Les nouveau-nés et jeunes enfants forment une population spécifique pour laquelle les études cliniques sont rares et difficiles à mettre en œuvre. La modélisation pharmacocinétique permet de réaliser des études non invasives et est donc particulièrement bien adaptée à cette population. Tout comme pour l'expérimentation scientifique qui nécessite plusieurs études pour conduire à un consensus, plusieurs bases de données pharmacocinétiques sont nécessaires afin d'arriver à un modèle optimal généralisable. Néanmoins, de nombreux modèles pharmacocinétiques sont publiés pour une même molécule, indépendamment les uns des autres, sans que l'on sache clairement lequel est le plus adapté. Inversement de nombreuses molécules ne sont pas ou peu étudiées et ne permettent pas de conduire à des recommandations fiables.L'objectif de ce travail de thèse était de rechercher une stratégie pour privilégier un modèle par rapport à un autre et ainsi, conduire à un modèle optimal permettant d'émettre des recommandations. Nous avons choisi de centrer notre intérêt sur les études pharmacocinétiques de population en néonatalogie et plus précisément sur deux molécules: la vancomycine et le phénobarbital. / Neonates and young infants are a specific population in which clinical studies are rare and difficult. Pharmacokinetic modeling allows to realize non invasive studies and is therefore particularly well suited for this population.As well as for scientific experimentation which requires several studies to lead to a consensus, several pharmacokinetic databases are needed to achieve a generalizable optimal model.Nevertheless, many pharmacokinetic models are published for the same molecule, independently of each other, without knowing clearly which is the most suitable. Conversely many molecules are not or little studied and can not lead to reliable recommendations.The aim of this thesis was to find a strategy to prefer a model over another, and thus lead to an optimal model to make recommendations. We chose to focus our interest on population pharmacokinetic studies in neonatology and more specifically on two molecules: vancomycin and phenobarbital.

O presente trabalho teve como objetivo avaliar a influência da cesariana eletiva no eritrograma de bezerros neonatos da raça Nelore durante os primeiros trinta dias de vida. Os neonatos foram alocados em 3 grupos 8 bezerros obtidos por parto espontâneo (PE), 8 bezerros nascidos por cesariana, sem indução do parto (CSI) e 8 bezerros nascidos por cesariana, com indução do parto (CCI) pelo uso de 25 mg de Dexametasona, 24 horas antes do procedimento cirúrgico, pela via intramuscular. Os nascimentos ocorreram entre 282 e 292 dias de gestação. As amostras de sangue para a determinação do quadro eritrocitário foram colhidas por punção da veia jugular externa, utilizando-se Sistema Vacutainer®. A determinação do eritrograma foi realizada no contador automatizado BC-2800 Vet Mindray®. Os resultados obtidos evidenciam que a cesariana eletiva não influenciou o número de hemácias, das taxas de hemoglobina e nos valores do volume globular a crase sanguínea de bezerros na primeira semana de vida. Na primeira semana de vida observou-se, independentemente do grupo experimental uma diminuição no número de hemácias, das taxas de hemoglobina e nos valores do volume globular. Até os 5 dias de vida, os valores de Volume Corpuscular Médio (VCM) e de Hemoglobina Corpuscular Média (HCM) dos bezerros nascidos de partos espontâneos por via vaginal são estatisticamente menores do que os obtidos em bezerros nascidos de cesariana eletiva. A partir de 10 dias de vida observou-se, nos animais obtidos de parto espontâneo, uma rápida recuperação nos valores de hemácias, hemoglobina e volume globular, enquanto nos animais nascidos por cesariana (induzida e não induzida) não ocorria essa recuperação nas taxas de hemoglobina e nos valores do volume globular. Nos bezerros obtidos por cesariana observou-se que no período entre 10 e 30 dias de vida o Volume Corpuscular Médio (VCM) e a Hemoglobina Corpuscular Média (HCM) diminuíram, passando a apresentarem microcitose e hipocromia quando comparados com bezerros obtidos por parto espontâneo. Entre todos os grupos, os animais nascidos por cesariana sem indução apresentaram os valores com menor magnitude do início ao fim do período experimental. Os animais nascidos a termo demonstram uma capacidade de recuperação fisiológica mais apurada que dos animais nascidos por cesariana. / This study aimed to evaluate the influence of elective cesarean section in erythrogram of newborn calves of the Nellore breed for the first thirty days of life. Neonates were divided into 3 groups 8 calves obtained by spontaneous birth (PE), 8 calves born by caesarean section without labor induction (CSI) and 8 calves born by cesarean section with induction of labor (CCI) by the use of 25 mg of dexamethasone 24 hours before the surgical procedure, by the intra-muscular). Births occurred between 282 and 292 days of gestation. Blood samples for determining the erythrocyte frame were collected by puncture of the external jugular vein using Vacutainer ™ System. The determination of erythrocyte was performed in the automated counter BC-2800 Vet Mindray®. The results show that elective caesarean section did no affect the number of red blood cells, hemoglobin rates and values of the globular volume in the first week of life of calves. At this first week of life was observed, regardless of the experimental group a decrease in the number of red blood cells, hemoglobin rates and values of the globular volume. Until 5 days of age, the mean corpuscular volume values (MCV) and Mean Corpuscular Hemoglobin (MCH) of calves born from spontaneous vaginal deliveries are statistically lower than those obtained in calves born from elective cesarean section. From 10 days of life was observed in animals obtained by from spontaneous delivery, a quick recovery in the values of erythrocytes, hemoglobin and packed cell volume, whereas in animals born via caesarean section (induced and uninduced) did not occur such recovery in the rates of hemoglobin and packed cell volume values. In calves obtained by Caesarean section it was observed that in the period between 10 and 30 days of life Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin (MCH) decreased, submitting a microcytic hypochromic when compared with calves obtained by spontaneous delivery. Among all groups, animals born by caesarean section without induction showed the values with lower magnitude from beginning to end of the trial period. Animals born at term show a more accurate physiological recovery capability in comparingson with animals born by Caesarean section.

Esta pesquisa avaliou o efeito da transferência passiva dos leucócitos do colostro na imunidade específica de bezerras recém-nascidas. Foram acompanhadas 20 bezerras Holandesas durante o período neonatal, distribuídas em dois grupos experimentais: grupo COL+ recebeu colostro fresco (4L) proveniente de suas respectivas mães; e grupo COL- recebeu colostro congelado e acelular (4L), oriundo de vacas doadoras de colostro. As avaliações foram realizadas antes da mamada do colostro (M0), 1-2 (M1), 7 (M2), 14 (M3), 21 (M4) e 28 dias pós-nascimento (M5). As bezerras foram submetidas ao exame clínico, seguido da colheita das amostras sanguíneas para realização de hemograma, imunofenotipagem e cultivo celular. Os dois grupos foram colostrados com colostro de igual qualidade com relação à concentração de imunoglobulinas (70-120 g/L). A concentração de células do colostro fresco fornecido ao grupo COL+ foi de 1.895.849 células/mL. Não foi possível encontrar diferenças para as funções vitais em relação aos grupos experimentais. O exame específico dos sistemas revelou um caso de broncopneumonia, três de inflamação umbilical e maior frequência de escore de fezes 3 no COL-. As alterações clínicas foram refletidas no eritrograma das bezerras, sendo encontrado menor valor médio para a taxa de hemoglobina (HGB) no COL- em M3. Em relação à idade, observou-se redução gradativa dos valores médios para He (hemácias), HGB, HCT (hematócrito) e índices hematimétricos no primeiro mês de vida. A frequência de bezerras anêmicas foi maior no grupo COL- nos momentos M4 e M5. Em relação ao leucograma, observou-se diferença entre os grupos para linfócitos no M0 e M2 com valores superiores no COL-. Em relação aos momentos foi possível detectar leucocitose por neutrofilia no M0 e M1, observando-se inversão da relação neutrófilo:linfócito a partir desses momentos. Os valores de CD45+CD45RO- foram maiores em M0 no COL-, além disso, observou-se aumento da expressão do marcador de memória celular CD45RO+ do M0 ao M1 nos dois grupos experimentais. O CD3+gamma-delta- aumentou no decorrer do estudo, em contrapartida as células CD3+gamma-delta+ foram menores em M5 com relação ao M0-M3. Foi detectado também aumento dos valores de CD14+MHCII+ no primeiro mês de vida indicando maturação das células apresentadoras de antígeno. Em relação à produção de citocinas pelas células mononucleares sanguíneas, foi possível identificar maior concentração de IFN-gamma em M4, quando as células do COL- foram estimuladas com S. aureus (1 mononuclear:10 bactérias inativadas). A concentração de IL-17 detectada a partir das células do COL+ foi maior em M3, quando as células foram estimuladas com ConA. Com base nos resultados obtidos, pode-se concluir que: a) bezerras COL- apresentaram maior frequência e intensidade de doenças que evoluíram para anemia da inflamação; b) bezerras COL- apresentaram maior número absoluto de linfócitos, representadas especialmente pela subpopulação CD3+gamma-delta+ nos episódios de maior frequência de diarreias; c) Linfócitos de memória CD45RO+ aumentaram após a colostragem em ambos os grupos, sugerindo que outros componentes acelulares do colostro podem apresentar papel fundamental no desenvolvimento da resposta imunológica de bezerras recém-nascidas; d) a subpopulação CD3+gamma-delta- e as células CD14+MHCII- e CD14+MHCII+ aumentaram durante o primeiro mês de vida, indicando maturação imunológica; e) as células mononucleares das bezerras não responderam ao Herpesvírus Bovino tipo 1, porém responderam aos estímulos bacterianos, especialmente para a Escherichia coli; a interpretação do leucograma em conjunto com a análise das variações apresentadas para as citocinas inflamatórias IFN-gamma e IL-17 permitem afirmar que as bezerras apresentaram resposta inflamatória retardada e de menor magnitude no COL-. / This study evaluated the effect of leukocytes passive transference from bovine colostrum in specific immunity of newborn calves. During neonatal period, 20 Holstein calves were followed. Animals were distributed in two experimental groups: COL+ which received fresh colostrum (4L) from their mothers, and COL- which received frozen and acellular colostrum (4L) that came from donor cows. The evaluations were performed in the following moments: before colostrum intake (M0), 1-2 (M1), 7 (M2), 14 (M3), 21 (M4) and 28 days after birth (M5). Heifers were submitted to clinical examination. Then, blood samples were harvested for hemogram, immunophenotyping and cell culture. Both groups were fed with the same quality of colostrum (immunoglobulin concentration 70-120 g/L). The cell concentration of fresh colostrum that was provided to COL+ group was 1.895.849 cells/mL. It was not possible to detect differences in vital functions concerning the experimental groups. The system specific examination reveled one case of bronchopneumonia, three cases of umbilical inflammation and major rates of diarrhea score 3 in group COL-. Clinical alterations were reflected in calves erythrogram. It was found lower mean value for hemoglobin (HGB) in M3 for COL-. Regarding age, a gradual reduction in mean values for erythrocytes, HGB, HCT (hematocrit) and hematimetric rates were observed in the first month of life. The frequency of anemic heifers was higher in COL- group at moments M4 and M5. Regarding leukogram, it was observed difference between groups for lymphocytes in M0 and M2 with higher values in COL-. Concerning moments, it was possible to detect leukocytosis by neutrophilia from M0 up to M1 and inversion of neutrophil:lymphocyte relation from this moment. Values of CD45+CD45RO- was higher in M0 for COL-, furthermore, increase of cellular memory marker expression CD45RO+ was observed from M0 to M1 in both groups. The CD3+gamma-delta- increased during the study. On the other hand, CD3+gamma-delta+ were lower in M5 in relation to M0-M3. Increase of CD14+MHCII+ values were also detected in the first month of life, indicating maturation of antigen presenting cells. Regarding cytokine production by mononuclear cells of heifers blood, it was possible to identify higher concentration of IFN-gamma in M4 when cells of COL- were stimulated with S. aureus (1 mononuclear: 10 inactivated bacteria). The concentration of IL-17 detected from COL+ cells was higher in M3, when cells were stimulated with ConA. Based on these results, it can be concluded that: a) COL- heifers presented higher frequency and intensity of diseases that evolved to anemia of inflammation; b) COL- heifers presented higher lymphocyte absolute number, represented specially by CD3+gamma-delta+ subsets in episodes of higher frequency of diarrhea; c) memory lymphocytes CD45RO+ increased after colostrum intake in both groups, suggesting that other acellular colostrum components can present fundamental role in development of immunological response in newborn heifers; d) the subset of CD3+gamma-delta- and the cells CD14+MHCII- and CD14+MHCII+ increased during the first month of life, indicating immunological maturation; e) heifers mononuclear cells did not respond for herpes virus bovine type 1, however, responded for bacterial stimulus, specially Escherichia coli. The interpretation of leukogram with the variation of presented analyses for inflammatory cytokines IFN-gamma and IL-17, allow to state that heifers presented delayed inflammatory response and of lesser magnitude in COL-.

O presente estudo avaliou o sistema oculomotor medido por movimentos oculares em bebês com diagnóstico de Displasia Broncopulmonar (DBP). Bebês com idade gestacional 37 semanas, dependentes de oxigênio em concentrações acima de 21% por mais de 28 dias foram incluídos no grupo DBP, bebês nascidos a Termo (idade gestacional > 37 semanas), não internados foram incluídos no grupo nascido a termo e bebês prematuros (idade gestacional < 37 semanas), que permaneceram internados e que não fizeram uso de oxigênio por mais de 10 dias foram incluídos no grupo prematuro. Os bebês dos três grupos tinham exame oftalmológico de biomicroscopia e de fundo de olho com resultados normais. Foram excluídos do estudo, bebês em uso de oxigênio sob ventilação mecânica e/ou drogas vasoativas, com diagnóstico de hemorragia intracraniana, retinopatia da prematuridade e malformações motoras e/ou neurológicas congênitas ou adquiridas identificadas no exame neonatal ou durante a estadia no berçário. Todos os bebês realizaram uma única avaliação binocular. As avaliações foram realizadas com os bebês sentados confortavelmente e eram compostas pela avaliação de quatro movimentos oculares: sacadas (SAC), perseguição lenta (PL), reflexo vestíbulo-ocular (RVO) e nistagmo optocinéticos (NOC). Os movimentos oculares foram transcritos em variável categórica (presente ou ausente) e para análise estatística foram feitas comparações entre o grupo DBP, grupo nascido a termo e grupo prematuro (Teste Cochran Q), para garantir a confiabilidade dos resultados apresentados durante a avaliação, 28% da amostra foi avaliada por três observadores e um teste de aderência X2 foi utilizado para medir a confiabilidade entre os três observadores. Durante o estudo foram avaliados 109 bebês, 107 foram incluídos no estudo, dois bebês, com IG < 37 semanas, foram excluídos por usarem oxigênio por um tempo igual há 15 dias. Dos 107 bebês avaliados, 23 foram inclusos no grupo DBP, 47 no grupo nascido a termo e 37 no grupo prematuro. Os bebês do grupo DBP tiveram IG média de 32 semanas ± 3 semanas, APGAR 1° minuto 6 ± 1, 5° minuto 8 ± 2, 37 dias em oxigênio ± 10 dias, na quantidade média de 2 L/min ± 0,5 L/min. O peso de nascimento, idade gestacional, APGAR NO 1° e 5° minutos do grupo nascido a termo, DBP e Prematuro diferem significativamente entre si (Teste Kruskal-Wallis p = 0.0000, 0.0000, 0.0000, 0.0013 e 0.0001, respectivamente). O grupo nascido a termo apresentou maiores valores quando comparado ao grupo DBP e prematuro. Bebês com DBP manifestam ausência de três dos quatro tipos de movimentos oculares medidos quando comparado com o grupo nascido a termo e prematuro (Teste Q Cochran onde Q > 2 e p, < 0,05) / This study evaluated the oculomotor system measured by eye movements in infants diagnosed with bronchopulmonary dysplasia (BPD). Infants 37 weeks gestational age, oxygen-dependent at concentrations above 21% for more than 28 days were included in the BPD group, term infants (gestational age > 37 weeks), not hospitalized were included in term groups and preterm infants (gestational age < 37 weeks), who remained hospitalized and did not use oxygen for more than 10 days were included in the premature group. The three groups of babies had eye examination and biomicroscopy of the fundus with normal results. Excluded from the study, babies on oxygen in mechanical ventilation and/ or vasoactive drugs; with a diagnosis of intracranial hemorrhage, retinopathy of prematurity, motor and/or neurological congenital or acquired malformations identified in neonatal or during the stay in the nursery. All infants made a single binocular assessment. The evaluations were conducted with babies seated comfortably and were composed by the evaluation of four eye movements: saccades (SAC), slow pursuit (PL), vestibuloocular reflex (VOR) and optokinetic nystagmus (NOC). Eye movements were transcribed into a categorical variable (present or absent) and statistical analysis were made between BPD group, term group and premature group (Cochran Q test) to ensure reliable of the results presented during the evaluation, 28 % of the sample was evaluated by three observers and an adherence X2 test was used to measure the reliable between three observers. During the study, 109 infants were evaluated, 107 were included in the study, two infants with GA < 37 weeks, were excluded by using oxygen for a time equal to 15 days. Of the 107 infants evaluated, 23 were included in the BPD group, 47 in the term group and 37 in the premature group. Babies in the BPD group had GA of 32 weeks ± 3 weeks, APGAR 1st minute 6 ± 1, 5th minutes 8 ± 2, 37 days ± 10 days in oxygen, in the median amount of 2 L / min ± 0.5 L / min. Birth weight, gestational age, APGAR score at 1st and 5th minutes from the term group, DBP and Premature differ significantly (Kruskal-Wallis test p = 0.0000, 0.0000, 0.0013 and 0.0001, respectively). The term group had higher values when compared to the BPD and premature. Babies with BPD manifest absence of three of the four types of eye movements measured when compared with the term group and preterm (Cochran Q test where Q > 2 and p < 0.05)

Dor em pediatria tem sido o foco de muitos estudos nas últimas décadas devido ao fato de que neonatos apresentam menor limiar a estímulos nocivos e inócuos em relação aos adultos. Desta forma, o uso de analgésicos é frequente para sedação e analgesia em UTIs pediátricas, e entre os opioides, a morfina é um dos mais utilizados. Adicionalmente, exposição a estímulos estressantes como a deprivação materna está entre os fatores ambientais relacionados a alterações no desenvolvimento neural. O estresse social ou a negligência do cuidado parental, e mais precisamente do cuidado materno em ratos, está associado a importantes alterações comportamentais na vida adulta. Entre estas alterações apresentadas ao longo da vida estão alterações na resposta ao estresse, e à alteração na sensibilidade a estímulos dolorosos, indexadas por hiperalgesia. Consquentemente, o estresse social gerado pela deprivação materna está relacionado a prejuízos cognitivos, emocionais e sociais, além de alterações neuroquímicas de longo prazo. Deste modo, é necessário atenção, prevenção e tratamento a esses eventos físicos, emocionais e comportamentais no período neonatal e durante a infância, uma vez que as bases neurobiológicas envolvidas nestes fenômenos ainda não foram completamente elucidadas. Considerando a relevância do tema, o objetivo deste estudo foi verificar os efeitos do tratamento com 5 μg de morfina, uma vez ao dia, do P8 ao P14 e a exposição a deprivação materna por 3 horas durante os primeiros 10 dias de vida em curto (P16), médio (P30) e longo prazo (P60), sobre o desenvolvimento dos reflexos neuromotores da prole por meio do Reflexo de Endireitamento, Geotaxia Negativa e Marcha; comportamento nociceptivo por meio dos testes Tail-Flick e Placa Quente, respectivamente. Um total de 58 filhotes foi utilizado. Os animais foram divididos em 5 grupos: controle total (C), que não recebeu nenhuma intervenção; salina (S), que recebeu solução salina; morfina (M), que recebeu morfina; deprivado salina (DS), que foram submetidos a deprivação maternal e receberam solução salina; e deprivado morfina (DM), que foram submetidos a deprivação maternal e receberam morfina. Em relação aos testes neuroquímicos, foram analisados níveis de BDNF, NGF, IL-1β e IL-4 em tronco e córtex cerebral que estão relacionados a fenômenos modulatórios em sistemas nervoso e imune. Os animais que receberam morfina e os deprivados maternos que receberam morfina apresentaram atraso no desenvolvimento dos reflexos iniciais. Alterações neuroquímicas também foram observadas. Os níveis de BDNF no tronco encefálico foram diminuídos em animais que receberam morfina e deprivação materna. Animais deprivados apresentaram um aumento nos níveis de NGF no tronco encefálico. Além disso, observou-se um aumento nos níveis de NGF do córtex cerebral em animais que receberam morfina, deprivados maternos e deprivados maternos que receberam morfina. Uma diminuição no limiar nociceptivo foi observada em animais que receberam morfina, deprivados maternos e os deprivados maternos que receberam morfina. Também houve interações em tronco encefálico e córtex cerebral nos níveis de BDNF, IL-1β e IL-4 entre as variáveis independentes: tratamento, deprivação e tempo, o que levou à modificação nos níveis centrais dos neuroimunomoduladores avaliados. Estes dados demonstram a importância de estudos focados nos efeitos do tratamento com morfina no período neonatal ao longo da vida, assim como na busca por alternativas terapêuticas que possam reverter possíveis alterações decorrentes da separação materna no período neonatal. / Pediatric pain has been the focus of many studies in the last decades due to the fact that neonates have a lower threshold for innocuous and noxious stimuli than for adults. Thus, the use of analgesics is frequent for sedation and analgesia in pediatric intensive care units, and among opioids, morphine is one of the most used. Additionally, exposure to stressful stimuli such as maternal deprivation is among the environmental factors related to changes in neural development. The social stress or neglect of parental care, and more precisely maternal care in rats, is associated with important behavioral changes in adult life. Among these changes presented throughout life are changes in the response to stress, and the change in sensitivity to painful stimuli, indexed by hyperalgesia. Consequently, the social stress generated by maternal deprivation is related to cognitive, emotional and social impairments, further to long-term neurochemical changes. Thus, attention, prevention and treatment are necessary to these physical, emotional and behavioral events in the neonatal period and during childhood, since the neurobiological bases involved in these phenomena have not yet been fully elucidated. Considering the relevance of the subject, the objective of this study was to verify the effects of treatment with 5 μg of morphine once a day from P8 to P14 and exposure to maternal deprivation for 3 hours during the first 10 days of short (P16), medium (P30) and long- term (P60), on the development of neuromotor reflexes of offspring through the righting Reflex, Negative Geotaxis and Gait; nociceptive behavior through the Tail-Flick and Hot Plate tests, respectively. A total of 58 puppies were utilized. The animals were divided in 5 groups: the total control group (C), which did not receive any intervention; saline group (S), which receive saline solution; morphine group (M), which receive morphine; deprived saline (DS), which were subjected to maternal deprivation and receive saline solution; and deprived morphine group (DM), which were subjected to maternal deprivation and receive morphine. In relation to the neurochemical tests, levels of BDNF, NGF, IL-1β and IL-4 were analyzed in the brainsteam and cerebral cortex and are related to modulatory phenomena of the nervous and immune systems. Animals that received morphine and deprived animais that received morphine showed a delay in the development of early reflexes. Neurochemical changes were also observed. BDNF levels in the brainstem were decreased in animals receiving morphine and maternal deprivation. Deprived animals had an increase in NGF levels in the brainstem. Besides, an increase in NGF levels of the cerebral cortex was observed in animals receiving morphine, maternal deprivation and maternal deprivation receiving morphine. A decrease in the nociceptive threshold was observed in animals receiving morphine, maternal deprivation, and maternal deprivation receiving morphine. There were also interactions in the brainstem and cerebral cortex in the levels of BDNF, IL-1β and IL-4 among the independent variables: treatment, deprivation and time, which led to the modification in the central levels of the neuroimmunomodulators evaluated. These data demonstrate the importance of studies focused on the effects of treatment with morphine in the neonatal period throughout life, as well as on the search for therapeutic alternatives that may reverse possible changes due to maternal deprivation in the neonatal period.

The concept of Electrical Bio Impedance prevails in this thesis. The EBI measurement which is used for obtaining the body composition is, by virtue of time becoming of great use as its one of the easiest method of finding out the body composition. In simple words, EBI is the opposition offered by the body to the current. It is just like another analysis tool. The result is only as good as the test is done. In this thesis, we have done the analysis on the neonatal EBI measurements of two kinds.In this work, 293 measurements are obtained from 12 babies and 230 measurements are obtained from 7 babies have been analyzed with the purpose of obtaining reference values for the spectrum of complex EBI. The analysis uses both statistical and model approach of obtaining reference values and in order to fit the given data, Cole model analysis is used.Filters were applied to get the highest degree of correctness. In the due course of the filtering, it was found that the measurements from some babies have been deleted. The Standard Error of Estimation (S.E.E.) is a parameter used for obtaining the further reliable and most probable output. The further analysis is done using MATLAB and the results are been compared to the previous analysis report.

This study was completed to better understand pronghorn antelope (Antilocapra americana) populations found throughout Idaho. Antelope were studied in three separate and distinct study areas. The Big Desert, Camas Prairie, and Little Lost and Pahsimeroi valleys were all selected as study sites. Idaho Department of Fish and Game (IDFG) is concerned with current pronghorn populations found throughout Idaho. Pronghorn are a valued big game species found in Idaho. Increasing pronghorn populations in Idaho is a focus of IDFG. We captured and VHF-collared pronghorn fawns found in our three study areas. Fawns were monitored daily with telemetry equipment for survival. Field necropsies were performed to determine cause of death for each fawn. We found that fawns across Idaho had acceptable survival rates compared to previous studies conducted on pronghorn. The highest cause of mortality on fawns was coyotes (Canis latrans). Other predators on pronghorn fawns were bobcats (Lynx rufus), golden eagles (Aquila chrysaetos), and black bear (Ursus americanus). We found that fawns radio-collared with a higher BMI (body mass index) were more likely to survive. We examined other relationships that could have an effect on fawn survival. Rabbits (Lepus californicus, Lepus townsendii Sylvilagus nuttallii, Brachylagus idahoensis, Lepus americanu) and ground squirrels (Urocitellus armatus, Urocitellus mollis, Urocitellus elegans, Urocitellus columbianus) were examined to see if there population numbers had an effect on pronghorn fawn survival. We found a relationship between rabbit density and fawn survival, as rabbit density increased pronghorn fawn survival increased. Ground squirrel density was found to have no effect. Coyote density was studied to see if coyote density effected pronghorn survival. No relationship was found between coyote density and pronghorn fawn survival. Habitat quality can impact animal populations. We examined habitat variables that could affect pronghorn fawn survival. NDVI (normalized difference vegetation index) was examined and we found no correlation in this study. Pronghorn fecal samples were collected and analyzed at a laboratory to look for diet quality correlation to pronghorn survival. We found a correlation between diet quality (DAPA) and pronghorn fawn survival. Diet quality can be linked to habitat quality, as habitat quality increases so does pronghorn fawn survival. Habitat quality, rabbits, and a fawn’s BMI all were linked to increased fawn survival. We recommend wildlife managers create and increase pronghorn habitat when possible to produce better pronghorn fawn survival.

O número de jovens infectados por HIV vem aumentando nas últimas décadas, o que salienta a necessidade de estratégias vacinais que sejam imunogênicas em fase precoce de vida capazes de induzir resposta de longa duração. A vacina quimérica LAMP-1/p55Gag, associa o gene que codifica a LAMP-1 (proteína de associação da membrana lisossomal) e o gene da gag do HIV-1, direciona o tráfego da proteína viralpara os compartimentos MIIC, possibilitando a apresentação dos peptídeos virais pela classe II do Complexo Principal de Histocompatibilidade (MHC II). Esta vacina quimérica é imunogênica em camundongos BALB/c adultos e neonatos e crucial para induzir resposta T e B de longa duração, com produção de elevados níveis de anticorpos. Contudo, os mecanismos imunológicos envolvidos na indução da resposta humoral da vacina LAMP/Gag, como a geração de células T auxiliares foliculares (TFH), ainda não são conhecidos no período neonatal. O objetivo do estudo foi avaliar a geração de células TFH, T citotóxicos e B foliculares em camundongos neonatos submetidos à imunização com as vacinas LAMP/Gag (LG) e Gag (G). Inicialmente,avaliamos a imunogenicidade das vacinas gênicas na imunização neonatal aos sete dias de idade em camundongos de linhagem C57BL/6. Os resultados mostram que a imunização neonatal com a vacina LG é capaz de aumentara frequência de células secretoras de IFN-&#978 aos peptídeos imunodominantes da região gag do HIV-1 e de células T CD8&#43IFN-&#978&#43 comparadas a vacina Gag. A imunização neonatal com a vacina LG também levou a produção de títulos elevados de anticorpos IgG1 anti-p24 e aumento da porcentagem de células secretoras de IgG1 Gag-específicas. O priming neonatal com LG é capaz de promover resposta celular e humoral anti-Gag de longa duração. Além disto, a imunização neonatal (ip) com LG foi capaz de induzir células TFH (CD4&#43CXCR5&#43PD-1&#43Bcl-6&#43), linfócitos T CD8&#43 foliculares (TFC) (CD8&#43CXCR5&#43) e formação de centro germinativo (CG) nos linfonodos mesentéricos, contudo, ambas as vacinas induziram células B foliculares (CD19&#43CXCR5&#43). Apesar da menor frequência de TFH dos neonatos em relação a adultos na imunização com LG, houve frequência similar de células TFC. A imunização intradérmica convencional induziu aumento do número de células TFH nos linfonodos inguinais já ao terceiro dia após o reforço vacinal, embora frequência similar de células TFH, TFC e B foliculares. Neste período foi possível observar que a vacina LG também induziu a geração de células B de CG. Outra peculiaridade da vacina LG neonatal foi o aumento da expressão gênica da enzima citidina deaminase (AID). Os resultados mostram que a vacina L/AMP/Gag é imunogênica na fase neonatal de camundongos C57BL/6 quanto à geração de resposta celular e humoral antígeno-específica e resposta de longa duração, similarmente aos camundongos BALB/c. Os dados mostraram que a vacina LG é eficaz na indução de células T foliculares, na maturação de tecidos linfoides com formação de CGs e na indução de transcritos para AID. No conjunto, os achados evidenciam que a estratégia da vacina quimérica L/AMP/Gag é eficaz neste período da vida, e possui importante papel adjuvante na maturação da resposta humoral. / The number of young people infected with HIV has been increasing in recent decades, which highlights the need for vaccine strategies that are immunogenic at early phase of life able of inducing long-term response. The chimeric LAMP-1/p55Gag vaccine, associates the gene encoding LAMP-1 (lysosomal associated membrane protein) and the HIV-1 gag gene, directs the traffic of viral protein to MIIC compartments, leading to presentation of the viral peptides through class II of the Major Histocompatibility Complex (MHC II). This chimeric vaccine is immunogenic in adult and neonatal BALB/c mice and crucial to induce long-term T and B responses, producing high levels of antibodies. However, the immunological mechanisms involved in the induction of the humoral response of the LAMP/Gag vaccine, such as the generation of T follicular helper cells (TFH), are unknown in the neonatal period. The objective of this study was to evaluate the generation of TFH, and follicular cytotoxic T cells and B cells in neonates submitted to immunization with the LAMP/Gag (LG) and Gag (G) vaccines. The results show that neonatal immunization at seven days-old in C57BL/6 mice strain with the LG vaccine is able to increasing the frequency of IFN-&#978-secreting cells to the immunodominant peptides of the HIV-1 gag region and of CD8&#43IFN-&#978&#43 T cells compared to the Gag vaccine. Neonatal immunization with the LG vaccine led to the production of high titers of anti-p24 IgG1 antibodies and the increased percentage of Gag-specific IgG1 secreting cells. Neonatal priming with LG is able to promote long-lasting anti-Gag humoral and cellular response. Moreover, neonatal (ip) immunization with LG was able to induce TFH cells (CD4&#43CXCR5&#43PD-1&#43Bcl-6 &#43), follicular CD8 &#43 T cells (TFC) (CD8&#43CXCR5&#43) and germinal center (GC) formation in the mesenteric lymph nodes, whereas both vaccines induced follicular B cells (CD19&#43CXCR5&#43). Despite the lower frequency of TFH in neonates in relation to adult counterpartin the immunization with LG, a similar percentage of TFC cells was observed. Conventional immunization by intradermal immunization induced an increased number of TFH cells in inguinal lymph nodes on the third day after booster vaccination, despite the similar frequency of follicular TFH, TFC and B cells. In this period the LG vaccine also induced generation of CG B cells. Another peculiarity of the LG neonatal vaccine was the increase in the gene expression of the enzyme activation-induced cytidine deaminase (AID).The results show that the LAMP/Gag vaccine is immunogenic in the neonatal phase of C57BL/6 mice for the generation of antigen-specific humoral and cellular response and long-term response, similar to BALB/c mice. The findings showed effective induction of follicular T cells, maturation of lymphoid tissues with formation of GCs and up-regulation oftranscripts for AID. Taken together, our findings demonstrate that the LAMP/Gag chimeric vaccine strategy is effective at this time in life, and has an important adjuvant role in the maturation of the humoral response.

La toxine de pertussis (PTX) est une A-B protéine considérée comme l’un des principaux facteurs de virulence de Bordetella pertussis, l’agent bactérien responsable de la coqueluche. Aujourd’hui, cette maladie représente encore un réel danger pour les nouveaux-nés et les nourrissons non ou partiellement immunisés. Actuellement, la coqueluche provoque encore la mort d’environ 350.000 individus par an. La toxicité de la PTX est liée à l’activité enzymatique de sa sous-unité A capable d’inhiber les voies de signalisation associées aux protéines Gi. La partie B, quant à elle, permet l’entrée de cette sous-unité A dans le cytoplasme des cellules cibles en se liant spécifiquement à son ou ses récepteurs membranaires toujours inconnus de nos jours. Des études réalisées chez la souris et chez l’homme ont montré que les vaccins anticoquelucheux combinés à différents antigènes vaccinaux étaient capables de moduler leurs réponses humorales spécifiques. Par ailleurs, la PTX est couramment qualifiée d’agent immunostimulant. En effet, des modèles murins de vaccination permirent d’identifier des propriétés adjuvantes de la PTX coadministrée avec des antigènes non relevants. Le travail développé dans ce manuscrit étudie les effets de la PTX sur 2 types cellulaires primordiaux sollicités lors d’une vaccination : la cellule dendritique (DC) et le lymphocyte T CD4+ naïf. Les DC sont les seules cellules présentatrices d’antigènes aptes à initier une réponse immune primaire. Dans un premier temps, nous avons montré que la PTX était capable d’activer des DC générées in vitro à partir de monocytes. En effet, elles acquièrent un phénotype mature caractérisé par une augmentation de l’expression membranaire des molécules costimulatrices et du CMH de classe II, démontrant un effet direct et spécifique de la PTX sur les DC myéloïdes. Parallèlement, ces DC produisent du TNF-a, de l’IL-12p40 et de l’IL-12p70 et activent NF-kappaB, un facteur de transcription essentiel au processus de maturation. Nous avons obtenu des résultats similaires avec une toxine génétiquement modifiée qui est enzymatiquement inactive. A partir de sang total incubé avec la PTX, nous avons par ailleurs observé que les DC circulantes du nouveau-né étaient déficientes dans leur maturation et leur sécrétion d’IL-12p70 comparées aux DC de l’adulte. D’autre part, il a été décrit précédemment que la PTX exerçait des effets mitogènes sur les lymphocytes T humains et murins. Cependant, le rôle qu’elle joue sur la population des lymphocytes T CD4 naïfs reste peu connu. A l’issue de notre second travail, nous pouvons dès lors affirmer que la PTX est également capable d’activer des lymphocytes T CD4+CD45RA+ naïfs isolés à partir des cellules mononuclées du sang périphérique, et ce indépendamment de son activité enzymatique. En effet, ces lymphocytes T CD4+ naïfs stimulés par la PTX prolifèrent, synthétisent des quantités non négligeables d'ARN messagers codant pour l’IL-2 et le TNF-a, augmentent l’expression membranaire des molécules CD40L, CD69 et CD25 et expriment la protéine Foxp3. Cette activation s’accompagne de la translocation nucléaire de NF-kappaB et NFAT. Parallèlement à l’adulte, la PTX active les lymphocytes T CD4 néonataux. Néanmoins, ceux-ci prolifèrent moins bien et expriment plus faiblement le CD40L à leur surface. Enfin, la PTX induit la sécrétion de taux importants d’IFN-g par des T CD4+CD45RA+ naïfs adultes mis en présence de DC autologues. Nous terminerons en proposant l’hypothèse suivante : La PTX pourrait exercer ses propriétés adjuvantes par l’intermédiaire de différents mécanismes comprenant notamment la maturation des DC d’origine myéloïde et l’activation des lymphocytes T CD4+CD45RA+ naïfs. Ces 2 populations cellulaires sont en effet les principaux protagonistes impliqués dans la réponse immune primaire.

Stem cell biologists creating cells and tissues for therapies, disease modeling, and drug screening have observed that differentiating pluripotent stem cells (PSCs) tend to produce cells at an embryonic stage of development but have difficulty maturing into adult definitive cells. A better understanding of developmental maturation will provide insights into embryogenesis and permit more accurate disease modeling. In the hematopoietic system, primitive and definitive cells are distinguished by functional transplantation assays, well characterized cell surface antigens, and gene expression signatures. We examined the transition in vivo in transplanted murine hematopoietic stem cells (HSCs) and in vitro in human PSC (hPSC) derived red blood cells (RBCs). We found that the hematopoietic microenvironment of the recipient significantly affects the outcome of HSC transplantation. The earliest embryonic HSCs perform better in neonatal recipients, whereas more mature adult-like HSCs perform better in adult recipients. The preference may be related to different active hematopoietic niches in neonates and adults, as we observed adult HSCs homing to different tissues in neonatal and adult recipients. Additionally, we found that proliferation may enhance the neonatal engraftment potential of adult-like HSCs. Our data highlight the importance of the host environment on transplantation outcomes, and point to the neonatal transplant model as a tool to functionally examine the earliest HSCs and primitive derivatives of PSCs.