, one medulloepithelioma, and one ependymoblastoma. Analyses identified genomic imbalances, including the gain of 16p and the nonsyntenic coamplification of MYCN and TERT loci. More detailed FISH analysis showed that coamplification of MYCN and TERT in one of the MBs manifested as dispersed nuclear speckling, consistent with the presence of double minute chromosomes. There was considerable cell-to-cell copy number heterogeneity present, but it was clear that both genes were amplified concordantly. The amplification of oncogenes seems to play an important role in the pathogenesis of MB, and the association between MYCN and TERT amplifications and poor prognosis has not been well recognized. The uncharacteristic pattern of genomic imbalances detected in MB tumors may be a reflection of the characteristics of these tumors occurring in South America.

Telomerase activity in prognostic histopathologic features of melanoma.

BACKGROUND: Telomerase activity (TA) is believed to play a role in the regulation of senescence and to limit the number of cell divisions. The deregulation of telomerase appears to contribute to oncogenesis and the formation of immortal cell lines. As a result, it is believed that it could be used as a prognostic marker in melanoma. METHODS: TA was assayed by the polymerase chain reaction PCR-ELISA-based telomeric repeat amplification protocol (TRAP assay). One hundred and eight samples were distributed in four histological groups: 30 samples from primary cutaneous melanomas, 24 from peritumoural skin sites, 28 from benign melanocytic lesions, and 26 from normal skin sites as a control. RESULTS: TA was different among the four tested groups (Kruskall-Wallis test p<0.001), and increasing values of TA were observed progressing from normal skin to benign and then to malignant lesions. Among melanoma samples, there was a significant association between TA and ulceration (p=0.025), TA and vascular invasion (p=0.018) and TA and mitotic rate (p=0.029) (Mann-Whitney test). A linear regression analysis showed significant associations between the increase of TA with Breslow thickness (p=0.004) and the presence of satellites (p=0.002). CONCLUSIONS: We observed that TA had increased from control skin to peritumoural skin, and then to benign melanocytic lesions and finally to melanoma, suggesting tumour progression. TA showed higher values in the presence of some important histopathologic parameters related to poor prognosis in cutaneous melanoma such as ulceration, vascular invasion, satellites, high rates of mitosis, and in thicker tumours.

Leucemia promielocítica aguda: caracterizacão de alteracões cromossômicas por citogenética tradicional e molecular ( FISH ) / Acute promyelocytic leukemia: characterization of chromosome abnormalities by classical cytogenetics and FISH

A leucemia promielocítica aguda (LPA) corresponde a 10 por cento -15 por cento das leucemias mielóides agudas (LMA). Este tipo de leucemia (LMA-M3 de acordo com a classificacão FAB) está associado, em cerca de 90 por cento dos casos, à translocacão t(15;17)(q22;q21), que resulta na fusão dos genes PML e RARalfa. A análise citogenética tradicional tem sido utilizada para confirmar o diagnóstico morfológico da LPA. Embora a t(15;17) não seja detectada em outros tipos de leucemia, podem ocorrer resultados "falso-negativos", decorrentes da análise de células que não pertencem ao clone neoplásico, da dificuldade de visualizacão da translocacão ou, até mesmo, da existência de rearranjos crípticos que mascaram a translocacão. Por outro lado, foram descritas alteracões cromossômicas alternativas em pacientes com LPA e, nesses casos, o tratamento com ATRA não é eficaz. No período de julho de 1993 a dezembro de 2002 foram encaminhados para análise citogenética 47 casos com suspeita e/ou diagnóstico clínico-morfológico de LPA. Trinta e quatro pacientes (72,3 por cento) apresentaram a t(15;17), detectada pela citogenética tradicional e/ou molecular. Em seis destes pacientes foram observadas alteracões cromossômicas adicionais ou rearranjos envolvendo um terceiro cromossomo. Em cinco (10 por cento) pacientes com características de LPA, a técnica de FISH não revelou a fusão PML/RARalfa, dado importante para a orientacão do diagnóstico e da conduta terapêutica desses pacientes. O presente trabalho foi realizado com o objetivo de avaliar a importância da análise citogenética tradicional e molecular no diagnóstico de pacientes com LPA.

Detection of micronuclei formation and nuclear anomalies in regenerative nodules of human cirrhotic livers and relationship to hepatocellular carcinoma.

Human cirrhosis is considered an important factor in hepatocarcinogenesis. The lack of substantial genetics and cytogenetics data in human cirrhosis led us to investigate spontaneous micronuclei formation, as an indicator of chromosomal damage. The analysis was performed in hepatocytes of regenerative, macroregenerative, and tumoral nodules from 30 cases of cirrhosis (paraffin-embedded archival material), retrospectively selected: cryptogenic, hepatitis C virus, and hepatitis C virus associated with hepatocellular carcinoma (HCC). Thirteen control liver samples of healthy organ donors were included. Micronucleated hepatocytes were analyzed with Feulgen-fast-green dyeing techniques. The spontaneous frequency of micronucleated hepatocytes in both regenerative and macroregenerative nodules of all cirrhotic patients was significantly higher than for the normal control group. There was no significant difference in frequency of micronucleated hepatocytes in regenerative nodules compared with macroregenerative nodules for all cases analyzed, whereas a significantly higher frequency of micronucleated hepatocytes was detected in tumoral nodules, compared with cirrhotic regenerative nodules and normal parenchyma. A higher frequency of the nuclear anomalies termed broken-eggs was observed in hepatitis C virus-related samples. Chromatinic losses and genotoxicity already existed in the cirrhotic regenerative nodules, which might predispose to development of HCC.

Comparative genomic hybridization analysis of pediatric adamantinomatous craniopharyngiomas and a review of the literature.

OBJECT: The purpose of this study was to examine chromosomal gains and losses in 11 pediatric adamantinomatous craniopharyngiomas by using comparative genomic hybridization (CGH), as well as to review the cytogenetic literature that has contributed to the characterization of these tumors. One source of confusion in the cytogenetic and CGH literature concerning craniopharyngioma is that the authors of most studies fail to distinguish between pediatric and later-onset forms of the disease. Thus, this study was focused on pediatric craniopharyngioma. METHODS: To determine an overview of the genetic events leading to the development of these tumors, 10 adamantinomatous craniopharyngiomas were analyzed using CGH; none of the tumor specimens demonstrated gains or losses of DNA sequence. CONCLUSIONS: In view of these findings as well as those published in the majority of previous cytogenetic studies of craniopharyngiomas, the authors conclude that the recurrent acquisition of chromosomal imbalances does not play a major role in tumorigenesis and that chromosomal gains and losses are a relatively rare event in primary tumors of pediatric origin.