Usefulness of collagen type IV in the detection of significant liver fibrosis in nonalcoholic fatty liver disease.

INTRODUCTION/AIMS: Liver fibrosis assessment is a key issue in the evaluation of nonalcoholic fatty liver disease (NAFLD) patients. In the present study, we aimed to validate a noninvasive marker panel to assess significant and advanced fibrosis in these patients. METHOD: 126 biopsy-proven NAFLD patients were included. NAFLD diagnosis was based on histological criteria. Fibrosis stages were determined according to NASH-Clinical Research Network criteria. Clinical and laboratorial data were collected during the interval of three months before or after liver biopsy. Histological fibrosis stages were classified as significant fibrosis (F2-F4) and advanced fibrosis (F3-F4). Five serum biomarkers [hyaluronic acid (HA), collagen type IV (cIV), procollagen type III (PC III), laminin (LN) and cholylglycine (CG)] were assessed by chemiluminescence immunoassays. RESULTS: Most patients were female (61.61%), mean age: 55.7 ±â€¯9.13 years old and mean BMI was 32.1 ±â€¯5.9 kg/m2. Prevalence of diabetes, dyslipidemia, arterial hypertension, and metabolic syndrome was 68.75%, 82.29%, 63.54% and 81.05%, respectively. Patients with cIV above 30 ng/mL had a 5.57-times (IC: 1.86-16.69) the chance of having significant fibrosis and 7.61-times (IC: 2.27-25.54) the chance of having advanced fibrosis versus patients with values below 30 ng/mL. HA, PC III, LN and CG did not detect the presence of significant and advanced fibrosis. The AUROC of clV for detection of significant (0.718) and advanced fibrosis (0.791) was better than that of other serum biomarkers. CONCLUSION: Type 4 collagen could predict the presence of significant and advanced fibrosis in NAFLD patients and it would be a useful tool in routine clinical practice.

Effects of atorvastatin on CYP3A4 and CYP3A5 mRNA expression in mononuclear cells and CYP3A activity in hypercholeresterolemic patients.

BACKGROUND: Variability of response to statins has been related to polymorphisms in genes involved in cholesterol homeostasis and statin metabolism, such as CYP3A4 and CYP3A5. We investigated the effects of atorvastatin on CYP3A4 and CYP3A5 mRNA expression in mononuclear cells and on CYP3A activity and their interactions with common variants. METHODS: Unrelated individuals (n=121) with hypercholesterolemia (HC) were treated with atorvastatin (10 mg/day/4 weeks). Ninety-two normolipidemic (NL) subjects were selected as a control group. Genotype analysis of CYP3A4*1B (rs2740574), CYP3A4*22 (rs35599367), CYP3A5*3C (rs776746), and CYP3A5*1D (rs15524) and mRNA levels in peripheral blood mononuclear cells (PBMCs) were estimated. CYP3A activity was phenotyped by the urinary cortisol to 6-beta-hydroxy-cortisol ratio. RESULTS: LDL cholesterol reduction in response to atorvastatin was positively correlated with change in CYP3A4 (R(2)=0.039, p=0.037) and CYP3A5 (R(2)=0.047, p=0.019) mRNA levels and negatively correlated with CYP3A activity (R(2)=0.071, p=0.022). CYP3A5*3C (AGT haplotype) was associated to lower basal CYP3A5 mRNA expression in HC (p<0.045), however none of the haplotype groups impacted treatment. CONCLUSION: It is likely that cholesterolemia status changes promoted by atorvastatin play a role in regulating CYP3A4 and CYP3A5 mRNA expression in PBMCs, as well as CYP3A activity. CYP3A5*3C (AGT haplotype) also contributes for the variability of CYP3A5 mRNA levels in PBMCs.

Uric acid biorhythm, a feature of long-term variation in a clinical laboratory database.

BACKGROUND: The biorhythm of serum uric acid was evaluated in a large sample of a clinical laboratory database by spectral analysis and the influence of the gender and age on uric acid variability. METHODS: Serum uric acid values were extracted from a large database of a clinical laboratory from May 2000 to August 2006. Outlier values were excluded from the analysis and the remaining data (n=73,925) were grouped by gender and age ranges. Rhythm components were obtained by the Lomb Scargle method and Cosinor analysis. RESULTS: Serum uric acid was higher in men than in women older than 13 years (p<0.05). Compared with 0-12 year group, uric acid increased in men but not in women older than 13 years (p<0.05). Circannual (12 months) and transyear (17 months) rhythm components were detected, but they were significant only in adult individuals (>26 years, p<0.05). Cosinor analysis showed that midline estimating statistic of rhythm (MESOR) values were higher in men (range: 353-368 µmol/L) than in women (range: 240-278 µmol/L; p<0.05), independent of the age and rhythm component. The extent of predictable change within a cycle, approximated by the double amplitude, represented up to 20% of the corresponding MESOR. CONCLUSIONS: Serum uric acid biorhythm is dependent on gender and age and it may have relevant influence on preanalytical variability of clinical laboratory results.

Hepatitis B marker seroprevalence and vaccination coverage in adolescents in the City of Itajaí, State of Santa Catarina, Southern Brazil, in 2008.

INTRODUCTION: Hepatitis B infection constitutes an important cause of morbidity and mortality worldwide. In Brazil, however, the current epidemiological situation is not clear. Considering the importance of establishing this prevalence, the aim of this study was to determine the prevalence of HBV markers in voluntary adolescents, junior high (secondary school) students, in the City of Itajaí, State of Santa Catarina, Brazil. METHODS: A seroepidemiological, transverse study was conducted with 353 randomly chosen adolescents from elementary school in 2008. Blood samples were analyzed for HBsAg, anti-HBc and anti-HBs. All analyses were conducted by automated microparticle enzyme immunosorbent assay (Abbott®, AxSYM system, Deerfield, IL, USA), according to the manufacturer's instructions. RESULTS: The prevalence of HBsAg was 0.6% (CI 95% 0.1 - 2.0), that of anti-HBc was 1.1% (CI 95% 0.3 - 2.9) and that of detectable anti-HBs was 83.6% (CI 95% 79.3 - 87.3). Hepatitis B vaccination coverage was 97.5% (CI 95% 95.2 - 98.8). CONCLUSIONS: These results demonstrate the success of the vaccination program against hepatitis B in the region studied and indicate that prevention strategies must be maintained and, if possible, expanded to contribute to the establishment of positive prevalence rates in all age groups.

Hepatitis B marker seroprevalence and vaccination coverage in adolescents in the City of Itajaí, State of Santa Catarina, Southern Brazil, in 2008 / Soroprevalência de marcadores da hepatite B e cobertura de vacinação em adolescentes no município de Itajaí, Estado de Santa Catarina Sul do Brasil, em 2008

INTRODUCTION: Hepatitis B infection constitutes an important cause of morbidity and mortality worldwide. In Brazil, however, the current epidemiological situation is not clear. Considering the importance of establishing this prevalence, the aim of this study was to determine the prevalence of HBV markers in voluntary adolescents, junior high (secondary school) students, in the City of Itajaí, State of Santa Catarina, Brazil. METHODS: A seroepidemiological, transverse study was conducted with 353 randomly chosen adolescents from elementary school in 2008. Blood samples were analyzed for HBsAg, anti-HBc and anti-HBs. All analyses were conducted by automated microparticle enzyme immunosorbent assay (Abbott®, AxSYM system, Deerfield, IL, USA), according to the manufacturer's instructions. RESULTS: The prevalence of HBsAg was 0.6% (CI 95% 0.1 - 2.0), that of anti-HBc was 1.1% (CI 95% 0.3 - 2.9) and that of detectable anti-HBs was 83.6% (CI 95% 79.3 - 87.3). Hepatitis B vaccination coverage was 97.5% (CI 95% 95.2 - 98.8). CONCLUSIONS: These results demonstrate the success of the vaccination program against hepatitis B in the region studied and indicate that prevention strategies must be maintained and, if possible, expanded to contribute to the establishment of positive prevalence rates in all age groups.

INTRODUÇÃO: A hepatite B constitui uma causa relevante de morbidade e mortalidade em todo o mundo. No Brasil, entretanto, a situação epidemiológica atual não é claramente conhecida. Considerando-se a importância do estabelecimento de tal prevalência, o objetivo deste estudo foi determinar a prevalência dos marcadores de HBV em adolescentes voluntários, estudantes do ensino fundamental na Cidade de Itajaí, Santa Catarina, Brasil. MÉTODOS: Trata-se de um estudo soroepidemiológico, transversal, conduzido com 353 adolescentes do ensino fundamental, randomicamente selecionados, em 2008. As amostras de sangue foram analisadas quanto à presença dos marcadores HBsAg, anti-HBc e anti-HBs. Todas as análises foram conduzidas em ensaio imunoenzimático de micropartículas automatizado (Abbott®, AxSYM system, Deerfield, IL, EUA), de acordo com as instruções do fabricante. RESULTADOS: A prevalência do HBsAg foi de 0,6% (CI 95% 0,1 - 2,0), do anti-HBc foi de 1,1% (CI 95% 0,3 - 2,9) e de títulos detectáveis de anti-HBs foi de 83,6% (CI 95% 79,3 - 87,3). A cobertura vacinal nos adolescentes estudados foi de 97,5% (CI 95% 95,2 - 98,8). CONCLUSÕES: Estes resultados demonstram o sucesso do programa de vacinação contra a hepatite B na região estudada e indicam que as estratégias de prevenção devem ser mantidas e, se possível, expandidas para que contribuam no estabelecimento de taxas de prevalência positivas em todas as faixas etárias.

Níveis séricos de 25-hidroxi vitamina D em pacientes portadores de insuficiência renal crônica em tratamento dialítico / 25-hydroxy vitamin D levels in chronic kidney disease patients on dialysis therapy

JUSTIFICATIVA E OBJETIVOS: A contribuição da deficiência de vitamina D para o desenvolvimento do hiperparatireoidismo secundário e da osteodistrofia renal motiva a racional suplementação de vitamina D. A sua deficiência tem sido associada,ainda, a elevado risco cardiovascular. O objetivo deste estudo foi descrever os níveis séricos de 25-hidroxi vitamina D em pacientes portadores de insuficiência renal terminal em terapia dialítica.MÉTODO: Dados clínicos e laboratoriais foram coletados em novembro de 2009 em 168 pacientes em terapia dialítica em um único centro. Os níveis séricos de 25-hidroxi vitamina D foram medidos utilizando o método automatizado de imunoensaio desenvolvido por DiaSorin. De acordo com o Kidney Disease Outcomes and Quality Initiative Guidelines, os pacientes foram classificados em três grupos: suficientes (> 30 ng/mL), insuficientes(15 a 30 ng/mL); e deficientes (< 15 ng/mL). A prevalência de deficiência ou insuficiência de vitamina D foi calculada. Para comparar as variáveis foram usados os seguintes testes: Qui-quadrado,Exato de Fisher e t de Student. O coeficiente de correlação de Spearman foi utilizado para variáveis contínuas. O valor de p < 0,05 foi considerado estatisticamente significativo.RESULTADOS: A média ± desvio padrão de 25-hidroxi vitaminaD foi 23,8 ± 13,3 ng/mL. A prevalência de deficiência devitamina D foi de 23,9% e de insuficiência foi de 47,8%. Quando se compararam as diferenças entre homens e mulheres, idade,albumina e fósforo foram inferiores nas mulheres (p < 0,05). Os níveis de 25-hidroxi vitamina D foram significantemente inferiores em pacientes em diálise peritoneal quando comparados aos em hemodiálise (p < 0,05). Na análise univariada houve pequena correlação positiva entre 25-hidroxi vitamina D e Kt/V.CONCLUSÃO: Os níveis de 25-hidroxi vitamina D foram baixos nesta população. Estudos prospectivos para avaliar o impacto desta deficiência se fazem necessários.(AU)

BACKGROUND AND OBJECTIVES: The contribution of vitamin D deficiency to the development of secondary hyperparathyroidism and renal osteodystrophy contributes the rationale for vitamin D supplementation. Vitamin D deficiency has been associated with elevated cardiovascular risk. This study aimed investigates the serum levels of vitamin D among patients undergoing dialysis.METHOD: In November 2009, we examined clinical and laboratory data from 168 patients in dialytic therapy in a unit. Serum level of vitamin D was included in laboratory data. Serum levels of vitamin D were measured using an automated, antibody and microparticle, chemiluminescence immunoassay method developedby DiaSorin. According to the Kidney Disease Outcomes and Quality Initiative guidelines, patients were assigned to thefollowing 3 groups: sufficient vitamin D serum level (> 30 ng/mL), insufficient level (15 to 30 ng/mL) and with severe deficiency(< 15 ng/mL). Prevalence of deficiency or insufficiency of vitamin D was calculated. To compare the variables were usedthe Chi-square, Fisher Exact or Student's t tests. Spearman correlation coefficient was used for continuous variables. A p value < 0.05 was considered statistically significant.RESULTS: Patient's serum vitamin D mean level was 23.8 ± 13.3 ng/mL. Prevalence of vitamin D deficiency was 23.9%. Prevalence of vitamin D insufficiency was 47.8%. When compared differences between men and women, age, albumin and phosphorus were inferior in women (p < 0.05). Levels of vitamin D were greatly inferior in peritoneal dialysis when compared with hemodialysis (p < 0.05). In the univariate analysis there were weak positive correlations between vitamin D and Kt/V.CONCLUSION: Serum vitamin D levels in this population were low. Prospective studies to assess the impact of low level of vitamin D in patients undergoing dialysis are warranted.(AU)

Real-time PCR-based quantification of Toxoplasma gondii in tissue samples of serologically positive outdoor chickens.

This study aimed to quantify Toxoplasma gondii in tissue samples of serologically positive chickens using real-time polymerase chain reaction (PCR). Of 65 chickens evaluated, 28 were positive for T. gondii antibodies. Brain and heart samples were collected from 26 seropositive chickens and DNA was extracted using Trizol® and amplified using real-time PCR with SYBR® Green. Parasite DNA was detected in 24 of the 26 samples analyzed; the number of positive tissue samples and the parasite quantity did not differ between tissue types. The results confirmed the analytical sensitivity of parasite detection in chicken tissue samples and demonstrated the possibility of using other molecular systems for genotypic analysis.

Real-time PCR-based quantification of Toxoplasma gondii in tissue samples of serologically positive outdoor chickens

This study aimed to quantify Toxoplasma gondii in tissue samples of serologically positive chickens using real-time polymerase chain reaction (PCR). Of 65 chickens evaluated, 28 were positive for T. gondii antibodies. Brain and heart samples were collected from 26 seropositive chickens and DNA was extracted using Trizol® and amplified using real-time PCR with SYBR® Green. Parasite DNA was detected in 24 of the 26 samples analyzed; the number of positive tissue samples and the parasite quantity did not differ between tissue types. The results confirmed the analytical sensitivity of parasite detection in chicken tissue samples and demonstrated the possibility of using other molecular systems for genotypic analysis.

Vitamin D and musculoskeletal health, cardiovascular disease, autoimmunity and cancer: Recommendations for clinical practice.

BACKGROUND: There is increasing evidence that, in addition to the well-known effects on musculoskeletal health, vitamin D status may be related to a number of non-skeletal diseases. An international expert panel formulated recommendations on vitamin D for clinical practice, taking into consideration the best evidence available based on published literature today. In addition, where data were limited to smaller clinical trials or epidemiologic studies, the panel made expert-opinion based recommendations. METHODS: Twenty-five experts from various disciplines (classical clinical applications, cardiology, autoimmunity, and cancer) established draft recommendations during a 2-day meeting. Thereafter, representatives of all disciplines refined the recommendations and related texts, subsequently reviewed by all panelists. For all recommendations, panelists expressed the extent of agreement using a 5-point scale. RESULTS AND CONCLUSION: Recommendations were restricted to clinical practice and concern adult patients with or at risk for fractures, falls, cardiovascular or autoimmune diseases, and cancer. The panel reached substantial agreement about the need for vitamin D supplementation in specific groups of patients in these clinical areas and the need for assessing their 25-hydroxyvitamin D (25(OH)D) serum levels for optimal clinical care. A target range of at least 30 to 40 ng/mL was recommended. As response to treatment varies by environmental factors and starting levels of 25(OH)D, testing may be warranted after at least 3 months of supplementation. An assay measuring both 25(OH)D(2) and 25(OH)D(3) is recommended. Dark-skinned or veiled individuals not exposed much to the sun, elderly and institutionalized individuals may be supplemented (800 IU/day) without baseline testing.

Allele frequencies of 15 STRs in a representative sample of the Brazilian population.

Allele frequencies for 15 short tandem repeat (STR) loci were obtained from a sample of 12,030 individuals undergoing paternity testing. This sample includes individuals from all States in Brazil, combined according to the current country division into five regions (North, Northeast, Central West, Southeast, and South). The most polymorphic loci were D2S1338 and D18S51. All the analysed loci meet Hardy-Weinberg equilibrium expectations. Combined power of discrimination and combined power of exclusion for the 15 tested STR loci were 0.999999999999990 and 0.9999992, respectively. Comparative analysis between populations from different Brazilian macroregions as well as between Brazil and other relevant populations are presented.

Efeitos quantitativos da estocagem de sangue periférico nas determinações do hemograma automatizado / Storage effects on peripheral blood samples as identified from automated hemograms

O presente trabalho avaliou as possíveis alterações em vários parâmetros do hemograma (contagem de eritrócitos totais, hematócrito, concentração de hemoglobina, volume corpuscular médio (VCM), hemoglobina corpuscular média (HCM), concentração da hemoglobina corpuscular média (CHCM), contagem total de leucócitos e contagem de plaquetas), frente a diferentes tempos de armazenamento da amostra em ambiente refrigerado a 4ºC e temperatura ambiente. As determinações foram realizadas através do contador automatizado Sysmex® XT2000i. As amostras sanguíneas foram obtidas de indivíduos sem alterações hematológicas diagnosticadas, sem clínica de doença, sendo considerados indivíduos com valores de referências hematológicos normais. Os resultados foram avaliados através de análise estatística descritiva e comparação de médias através da análise de variância (ANOVA). Os resultados dos parâmetros CHCM e contagem de plaquetas mostraram diferenças estatisticamente significativas para ambas as temperaturas de estocagem. Na temperatura ambiente, os parâmetros que também apresentaram diferença estatística significante foram para o hematócrito, VCM e índice de variação do tamanho dos eritrócitos ( RDW). Conclui-se, portanto, que os resultados dos hemogramas liberados pelo aparelho analisado podem ser satisfatórios quando realizados entre 12 a 24 horas após a coleta da amostra para a maioria dos parâmetros avaliados.

This study evaluated possible alterations in different hematologic parameters [total erythrocyte count, hematocrit, hemoglobin concentration, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MHCM), total leukocyte count and platelet count], of blood samples submitted to varied storage times at both 4ºC and at room temperature. The analyses were performed using a Sysmex XT2000i automated hematology analyzer. Written consent was obtained from donors with no hematological disorders or clinical abnormalities and thus from individuals with apparently normal laboratory reference values for healthy adults. The results were analysed by descriptive statistics with comparisons of the means being achieved by analysis of variance (ANOVA). The means of the MCHC parameters and platelet count showed significant statistical differences for both storage temperatures. At room temperature the hematocrit, MCV and red cell distribution width (RDW) readings also presented statistically significant differences. Therefore, it can be concluded that hemogram results performed by automated blood analyzers are satisfactory when carried out within 24 hours after drawing the blood sample for most of the parameters evaluated.

15 STR loci frequencies in the population from Paraná, Southern Brazil.

Allele frequencies for 15 short tandem repeats (STR) loci were obtained from a sample of 4076 unrelated individuals undergoing paternity testing. The population is from Paraná, Southern Brazil. The loci are the most commonly used in forensic and paternity testing, being analyzed by the AmpFlSTR((R)) Identifiler (Applied Biosystems) commercial kit. The most polymorphic loci were D2S1338 and D18S51. Excepting the D13S317, all loci were in Hardy-Weinberg equilibrium. Comparative analyses between our population data and other populations are presented.

Validação do método de detecção de Chlamydia trachomatis por Reação em Cadeia da Polimerase em tempo real / Validation of a Chlamydia trachomatis detection method by Real Time Polymerase chain reaction

Infecções causadas por Chlamydia trachomatis são recentemente reconhecidas como as mais prevalentes e estão entre as mais prejudiciais doenças sexualmente transmissíveis (DST) no mundo. A ausência de sintomas dificulta o diagnóstico clínico das infecções clamidiais. Um dos desafios na prevenção das infecções causadas por C. trachomatis é a disponibilidade de diagnósticos laboratoriais com alta sensibilidade, especificidade e reprodutibilidade. No presente estudo, foi comparado o desempenho do testes de Reação em Cadeia da Polimerase (PCR) COBAS Amplicor® Chlamydia trachomatis (Roche) e PCR em tempo real (SYBR® – Green), usados para detecção de Chlamydia trachomatis em espécimes de urina e secreções urogenitais. O COBAS Amplicor® (Roche) é umteste de PCR convencional e foi usado como ferramenta para controle e teste de padrão ouro para diagnóstico no presente trabalho. Um total de 136 amostras de secreção endocervical (61), secreção uretral (14) e urina (61), foram obtidas de pacientes submetidos a testes de detecção de Chlamydia trachomatis no Laboratório Alvaro S/A. Os métodos de PCR em tempo real e COBAS Amplicor® (Roche)foram igualmente sensíveis para o diagnóstico de C. trachomatis em amostras de secreção uretral. Entretanto, em análises de urinae secreção endocervical o teste COBAS Amplicor® (Roche) mostrou 57% e 33% de positividade, respectivamente, contra 47% e 24% de positividade obtidos para as mesmas amostras usando a PCR em tempo real. A maior sensibilidade (100%) obtida usando-seo ensaio de PCR em tempo real, foi em análises das amostras de secreção endocervical. A PCR em tempo real, comparada com o teste padrão ouro, mostrou uma alta especificidade (100%) e um valor preditivo positivo de 100% para as diferentes amostras clínicas estudadas. Em conclusão, os dados apresentados neste estudo indicam que a PCR em tempo real mostrou um alto desempenho para detecção de C. trachomatis em diferentes espécimes clínicos...

Infections caused by Chlamydia trachomatis are now recognized as the most prevalent and are among the most damaging of all sexually transmitted diseases (STD) worldwide. Lack of symptoms makes clinical diagnosis of chlamydial infection difficult. One of the challenge in the prevention of C. trachomatis infections is the availability of laboratory diagnostics with high sensitivity, specificity and reliable. In the present study, the performance of conventional polymerase chain reaction (PCR) and Real-time PCR (SYBRR –Green) tests used for detection of Chlamydia trachomatis in urine and urogenital specimens were compared. The COBAS AmplicorR (Roche) is a conventional PCR and was used as a tool of control and gold standard test for diagnosis in this work. A total of 136 samples of endocervical secretion (61), uretral secretion (14) and urine (61), were obtained from patients submitted of Chlamydia trachomatis detection tests of Laboratório Alvaro S/A. Real-time PCR (SYBRR –Green) and COBAS AmplicorR Chlamydia trachomatis Test (Roche) methods were equally sensitivity for diagnostic of C. trachomatis in uretral secretion samples. However, in analyses of the urine and endocervical secretion samples the COBAS AmplicorR test displayed 57% and 33% of positivity, recpectively, against 47% and 24% of positivity obtained for the same samples using Real-time PCR. The highest sensibility (100%) using Real-timePCR assay was obtained in analyses of endocervical secretion samples. Real-time PCR, comparing with our gold standard showed a high spedificity (100%) and a positive predictive value of 100% for different clinical samples studied. In conclusion, the data presented in this study indicate that PCR - Real time have shown a high performance for detection of C. trachomatis in different clinical specimens. Moreover, this method can be used as a powerful tool in diagnostic and epidemiological investigation of infection associated with C. trachomatis.

Uremic hyperhomocysteinemia: a randomized trial of folate treatment for the prevention of cardiovascular events.

Homocysteine is a risk factor for atherosclerosis in the general population, and serum homocysteine levels are almost universally elevated in chronic renal failure patients. When such patients are treated with dialysis, cardiovascular disease accounts for more than 50% of their mortality, which, in some proportion, may be pathophysiologically related to the elevated serum homocysteine levels. From April 2003 to March 2005, we conducted a 2-year, double-blind, randomized, placebo-controlled trial of 186 patients with end-stage kidney disease due to any cause, who were older than 18 years and stable on hemodialysis. Patients were assigned to receive either oral folic acid 10 mg 3 times a week immediately after every dialysis session under nurse supervision or an identical-appearing placebo for the entire study. On admission, plasma total homocysteine (tHcy) levels were above 13.9 micromol/L in 96.7% of patients (median 25.0 micromol/L, range 9.3-104.0 micromol/L). In the placebo group, tHcy levels remained elevated at 6, 12, and 24 months, while oral folate significantly decreased tHcy to a median value of 10.5 (2.8-20.3) micromol/L, (p<0.01). During the study, 38 patients (folic acid group 17 vs. placebo group 21; p=0.47) died from cardiovascular disease. Kaplan-Meier life table analysis dealing with the incidence of cardiovascular events, both fatal and nonfatal (myocardial infarction, arrhythmias, angina, heart failure, cerebrovascular accident), showed that 2 years of folic acid treatment and the lowering of the homocysteine blood levels had no effect on cardiovascular events (p=0.41; hazard ratio 1.24, 95% CI 0.74-2.10). However, the carotid artery intima-media wall thickness measured in a blinded fashion decreased from 1.94 +/- 0.59 mm to 1.67 +/- 0.38 mm (p<0.01) after 2 years of folate therapy. In this short-term study of uremic patients, 2 years of folic acid supplementation normalized the tHcy blood levels in 92.3% of patients but did not change the incidence of cardiovascular events compared with the control group. However, ultrasonography of the common carotid arteries performed at entry and 24 months later showed a significant decrease in intima-media thickness with folate supplementation. This suggests that early folate supplementation may benefit patients with chronic renal failure by preventing cardiovascular deterioration.

Desenvolvimento e avaliação de um sistema automatizado biosseguro para o tratamento, reciclagem e descarte de resíduo de microbiologia clínica / Development and evaluation of a biosafe automatized system for treatment, recycling and discarding of clinical microbiology residues

No Brasil, diariamente, são descartadas 2,3 toneladas de meios de cultura potencialmente contaminados com microorganismos. A resolução RDC No 306/2004 da Agência Nacional de Vigilância Sanitária preconiza que os resíduos devem ser tratados antes do descarte, visando à redução da carga microbiana. Foram desenvolvidos 2 métodos para avaliar a sensibilidade e a eficiência, assim como a concentração ideal, de um agente químico (biocida) contra microorganismos contaminantes. O método de difusão com Perfurador Circular de Ágar (PCA) e o método com Perfurador Linear de Ágar (PLA) foram testados com 13 cepas de microorganismos. O biocida avaliado foi a combinação de hipoclorito de sódio (NaClO) com ácido acético (CH3COOH). A partir destes resultados, foi desenvolvido um equipamento automatizado para processar a redução da carga microbiana (SADEMC) dos meios de cultura contaminados. A redução da carga microbiana foi avaliada pelo método quantitativo da reação da transcriptase reversa com detecção em tempo real. O método PCA mostrou ser reprodutível e eficiente para medir a inibição do crescimento bacteriano de um biocida. A concentração mínima de biocida capaz de reduzir o crescimento microbiano foi de 250 ppm para a solução aquosa de NaClO a 0,25% e de 200 ppm para a de CH3COOH a 0,2%. No SADEMC, foi possível processar 4,6kg de meios de culturas em 100 litros da concentração mínima eficaz do biocida por 15 minutos, e atingir uma redução da carga microbiana de, aproximadamente, 1,4E10 unidades formadoras de colônias. Podemos concluir que o SADEMC promove uma redução de carga microbiana compatível com os níveis exigidos pela RDC No. 306; fornece biossegurança na sua manipulação e que resulta em plástico reciclável

In Brazil, daily, 2.3 tons of potentially contaminated cultured medium with microorganisms are discarded. The RDC 306 resolution from the Brazilian National Health Department rules out that residue must be treated prior to discart in order to reduce microbial load. Two methods were developed to evaluate the sensitivity, efficiency and ideal concentration of a chemical agent (biocide) against microorganisms. The Ágar's Diffusion Method by Circular Perforator (PCA) and by Linear Perforator (PLA) were tested with 13 microorganism lines and the biocide composed by Sodium Hypochlorite (NaClO) and its combination with Acetic Acid (CH3COOH). The microbial load reduction was evaluated by the real time reverse transcription-polymerase chain reaction. From the in vitro data, an automatic equipment to process the potentially contaminated culture media (SADEMC) was developed. The PCA method was reproductive and efficient to measure the bacterial growth inhibition induced by the biocide. The minimum biocide concentration capable to reduce the microbial growth was a solution of 0.25% NaClO (250 ppm) and 0.2% CH3COOH (200 ppm). In the SADEMC, the direct exposition of 4.6 kg of culture media in 100 liters of biocide for a period of 15 minutes is capable to reduce the microbial load in approximately 1,4E10 of colony-forming unit. We may conclude that the SADEMC is able to promote a microbial load reduction more intense than the one demanded by the RDC 306 resolution. In addition to that, the SADEMC contemplates personnel safety and allows recycling the plastic residues

Estudo do perfil de hemoglobinas em 9, 189 testes realizados no Alvaro Centro de Análises e Pesquisas Clínicas / The characteristic of 9, 189 tests of hemoglobinopathy diagnostic from the Alvaro Center of Clinical Chemistry and Research in the 2004 year

As hemoglobinopatias são um grupo heterogêneo de distúrbios caracterizados por alterações que envolvem genes estruturais e promovem a formação de moléculas de hemoglobinas com variações polimórficas características. Essas alterações, denominadas variantes, possuem freqüências diversificadas, de acordo com distribuição geográfica e/ou étnica. Avaliou-se o perfil hemoglobínico dos testes realizados no Alvaro Centro de Análises e Pesquisas Clínicas, durante o ano de 2004. Foram analisados 9189 testes, realizados por cromatografia líquida de alta performance (HPLC), provenientes de todas as regiões do Brasil. Das amostras analisadas, 81,02% apresentaram perfil hemoglobínico normal, enquanto foi observado alteração em 18,98%. Destas, os perfis hemoglobínicos identificados foram: Hb AS 9,11%; sugestivo de beta talassemia 5,50%; Hb AC 2,47%; Hb SS 0,76%; Hb SC 0,39%; Hb CC 0,20%; PHHF 0,19%; Hb AD 0,18%; e outras formas raras de hemoglobinas em 0,18%. Observou-se que houve predomínio da prevalência de Hb AS (traço falciforme) entre os testes com perfil hemoglobínico alterado. Ainda, houve uma freqüência relativamente elevada de testes sugestivos de beta talassemia.

The hemoglobinopathies are a heterogeneous group of haemoglobin disturbs characterized by the structural genes mutations which results in polymorphic haemoglobin molecules. These abnormal hemoglobins are called “hemoglobin variants”. Their frequencies vary accordingly to the geographic region and/or ethnic group distribution. Quantification of the various hemoglobinic structures was performed in 9189 samples from all over Brazil. The variants hemoglobin frequency and beta-thalassemia were analyzed. The normal hemoglobinic structure and its variants were detected by High Performance Liquid Chromatography (HPLC). We found 7445 (81.02%) samples presenting normal hemoglobin structure, Hb AA; whereas 1744 (18.98%) samples revealed some abnormality. The following characteristics were observed among the abnormal hemoglobins: Hb AS (9.11%); suggestive of beta thalassemia (5.50%); Hb AC (2.47%); Hb SS (0.76%), Hb SC (0.39%); Hb CC (0.20%); HPFH (0.19%); Hb AD (0,18%); and some other rare hemoglobins (0.18%). From our study, we were able to identify a major prevalence of sickle cell trait (Hb AS) among the various haemoglobin variants. In addition to that, we were able to identify a higher relative frequency of suggestive of beta-thalassemia tests in this population.

Comparação de dois métodos de PCR para detecção da mutação de Leiden no fator V / Comparison of two PCR methods for the detection of the factor V Leinden mutation

Um importante fator de predisposição genética para trombose venosa em humanos é uma mutação de G->A no nucleotídeo 1691 do gene que codifica o fator V da coagulação, conhecida como a mutação de Leiden. Técnicas de biologia molecular como a reação em cadeia da polimerase (PCR) tem demonstrado ser de grande importância na detecção dessa mutação. O objetivo do presente estudo é comparar os resultados de dois diferentes métodos, baseadas na técnica de PCR, na detecção da mutação de Leiden no gene do fator V. O primeiro método foi realizado usando o kit comercial AMS50 (Clonit), enquanto o segundo, um método publicadopor Hezard et al., 1997. Ambos os métodos detectam o alelo selvagem e o alelo mutado, e permitem classificar o genótipo como sendo homozigoto selvagem, heterozigoto ou homozigoto mutado. A análise do DNA extraído do sangue total de 38 indivíduos usando esses dois métodos de PCR forneceu os mesmos resultados, e permitiu identificar 32 indivíduos homozigotos selvagens e 6 indivíduos heterozigotos para a mutação de Leiden. Esses resultados confirmam a importância desses dois métodos, baseados na PCR, na detecção dessa mutação em rotina clínica laboratorial

Análise crítica da pseudosepticemia e falso negativo: valor diagnóstico das hemoculturas / Critical analysis of the pseudosepticemia and false negative: diagnostic value of the hemoculturas

A septicemia é uma das causas de morbidade e mortalidade em crianças e adultos. A dificuldade diagnóstica está associada tanto à obtenção de amostras, assim como ao sistema utilizado na cultura, seja automatizada ou manual. O objetivo foi determinar a freqüência de pseudosepticemia e falso negativo nos procedimentos automatizados e manuais em hemoculturas de dois centros diagnósticos: Hospital Universitário e um Centro de Análises e Pesquisas Clínicas. Utilizou-se Bactec 9240 e 9050 para hemocultura de 973 amostras, seguido de cultura em Agar sangue, Agar chocolate, MacConkey e Sabouraud. A identificação bacteriana foi realizada pelo sistema Vitek (automatizado) e manual utilizando provas bioquímicas em tubos. Estudo comparativo entre o sistema automatizado e o manual foi realizado utilizando 41 amostras de sangue. Os resultados mostram 12,33 porcento de positividade no total em ambos os centros. Das 932 culturas realizadas 3,86 porcento foram consideradas pseudo septicemias, e 8,47 porcento septicemias. As pseudo septicemias foram representadas por Staphylococcus coagulase negativa e as septicemias por: Staphylococcus aureus (19,0 porcento), Escherichia coli (13,9 porcento), Staphylococcus coagulase negativa (11,4 porcento), Acinetobacter baumanii (8,9 porcento ), Enterecoccus faecalis (7,6 porcento), Klebsiella pneumoniae (7,6 porcento), Enterobacter cloacae (5,1 porcento), Streptococcus pneumoniae (5,1 porcento ), outros (11,4 porcento) e leveduras (10 porcento ). Nas 41 culturas realizadas pelos sistemas automatizado e manual detectaram-se 5 e 3 amostras positivas, respectivamente, com resultado falso negativo de 40 porcento para o sistema manual. O percentual de hemoculturas positivas foi semelhante nos dois centros mostrando representatividade de amostra. A elevada incidência de pseudo septicemia enfatiza os cuidados pré-analíticos e a evidência de um alto percentual de resultados falsos negativo portanto não recomendando o uso do sistema manual para a rotina de hemoculturas.

Cistinúria - relato de caso e revisäo de literatura / Cystinuria - report of case and revision of literature

Cistinúria é uma doença genética autossômica recessiva, que na forma heterozigota leva a moderadas excreçöes dos aminoácidos cistina, lisina, arginina e ornitina na urina, e na forma homozigota leva à perda maciça dos quatro aminoácidos na urina e alto risco de formaçäo de cálculos renais. Descrevemos aqui o caso de uma criança, com história familiar de formaçäo de cálculos renais, que começou a eliminá-los aos 2 meses de idade, sendo diagnosticada e tratada 4 meses depois. Atualmente, com 4 anos de idade, apresenta funçäo renal normal, teve um episódio de infecçäo urinária e eliminou 10 cálculos após o início do tratamento. Foi submetida a procedimemto cirúrgico para retirada de um cálculo. Nos últimos 3 anos, sob tratamento, näo houve eliminaçäo ou evidência de formaçäo de cálculos renais.(au)