Btk Mutations Selectively Regulate Btk Expression And Upregulate Monocyte Xbp1 Mrna In Xla Patients.

Mutations in the Bruton agammaglobulinemia tyrosine kinase (BTK) gene are responsible for X-linked agammaglobulinemia (XLA). Unfolded or misfolded proteins can trigger stress pathways in the endoplasmic reticulum (ER), known as unfolded protein response (UPR). The aim was to clarify the involvement of UPR in XLA pathophysiology. By reverse transcription-quantitative PCR, we evaluated the expression of BTK and 12 UPR-related genes in eight patients. Moreover, we assessed the BTK protein expression and pattern in the patients' monocytes by flow cytometry and fluorescence immunocytochemistry. We found a reduced BTK expression in patients with stop codon mutations (P < 0.02). However, missense mutations did not affect BTK expression. Flow cytometry showed a reduction of BTK in patients which was corroborated by an absent or nonfunctional protein synthesis revealed by immunocytochemistry. In contrast with the other UPR-related genes, X-box binding protein 1 (XBP1) was markedly upregulated in the patients (P < 0.01), suggesting Toll-like receptor (TLR) activation since BTK directly interacts with TLRs as a negative regulator and XBP1 can be activated in direct response to TLR ligation. Different BTK mutations can be identified by the BTK expression. Inasmuch as UPR-related genes were downregulated or unaltered in patients, we speculate the involvement of the TLRs-XBP1 axis in the XLA pathophysiology. Such data could be the basis for further studies of this novel pathomechanism concerning XLA.3171-18

Vacinas pertussis acelular e pertussis "low": eventos adversos e o papel das mutações

Objective: to discuss the current PAHO recommendation that does not support the substitution of traditional cellular DTP vaccine by acellular DTP, and the role of mutations, in humans, as the main cause of rare adverse events, such as epileptic-like convulsions, triggered by pertussis vaccine. Data review: the main components related to toxic effects of cellular pertussis vaccines are the lipopolysaccharide of bacterial cell wall and pertussis toxin. The removal of part of lipopolysaccharide layer has allowed the creation of a safer cellular pertussis vaccine, with costs comparable to the traditional cellular vaccine, and which may be a substitute for the acellular vaccine. Conclusion: The new methodology introduced by Instituto Butantan allows for the development of a new safer pertussis vaccine with low LPS content (Plow), and the use of the lipopolysaccharide obtained in the process in the production of monophosphoryl lipid A. This component has shown potent adjuvant effect when administered together with influenza inactivated vaccine, making possible to reduce the antigen dose, enhancing the production capacity and lowering costs.Objetivo: Discutir as recomendações da WHO-OPAS que não consideram indicada a substituição da vacina DTP celular clássica pela DTP acelular e o papel de mutações, em humanos, como principal causa dos raros eventos de convulsões epileptiformes desencadeadas pela vacina pertussis. Revisão dos dados: Os principais componentes relacionados aos efeitos tóxicos da vacina pertussis celular são o lipopolissacarídio da parede celular da bactéria e a toxina pertussis. A remoção de parte da camada lipopolissacarídica permitiu a criação de uma vacina pertussis celular, mais segura e de custo comparável ao da vacina celular tradicional, podendo substituir a vacina pertussis acelular. Conclusão: A nova vacina pertussis, com baixo teor de LPS (Plow) desenvolvida pelo Instituto Butantan, além de oferecer uma vacina mais segura, permite o aproveitamento do lipopolissacarídeo para a produção de monofosforil lipídeo A. Esse componente mostrou-se potente como adjuvante e altamente eficiente quando administrado com a vacina de influenza, levando à possibilidade de se reduzir a dose de antígeno, aumentando a capacidade de produção e redução dos custos

Research On Zinc Blood Levels And Nutritional Status In Adolescents With Autoimmune Hepatitis.

Zinc deficiency in children and adolescents impairs their growing, development and immune system. To verify the existence of plasma and leukocyte zinc deficiency in adolescents with autoimmune hepatitis. The study comprised 23 patients with autoimmune hepatitis, aged 10-18 years, assisted at the Ambulatory Service of Pediatric Hepatology of the University of Campinas Teaching Hospital, Campinas, SP, Brazil, and adolescents with ages compatible with the patients' ages comprised the control group. Sample of blood in both groups was collected for the analyses of plasma zinc and leukocyte zinc by atomic absorption spectrophotometry, beyond the nutritional status was evaluated in each adolescent. The following statistical tests were used: Mann-Whitney, Spearman's correlation and interclass concordance analysis. The significance level adopted was 5%. The average zinc level in plasma in patients was 71.91 ± 11.79 µg/dL and, in the control group, it was 80.74 ± 10.92 µg/dL, showing a significant difference (P = 0.04). The leukocyte zinc level in patients was 222.33 ± 166.13 pmol/10⁶ cells and, in the control group, it was 226.64 ± 217.81 pmol/10⁶ cells; there was no statistical significance between them (P = 0.45). The evaluation of the nutritional status showed that eutrophy is prevalent in patients, and they presented a higher body fat value than the control group, with a significant difference. More research is needed with adolescents with autoimmune hepatitis regarding levels of essential micronutrients, such as zinc, because a good nutritional status can improve the prognostic of liver disease.4862-

Evolution of nutritional status of infants infected with the human immunodeficiency virus

CONTEXT: There are today only a limited number of studies defining growth parameters and nutritional status for HIV children. OBJECTIVE: To study the nutritional status of infants infected with the human immunodeficiency virus. TYPE OF STUDY: Longitudinal study. SETTING: Department of Pediatrics, Faculty of Medical Sciences, UNICAMP, Campinas, Brazil. PARTICIPANTS: One hundred and twenty-four children born to HIV infected mothers were evaluated from birth until the age of two years. They were subdivided into two groups: 71 infected children and 53 non-infected children. MAIN MEASUREMENTS: Growth was evaluated in both groups by comparing Z-scores for weight/age (w/a), length/age (H/a) and weight/length (w/H) (using the NCHS curves as reference). RESULTS: The Z-score analyses showed that there was a significant difference between the two groups for all the variables studied, except for the H/a value at 3 months of age and the W/H value at 21 months of age, which showed P > 0.05. CONCLUSIONS: The growth of infected infants was observed to be severely affected in comparison with that of seroreversed infants in the same age groups. Although clinical manifestations may take time to appear, the onset of growth changes begin soon after birth.CONTEXTO: Atualmente há estudos limitados que definem parâmetros de crescimento e estado nutricional em crianças com HIV. OBJETIVO: Estudar o estado nutricional de crianças infectadas com o vírus da imunodeficiência humana. TIPO DE ESTUDO: Estudo longitudinal. LOCAL: Serviço de imunodeficiência pediátrica da Universidade Estadual de Campinas, Campinas, Brasil. PARTICIPANTES: 124 crianças nascidas de mulheres infectadas com o HIV foram avaliadas desde o nascimento até dois anos de idade. Elas foram subdivididas em dois grupos: 71 crianças infectadas e 53 crianças não-infectadas. PROCEDIMENTOS: A avaliação do estado nutricional foi realizada pela comparação do Z-score para peso/idade(P/I),altura/idade(A/I) e peso/altura(P/A),usando as curvas do NCHS como referência. Para a comparação do Z-score foram utilizados os testes T de Student e U de Mann-Whitney. Os testes foram conduzidos ao nível de significância=5%. RESULTADOS: A análise do Z-score revelou que houve evidência de diferença entre os dois grupos, para todas as variáveis estudadas, exceto o índice de A/I aos 3 meses e P/A aos 21 meses. CONCLUSÕES: O estado nutricional das crianças infectadas foi gravemente afetado em comparação com as crianças sororreversoras no mesmo grupo de idade. Embora as manifestações clínicas possam demorar a aparecer, as alterações no crescimento surgem logo após o nascimento.Pontifícia Universidade Católica de Campinas Department of NutritionUniversidade Metodista de PiracicabaState University of Campinas Faculty of Medical Sciences Department of PediatricsFederal University of São Paulo Department of PediatricsUNIFESP, Department of PediatricsSciEL

Lack Of Association Between Nutritional Status And Change In Clinical Category Among Hiv-infected Children In Brazil.

Malnutrition is common among HIV-infected children. Our objective was to study the occurrence of malnutrition and its relationship with changes in clinical category among HIV-infected children. Longitudinal study, at the Pediatrics Department and Pediatrics Investigation Center (CIPED), Faculdade de Ciências Médicas da Universidade Estadual de Campinas (Unicamp). We reviewed the hospital records of 127 vertically HIV-infected children. Anthropometric measurements were obtained at the beginning of follow-up, at clinical category change and five months later. These were converted to z-scores of weight/age, height/age and weight/height. Data were presented as means, standard deviations, frequency counts and percentages. The Wilcoxon and Kruskal-Wallis tests and odds ratios were used in the analysis. We found that 51 (40.2%) were undernourished and 40 (31.5%) were stunted, with higher risk of being included in clinical category C. There was an association between nutritional condition and the clinical categories of the Centers for Disease Control classification (1994), and with age at symptom onset (except for height z-score). During follow-up, 36 patients (28.4%) changed their clinical category, which occurred early among the undernourished patients. The group that changed its clinical category maintained the same z-score distribution for weight, height and weight/height throughout follow-up. Aids manifestation severity was associated with nutritional status and with age at symptom onset, but change in clinical category was not followed by worsening of nutritional status.12362-

Whey And Soy Protein Supplements Changes Body Composition In Patients With Crohn's Disease Undergoing Azathioprine And Anti-tnf-alpha Therapy.

Crohn´s disease (CD) is a chronic transmural inflammation of the gastrointestinal tract of unknown cause. Malnutrition associated with active CD has been reduced although obesity has increased. Dietary strategies such as those with high-protein have been proposed to reduce body fat. This study compares the effects of two supplements on the nutritional status of CD patients. 68 CD patients were randomized in two groups: whey protein group (WP) and soy protein group (SP). Using bioimpedance analysis, anthropometry and albumin and pre-albumin dosages the nutritional status was measured before starting the intervention and after 8 and 16 weeks. The disease activity was determined by Crohn's Disease Activity Index and serum C-reactive protein dosage and dietary intake by 24h dietary recalls. Forty-one patients concluded the study and both supplements changed body composition similarly. Triceps skin fold thickness (p< 0.001) and body fat percentage (p=0.001) decreased, whereas mid-arm muscle circumference (p=0.004), corrected arm muscle area (p=0.005) and body lean percentage (p=0.001) increased. For Crohn's disease patients undergoing anti TNF-alpha and azatioprine therapies, supplementation with whey and soy proteins changes body composition through reduction of body fat and thus contributes to control inflammation.311603-161

Acceptability Of An Alimentary Supplement Of Whey-protein Concentrate And Tgf- β In Patients With Crohn's Disease.

The objective of this study was to evaluate the acceptability of an alimentary supplement of bovine whey-protein concentrate (WPC) and TGF- β , unavailable commercially, by patients with Crohn's disease (CD) and determine the chemical composition, solubility, and total amino acids content. The supplement was diluted in water, and an acceptance test was done to evaluate the aroma, flavour, and viscosity of the product using facial hedonic scale (nine-point scale), applied on 54 CD patients. The supplement composition indicated 73.3% protein, 10.5% fat, 2.2% ash, 6.3% water, and 7.7% carbohydrate. The supplement is presented as a good protein source and high content of essential amino acids. The average acceptance for all the attributes was between 5.0 and 6.0, and the flavour was mainly associated with soybean/grain, sour milk, and sweet/vanilla flavour. The results indicated that the supplement provided important nutritional properties for CD patients; however, for a large number of individuals to be encouraged to perform supplementation, it is essential to improve the sensory quality of the product. In order to do so, additional research is necessary to prevent the formation of volatiles which cause off-flavours or to mask undesirable aromas/flavours found in it.201394786

Hepatotoxicity In Hiv-infected Children And Adolescents On Antiretroviral Therapy.

Adverse drug reactions are a significant problem in patients on antiretroviral therapy (ART). We determined liver enzyme elevation frequencies in HIV-infected children and adolescents receiving ART, and their association with risk factors. Cross-sectional study, at the Pediatrics Immunodeficiency Division, University Hospital, Universidade Estadual de Campinas. Medical records of 152 children and adolescents (54.6% male; median age 7.48 years) were analyzed, with a mean of 2.6 liver enzyme determinations per patient. Clinically, patients were classified in categories N (6), A (29), B (78) and C (39). Serum levels of aspartate aminotransferase and alanine aminotransferase were evaluated. Hepatotoxicity was scored as grade 1 (1.1-4.9 times upper limit of normality, ULN), grade 2 (5.0-9.9 times ULN), grade 3 (10.0-15.0 times ULN) and grade 4 (> 15.0 times ULN). To assess hepatotoxicity risk factors, odds ratios (OR) and adjusted odds ratios (aOR) for age, gender, TCD4+ cell count, viral load and medication usage were calculated. We observed grade 1 hepatotoxicity in 19.7 % (30/152) patients. No cases of grade 2, 3 or 4 were detected. There was a significant association between hepatotoxicity and use of sulfonamides (OR, 3.61; 95% confidence interval (CI), 1.50-8.70; aOR, 3.58; 95% CI, 1.44-8.85) and antituberculous agents (OR, 9.23; 95% CI, 1.60-53.08; aOR, 9.05; 95% CI, 1.48-55.25). No toxicity was associated with ART. One fifth of patients experienced mild hepatotoxicity, attributed to antituberculous agents and sulfonamides. Our results suggest that ART was well tolerated.125205-

Clinical trials field strategies with novel vaccines produced in Brazil

OBJECTIVE: To report field strategies applied to clinical trials with vaccines developed by Instituto Butantan in Campinas, Brazil, in 2004 and 2006. METHODS: This report describes the planning and the operational issues of two clinical trials conducted to evaluate immunogenicity and safety of recombinant Hepatitis B vaccine combined with BCG vaccine (BCG/VrHB-IB) and quadrivalent diphteria-tetanus-Haemophilus influenzae type b-cellular pertussis vaccine with low lipopolysaccharide (DTPm/Hib). RESULTS: The main field strategies applied were: a) Partnership between the researchers and managers from Municipal Health Department and b) Research procedures at home or in Health Centers attended by participants. In the first study, BCG vaccine and VrHB-IB (combined or separated) were given to 552 newborns in the maternity, followed by two subsequent doses of VrHB-IB vaccine in households. The second study included 241 infants at Health Centers, which were vaccinated with DTPm/Hib vaccines concomitantly to the others recommended by the National Immunization Program. In both studies, blood samples were taken at home. No adverse events occurred during the experimental period. The field strategies used in those clinical trials allowed adherence by 90.2 and 93.8% of the participants of the first and second study, respectively. The vaccines were given according to the recommendation of National Immunization Program and blood samples were obtained according to the protocol schedules. CONCLUSIONS: The field strategies were important to guarantee enrollment and protocol compliance, causing little interference in families' daily routine, pediatrics appointments and children's vaccine.OBJETIVO: Relatar as estratégias de campo utilizadas em dois ensaios clínicos com vacinas desenvolvidas pelo Instituto Butantan, em 2004 e 2006. MÉTODOS: Estudo do tipo relato de experiência, em que se descreve o planejamento e a operacionalização dos ensaios clínicos, que avaliaram a imunogenicidade e a segurança da vacina BCG combinada com a vacina da hepatite B (VrHB-IB) e da tetravalente bacteriana modificada pela extração do lipopolissacarídeo (LPS) do componente pertussis (DTPm/Hib). RESULTADOS: As principais estratégias de campo utilizadas foram: a) Parceria entre os pesquisadores e os gestores da Secretaria Municipal de Saúde e b) Realização dos procedimentos da pesquisa nos domicílios ou nos Centros de Saúde frequentados pelos participantes. No primeiro estudo, foram vacinados 552 recém-nascidos na maternidade com a BCG/VrHB-IB (combinadas ou separadas) e nos domicílios, com as duas doses subsequentes de VrHB-IB. O segundo estudo incluiu 241 lactentes em Centros de Saúde da rede municipal, vacinados com tetravalente bacteriana (com componente pertussis total ou modificado). Em ambos os estudos, amostras de sangue foram colhidas nas residências. Não houve relatos de eventos adversos. A adesão foi de 90,2% para o primeiro estudo e 93,8%, para o segundo. As vacinas foram administradas nas datas preconizadas pelo Programa Nacional de Imunizações e as coletas de sangue, de acordo com o cronograma dos estudos. CONCLUSÕES: As estratégias utilizadas facilitaram o recrutamento das crianças e garantiram cumprir o protocolo da pesquisa com alta adesão, sem interferir no vínculo da família com o Serviço de Saúde, no calendário vacinal ou no seguimento pediátrico dos participantes.OBJETIVO: Relatar las estrategias de campo utilizadas en dos ensayos clínicos con vacunas desarrolladas por el Instituto Butantan, en 2004 y 2006. MÉTODOS: Estudio de tipo relato de experiencia, en que se describe la planificación y la operacionalización de los ensayos clínicos, que evaluaron la imunogenicidad y la seguridad de la vacuna BCG combinada con la vacuna de la hepatitis B (VrHB-IB) y de la tetravalente bacteriana modificada por la extracción del lipopolisacárido (LPS) y del componente pertussis (DPTm/Hib). RESULTADOS: Las principales estrategias de campo utilizadas fueron: a) Colaboración entre los investigadores y los gestores de la Secretaría Municipal de Salud y b) Realización de los procedimientos de la investigación en los domicilios o Centros de Salud frecuentados por los participantes. En el primero estudio, se vacunaron 552 recién nacidos en la maternidad con la BCG/VrHB-IB (combinadas o separadas), y en los domicilios, con las dos dosis subsiguientes de VrHB-IB. El segundo estudio incluyó a 241 lactantes en Centros de Salud de la red municipal, vacunados con tetravalente bacteriana (con componente pertussis total o modificado). En ambos estudios, muestras de sangre fueron recogidas en las residencias. No hubo relato de eventos adversos. La adhesión fue de 90,2% para el primero estudio y de 93,8% para el segundo. Las vacunas fueron administradas en las fechas preconizadas por el Programa Nacional de Inmunizaciones y las muestras de sangre fueron recogidas conforme al cronograma de estudios. CONCLUSIONES: Las estrategias utilizadas facilitaron el reclutamiento de los niños y garantizaron cumplir con el protocolo de la investigación con alta adhesión, sin interferir en el vínculo de la familia con el Servicio de Salud, en el calendario de vacunas o en el seguimiento pediátrico de los participantes.20220