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Design and development / Respiratory distress syndrome (RDS) is a major cause of mortality and long-term morbidity annually affecting 14% preterm infants worldwide. Therapies have been developed to overcome this common disorder; however, limitations exist with these treatments that often lead to complications including bronchopulmonary dysplasia (BPD). One approach to address RDS is to implement a microfluidic oxygenator that serves as a respiratory support system for preterm neonates while the lungs fully develop, extra-uterine. This artificial lung assist device (LAD) is characterised by its non-invasiveness (given that it is connected via umbilical vessels), pumpless configuration, ambient air operation, portability and low priming volume. Furthermore, the LAD is formed by single oxygenator units (SOU) that are stacked in a parallel array which allows for usage on different body weights. The objective of this thesis is to design an electrochemical system to provide an in-situ enriched O2 environment able to supply 1.9 ml O2/min for use in the SOU while maintaining the simplicity of operation of the oxygenator. An inexpensive, electrically powered and compact device was envisioned allowing for a higher permeation flux to fully oxygenate the blood. Moreover, the system would be easy to manufacture, low maintenance and avoid the risk of gas contamination. In the initial work, different designs of electrolytic cells were developed and tested. The two- chamber design connected by a gel membrane showed an O2 production 10 times higher than with previous designs with 42 mg O2/L. Subsequently, different supporting electrolytes were tested. NaOH demonstrated a better performance and no degradation of the electrode in contrast to NaCl and Na2SO4. Stainless steel mesh (SSM) and graphite sheet electrodes were then tested; it was observed that stainless steel produced 3.4 times more dissolved oxygen (DO) than graphite with 28.3 mg O2/L. Experimentation with electrolysis of water showed that the DO in water reached stability 3 min after the electrolysis process was initiated measuring a change of DO of 29 mg/L at 3 A. Furthermore, an active oxygenation (AO) system was developed for in-vitro experiments via electrolysis of water and compared to a passive oxygenation (PO) system exposing blood to enriched O2 air and ambient air, respectively. It was demonstrated that AO provided 300% greater oxygenation to blood than PO. The electrolysis chamber designed for the microfluidic oxygenator allows the oxygenator to maintain its essential characteristics of simplicity and low cost while increasing the rate of oxygenation of blood. Preterm neonates suffering from RDS need an artificial lung that can partially support the oxygenation of their blood. Thus, combining the oxygenator with the O2 generation in-situ system enables a greater blood O2 uptake of 300% making possible the development of an efficient artificial lung. / Thesis / Master of Applied Science (MASc)

Yes / Aim and Objectives: To explore fathers' views and experiences of providing Kangaroo Care (KC) to their baby cared for in a Neonatal Intensive Care Unit (NICU). Kangaroo Care has been known to improve the health outcome for preterm, low birth weight and medically vulnerable term infants and achieve the optimal perinatal health wellbeing for parents and infants. Historically, mothers are considered as the dominant KC providers, whereas fathers are spectators and have been overlooked. Little is known about the fathers' perspectives in providing KC in NICUs. Methods: Individual semi-structured interviews were conducted with 10 fathers who delivered KC to their baby when in the NICU. Data were analysed using Braun and Clarke's six-phase thematical framework. The Consolidated Criteria for Reporting Qualitative Research (COREQ) checklist was followed to report this qualitative study. Findings: Fathers in this study identified they were passing a silent language of love and connecting with their baby by the act of KC in a challenging environment. Three themes emerged: ‘Positive psychological connection’, ‘Embracing father-infant Kangaroo Care’ and ‘Challenges to father-infant Kangaroo Care’. Conclusion: The findings of this study show KC enhances the bonding and attachment between fathers and infants. The conceptualisation of the paternal role in caregiving to a newborn is evolving as a contemporary practice. Further research is warranted to confirm or refute the study findings. Policies and facilities should be modified to include father–infant KC within the fields of neonatal care. Relevance to Clinical Practice: It is important for nurses and other health professionals to support and enable fathers to give KC. Father–infant KC is recommended in neonatal care settings. / Open access publishing facilitated by University of South Australia, as part of the Wiley - University of South Australia agreement via the Council of Australian University Librarians.

Yes / . Kangaroo care (KC) has been used widely in neonatal care to promote bonding/attachment and neurodevelopment for preterm and term infants. However, current literature suggests that research mainly focuses on infants' and mothers' experiences. The role of fathers in caring for their infant/child is changing and evolving in many countries around the globe. Yet little is known about fathers' experiences of KC in neonatal units. This review, therefore, aims to scope the current evidence of Father-infant KC (Father KC) in Neonatal Intensive Care Units (NICUs). Research question. What impact does KC have upon fathers when their baby is cared for in NIUCs? Search method. A scoping review was conducted and guided by the framework of Arksey and O'Malley (2005). The data sources consisted of Medline, Embase, America Psychological Association (APA) PsycInfo, Emcare, Cochrane Central, Web of Science, Google Scholar and ProQuest. The study inclusion criteria were: (1) studies involving fathers who had experience of KC with their baby whilst in Neonatal Intensive Care Units (NICUs), and other neonatal care settings, such as Special Care Baby Nursery (SCBU), delivery/labour room and postnatal ward; (2) literature published from 2000 to 2020; (3) primary studies including qualitative, quantitative, and mixed methods studies; (4) studies published in English. Results. The total number of studies identified were 13. Seven studies were qualitative and six were quantitative. None were mixed methods. Studies reported several positive KC benefits on fathers, such as reduced stress, promotion of paternal role and enhanced father-infant bond. It was highlighted that KC could be time-consuming for fathers and challenging to practise when balancing work and family life commitments. Conclusion. This review provides evidence that KC practice has health and wellbeing benefits for fathers and infants in NICUs and other relevant neonatal care settings, The findings of this review support the justification to promote Father KC in NICU environments, and guide policies to include father involvement. Implementing Father KC in NICU settings will assist fathers to care and connect with their baby. Further research is needed to explore how to facilitate and evaluate KC education for fathers from diverse s and cultures.

L’activation des lymphocytes T nécessite un double signal. Le premier est antigénique et permet la reconnaissance d’un peptide spécifique présenté à la surface de cellules présentatrices d’antigène (APC). Le second signal est co-stimulateur et implique l’interaction avec des molécules activatrices exprimées par les APC et la présence de cytokines proinflammatoires. Les cellules dendritiques (DC) sont les uniques APC capables de délivrer ce double signal et d'activer les lymphocytes T naïfs, initiant ainsi les réponses immunes primaires. L’immaturité du système immunitaire du nouveau-né est responsable d’une plus grande susceptibilité aux maladies infectieuses ainsi qu’une faible réponse vaccinale. Des déficiences tant au niveau de l’immunité innée que de l’immunité acquise participe à une faible défense face aux agressions. A la naissance, les DC expriment des niveaux faibles de molécules co stimulatrices et présentent un défaut majeur de synthèse d'IL 12, cytokine cruciale pour l’établissement de réponses de type Th1. Le but de ce travail est d’évaluer la capacité des DC du nouveau-né humain à activer les lymphocytes T CD8+. Dans une première approche, nous avons utilisé un modèle unique d’induction de réponse primaire in vitro qui permet d'étudier l'activation de lymphocytes T CD8+ spécifiques de l’antigène Melan-A, une protéine du soi exprimée par les mélanocytes. Ces lymphocytes existent à des fréquences particulièrement élevées chez les individus sains HLA-A2 et présentent les caractéristiques de lymphocytes T naïfs. Dans ce modèle, nous avons d’abord analysé les capacités immunostimulatrices de différentes populations de DC différenciées in vitro. Nous avons observé que les DC différenciées par la culture de monocytes purifiés en présence d'IL-3 et d’IFN-beta sont capables d’initier une réponse fonctionnelle des lymphocytes T CD8+, analogue à celle induite par les DC différenciées en présence de GM-CSF et d’IL-4. Ce même modèle nous a permis de démontrer que, en dépit de leur défaut de production d’IL 12, les DC du nouveau-né sont capables d'induire efficacement une réponse lymphocytaire T CD8+ cytotoxique. Afin dévaluer la relevance in vivo de nos observations, nous avons étudié le phénotype et la fonction des DC circulantes chez des nouveau-nés infectés par le cytomégalovirus (CMV). L’infection par le CMV au cours de la vie fœtale représente une situation clinique où le nouveau-né développe une réponse mature et fonctionnelle des lymphocytes T CD8+, alors que celle des lymphocytes T CD4+ est déficiente. Ces expériences ont montré que le phénotype, la fonction et la réponse à différents stimuli des APC présentes en périphérie ne sont pas affectés par l’infection congénitale par le CMV. Ces résultats suggèrent que l’observation des DC circulantes des nouveau-nés infectés par le CMV ne permet pas d’analyser l’influence du virus sur la fonction des DC néonatales. Dans ce but, nous avons reproduit un modèle d’infection in vitro de DC par une souche primaire du CMV. L’utilisation de micropuces à ADN nous a permis de comparer l’expression de gènes différentiellement induits par l’infection des DC d’adultes et de nouveau-nés. Nous avons ainsi révélé une proportion importante de gènes différentiellement induits, parmi lesquels celui de l’IFN-beta. Nous avons confirmé ce défaut au niveau protéique et mis en évidence une production d’IL 12 déficiente en réponse à l’infection par CMV. L’ensemble de nos résultats indique que malgré leur immaturité, les DC du nouveau-né sont capables, dans certaines circonstances, d’induire une réponse lymphocytaire T CD8+ cytotoxique. Cependant, le défaut de production de certaines cytokines co-stimulatrices pourrait être impliqué dans la faible réponse des lymphocytes T CD4+ à l’infection par CMV. Ces observations ont d’importantes implications pour la compréhension de l’induction de réponses cytotoxiques au cours d’infections virales et pour l’élaboration de stratégies vaccinales en début de vie.

As afecções dos componentes umbilicais merecem destaque no período neonatal, pois são comumente encontradas nos bezerros, podendo levar a graves complicações. O diagnóstico das doenças umbilicais muitas vezes não é totalmente esclarecido pela palpação abdominal bimanual, uma vez que, em parcela dos casos, esse método semiológico não é sensível para determinar o grau de acometimento das estruturas umbilicais intra-abdominais. Assim, a ultrassonografia apresenta-se como um valioso exame complementar para o estabelecimento de um diagnóstico preciso sobre a localização e extensão das onfalopatias intra-abdominais. Diante dessas limitações e da raridade de pesquisas com estabelecimento de padrões da ultrassonografia do umbigo, a proposta desse estudo foi avaliar o processo de involução fisiológica dos componentes umbilicais de bezerros sadios, considerando o uso de antisséptico clássico, a tintura de iodo, em concentrações de 2% e 5%, usado na cura do umbigo nos primeiros dias após o nascimento. A avaliação foi realizada por meio do exame ultrassonográfico dos componentes umbilicais, em diferentes posições, caracterizando as modificações ocorridas durante o processo de involução, em relação ao aspecto das imagens, com medidas de diâmetro e espessura dos componentes vasculares e úraco, assim como às peculiaridades decorrentes dos dois tipos de desinfecção da região. Foram avaliados 23 bezerros da raça Holandesa, machos, oriundos de propriedade leiteira localizada no Estado de São Paulo, desde o nascimento até os 30 dias de vida. Os resultados obtidos, evidenciaram que a veia e as artérias umbilicais perdem as suas características de vasos, assumindo aspecto de ligamento devido à proliferação de tecido fibroso. Esse processo caracterizou-se e seguiu um padrão, no qual o tecido fibroso inicialmente estava presente na região interna da parede do vaso, seguindo, com a involução, em direção ao centro da luz vascular. Esse processo de involução é mais precoce em porções dos vasos mais distantes do umbigo externo, não havendo distinção de comportamento determinada por uso das diferentes concentrações do antisséptico. Além das imagens, foram também padronizadas as medidas do diâmetro dos componentes umbilicais e da espessura de suas paredes, ao longo do processo de involução fisiológica, durante os primeiros 30 dias de vida dos bezerros, comprovando-se a precisão do exame ultrassonográfico para essa avaliação e estabelecendo-se referências ultrassonográficas para fundamentar o diagnóstico e escolha do tratamento das onfalopatias / Diseases of the umbilical components are very important in the neonatal period. Commonly found in calves, they can lead to serious complications. Diagnosis of umbilical diseases is often not fully enlightened by bimanual abdominal palpation, since this semiotic method is not sensitive enough to determine the extent of involvement of the umbilical intra-abdominal structures in part of the cases. Thus, ultrasonography is a valuable complementary test for establishing an accurate diagnosis on the location and extent of intra-abdominal omphalitis. Because of these limitations and the scarce research on the establishment of the standards of navel ultrasound, the purpose of this study was to evaluate the physiological involution process of umbilical components of healthy calves, considering the use of classic antiseptic, iodine tincture in concentrations 2% and 5%, used in navel treatment during the first days after birth. Evaluation was performed by ultrasonography of umbilical components in different positions and the changes were characterized during the process of involution regarding the appearance of the images, the measures of the diameter and thickness of vascular and urachus components, as well as the peculiarities from the two types of disinfecting. Twenty-three Holstein male calves, reared in dairy property located in the State of São Paulo, were evaluated from birth to 30 days old. Results obtained showed that the vein and umbilical arteries lose their blood vessels characteristics, assuming a ligament aspect due to fibrous tissue proliferation. This process was characteristic and followed a pattern, in which the fibrous tissue was initially present in the inner part of the vessel wall, following with involution, toward the center of the vessel lumen. This process of involution was earlier in the parts of the blood vessels that were farthest from the external navel, with no particular behavior distinct by the use of different antiseptic concentrations. Besides the images, the measurements of the diameter of the umbilical components and the thickness of their walls were also standardized along the physiological involution process during the first 30 days of life the calf, confirming the accuracy of ultrasonography for such assessment and establishing references to improve the diagnosis and the choice of treatment of umbilical diseases

Submitted by Luanna Matias (lua_matias@yahoo.com.br) on 2015-03-02T15:21:07Z No. of bitstreams: 2 Dissertação - Angela Ferreira Lopes de Teive e Argolo - 2010.pdf: 1932564 bytes, checksum: 559ed3fd1d33b67d918a9b0e761005ea (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-03-04T10:57:42Z (GMT) No. of bitstreams: 2 Dissertação - Angela Ferreira Lopes de Teive e Argolo - 2010.pdf: 1932564 bytes, checksum: 559ed3fd1d33b67d918a9b0e761005ea (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-03-04T10:57:42Z (GMT). No. of bitstreams: 2 Dissertação - Angela Ferreira Lopes de Teive e Argolo - 2010.pdf: 1932564 bytes, checksum: 559ed3fd1d33b67d918a9b0e761005ea (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2010-12-21 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The research about the epidemiology and the pathogeneses of dengue virus during the gestational period is essential to the understanding of clinical dengue severity among women and their offspring. It has been postulated that children born from women with antidengue antibodies may develop severe dengue during the first viral dengue exposure. The objectives were (a) to determine the prevalence and incidence of dengue infection in pregnant women and their neonates in two public hospitals in the city of Goiania, Goias State, Central Brazil, (b) to assess the percent of newborns with maternal IgG and DENV vertical transmission, (c) to evaluate the association of maternal recent dengue infection (IgM) with the characteristics of the delivery and neonates. A cross sectional study was conducted to assess recent (IgM) and previous (IgG) markers of dengue infection among pregnant women and newborns. The survey was conducted in two public hospitals in the city of Goiania (~1.2 million inhabitants), Central Brazil, from December 2009 to May 2010. We recruited women after giving birth. Women were interviewed to collect data about previous dengue infection, symptoms of dengue during gestational period, pregnancy period, delivery mode and outcomes pregnancy. Blood samples were collected from women and neonates (umbilical cord). Previous infection was defined by the positivity to IgG serology (PAN BIO) and recent infection by IgM (PAN BIO). The detection of NS1Ag and viral nucleic acid (RT-PCR) were performed in the subsample of participants who reported specific symptoms of dengue fever ten days previous to the delivery time. Prevalence and incidence estimates were calculated with 95% Confidence Intervals (95% CI). The agreement between the positive and negative serological results of the pair of women and their concepts was calculated (Kappa Index). Positive and Negative Predictive values between the history of previous dengue and the results of serological markers were calculated. A case-control analysis was performed to assess risk factors among IgM positive women (cases) and the outcomes of pregnancy compared to the seronegative women (controls). Analysis was performed using SPSS 17.0 e o Epi Info 6.04. The investigation was approved by the regional Ethical Committee and all participants signed informed consent according to the Brazilian regulation. A total of 505 women and 505 neonates were investigated. The mean age of women was 25.8 (SD = 6.4), 83.6% of ABSTRACT xv deliveries were between 37 and 41 weeks of pregnancy. The prevalence of IgG dengue antibody was 53.9% (95% IC 49.3% - 58.0%) among women. Maternal antibodies were also found in 99.2% of the offspring (Kappa index = 0.96). NS1 antigen was negative among women and one case of DENV-2 was detected. The incidence of recent dengue infection (IgM positivity and RT-PCR) was 8.9% (95% CI 6.6% - 11.8%) among women and 1.6% (95% IC 0.7%-3.2%) among children. Eight neonates had IgM antibodies with evidence of vertical DENV transmission. The positive predictive value of the previous history of dengue related to seropositivity (IgG) was 88%. The outcomes of pregnancy were similar among seropositive and seronegative women in the case-control analysis. In our setting, half of the pregnant women had dengue antibodies. There was high agreement rates between the serological markers detected among the pair of women and their concepts. Acute dengue infection among pregnant women was not associated with the outcomes unfavorable of pregnancies. / A pesquisa sobre a epidemiologia e patogênese dos vírus dengue durante o período gestacional é essencial para compreender a gravidade clínica da dengue entre mulheres e seus filhos. Postula-se que crianças nascidas de mulheres com anticorpos antidengue podem desenvolver formas graves da doença durante a primeira exposição aos vírus dengue. Os objetivos foram: (a) determinar prevalência e incidência da infecção de dengue em gestantes e seus neonatos em dois hospitais públicos na cidade de Goiânia - Goiás, Centro-Oeste Brasil, (b) avaliar o percentual de recém-nascidos com anticorpos antidengue IgG maternos e a transmissão vertical dos DENV, (c) avaliar a associação de infecção materna recente por DENV (IgM) com as características do parto e dos recém-nascidos. Um estudo transversal foi realizado para avaliar marcadores de infecção recente (IgM) e prévia (IgG) de dengue em gestantes e recém-nascidos. A pesquisa foi realizada em dois hospitais públicos na cidade de Goiânia (1,2 milhões de habitantes), Centro-Oeste do Brasil, de dezembro de 2009 a maio de 2010. Foram recrutadas mulheres após o parto. Estas foram entrevistadas para coletar dados sobre infecção prévia de dengue, sintomas de dengue durante o período gestacional, tipo de parto e desfechos na gravidez. Amostras de sangue foram coletadas das mulheres e recém-nascidos (sangue de cordão umbilical). Infecção prévia por dengue foi definida pela positividade sorológica de IgG (PANBIO) e infecção recente por IgM (PANBIO). A pesquisa de antígeno NS1 e ácido nucléico viral (RT-PCR) foram realizadas em uma subamostra das participantes que relataram sintomas específicos de dengue em até dez dias do parto. As estimativas de prevalência e incidência foram calculadas com seus respectivos intervalos de 95% de confiança (IC 95%). A concordância entre os resultados sorológicos positivos e negativos entre os pares de gestantes e seus conceptos foi calculada (Índice Kappa), assim como os Valores Preditivos (positivo e negativo) entre história prévia de dengue e resultados dos marcadores sorológicos. Uma análise do tipo caso-controle foi realizada para avaliar os fatores de risco no desfecho da gravidez entre mulheres IgM positivo (casos) comparadas às mulheres soronegativas (controles). A análise foi realizada usando SPSS 17.0 e Epi Info 6.04. O estudo foi aprovado pelo Comitê de Ética regional e todas as participantes assinaram o termo de consentimento informado de acordo com a regulamentação brasileira. Um total xiii de 505 mulheres e 505 recém-nascidos foram investigados. A idade média das mulheres foi 25,8 (dp = 6,4), 83,6% dos partos ocorreram entre 37 e 41 semanas de gestação. A prevalência de anticorpos antidengue IgG foi de 53,9% (IC95% 49,3% - 58,0%) entre as mulheres. Os anticorpos maternos também foram encontrados em 99,2% dos recémnascidos (Índice Kappa = 0,96). A pesquisa do antígeno NS1 foi negativa entre as mulheres e um caso de DENV-2 foi detectado. A incidência de infecção recente por dengue (IgM positivo e RT-PCR) foi de 8,9% (IC95% 6,6% - 11,8%) entre as mulheres e 1,6% (IC95% 0,7% -3,2%) entre as crianças. Oito recém-nascidos apresentaram anticorpos IgM com evidência de transmissão vertical de DENV. O Valor Preditivo Positivo da história prévia de dengue relacionados à soropositividade (IgG) foi de 88%. Os desfechos da gravidez foram semelhantes entre as mulheres soropositivas e soronegativas na análise caso-controle. Nesse estudo, metade das gestantes possuía anticorpos antidengue. Houve altas taxas de concordância de marcadores sorológicos detectados entre os pares de mulheres e neonatos. A infecção aguda por dengue em gestantes não foi associada com desfechos desfavoráveis na gravidez.

Die Besiedlung des kindlichen Darms beginnt unmittelbar mit der Geburt. Hier findet der erste Kontakt mit verschiedensten Bakterien statt. Deshalb war die zentrale Fragestellung unserer Untersuchung, wie sich die Bakterienarten bei den Geburtsmodi unterscheiden und wie sich die bakterielle Zusammensetzung im Stuhl von Neugeborenen im Laufe der Jahre verändert hat. Wir untersuchten die Stuhlproben von 42 Neugeborenen, wovon 22 vaginal und 20 per Sectio entbunden wurden, am ersten, zweiten und dritten Lebenstag auf Bakterienwachstum. Es wurden die aufgetretenen Bakteriengattungen mittels Kultur bestimmt und von den aufgetretenen Staphylokokken und Enterokokken die Arten und deren Resistenzen mittels des Phönix-Vollautomaten untersucht. Wir konnten zeigen, dass im Gegensatz zu früheren Untersuchungen heutzutage vermehrt Staphylokokken den Darm von Neugeborenen besiedeln. Außerdem fiel auf, dass per Sectio entbundene Kinder eine verminderte Heterogenität in ihrer Stuhlflora haben und erst später besiedelt werden als vaginal entbundene Kinder.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A síndrome da imunodeficiência adquirida (AIDS), causada pelo vírus da imunodeficiência humana (HIV), é uma das mais destrutivas epidemias do mundo, e a infecção pelo HIV em mulheres jovens vem aumentando rapidamente nos dias atuais. Esse fato tem um impacto importante na transmissão vertical do vírus. Apesar da grande maioria dos casos de aids pediátrica em todo mundo resultar da transmissão vertical, aproximadamente dois terços das crianças expostas ao HIV durante a vida fetal não são infectadas pelo vírus. Muitos trabalhos sugerem que durante a gestação doenças infecciosas maternas podem ter consequências complexas para o desenvolvimento do feto, e poucos trabalhos têm explorado o impacto da exposição ao HIV sobre a responsividade imunológica de crianças não infectadas a diferentes estímulos, particularmente na era das drogas antirretrovirais. Portanto, esse trabalho teve como objetivo avaliar eventos imunes em neonatos não-infectados expostos ao HIV-1 nascidos de gestantes que controlam (G1) ou não (G2) a carga viral plasmática, usando neonatos não expostos como controle. Para tanto, sangue do cordão umbilical de cada neonato foi coletado, plasma e células mononucleares foram separados e a linfoproliferação e o perfil de citocinas foram avaliados. Os resultados demonstraram que a linfoproliferação in vitro induzido por ativadores policlonais foi maior nos neonatos do G2. Entretanto, nenhuma cultura de célula respondeu a um conjunto de peptídeos sintéticos do envelope do HIV-1. A dosagem de citocinas no plasma e nos sobrenadantes das culturas ativadas policlonalmente demonstrou que, enquanto a IL-4 e IL-10 foram as citocinas dominantes produzidas nos grupos G1 e controle, a secreção de IFN-γ, IL-1, Il-6, IL-17 e TNF-α foi significativamente superior nos neonatos G2. Níveis sistêmicos de IL-10 observados dentre os neonatos G1 foram maiores naqueles nascidos de mães tratadas com drogas inibidoras da transcriptase reversa do vírus. Por outro lado, níveis superiores de citocinas inflamatórias foram observados dentre estes nascidos de gestantes tratadas com terapia antirretroviral de alta eficácia. Em resumo, nossos resultados indicam uma responsividade imune alterada em neonatos expostos in utero ao HIV-1 e reforça o papel do tratamento materno anti-viral com drogas menos potentes em atenuar tais distúrbios. / The acquired immunodeficiency syndrome (AIDS), caused by human immunodeficiency virus (HIV), is one the most destructive epidemic in the world, and the HIV-infection in young women has been increasing fast in the recent times. This fact has an important impact on vertical virus transmission. Although the great majority of the pediatric AIDS cases all over the world results from vertical transmission, approximately two thirds of the children exposed to the HIV during the fetal life are not infected by the virus. Many works suggest that during pregnancy, maternal infectious diseases could have complex consequences to the fetus development, and few works have explored the impact of HIV exposition on immunological responsiveness of uninfected children to different stimuli, particularly in the era of the anti-retroviral drugs. Therefore, this work aimed to evaluate immune events in HIV-1-exposed uninfected neonates born from pregnant women who controlled (G1) or not (G2) the plasma viral load, using unexposed neonates as control. Cord blood from each neonate was collected, plasma and mononuclear cells were separated and the lymphoproliferation and cytokine pattern were evaluated. The results demonstrated that the in vitro lymphoproliferation induced by polyclonal activators was higher in the G2 neonates. Nevertheless, no cell culture responded to poll synthetic HIV-1 envelope peptides. The cytokine dosage in the plasma and supernatants of polyclonally-activated cultures demonstrated that, while IL-4 and IL-10 were the dominant cytokines produced in G1 and control groups, the secretion of IFN-γ, IL-1, Il-6, IL-17 and TNF-α was significantly higher in G2 neonates. Systemic levels of IL-10 observed among the G1 neonates were higher in those born from mothers treated with viral transcriptase reverse inhibitors drugs. On the other hand, higher levels of inflammatory cytokines were observed among them born from pregnant women treated with highly active anti-retroviral therapy. In summary, our results indicate an altered immune responsiveness in neonates exposed in utero to HIV and support the role of maternal anti-viral treatment with less potent drugs to attenuate it.

Staphylococcus coagulase-negativo (SCN) estão frequentemente envolvidos em infecções nosocomiais associadas com o uso de cateteres e outros procedimentos médicos invasivos. A habilidade de aderir às superfícies abióticas e de produzir biofilme tem sido reconhecida entre os principais fatores de virulência dos SCN, especialmente de S. epidermidis, a principal espécie responsável por infecções relacionadas à assistência a saúde - IRASs. Dentre as demais espécies de SCN capazes de produzir biofilme, S. haemolyticus tem sido relacionado com quadros de infecções em recém-nascidos (RNs). O presente estudo teve como objetivo principal investigar aspectos microbiológicos e epidemiológicos dos processos infecciosos invasivos relacionados com SCN em neonatos internados em unidade de terapia intensiva neonatal (UTIN) de um hospital universitário do município do Rio de Janeiro (2008-2010). A técnica de PCR multiplex-mPCR foi empregada na determinação das espécies de 40 amostras de SCN isoladas de hemoculturas de RNs fazendo uso de cateteres intravenosos e submetidos à terapia antimicrobiana empírica com vancomicina e/ou gentamicina. A fenotipagem foi realizada por três métodos distintos: Simplificado em microplaca, Vitek 2 e API-Staph. Os perfis de resistência aos antimicrobianos foram verificados através do teste de disco-difusão, determinação de CIM (Oxacilina) e presença do gene mecA. A capacidade de produção de biofilme foi investigada pelos testes do Ágar Vermelho do Congo e ensaios de aderência em superfícies abióticas (poliestireno e vidro) além da PCR para os genes icaAB, atlE e aap. O perfil genômico dos micro-organismos foi determinado pela técnica de PFGE. Os resultados demonstraram o isolamento de S. haemolyticus (77%), S. epidermidis (15%), S. captis (5%) e S. warneri (3%). A análise comparativa dos resultados obtidos pelo m-PCR com métodos fenotípicos demonstrou uma concordância de 97,5% com o esquema simplificado e de ~40% Vitek 2 e o API Staph. A maioria (82,5%) das amostras apresentou perfis variados de multiresistência aos 16 antimicrobianos testados e resistência a oxacilina, apesar de 25% destas não apresentarem o gene mecA. Apesar da maioria das amostras de SCN ter apresentado capacidade de produzir slime e/ou biofilme não foi observada total correlação com a presença dos genes mecA, icaAB, aap, atlE, enfatizando a natureza multifatorial da produção de biofilme de SCN. Diferente do observado para as demais espécies, algumas amostras de S. haemolyticus foram incapazes de aderir ao vidro e ao poliestireno e/ou apresentaram os genes aap (38,7%), atlE (42%) além de icaAB (71%). Na UTIN foi detectada a presença de seis diferentes tipos clonais da espécie prevalente, indicando a disseminação de S. haemolyticusnesta unidade hospitalar e a endemicidade em nossa comunidade. / Coagulase-negative staphylococci (CNS) are often involved in nosocomial infections associated with catheters and other invasive medical procedures. The ability to adhere to abiotic surfaces and produce biofilms has been recognized among the major virulence factors of the SCN, especially Staphylococcus epidermidis, the main species responsible for infections related to health care - IRASs.Among the other species capable of producing biofilm, Staphylococcus haemolyticus has been associated with cases of infections in neonates.Here in, we investigated the microbiological and epidemiological aspects of invasive infections related to CNS in the neonatal intensive care unit (NICU) of a university hospital located at Rio de Janeiro metropolitan area (2008-2010).PCR multiplex-mPCR was used in the determination of the species of 40 CNS strains isolated from blood cultures of neonates making use of intravenous catheters and subjected to empirical antimicrobial therapy with vancomycin and / or gentamicin.Phenotyping was performed by three different methods: Simplified scheme in microplates, Vitek 2 and API-Staph. Antimicrobial resistance profiles were verified by the disk diffusion test, oxacillin MIC determinationand PCR for mecA gene. Evaluation of biofilm production was performed by PCR for genes icaAB, atlE and aap and the tests on Congo Red Agar plates and abiotic surfaces (polystyrene and glass).Clonality was determined by PFGE technique. Data revealed the isolation of S. haemolyticus (77%), S. epidermidis (15%), S. captis (5%) and S. warneri (3%).The comparative analysis of the results obtained by mPCR and phenotypic methods showed 97.5% concordance with the simplified scheme and ~40% with the Vitek 2 and API Staph systems. Different multidrug resistance profiles to 16 antimicrobials, including resistance to oxacillin was observed for 82.5% of the CNS isolates, although the mecA gene was not detected in 25% of these strains.Although most of the CNS isolates showed the ability to produce slime and/or biofilm, a complete correlation with the presence of mecA, icaAB, aap, atlE genes was not observed, emphasizing the multifactorial nature of biofilm production of SCN.Different from other CNS species, some strains of S. haemolyticus were unable to adhere to glass and polystyrene surfaces and/or to exhibit aap (38.7%), atlE (42%), icaAB (71%) genes. PFGE analysis revealed the presence of six different clonal types of the prevalent species, indicating the dissemination in this hospital and endemicity in our community of S. haemolyticus.