Dissertations / Theses on the topic 'Jaundice in the Neonate'

A icterícia por hiperbilirrubinemia indireta afeta mais de 60% dos recém-nascidos a termo. O tratamento, quando necessário, é realizado através da fototerapia. Não existem estudos na literatura avaliando os efeitos da fototerapia na função dos neutrófilos de recém-nascidos. O melhor entendimento da função dos neutrófilos nos recém-nascidos antes e após a fototerapia seria importante para avaliar as possíveis repercussões na expressão dos neutrófilos desencadeadas pelo tratamento fototerápico. O objetivo deste estudo foi avaliar e comparar a função dos neutrófilos, através da mensuração pela citometria de fluxo da expressão dos principais marcadores de superfície em recémnascidos ictéricos, antes e após 24 horas de fototerapia. Metodologia: Foram incluídos recém-nascidos com idade gestacional ≥ 35 semanas e peso de nascimento ≥ 2000g, que possuiam critérios da Academia Americana de Pediatria para tratamento fototerápico. Os critérios de exclusão foram: mal-formações congênitas, síndromes com alterações cromossômicas, erro inato do metabolismo, infecções do grupo STORCH, asfixia neonatal, sepse ou suspeita de sepse, exsanguineotransfusão, transfusão de hemocomponentes e uso de imunoglobulina. Foi realizada a avaliação de expressão da intensidade média de fluorescência (IMF) de CD10, CD11b, CD11c, CD15, CD16, CD18, CD62L, CD64 e CD66, antes do início e após 24 horas do início da fototerapia. Foram utilizados o teste T de Student para análise dos dados. Resultados: Foram incluídos 25 recém-nascidos no estudo, com idade mediana de 53 (27.5-75.5) horas de vida e bilirrubina média de 13.6±2.85 mg/dL. Não houve diferença estatística na expressão de CD11b, CD15, CD18, CD62L, CD64 e percentual de neutrófilos antes e após 24 horas de fototerapia. Ocorreu aumento da expressão de CD10 8 (p=0.038) e CD16 (p=0.017) e redução da expressão de CD11c (p=0.023) e CD66acde (p=0.004) após 24 horas de fototerapia. Conclusão: Os recém-nascidos submetidos ao tratamento fototerápico apresentaram aumento da expressão de CD10 e de CD16 e diminuição da expressão de CD11c e de CD66acde após 24 horas de exposição, que pode estar relacionado a um efeito antiinflamatório da fototerapia nos recém-nascidos expostos a este tratamento.<br>Jaundice due to indirect hyperbilirubinemia affects more than 60% of term neonates. The treatment when necessary is carried out using phototherapy. There are no studies in the literature evaluating the effect of phototherapy on the function of neonates' neutrophils. A better understanding of the function of neutrophils in neonates before and after phototherapy would be important in order to assess potential effects on the expression of neutrofils triggered by the phototherapy treatment. The aim of this study was to assess and compare the function of neutrophils by measuring the expression of the main surface markers in icteric neonates, using flow cytometry, before and after 24 hours of phototherapy. Methodology: Neonates at a gestational age ≥ 35 weeks and at a birth weight ≥ 2000g who met the criteria of the American Academy of Pediatrics for phototherapy were included. The exclusion criteria were: congenital malformations, syndromes with chromosomal alterations, inborn errors of metabolism, infections of the STORCH group, neonatal asphyxia, sepsis or suspicion of sepsis, exchange transfusion, transfusion of blood components, and use of immunoglobulin. The evaluation of the MFI expression of CD10, CD11b, CD11c, CD15, CD16, CD18, CD62L, CD64 and CD66 was performed before and 24 hours after the initiation of phototherapy. The chi-square and Student T tests were used for data analysis. Results: Twenty-five neonates were included in the study at the mean age of 53 (27.5- 75.5) hours of life and with a mean bilirubin level of 13.6±2.85 mg/dL. There was no statistical difference in the expression of CD11b, CD15, CD18, CD62L, CD64 and percentage of neutrophils before and after 24 hours of phototherapy. There was an increase in the expression of CD10 (p=0.038) and CD16 (p=0.017) and a reduction in 10 the expression of CD11c (p=0.023) and CD66acde (p=0.004) after 24 hours of phototherapy. Conclusion: The newborns submitted to phototherapy had increased expression of CD10 and CD16 and decreased expression of CD11c and CD66acde after 24 hours of exposure, which may be related to an anti-inflammatory effect of phototherapy on the neonates exposed to this treatment.

Made available in DSpace on 2014-10-09T12:26:28Z (GMT). No. of bitstreams: 0<br>Made available in DSpace on 2014-10-09T14:09:41Z (GMT). No. of bitstreams: 0<br>Tese (Doutoramento)<br>IPEN/T<br>Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

A icterícia neonatal é geralmente benigna, mas desfechos desfavoráveis podem ocorrer e a identificação dos casos de maior risco seria muito útil. Alguns fatores de risco já conhecidos são prematuridade, desidratação, aleitamento materno, deficiência de G6PD e incompatibilidade sanguínea. As alterações na conjugação hepática de bilirrubina devido a polimorfismos da UGT1A1 também podem contribuir para esse maior risco. O objetivo deste estudo foi estimar a freqüência da deficiência de G6PD e/ou das variantes polimórficas da UGT1A1 como fatores de risco para hiperbilirrubinemia grave em neonatos com mais de 35 semanas de idade gestacional e peso superior a 2000g em uma Unidade Neonatal do Sul do Brasil. Estudo prospectivo, observacional, de casos e controles, que incluiu 243 recémnascidos admitidos para fototerapia no HCPA e 247 controles, entre março e dezembro de 2007. Foi realizada dosagem da atividade da G6PD e análises genético-moleculares do respectivo gene. Foi também realizado PCR para a UGT1A1 com eletroforese capilar em analisador genético ABI 3130xl e análise no programa GeneMapper®. Foram detectados genótipos polimórficos da UGT1A1 em 16% dos pacientes, com prevalência nos ictéricos de 13,5% e nos normais de 18,2%, diferença não significativa. Identificada maior prevalência dos polimorfismos em negros e pardos (25%) em relação aos brancos (13%) (p=0,014). A prevalência da deficiência de G6PD foi 4,6%, sem mostrar correlação com a icterícia. Concluímos que nesta amostra de recém-nascidos do sul do Brasil nem as variantes da UGT1A1, nem a deficiência de G6PD foram associadas à hiperbilirrubinemia grave, com prevalências semelhantes às verificadas em outras populações. Considerando a grande miscigenação presente nessa região, outros fatores e interações gênicas devem ser procurados, incluindo possivelmente o estudo de outros polimorfismos, identificando fatores de risco para explicar a doença, um importante problema de saúde a merecer a atenção dos pesquisadores.<br>Neonatal jaundice is usually benign, but unfavorable outcomes may happen; therefore, the identification of high-risk cases would be very useful. Some risk factors already known are prematurity, dehydration, breastfeeding, G6PD deficiency and blood incompatibility. Alterations in the hepatic conjugation of bilirubin due to UGT1A1 polymorphisms may also contribute to this higher risk. The objective of this study was to estimate the frequency of G6PD deficiency and the promoter region of UGT1A1 gene variants as risk factors to severe hyperbilirubinemia in newborns of over 35 weeks of gestational age and weighing above 2,000g in a Neonatal Service in Southern Brazil. This is a prospective and observational study of cases and controls which included 243 newborns admitted for phototherapy at HCPA and 247 controls, between March and December, 2007. G6PD activity was determined and the deficient cases were investigated by genetic analysis. PCR for the UGT1A1 variants was also performed, followed by capillary electrophoresis in genetic analyzer ABI 3130xl and the analysis in GeneMapper® program. Polymorphic genotypes were detected in 16% of the patients, prevalence in icteric patients was 13,5% and in normal individuals was 18,2%, a difference which was not significant. A higher prevalence of polymorphisms in blacks and mulattos (25%) was identified when compared to whites (13%) (p=0,014). A prevalence of 4,6% of G6PD deficiency was found, without association to jaundice. We concluded that in this sample of newborns from the South of Brazil, polymorphic variants of UGT1A1 were not associated to severe hyperbilirubinemia as well as G6PD deficiency; being the prevalence similar to those found in other populations. Considering the high miscegenation that occurs in this area of Brazil, perhaps other factors and genic interactions should be sought in order to identify genetic risk factors, possibly including the study of further polymorphisms, as neonatal jaundice remains an important health problem to be approached by investigators.

Orientador: Aldo Eloizo Job<br>Resumo: As vantagens do uso de dispositivos ópticos na área das ciências da vida são amplamente exploradas, especialmente para o diagnóstico médico e protocolos de tratamento em uma prática de cuidados de saúde. Além disso, alguns pesquisadores têm atraído muita atenção ao desenvolvimento de materiais biocompatíveis e vestíveis para melhorar a segurança de dispositivos médicos, bem como para estabelecer novos procedimentos para monitorar a resposta de terapia e diagnóstico clínico. O presente trabalho tem como objetivo desenvolver uma membrana biocompatível-luminescente não invasiva para reduzir os erros mais comuns relatados em fototerapia convencional de luz azul para icterícia neonatal. Dentre estes erros destacam-se: intensidade de irradiância que chega ao recém-nascido bem como seu posicionamento frente a fonte de radiação e qualidade das lâmpadas dos aparelhos fototerápicos. Neste contexto, a membrana foi confeccionada como um filme de sensor de luz azul, fluorescente e colorimétrico a partir de polímeros: poli [2-metoxi, 5-(2'etilhexyloxy)-pfenilenovileno (MEH-PPV) em uma matriz de borracha natural. Para tal, as membranas de borracha natural (BN) foram obtidas utilizando o látex, extraído das árvores Hevea brasiliensis (clone RRIM 600), por casting, com posterior tratamento térmico em estufa. Em seguida, também por casting, a solução de MEH-PPV/BN, razão igual a 4000, foi depositada na membrana BN para fazer uma camada fina de material luminescente. A membrana de MEH-PPV/BN fo... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: The advantages of optical devices in the life sciences are widely exploited, especially for medical diagnosis and treatment protocols in a health care practice. In addition, some researchers have attracted much attention to development flexible and biocompatible materials to improve security of medical devices, as well as to establish new procedures for monitoring therapy response and clinical diagnosis. The present paper aims to develop a new noninvasive biocompatible-luminescent membrane to reduce the most common user errors with conventional blue-light phototherapy of neonatal jaundice. The membrane was design based on a polymer-based fluorescent and colorimetric blue-light sensor film. This material was based on poly[2-methoxy,5-(2'etilhexyloxy)-p-fenilenovileno (MEH-PPV) in a natural rubber matrix. Natural rubber membranes (BN) were obtained using latex from Hevea brasiliensis trees (clone RRIM 600) by casting, with the time and temperature of thermal treatment fixed on 65°C for 10h. Then, also by casting, MEH-PPV/NR solution, ratio equal to 4000, was deposited on NR membrane for make thin-layer of luminescent material. The MEH-PPV/NR membrane was submitted to the same conditions used for neonatal hyperbilirubinemia treatment (Blue light phototherapy, 40 µW/m2 /nm, 460nm). The proprieties of membranes were characterized by optical (UV-Vis, PL) and infrared (FTIR) spectroscopy confocal microscopy (MC), scanning electron microscopy (SEM) and atomic force microscopy (AFM) a... (Complete abstract click electronic access below)<br>Doutor

Introduction : La variabilité du rythme cardiaque est étudiée à partir des variations de durée des cycles cardiaques (intervalle R-R de l’électrocardiogramme). Ces variations peuvent être analysées par des méthodes linéaires (temporelles et fréquentielles) et non linéaires (théorie de l’information ou des fractales) de quantifications mathématiques et statistiques qui donnent des informations innovantes sur les signaux analysés. L’application de ces méthodes d’étude en néonatologie a démontré un intérêt pour le diagnostique précoce de l’infection néonatale tardive du prématuré mais n’avait pas été étudié dans l’infection néonatale précoce du nouveau-né à terme, dans le contexte des évènements cardio-respiratoires suivant la primo-vaccination des prématurés ou pour évaluer un effet neurologique de l’hyperbilirubinémie dans l’ictère néonatal. Notre hypothèse dans ce travail était qu’il était possible de : (i) caractériser la variabilité du rythme cardiaque en cas d’infection materno fœtale ou de méningite néonatale, (ii) mettre en évidence des facteurs prédisposant à la survenue d’évènements cardio-respiratoires post-vaccinaux, (iii) Identifier un éventuel retentissement neurologique de l’ictère néonatal par étude de la variabilité du rythme cardiaque<br>The heart rate variability measures permitted to evaluate equilibrium state and perturbation in the regulation of cardio-vascular system. These tools, based on heart rate variability analysis, helped to recognize associated disease state as early onset neonatal sepsis and non-infectious inflammatory response induced to immunization. An increase in global variability (SD), long term variability (SD, LF) and low approximated entropy (ApEn) were observed in the proven-sepsis full term infants. Importance of decrease in ApEn was correlated to the severity of sepsis assessed by blood markers. These suggest an association of sepsis with uncoordinated sympatho-vagal coactivation together with loss of adaptability. In premature infants, the risk of increase in cardio-respiratory events after the first immunization was associated with a specific pre-immunization profile: sympathetic predominance in heart rate control (high LF/HF ratio), abnormal oversimplification of heart rate variability and persistence rhythm control immaturity. Increased ApEn after immunization reflects a marginal result from adaptability of the heart rate to environmental changes without possibility to reserve in case of severe infection

2006/2007<br>INTRODUZIONE A basse concentrazioni la bilirubina non coniugata (unconjugated bilirubin, UCB) prodotta dalla degradazione dell’emoglobina, sembra essere un potente anti-ossidante, mentre è estremamente dannosa ad alte concentrazioni, causando encefalopatia nei neonati con severo ittero. Il 70% dei bambini che presentano kernittero muoiono entro sette giorni di vita, mentre il 30% dei sopravvissuti manifesta irreversibili conseguenze come sordità, ritardo mentale e danni cerebrali permanenti. L’encefalopatia dovuta ad alti livelli di bilirubina rappresenta oggi la maggior causa di riammissione ospedaliera nei neonati entro il primo mese di vita. Storicamente gli studi riguardanti le modalità di ingresso della bilirubina nel sistema nervoso centrale si sono concentrati sulla barriera emato-encefalica (blood brain barrier, BBB), costituita dai microvasi e dai capillari del cervello (micro vessels, MV). Tali studi hanno dimostrato come solamente la bilirubina non coniugata e non legata all’albumina del sangue, definita “bilirubina libera” (free bilirubin, Bf) sia capace di attraversare le membrane cellulari e diffondere nel tessuto. Tuttavia i microvasi non sono l’unica interfaccia sangue-tessuto presente nel cervello. Una seconda barriera è costituita dai plessi coroidei (CP). Questi, collocati nei ventricoli del cervello, mediano il passaggio delle molecole dal sangue al liquido cefalorachidiano e viceversa, posseggono una ampia superficie di scambio, il più alto flusso sanguigno del sistema nervoso centrale ed un fenotipo barriera meno restrittivo rispetto ai microvasi del parenchima. L’ingresso della bilirubina nel cervello sembra essere attivamente controllato da due trasportatori appartenenti alla famiglia delle “ATP dependent transporters”, Mrp1 e Pgp. Tali trasportatori potrebbero mantenere bassa la concentrazione della bilirubina limitandone l’ ingresso a livello di barriere o agendo direttamente a livello delle cellule del parenchima. Nonostante l’ impatto di questi trasportatori sulla disponibilità nel sistema nervoso centrale non solo della bilirubina ma egualmente di atre molecole potenzialmente tossiche, così come dei principi attivi, la loro espressione e localizzazione nelle interfacce sangue-cervello non sono del tutto chiare. Per tali motivi il lavoro di questi tre anni di tersi è stato incentrato a: Ia) chiarire il livello di espressione proteica relativa di Mrp1 e Pgp nelle due principali barriere cerebrali, la BBB (blood brain barrier, barriera emano encefalica) e la BCSFB (blood CerebroSpinal Fluid barrier, barriera emato liquorale); Ib) Definire l’andamento della loro espressione nel corso dello sviluppo post-natale in situazione fisiologica. II) Valutare l’effetto di elevati livelli serici di bilirubina sull’espressione di Mrp1 e Pgp nelle barriere emato encefaliche, come prima linea di difesa verso la bilirubina nel kernittero. Per raggiungere questo secondo obiettivo abbiamo utilizzato il ratto Gunn, considerato il modello in vivo per la sindrome di Crigler-Najjar e il kernittero. I ratti Gunn presentano elevati livelli di bilirubina serica ed un quadro clinico simile a quanto si riscontra nell’uomo. L’iperbilirubinemia, nel ratto, è dovuta ad una mutazione nell’enzima responsabile della coniugazione del pigmento, passaggio fondamentale per la sua successiva eliminazione. Nell’omozigote (jj) la bilirubina totale nel sangue (TBS) è molte volte più alta che nell’eterozigote (Jj) in cui l’allele non mutato codifica per l’enzima nella sua forma attiva, sufficiente a mantenere livelli di bilirubina normali. RISULTATI Ia) Attraverso una quantificazione relativa dell’ espressione proteica, ottenuta tramite Western blot, abbiamo dimostrato una espressione speculare dei due trasportatori nelle interacce sangue cervello. Mentre i microvasi sono caratterizzati dalla forte espressione di Pgp, ed i livelli di Mrp1 sono 15-20 volte inferiori rispetto ai plessi, questi ultimi presentano una elevata espressione di Mrp1 ed una quasi completa assenza di Pgp. Per quanto riguarda l’espressione di Mrp1 nei plessi coroidei (CP), abbiamo potuto evidenziare una differenza, con la massima espressione nel plesso del 4° ventricolo rispetto ai ventricoli laterali. Tramite immunofluorescenza abbiamo poi evidenziato per entrambe i trasportatori una localizzazione lato sangue, con Pgp luminale nei vasi e Mrp1 baso-laterale nel plessi coroidei. Ib) Anche l’andamento dell’espressione durane lo sviluppo post-natale differisce. Mentre Mrp1 è sin dalla nascita (2 giorni di vita) altamente espresso in entrambe le barriere, Pgp è inizialmente espresso a livelli più bassi (4,6 volte meno) rispetto all’ adulto (60 giorni). Contemporaneamente anche la densità dei vasi nel parenchima aumenta. II) Nel modello iperbilirubinemico rappresentato dal ratto Gunn, la TBS (jj) e molte volte più alta che nell’ eterozigote (Jj) e tale differenza permane per tutto l’arco di tempo esaminato (0-60 giorni dalla nascita). Al contrario la bilirubina libera (calcolata) è elevata solo nelle prime due settimane di vita, quando il rapporto bilirubina-albumina nel sangue è superiore all’unità. Poi, il rapido aumento della concentrazione ematica di albumina determina un significativo calo della Bf. Mentre l’analisi degli effetti (macroscopici) dell’iperbilirubinemia sullo sviluppo degli emisferi cerebrali non evidenzia differenze tra Jj e jj; in questi ultimi la crescita del cervelletto è severamente inibita. Già a 17 giorni di vita l’ipoplasia del cervelletto si manifesta con una differenza nel peso del 50% nei jj rispetto agli animali normo bilirubinemici di pari età. Durante tale periodo anche l’espressione dei due trasportatori nelle barriere è modificata. L’espressione proteica di Pgp nella BBB degli animali iperbilirubinemici è aumentata ad ogni età presa in esame. Tuttavia tale incremento non modifica in maniera importante la quantità del trasportatore nei MV durante lo sviluppo post natale, rimanendo quindi poco espresso (5 volte meno rispetto all’adulto) almeno fino ai 17 giorni di vita. Contemporaneamente la presenza Mrp1 nella BCSFB è inibita. Già a 9 giorni nel plesso del 4° ventricolo Mrp1 è il 50% rispetto al controllo (pari età, Jj). Anche se nei plessi dei ventricoli laterali l’inibizione dell’espressione è inferiore, nell’insieme la quantità di Mrp1 è fortemente ridotta negli animali iperbilirubinemici. Contrariamente, nei ratti Jj, Mrp1 ha un andamento simile a quello descritto nella sezione (Ib). CONCLUSIONI I risultati da noi ottenuti sottolineano importanti differenze tra le due barriere. La barriera emato-encefalica si sviluppa durante il primo periodo post-natale, in un ambiente caratterizzato dalla forte presenza di membrane cellulari. Similarmente l’espressione di Pgp è inizialmente bassa ed incrementa molto durante lo sviluppo post-natale. Al contrario I plessi coroidei appaiono precocemente in età embrionale, contribuiscono allo sviluppo del cervello e posseggono il più alta espressione di enzimi di fase II, coinvolti nel metabolismo di potenziali sostanze tossiche, del cervello. Un alto livello di Mrp1 sin dalla nascita suggerisce un suo coinvolgimento nel trasporto di qualche sostanza importante nello sviluppo del cervello o in un suo precoce coinvolgimento nel mantenimento dello stato ossido riduttivo, o nell’eliminazione di metabolici dal sistema nervoso centrale. Elevati livelli di bilirubina, come nel modello Gunn, modulano sia l’espressione di Pgp nella BBB, che di Mrp1 nella BCSFB. Tuttavia l’incremento nell’espressione di Pgp nei microvasi non sembra essere sufficiente a contrastare efficacemente l’ingresso della bilirubina libera, molto elevata fino al 17 giorno di vita. La simultanea riduzione di Mrp1 nei plessi coroidei, può facilitare l’ingresso o ridurre l’efflusso della bilirubina nel liquido cefalo rachidiano, consentendo l’accumulo e conseguente danno dei tessuti esposti.<br>................................................................ ....................... .. .BACKGROUND The unconjugated bilirubin (UCB), a heme degradation product, has been suggested to be a potent antioxidant at low concentration while it seems to be extremely dangerous at higher concentrations, causing encephalopathy in severely jaundiced neonates. Around 70% of children with kernicterus die within seven days, while the 30% survivors usually suffer irreversible sequels, including hearing loss, paralysis of upward gaze, mental retardation, and cerebral palsy with athetosis. Bilirubin encephalopathy is actually the leading cause of hospital readmission of newborns within the first month after birth. Historically the studies concerning the bilirubin entry the central nervous system have focused on the blood brain barrier (BBB), located at the level of the endothelial cells forming the brain micro vessels (MV), leading to the “free bilirubin theory”. It consists in the idea that only the free unconjugated bilirubin, the part of bilirubin exceeding the binding ability of the serum albumin, is able to cross the cell membranes and diffuse in tissue. In brain a second blood brain barrier is present. It is located at the level of the epithelial cells forming the choroids plexuses, between the blood and the cerebrospinal fluid (blood-cerebrospinal fluid barrier, BCSFB). Despite the largest surface area available for the exchanges, the high blood flux, the strategically position between two circulating fluids and the more leaky phenotype, limited studies have been made concerning its role in limiting the bilirubin entry the brain. Two ATP dependent transporters, the Multidrug Resistance-associated Protein 1 (Mrp1) and the MultiDrug resistance Protein (Pgpor MDR1), appear to be actively involved in UCB trafficking. The transporters play an important role in keeping extra cellular bilirubin concentration, such as potentially toxic compounds, below toxic levels by limiting the entry of UCB from blood to brain, or else in controlling intracellular bilirubin levels in parenchyma cells. Despite the importance of Mrp1 and Pgp on BBI their pattern of expression and cellular localization remains still unsettled. Based on these considerations - The first aim of the thesis was clarify the relative protein expression of these transporters at the two major BBI protecting the brain from toxic insults (Ia), and to identify their post-natal developmental profile of expression and cellular localisation (Ib). Similarly, no data about the Mrp1 and Pgp expression on BBI during the bilirubin encephalopathy are available. - The second aim of the thesis was investigate a relation between the high level of blood bilirubin and Mrp1 and Pgp expression in brain barriers in vivo using the Gunn rat (II), in witch the symptoms closely correlate to the human kernicterus and Crigler-Najjar syndrome type I. In this animal model, a mutation in the enzyme responsible for the conjugation and subsequent elimination of bilirubin, leads to the total absence of the enzymatic activity in the homozygous animals (jj), causing a severe life long hyperbilirubinemia. In the heterozygous Gunn rats (Jj), the enzymatic activity, until if reduced, is present and result in normal serum bilirubin levels. RESULTS Ia) By quantitative Western blot, we have demonstrated a mirroring expression of the two transporters at the blood brain interfaces in the adult rat. On the BBB the Pgp is strongly expressed and the Mrp1 amount is 15-20 times lower than in CPs. At the contrary, the CPs are characterized by the high expression of Mrp1, with a difference between the lateral ventricle (LV) and the 4th ventricle (4thV) CP, the former being a lower Mrp1 expression than in the last. In both LV and 4thV CPs, Pgp is virtually absent. By immunofluorescence we revealed that both ABC transporters are located at the blood side, the Pgp luminal on MV, and Mrp1 basal on CPs. Ib) With respect to the post-natal development, the Mrp1 expression is high since the early post-natal age and do not change significantly from birth to adult life in both barriers. By contrast, Pgp expression is weak a P9 and increase 4.6 fold with maturation on MV. Synchronously the density of Pgp stained MV in parenchyma seems increasing. II) In the homozygous Gunn rat (jj) the total bilirubin in serum is several time higher than in the heterozygous (Jj) animals all life long. By contrast the (calculated) free bilirubin is extremely elevated until the first week of life, when the bilirubin-albumin ratio exceed the unit, then drop due to the developmental increasing albumin concentration in blood. While no differences in Cx weight have been found between Jj and jj rat at every postnatal age, the cerebellum development is strongly impaired by the bilirubin toxic effect, displaying a Summarymarked hypoplasia, with about the 50% of weight loss respect the Jj control at 17 days after the birth. Concerning the ABC transporters, the differential pattern of expression between blood brain interfaces is maintained. But, in jj Gunn rats, the Pgp expression at the BBB is up-regulated at every post natal age analysed, also if this increase do not seems to be sufficient to confer protection at list until P17, when the amount of the transporter in the MV is about 5 times lower than in adult (P60). At the same time the Mrp1 expression on the BCSFB is down regulated. Since P9 the amount of Mrp1 in the 4thV CP of jj rats drops around to the 50% respect the amount in the littermates. In the LV CP the decrease is less marked, but in any case the Mrp1expression in both CPs is strongly impaired. This down regulation seems to be post-transcriptional. In the Jj animals, the Mrp1 relative expression is already high in both plexuses at early postnatal stages. A significant difference was noted only between LV CP (76%) and 4thV CP at P60 (100%). CONCLUSIONS All together these results indicate that the two barriers differ: The BBB develops after the birth and is surrounded by the lipid rich parenchyma environment, in agreement with the transporter preference for the lipid compounds and the strong post-natal developmental increase of the Pgp amount on MV. The CPs develops early in the foetal life, are involved in the guidance of the brain development and posses the highest phase II metabolising enzymes in the brain. The Mrp1 amount in CPs is similar to the adult level since the birth and may be involved in the transport of some compounds important in the brain development, in the detoxification or in the maintenance of the redox state (GS- sulfo- conjugates, LC4, etc.). In Gunn rats, as model for Kernicterus and Crigler-Najjar syndrome type I, the Pgp offered protection is not sufficiently modulate until P17, when the amount of the free bilirubin is elevated and could cross the brain barriers. The simultaneously down regulation of Mrp1 at the BCSFB may facilitate the entry of the bilirubin or strongly impair their clearance in the central nervous system, leading to the accumulation in brain and subsequent damage of tissue.<br>1974

Objetivos: A deficiência de glicose-6-fosfato desidrogenase (G6PD) está associada a um maior risco de encefalopatia bilirrubínica e de crise hemolítica aguda grave desencadeada por drogas como a primaquina e a dapsona. Conhecer a prevalência dessa deficiência enzimática em área onde a malária e a hanseníase ainda estão presentes e conhecer a prevalência das principais mutações traz subsídios para planejamento de estratégias com vistas à redução de riscos associados a esta deficiência enzimática. Métodos: Estudo descritivo transversal conduzido em uma região do centro-oeste do Brasil. Exame de triagem para deficiência de G6PD foi realizado em 3573 recémnascidos. Exame confirmatório foi necessário em 188 crianças triadas como possíveis portadores de deficiência. Nas crianças em que foi confirmada a deficiência de G6PD foi feita pesquisa das mutações G202A (G6PD A-) e C563T (G6PD Mediterrâneo) por PCR. Resultados: A deficiência de G6PD foi confirmada em 63 crianças, sendo 60 meninos (95,2%) e três meninas (4,8%). O percentual de exames falso-positivos na fase de triagem foi de 66,5%, estando o percentual de falso-positivos associado à temperatura e tempo de transporte das amostras. Entre as crianças que confirmaram deficiência de G6PD, foi mais frequente a história de anemia em familiares e de icterícia neonatal. Houve associação entre hematócrito baixo e deficiência enzimática, mas não com hemoglobina, contagem de reticulócitos ou neutrófilos. A prevalência da deficiência de G6PD (IC95%) foi de 1,76% (1,37; 2,24) entre os recém-nascidos triados e de 3,34% entre os meninos (2,58; 4,25). A mutação C563T não foi identificada em nenhuma criança, mas a mutação G202A estava presente em 58 crianças - 92,06% (IC95%: 83,29 - 97,03): 56/60 meninos e em 2/3 meninas homozigotas. Foi identificado um menino com Kernicterus portador da mutação G202A em hemizigose. Conclusão: O elevado percentual de falso-positivos na etapa de triagem, o tempo necessário entre coleta e confirmação da presença de deficiência enzimática, associado ao alto custo da triagem universal, não apoiam a inclusão da triagem de deficiência de G6PD no programa de triagem neonatal brasileiro. Na região avaliada, a prevalência observada em meninos indica que a triagem de deficiência de G6PD deva ser realizada antes do uso de drogas como a primaquina e a dapsona somente em meninos. Foi elevada a prevalência da mutação G202A, de classe III, sendo esta mutação associada a uma menor morbidade. A identificação de um menino com Kernicterus com deficiência de G6PD indica que há necessidade de se planejar estratégias para minimizar o risco dessa morbidade associada à deficiência enzimática<br>Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is associated with an increased risk of bilirubin encephalopathy in neonates and acute hemolytic crisis triggered by drugs such as primaquine and dapsone. In an area where malaria and Hansen\'s disease are still present, knowing the prevalence of this enzyme defect and determining the prevalence of major mutations is important for planning strategies for reducing the risks associated with this enzyme deficiency. Methods: Sectional study was conducted in a Midwestern region of Brazil. Screening for G6PD deficiency was performed in 3,573 neonates. Confirmatory tests were necessary for 188 positively screened children. After confirmation, PCR investigation was utilized to identify the mutations. Results: G6PD deficiency was confirmed in 63 children: 60 boys (95.2%) and 3 girls (4.8%). The percentage of false-positive cases in the screening phase, 66.5% and was associated with the temperature and transportation time of the samples. Family history of anemia and jaundice was more frequent among the children with confirmed G6PD deficiency. An association between a low hematocrit and enzyme deficiency was observed. However, there was no association with hemoglobin reticulocyte or neutrophils counts. The prevalence of G6PD deficiency (CI95%) was 1.76% (1.37; 2.24) among all screened neonates and 3.34% (2.58; 4.25) among male children. The C563T mutation was not identified in any child. The G202A mutation was present in 58 children - 92.06% (CI95%: 83.29 - 97.03), 56/60 boys and 2/3 homozygous girls. One boy with a hemizygous G202A mutation was identified as having Kernicterus. Conclusion: The high percentage of false-positive results when first screening for G6PD deficiency; the long delay time between the test and result; along with the high cost of the this screening test, are all factors that do not support adding this test to the already established Brazilian neonatal screening programs. The prevalence observed among boys does indicate that screening for G6PD deficiency should be performed in this region before the use of drugs such as primaquine and dapsone only in boys. This study found a high prevalence of the G202A mutation, a Class III variant associated with lower morbidity. The identification of a G6PD deficient boy with Kernicterus reinforces the necessity for strategies to abolish the morbidity associated with this enzyme deficiency

I neonati nel reparto di terapia intensiva neonatale, specialmente quelli nati pretermine e con basso peso alla nascita, possono aver bisogno di nutrizione enterale e parenterale (NP) con lipidi. I lipidi forniti con la dieta sono essenziali per raggiungere i requisiti nutrizionali ed energetici necessari per una corretta crescita. Uno stato di malnutrizione nella vita post-natale e una scarsa crescita intra-ospedaliera sono state associate ad un ritardo di crescita a lungo termine e del neurosviluppo. Questa tesi descrive studi sul metabolismo lipidico di neonati alimentati per via enterale e parenterale. I nostri dati suggeriscono che la placenta umana limita la disponibilità di fitosteroli (steroli di origine naturale presenti in prodotti derivati da piante come gli oli vegetali) al feto. Il metabolismo dei fitosteroli nel neonato pretermine in NP è ridotto rispetto a quello del neonato a termine e dell’adulto: più bassa esterificazione e lenta eliminazione dal torrente sanguigno. La difficoltà di metabolizzare i fitosteroli è marcatamente ridotta in caso di colestasi. L’assunzione orale/enterale di alimenti con elevato contenuto di fitosteroli (es. formule per l’infanzia a base di oli vegetali) è associata ad alti livelli di fitosteroli plasmatici nei neonati pretermine e a termine. Una dieta con basso contenuto di fitosteroli è preferibile per i neonati. L’olio di pesce assunto per via intravenosa non influenza negativamente la crescita ponderale dei pretermine. Le emulsioni con olio di pesce sembrano essere sicure anche se il loro effetto sulla crescita di organi come cervello e polmoni richiede ulteriori studi. L’ipertrigliceridemia è associata ad una riduzione degli apporti dei lipidi intravenosi ma non a ridotta crescita e neurosviluppo nei pretermine. Le dosi raccomandate di lipidi intravenosi potrebbero essere in eccesso rispetto a quelle necessarie per una corretta crescita. I benefici di lipidi extra in caso di ipertrigliceridemia restano da chiarire<br>Infants admitted to neonatal intensive care unit, especially those born prematurely and with low birth weight, may require enteral and parenteral nutrition (PN) with lipids. Dietary lipids are essential for infants to achieve nutrient and energy requirements to ensure an adequate growth. Early life malnutrition and poor in-hospital growth have been associated with later impaired long-term growth and neurodevelopment. This thesis describes a number of investigations on lipid metabolism in enterally and parenterally fed infants. Our data suggest that human placenta tends to limit the availability of phytosterols (naturally occurring sterols found in plant-derived products, such as vegetable oils) to the foetus. Phytosterol metabolism in preterm infants on routine PN with vegetable oils is reduced in comparison with term infants and adults: lowest esterification and slow elimination from the bloodstream. In case of cholestasis, the ability of preterm infants to manage phytosterols in intravenous (IV) lipid emulsions (LE) is markedly low. A high enteral phytosterol intake as it occurs in infant milk formula-fed infants results in elevated plasma phytosterol levels both in preterm and term infants. A low phytosterol diet should be preferred for infants. We found that IV fish oil does not negatively affect weight gain in small preterm infants. New generation IV LE containing fish oil appear to be safe for preterm infants but their effect on organ growth (such as brain and lungs) deserves further studies. In a case-control study, we found that hypertriglyceridemia (HiTG) affects IV LE intakes in small preterm infants on routine PN. We did not find any association between HiTG and reduced growth and poorer neurodevelopment. The recommended doses of IV LE may still be in excess compared to those effectively required for adequate growth and metabolism of small preterm infants. The benefits of the extra IV LE during HiTG remain to be clarified.

Digoxin, a steroidal glycoside that inhibits Na⁺/K⁺-ATPase, is the most commonly prescribed cardiac medication in North America. Blood levels of this drug are routinely measured to reduce the risks of toxicity. Reports questioning the specificity of antisera used in radioimmunoassays for serum digoxin measurements began to appear after 1975¹ when plasma from patients with renal failure, not on glycoside therapy, showed false-positive digoxin levels. Since then, digoxin-like immunoreactive substances (DLIS) have been found in sera from patients with hepatic failure, hypertension, pre-eclampsia, in amniotic fluid and cord blood. Some of the highest values for DLIS have been detected in premature infants, where levels have often exceeded the therapeutic range (0.2-2.0 µg/L) for digoxin. Cord blood has been identified as a rich source of DLIS. Dahl et al² were the first to suggest that a circulating saluretic substance "endoxin", may cause hypertension in salt sensitive rats. Gruber et al³ reported on the existence of digoxin-like factor(s) in the plasma of volume-expanded dogs. Plasma from these dogs inhibited Na⁺/K⁺ATPase activity. A number of other studies have supported the concept that such digoxin-like factors may be of etiological significance in hypertension⁴. In view of these observations, a study was undertaken to isolate and fractionate DLIS from mixed cord blood and determine whether or not any of this digoxin-like material possessed Na⁺/K⁺-ATPase inhibitory properties. Cord blood collected in the Grace Hospital Maternity Unit (Vancouver, BC), was pooled and DLIS extracted using C₁₈,R-Sep Paks. Extracts were resolved by high performance liquid chromatography (HPLC) into several fractions containing digoxin equivalent immunoactivity as measured by radioimmunoassay (RIA). A number of steroids and bile acids (dehydroepi-androsterone-sulfate, cortisone, Cortisol, deoxycortisone, ∆⁴androstene-dione, progesterone and glycochenodeoxycholic acid) cross-reacted with digoxin antisera and had HPLC retention times similar to DLIS-containing fractions. The ability of HPLC generated DLIS positive cord blood fractions to inhibit Na⁺/K⁺-ATPase activity was determined in three different assay systems; red cell ⁸⁶Rb uptake canine kidney-Na⁺/K⁺-ATPase and red cell membrane-Na⁺/K⁺-ATPase. At least six fractions contained DLIS and inhibited Na⁺/K⁺-ATPase activity. Inhibition varied with the assay system used but none of the fractions inhibited ⁸⁶Rb uptake by erythocytes. One fraction (which eluted at 29 minutes) contained progesterone; 72% of the inhibitory activity present in this fraction was attributable to this steroid. Another inhibitory fraction co-eluted with dehydroepiandrosterone-sulfate (DHEAS-S). The only fractions found to inhibit both the red cell membrane and canine kidney Na⁺ /K⁺-ATPase enzymes eluted at 7 and 29 minutes. In summary, a number of digoxin-like immunoreactive substances were isolated from cord blood by HPLC fractionation and found to inhibit Na⁺/K⁺-ATPase activity. Inhibition varied with the assay system used. There was no apparent correlation between inhibition and digoxin immunoreactivity. Very large quantities (500 mL) of cord blood were extracted to demonstrate these properties. It remains to be determined whether or not DLIS isolated during the perinatal period is of physiological significance.<br>Medicine, Faculty of<br>Pathology and Laboratory Medicine, Department of<br>Graduate

This item is only available in print in the UCF Libraries. If this is your Honors Thesis, you can help us make it available online for use by researchers around the world by following the instructions on the distribution consent form at http://library.ucf.edu/Systems/DigitalInitiatives/DigitalCollections/InternetDistributionConsentAgreementForm.pdf You may also contact the project coordinator, Kerri Bottorff, at kerri.bottorff@ucf.edu for more information.<br>Bachelors<br>Arts and Sciences<br>Nursing

A Thesis submitted to The University of Arizona College of Medicine - Phoenix in partial fulfillment of the requirements for the Degree of Doctor of Medicine.<br>There are several, predictable complications of prematurity that are faced by premature infants. Patent ductus arteriosus is among them. This study is an evaluation of a near infrared spectroscopy device to see if the premature infants can be monitored in order to detect significant ductal steal secondary to symptomatic PDA. We hypothesize that in these cases, there would be a difference between renal and cerebral regional oxygenation. To evaluate a difference, it is essential to first establish baseline regional oxygenation values, which are scarce in current literature. There are many pathophysiologic states experienced by premature infants, such as hypoxic spells and intraventricular hemorrhage, which may affect the data. Therefore, the effects of abnormal brain activity on cerebral monitoring are also evaluated. Our preliminary data shows a mean cerebral reading of 77.5189 and a mean renal reading of 70.9105, both without any linear trends. Now that normative data for regional oxygen saturation in the preterm neonate has been established, this opens up a pathway for the study of additional states, such as the left to right shunt of sPDA, since there is now a standard for comparison.

As technology and interventional techniques continue to evolve, both the volume and complexity of cardiac catheterizations will increase, leading to a rise in the number of complications. One of the most morbid complications of cardiac catheterization is vascular injury. We report the case of a 31-day-old, 3.0-kg infant with hypoplastic left heart syndrome who experienced a left common iliac artery disruption during cardiac catheterization resulting in a retroperitoneal hemorrhage. The extent of the vascular injury combined with the vessel caliber posed a technically challenging surgical scenario. Ultimately, the vascular supply to the left lower extremity was reconstructed by the plastic surgery team with a reverse autologous vein graft. To our knowledge, this multidisciplinary approach with the involvement of plastic surgery represents a unique case.

This item is only available in print in the UCF Libraries. If this is your Honors Thesis, you can help us make it available online for use by researchers around the world by following the instructions on the distribution consent form at http://library.ucf.edu/Systems/DigitalInitiatives/DigitalCollections/InternetDistributionConsentAgreementForm.pdf You may also contact the project coordinator, Kerri Bottorff, at kerri.bottorff@ucf.edu for more information.<br>Bachelors<br>Nursing<br>Nursing

Two studies were conducted to investigate factors involved in absorption of immunoglobulins in the preruminant neonate. In the first study 26 Angus cows were fed 57% or 100% of their NRC recommended requirements for protein and energy for the last third of gestation in each of 2 years. Resulting calves were fed measured amounts of colostrum from their dam or from a cow of the reciprocal nutritional treatment group. Cows from the restricted intake treatment lost weight and body condition (P < .05). Gestation length, birth weight, calving ease, days open end weaning weight, however, were not affected by treatment. Calves born to restricted fed dame had higher serum cortisol and lower serum T₃ concentration (P < .05), but absorption of IgG was not affected. Calves fed colostrum from restricted cows did have lower serum IgG concentrations (P < 0.2); although, none of the calves were considered hypogammaglobunemic. In the second study, 2 trials were conducted to evaluate the effect of varying cortisol concentrations on Ig absorption in lambs. Treatments consisted of control (CO), high cortisol (HC), single peak of cortisol (SP) end low cortisol (LC). Lambs in trial 1 were obtained on d 136 to 138 of gestation by caesarean operation. HC and SP lambs tended to have a faster rate of Ig absorption through 24 h, but did not differ from CO lambs in serum Ig concentration by 36 h. Precocious closure to Ig absorption had occurred for LC lambs by 20 h end they had lower Ig concentrations at 36 and 48 h (P < 0.05). In trial 2, lambs were obtained on d 140 to 142 of gestation. Lambs had a shorter Ig absorptive period than trial 1 (24 vs 36 h for CO]. Premature closure for the LC lambs occurred by 16 h postpartum (P < 0.05) and they tended to have depressed Ig concentrations post 24 h.<br>Ph. D.

Гипербилирубинемии новорожденных на протяжении многих лет занимают одно из ведущих заболеваний новорожденных. За последние десять лет отмечается рост количества неонатальных желтух у доношенных новорожденных: в 1996 г. они встречались с частотой 4,8%, в 2001 г. – 8,6 %, а в 2006 г. – 11,7 %. Это послужило поводом для детального изучения возможных причин их появления. При цитировании документа, используйте ссылку http://essuir.sumdu.edu.ua/handle/123456789/5130