MAMMALIAN TARGET OF RAPAMYCIN COMPLEX 1 IS INVOLVED IN DIFFERENTIATION OF REGENERATING MYOFIBERS IN VIVO

This work was undertaken to provide further insight into the role of mammalian target of rapamycin complex 1 (mTORC1) in skeletal muscle regeneration, focusing on myofiber size recovery. Rats were treated or not with rapamycin, an mTORC1 inhibitor. Soleus muscles were then subjected to cryolesion and analyzed 1, 10, and 21 days later. A decrease in soleus myofiber cross-section area on post-cryolesion days 10 and 21 was accentuated by rapamycin, which was also effective in reducing protein synthesis in these freeze-injured muscles. The incidence of proliferating satellite cells during regeneration was unaltered by rapamycin, although immunolabeling for neonatal myosin heavy chain (MHC) was weaker in cryolesion+rapamycin muscles than in cryolesion-only muscles. In addition, the decline in tetanic contraction of freeze-injured muscles was accentuated by rapamycin. This study indicates that mTORC1 plays a key role in the recovery of muscle mass and the differentiation of regenerating myofibers, independently of necrosis and satellite cell proliferation mechanisms. Muscle Nerve 42: 778-787,2010

Mucosal and systemic anti-GAG immunity induced by neonatal immunization with HIV LAMP/gag DNA vaccine in mice

Vaccines capable of inducing mucosal immunity in early postnatal life until adulthood, protecting early sexual initiation, should be considered as strategies to vaccination against HIV. The HIV-1 GAG protein as a chimera with the lysosome-associated membrane protein (LAMP/gag), encoded by a DNA vaccine, is targeted to the endosomal/lysosomal compartment that contains class II MHC molecules and has been shown to be immunogenic in adult mice. Assuming that one such strategy could help to overcome the immunological immaturity in the early postnatal period, we have evaluated the systemic and mucosal immunogenicity of LAMP/gag immunization in neonatal mice. Intranasal immunization with LAMP/gag vaccine induced higher levels of sIgA and IgG anti-GAG antibodies in intestinal washes than did the gag vaccine. The combination of ID injections and the IN protocol with the chimeric vaccine promoted the increase of Ab levels in sera. Both vaccines induced splenic IFN-gamma- secreting cells against GAG peptide pools, as well as in vivo cytotoxic T lymphocyte (CTL) function, and increased the percentage of CD8+ T cells to the immunodominant class I peptide in gut and spleen. However, only the chimeric vaccine was able to enhance Th1/Th2 cytokine secretion in response to class II GAG peptide and to enhance IL-4-secreting cells against GAG peptides and p24 protein stimuli. Long-lasting humoral and cellular responses were detected until adult age, following neonatal immunization with the chimeric vaccine. The LAMP/gag vaccination was able to induce potent GAG-specific T and B cell immune responses in early life which are essential to elicit sustained and long-lasting mucosal and systemic humoral response. (C) 2010 Elsevier GmbH. All rights reserved.

On the aetiology of bovine neonatal pancytopenia (BNP)

Bovine Neonatal Pancytopenia (BNP) is a novel haemorrhagic disease in sucking calves, characterised by bleeding, haematological changes and high mortality. Dams that gave birth to BNP affected calves were immunized with PregSure® BVD, a highly adjuvanted vaccine against Bovine Viral Diarrhoea (BVD). We can show that bioprocess impurities in the vaccine, originating from the cell line used for vaccine production induces alloantibodies in vaccinated cattle. Via flow cytometry and immunoprecipitation we can demonstrate that PregSure® BVD immunization leads to BNP alloantibody production. BNP alloantibodies target highly polymorphic bovine MHC-I molecules (BoLA I). We sequenced eight BoLA I variants expressed by the production cell line and identified three alleles which are responsible for the majority of PregSure® BVD induced BoLA I reactivity. The BoLA I alleles of BNP unaffected calves are not recognized by the BNP associated alloantibodies of their respective dams. We also examined whether BNP alloantibodies cross-react with human cells, thus being a potential hazard for human colostrum consumers and could show that BNP alloantibodies are cross-reactive to human MHC-I and can even be found in commercial colostrum powder manufactured from cows immunized with PregSure® BVD. Overall we can demonstrate that BNP is a vaccine induced alloimmune disease.

Neonatal Outcomes Of Late Preterm And Early Term Birth.

To compare neonatal deaths and complications in infants born at 34-36 weeks and six days (late preterm: LPT) with those born at term (37-41 weeks and six days); to compare deaths of early term (37-38 weeks) versus late term (39-41 weeks and six days) infants; to search for any temporal trend in LPT rate. A retrospective cohort study of live births was conducted in the Campinas State University, Brazil, from January 2004 to December 2010. Multiple pregnancies, malformations and congenital diseases were excluded. Control for confounders was performed. The level of significance was set at p<0.05. After exclusions, there were 17,988 births (1653 late preterm and 16,345 term infants). A higher mortality in LPT versus term was observed, with an adjusted odds ratio (OR) of 5.29 (p<0.0001). Most complications were significantly associated with LPT births. There was a significant increase in LPT rate throughout the study period, but no significant trend in the rate of medically indicated deliveries. A higher mortality was observed in early term versus late term infants, with adjusted OR: 2.43 (p=0.038). LPT and early term infants have a significantly higher risk of death.

Effects Of Therapeutic Approach On The Neonatal Evolution Of Very Low Birth Weight Infants With Patent Ductus Arteriosus.

To analyze the effects of treatment approach on the outcomes of newborns (birth weight [BW] < 1,000 g) with patent ductus arteriosus (PDA), from the Brazilian Neonatal Research Network (BNRN) on: death, bronchopulmonary dysplasia (BPD), severe intraventricular hemorrhage (IVH III/IV), retinopathy of prematurity requiring surgical (ROPsur), necrotizing enterocolitis requiring surgery (NECsur), and death/BPD. This was a multicentric, cohort study, retrospective data collection, including newborns (BW < 1000 g) with gestational age (GA) < 33 weeks and echocardiographic diagnosis of PDA, from 16 neonatal units of the BNRN from January 1, 2010 to Dec 31, 2011. Newborns who died or were transferred until the third day of life, and those with presence of congenital malformation or infection were excluded. Groups: G1 - conservative approach (without treatment), G2 - pharmacologic (indomethacin or ibuprofen), G3 - surgical ligation (independent of previous treatment). Factors analyzed: antenatal corticosteroid, cesarean section, BW, GA, 5 min. Apgar score < 4, male gender, Score for Neonatal Acute Physiology Perinatal Extension (SNAPPE II), respiratory distress syndrome (RDS), late sepsis (LS), mechanical ventilation (MV), surfactant (< 2 h of life), and time of MV. death, O2 dependence at 36 weeks (BPD36wks), IVH III/IV, ROPsur, NECsur, and death/BPD36wks. Student's t-test, chi-squared test, or Fisher's exact test; Odds ratio (95% CI); logistic binary regression and backward stepwise multiple regression. Software: MedCalc (Medical Calculator) software, version 12.1.4.0. p-values < 0.05 were considered statistically significant. 1,097 newborns were selected and 494 newborns were included: G1 - 187 (37.8%), G2 - 205 (41.5%), and G3 - 102 (20.6%). The highest mortality was observed in G1 (51.3%) and the lowest in G3 (14.7%). The highest frequencies of BPD36wks (70.6%) and ROPsur were observed in G3 (23.5%). The lowest occurrence of death/BPD36wks occurred in G2 (58.0%). Pharmacological (OR 0.29; 95% CI: 0.14-0.62) and conservative (OR 0.34; 95% CI: 0.14-0.79) treatments were protective for the outcome death/BPD36wks. The conservative approach of PDA was associated to high mortality, the surgical approach to the occurrence of BPD36wks and ROPsur, and the pharmacological treatment was protective for the outcome death/BPD36wks.

Hypothermia And Early Neonatal Mortality In Preterm Infants.

To evaluate intervention practices associated with hypothermia at both 5 minutes after birth and at neonatal intensive care unit (NICU) admission and to determine whether hypothermia at NICU admission is associated with early neonatal death in preterm infants. This prospective cohort included 1764 inborn neonates of 22-33 weeks without malformations admitted to 9 university NICUs from August 2010 through April 2012. All centers followed neonatal International Liaison Committee on Resuscitation recommendations for the stabilization and resuscitation in the delivery room (DR). Variables associated with hypothermia (axillary temperature <36.0 °C) 5 minutes after birth and at NICU admission, as well as those associated with early death, were analyzed by logistic regression. Hypothermia 5 minutes after birth and at NICU admission was noted in 44% and 51%, respectively, with 6% of early neonatal deaths. Adjusted for confounding variables, practices associated with hypothermia at 5 minutes after birth were DR temperature <25 °C (OR 2.13, 95% CI 1.67-2.28), maternal temperature at delivery <36.0 °C (OR 1.93, 95% CI 1.49-2.51), and use of plastic bag/wrap (OR 0.53, 95% CI 0.40-0.70). The variables associated with hypothermia at NICU admission were DR temperature <25 °C (OR 1.44, 95% CI 1.10-1.88), respiratory support with cold air in the DR (OR 1.40, 95% CI 1.03-1.88) and during transport to NICU (OR 1.51, 95% CI 1.08-2.13), and cap use (OR 0.55, 95% CI 0.39-0.78). Hypothermia at NICU admission increased the chance of early neonatal death by 1.64-fold (95% CI 1.03-2.61). Simple interventions, such as maintaining DR temperature >25 °C, reducing maternal hypothermia prior to delivery, providing plastic bags/wraps and caps for the newly born infants, and using warm resuscitation gases, may decrease hypothermia at NICU admission and improve early neonatal survival.

[factors Impacting The Growth And Nutritional Status Of Cystic Fibrosis Patients Younger Than 10 Years Of Age Who Did Not Undergo Neonatal Screening.]

The aim of this study was to evaluate by clinical and laboratory parameters how cystic fibrosis (CF) affects growth and nutritional status of children who were undergoing CF treatment but did not receive newborn screening. A historical cohort study of 52 CF patients younger than 10 years of age were followed in a reference center in Campinas, Southeast Brazil. Anthropometric measurements were abstracted from medical records until March/2010, when neonatal screening program was implemented. Between September/2009 and March/2010, parental height of the 52 CF patients were also measured. Regarding nutritional status, four patients had Z-scores ≤ -2 for height/age (H/A) and body mass index/age (BMI/A). The following variables were associated with improved H/A ratio: fewer hospitalizations, longer time from first appointment to diagnosis, longer time from birth to diagnosis and later onset of respiratory disease. Forced vital capacity [FVC(%)], forced expiratory flow between 25-75% of FVC [FEF25-75(%)], forced expiratory volume in the first second [FEV1(%)], gestational age, birth weight and early respiratory symptoms were associated with IMC/A. Greater number of hospitalizations, diagnosis delay and early onset of respiratory disease had a negative impact on growth. Lower spirometric values, lower gestational age, lower birth weight, and early onset of respiratory symptoms had negative impact on nutritional status. Malnutrition was observed in 7.7% of cases, but 23% of children had nutritional risk.

Candidemia neonatal, em hospital público do Mato Grosso do Sul

O objetivo de nosso estudo foi realizar tipagem molecular de 25 amostras clínicas de Candida spp, isoladas de crianças com candidemia, internadas na unidade de terapia intensiva neonatal de um Hospital Universitário entre 1998 a 2006. Dados demográficos e clínicos foram obtidos de prontuários para conhecimento dos aspectos clínicos e epidemiológicos. Identificação das leveduras foi feita por método convencional e a susceptibilidade antifúngica por método de microdiluição. O perfil genético foi determinado pela técnica de RAPD-PCR. Candida albicans (11; 44%) e Candida parapsilosis (10; 40%) foram as mais isoladas. Dezessete (68%) dos recém-nascidos tinham peso inferior a 1.500g. Prematuridade (92%), uso de cateter venoso central (100%), foram as condições de risco mais associados. Dezenove (76%) pacientes foram a óbito. Apenas uma cepa de Candida parapsilosis, mostrou ser sensível dose dependente ao fluconazol. Na análise molecular, foram observados 11 padrões genéticos distintos. Somente em dois casos foi observada relação epidemiológica, sugerindo mesma fonte de infecção.

Sobrevida e fatores de risco para mortalidade neonatal em uma coorte de nascidos vivos de muito baixo peso ao nascer, na Região Sul do Município de São Paulo, Brasil

Estudos populacionais sobre mortalidade neonatal de nascimentos de muito baixo peso ao nascer contribuem para identificar sua complexa rede de fatores de risco. Foi estudada uma coorte de 213 recém-nascidos com peso inferior a 1.500g (112 óbitos neonatais e 101 sobreviventes) na Região Sul do Município de São Paulo, Brasil, em 2000/2001. Foram realizadas entrevistas domiciliares e obtidos dados de prontuários hospitalares. Foi realizada análise de sobrevida e empregada regressão múltipla de Cox. A elevada mortalidade na sala de parto, no primeiro dia de vida e ausência de sobreviventes < 700g dos nascimentos < 1.000g e com menos de 28 semanas sugere que condutas mais ativas destinam-se a nascituros de maior viabilidade. Mães residentes em favela, com história anterior de cesárea e aborto provocado, adolescentes, com sangramento vaginal e ausência de pré-natal aumentaram o risco de óbito neonatal. Partos cesarianos e internação em berçários mostraram efeito protetor. O peso ao nascer abaixo de 1.000g e Apgar menor que 7 foram risco. A elevada mortalidade está associada às condições de vida, características maternas e dos nascimentos e condições assistenciais. A melhoria da atenção pré-natal e ao recém-nascido pode atuar na redução da mortalidade.

Morte neonatal precoce segundo complexidade hospitalar e rede SUS e não-SUS na Região Metropolitana de São Paulo, Brasil

O objetivo foi analisar o perfil dos recém-nascidos, mães e mortalidade neonatal precoce, segundo complexidade do hospital e vínculo com o Sistema Único de Saúde (SUS), na Região Metropolitana de São Paulo, Brasil. Estudo baseado em dados de nascidos vivos, óbitos e cadastro de hospitais. Para obter a tipologia de complexidade e o perfil da clientela, empregaram-se análise fatorial e de clusters. O SUS atende mais recém-nascidos de risco e mães com baixa escolaridade, pré-natal insuficiente e adolescentes. A probabilidade de morte neonatal precoce foi 5,6‰ nascidos vivos (65% maior no SUS), sem diferenças por nível de complexidade do hospital, exceto nos de altíssima (SUS) e média (não-SUS) complexidade. O diferencial de mortalidade neonatal precoce entre as duas redes é menor no grupo de recém-nascidos < 1.500g (22%), entretanto, a taxa é 131% mais elevada no SUS para os recém-nascidos > 2.500g. Há uma concentração de nascimentos de alto risco na rede SUS, contudo a diferença de mortalidade neonatal precoce entre a rede SUS e não-SUS é menor nesse grupo de recém-nascidos. Novos estudos são necessários para compreender melhor a elevada mortalidade de recém-nascidos > 2.500g no SUS.

Características dos nascidos vivos, das mães e mortalidade neonatal precoce na Região Metropolitana de São Paulo, Brasil

O objetivo foi descrever as características do recém-nascido, da mãe e da mortalidade neonatal precoce, segundo local de parto, na Região Metropolitana de São Paulo, Brasil. Utilizou-se coorte de nascidos vivos vinculados aos respectivos óbitos neonatais precoces, por técnica determinística. Identificou-se o parto domiciliar a partir da Declaração de Nascido Vivo e os ocorridos em estabelecimentos a partir da vinculação com o Cadastro Nacional de Estabelecimentos de Saúde. Foram estudados 154.676 nascidos vivos, dos quais 0,3% dos nascimentos ocorreram acidentalmente em domicílio, 98,7% em hospitais e menos de 1% em outro serviço de saúde. A mortalidade foi menor no Centro de Parto Normal e nas Unidades Mistas de Saúde, condizente com o perfil de baixo risco obstétrico. As taxas mais elevadas ocorreram nos prontos-socorros (54,4 óbitos por mil nascidos vivos) e domicílios (26,7), representando um risco de morte, respectivamente, 9,6 e 4,7 vezes maior que nos hospitais (5,6). Apesar da alta predominância do parto hospitalar, há um segmento de partos acidentais tanto em domicílios como em prontos-socorros que merece atenção, por registrar elevadas taxas de mortalidade neonatal precoce.

Matched case-control study evaluating the frequency of the main agents associated with neonatal diarrhea in piglets

A case-control study was carried out in litters of 1 to 7-day-old piglets to identify the main infectious agents involved with neonatal diarrhea in pigs. Fecal samples (n=276) from piglets were collected on pig farms in the State of Rio Grande do Sul, Brazil, from May to September 2007. Litters with diarrhea were considered cases (n=129) and normal litters (n=147) controls. The samples were examined by latex agglutination test, PAGE, conventional isolating techniques, ELISA, PCR, and microscopic methods in order to detect rotavirus, bacterial pathogens (Escherichia coli, Clostridium perfringens type A and C, and Clostridium difficile), and parasites (Coccidian and Cryptosporidium spp.). Outbreaks of diarrhea were not observed during sampling. At least one agent was detected in fecal samples on 25 out of 28 farms (89.3%) and in 16 farms (57.1%) more than one agent was found. The main agents diagnosed were Coccidia (42.86%) and rotavirus (39.29%). The main agents identified in litters with diarrhea were Clostridium difficile (10.6%), Clostridium perfringens type A (8.8%) and rotavirus (7.5%); in control litters, Clostridium difficile (16.6%) and Coccidian (8.5%). Beta hemolytic Escherichia coli and Clostridium perfringens type C were not detected. When compared with controls, no agent was significantly associated with diarrhea in case litters. These findings stress the need for caution in the interpretation of laboratorial diagnosis of mild diarrhea in neonatal pigs, as the sole detection of an agent does not necessarily indicate that it is the cause of the problem.

Uso de hidrolizado proteico en los pacientes de una unida de terapia intensiva neonatal

Introducción. La leche humana es el primer alimento escogido para alimentar a los recién nacidos (RN), especialmente a los prematuros (RNPT y a los de peso muy bajo (RNMBP). Cuando esta leche no está disponible, deben ser recetadas fórmulas especializadas, que promuevan un crecimiento y desarrollo adecuados. El uso de los hidrolizados proteicos (HP) en la UTI neonatal ha sido promovido, debido al riesgo potencial que los RNPT/MBP presentan, por la inmadurez gastrointestinal, problemas de absorción e intolerancia alimentaria. Sin embargo, tales formulaciones no fueran elaboradas para atender las necessidades específicas de estos niños, y pueden, cuando son utilizadas, ocasionar un déficit nutricional. Objetivo. Comparar la progresión de la nutrición enteral (NE) (a través de la oferta de volumen, calorías y proteínas), la evolución ponderal, y las complicaciones clínicas entre recién nacidos de peso muy bajo (RNPMB) alimentados con fórmula para prematuros (FP) e hidrolizado proteico (HP) en la UTI neonatal. Casuística y métodos. Fueran estudiados dos grupos de RNPMB: grupo 1:13RNPMB con edad gestacional media de nacimiento de 30 semanas, peso medio de nacimiento de 1 167 gramos, que recibieron FP. Grupo 2:8 RN con edad gestacional media de nacimiento de 29 semanas, peso medio de nacimiento de 1 141 gramos, alimentados con HP. Las ofertas de volumen, la de calorías y la de proteínas fueran diariamente calculadas para todos los niños en ambos grupos. La nutrición parenteral (NP) que complementaría a la terapia nutricional en las primeras semanas, también fue calculada. Los RNMBP fueron pesados diariamente, por la mañana, en balanza digital (escala de 10 g) debidamente calibrada. El peso medio diario fue calculado hasta el dia 15 de vida. Fueron construidas curvas de crescimento, a través del polinomio de segundo grado para ambos grupos. Para el análisis estadístico se utilizó el test de medias. Resultados. ) La oferta media de volumen, calorías y proteínas fue mayos en la primera semana en el grupo 2, pero de una forma no significativa (p=0.38; 0.34 y 0.05, respectivamente). En las semanas subsequentes, no hubo diferencias significativas en la progresión de NE entre los grupos. No fueron observadas complicaciones clínicas, sólo ocurrieron dos casos de distensión abdominal en el grupo 1, pero no fueron caracterizados como enterocolitis necrosante (ECN). No hubo diferencias estadísticas relacionadas con el aumento de peso entre los dos grupos durante el periodo de estudio; sin embargo, el grupo 1 presentó una tendencia de mejor evolución. conclusión. La prograsión de la NE fue semejante con la utilización de los dos tipos de fórmulas, no feron encontradas diferencias en el aumento de peso de los grupos, y no hubo complicaciones clínicas en ninguno de ellos. No hubo ventajas en el uso del HP en la UTI neonatal; por tanto, no está indicado para RNPMB, sin disfunción intestinal, debido a que su composición nutricional no prevé las necesidades de este grupo de niños

Maternal immunization with ovalbumin prevents neonatal allergy development and up-regulates inhibitory receptor Fc gamma RIIB expression on B cells

Background: Preconception allergen immunization prevents neonatal allergen sensitization in mice by a complex interaction between regulatory cells/factors and antibodies. The present study assessed the influence of maternal immunization with ovalbumin (OVA) on the immune response of 3 day-old and 3 week-old offspring immunized or non-immunized with OVA and evaluated the effect of IgG treatment during fetal development or neonatal period. Results: Maternal immunization with OVA showed increased levels of Fc gamma RIIb expression in splenic B cells of neonates, which were maintained for up to 3 weeks and not affected by additional postnatal OVA immunization. Maternal immunization also exerted a down-modulatory effect on both IL-4 and IFN-gamma-secreting T cells and IL-4 and IL-12-secreting B cells. Furthermore, immunized neonates from immunized mothers showed a marked inhibition of antigen-specifc IgE Ab production and lowered Th2/Th1 cytokine levels, whereas displaying enhanced Fc gamma RIIb expression on B cells. These offspring also showed reduced antigen-specific proliferative response and lowered B cell responsiveness. Moreover, in vitro evaluation revealed an impairment of B cell activation upon engagement of B cell antigen receptor by IgG from OVA-immunized mice. Finally, in vivo IgG transference during pregnancy or breastfeeding revealed that maternal Ab transference was able to increase regulatory cytokines, such as IL-10, in the prenatal stage; yet only the postnatal treatment prevented neonatal sensitization. None of the IgG treatments induced immunological changes in the offspring, as it was observed for those from OVA-immunized mothers. Conclusion: Maternal immunization upregulates the inhibitory Fc gamma RIIb expression on offspring B cells, avoiding skewed Th2 response and development of allergy. These findings contribute to the advancement of prophylactic strategies to prevent allergic diseases in early life.