<p>Bilirubin is a degradation product of haem, which is constantly formed in all</p><p>mammals. Increased levels of bilirubin in humans lead to jaundice, a condition</p><p>that is very common during the first days after birth. This neonatal</p><p>jaundice can routinely be treated by phototherapy without any serious side</p><p>effects. During this treatment, bilirubin undergoes a photoreaction to isomers</p><p>that can be excreted. The most efficient photoreaction is the isomerisation</p><p>around a double bond (Z-E-isomerisation), which results in more soluble</p><p>photoproducts.</p><p>The work presented in this thesis shows results of a femtosecond optical</p><p>spectroscopy study, combined with quantum-mechanical investigations, of</p><p>the mechanism of isomerisation of bilirubin. The spectroscopic research was</p><p>conducted with bilirubin in organic solvents, and in buffer complexed by</p><p>human serum albumin. This albumin complex is present in the blood, and</p><p>has thus medical importance. Quantum-chemical calculations (CASSCF) on</p><p>a bilirubin model were used to explain experimental results.</p><p>The fluorescence decay observed with femtosecond spectroscopy shows an</p><p>ultrafast component (~120 fs), which is explained by exciton localisation,</p><p>followed by processes with a lifetime of about 1-3 ps. These are interpreted</p><p>as the formation of a twisted intermediate, which decays with a lifetime of</p><p>10-15 ps back to the ground state, as observed by absorption spectroscopy.</p><p>CASSCF calculations, in combination with the experimental results, suggest</p><p>the ca. 1-3 ps components to be relaxation to the twisted S1 minimum, followed</p><p>by the crossing of a barrier, from where further relaxation takes place</p><p>through a conical intersection back to the ground state.</p><p>Time-dependent DFT calculations were utilised to analyse the absorption</p><p>spectrum of bilirubin. Good agreement with the measured spectrum was</p><p>achieved, and low-lying states were observed, that need further investigation.</p><p>The theoretically obtained CD spectrum provides direct evidence that</p><p>bilirubin preferentially binds to human serum albumin in the enantiomeric</p><p>P-form at neutral pH.</p> / <p>Bilirubin är en nedbrytningsprodukt av hem som ständigt bildas hos alla</p><p>däggdjur. En förhöjd bilirubinkoncentration i den mänskliga kroppen kan</p><p>leda till gulsot, något som är mycket vanligt under de första dagarna efter</p><p>födelsen (neonatal gulsot). Fototerapi används rutinmässigt som säker behandlingsmetod,</p><p>under vilken bilirubin genomgår en fotoreaktion till en</p><p>isomer som kan utsöndras. Den mest effektiva fotoreaktionen är en Z-Eisomerisation,</p><p>vilken leder till lösligare fotoprodukter.</p><p>Arbetet som presenteras i denna avhandling visar resultaten av en kombinerad</p><p>femtosekund optisk-spektroskopisk och kvantmekanisk undersökning</p><p>av mekanismen bakom bilirubins isomerisation. Den spektroskopiska</p><p>studien genomfördes med bilirubin, löst i organiska lösningsmedel och i</p><p>buffert i komplex med humant serumalbumin. Detta albuminkomplex finns i</p><p>blodet, och är därför av medicinskt intresse. Kvantmekanistiska CASSCFberäkningar</p><p>på en bilirubinmodell användes för att förklara de experimentella</p><p>resultaten.</p><p>Det uppmätta fluorescence sönderfallet visar ultrasnabba komponenter</p><p>(~120 fs). Dessa tolkas som excitonlokalisering, som följs av bildandet av</p><p>ett vridet intermediat med en hastighetskonstant på ca. 1 ps-1(beroende på</p><p>lösningsmedlet). Absorptionsmätningar visar att detta intermediat sönderfaller</p><p>tillbaka till grundtillståndet med en livstid på 10-15 ps.</p><p>CASSCF beräkningar, i kombination med de experimentella resultaten, tyder</p><p>på att sönderfallet med livslängden på ca. 1 ps är en relaxation till det</p><p>vridna S1-tillståndet. Reaktionsvägen därifrån antas passera en barriär till en</p><p>konisk genomskärning, som möjliggör snabb relaxation till grundtillståndet.</p><p>Tidsberoende DFT-beräkningar användes för att analysera bilirubins absorptionsspektrum,</p><p>vilket gav bra överensstämmelse med uppmätta data. Dessutom</p><p>hittades ett tillstånd med låg excitationsenergi, som kräver ytterligare</p><p>studier. Med hjälp av det beräknade CD-spectret kunde det visas att bilirubin</p><p>binder till albumin i P-formen vid neutralt pH.</p>

Bilirubin is a degradation product of haem, which is constantly formed in all mammals. Increased levels of bilirubin in humans lead to jaundice, a condition that is very common during the first days after birth. This neonatal jaundice can routinely be treated by phototherapy without any serious side effects. During this treatment, bilirubin undergoes a photoreaction to isomers that can be excreted. The most efficient photoreaction is the isomerisation around a double bond (Z-E-isomerisation), which results in more soluble photoproducts. The work presented in this thesis shows results of a femtosecond optical spectroscopy study, combined with quantum-mechanical investigations, of the mechanism of isomerisation of bilirubin. The spectroscopic research was conducted with bilirubin in organic solvents, and in buffer complexed by human serum albumin. This albumin complex is present in the blood, and has thus medical importance. Quantum-chemical calculations (CASSCF) on a bilirubin model were used to explain experimental results. The fluorescence decay observed with femtosecond spectroscopy shows an ultrafast component (~120 fs), which is explained by exciton localisation, followed by processes with a lifetime of about 1-3 ps. These are interpreted as the formation of a twisted intermediate, which decays with a lifetime of 10-15 ps back to the ground state, as observed by absorption spectroscopy. CASSCF calculations, in combination with the experimental results, suggest the ca. 1-3 ps components to be relaxation to the twisted S1 minimum, followed by the crossing of a barrier, from where further relaxation takes place through a conical intersection back to the ground state. Time-dependent DFT calculations were utilised to analyse the absorption spectrum of bilirubin. Good agreement with the measured spectrum was achieved, and low-lying states were observed, that need further investigation. The theoretically obtained CD spectrum provides direct evidence that bilirubin preferentially binds to human serum albumin in the enantiomeric P-form at neutral pH. / Bilirubin är en nedbrytningsprodukt av hem som ständigt bildas hos alla däggdjur. En förhöjd bilirubinkoncentration i den mänskliga kroppen kan leda till gulsot, något som är mycket vanligt under de första dagarna efter födelsen (neonatal gulsot). Fototerapi används rutinmässigt som säker behandlingsmetod, under vilken bilirubin genomgår en fotoreaktion till en isomer som kan utsöndras. Den mest effektiva fotoreaktionen är en Z-Eisomerisation, vilken leder till lösligare fotoprodukter. Arbetet som presenteras i denna avhandling visar resultaten av en kombinerad femtosekund optisk-spektroskopisk och kvantmekanisk undersökning av mekanismen bakom bilirubins isomerisation. Den spektroskopiska studien genomfördes med bilirubin, löst i organiska lösningsmedel och i buffert i komplex med humant serumalbumin. Detta albuminkomplex finns i blodet, och är därför av medicinskt intresse. Kvantmekanistiska CASSCFberäkningar på en bilirubinmodell användes för att förklara de experimentella resultaten. Det uppmätta fluorescence sönderfallet visar ultrasnabba komponenter (~120 fs). Dessa tolkas som excitonlokalisering, som följs av bildandet av ett vridet intermediat med en hastighetskonstant på ca. 1 ps-1(beroende på lösningsmedlet). Absorptionsmätningar visar att detta intermediat sönderfaller tillbaka till grundtillståndet med en livstid på 10-15 ps. CASSCF beräkningar, i kombination med de experimentella resultaten, tyder på att sönderfallet med livslängden på ca. 1 ps är en relaxation till det vridna S1-tillståndet. Reaktionsvägen därifrån antas passera en barriär till en konisk genomskärning, som möjliggör snabb relaxation till grundtillståndet. Tidsberoende DFT-beräkningar användes för att analysera bilirubins absorptionsspektrum, vilket gav bra överensstämmelse med uppmätta data. Dessutom hittades ett tillstånd med låg excitationsenergi, som kräver ytterligare studier. Med hjälp av det beräknade CD-spectret kunde det visas att bilirubin binder till albumin i P-formen vid neutralt pH.

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. / 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.

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. / 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.

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. / 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.

Introduction: High levels of bilirubin in newborns can cause permanent neurodevelopmental disabilities, and it is crucial to keep the incidence low. However, the Swedish Neonatal Register revealed a high incidence of extreme neonatal hyperbilirubinemia (bilirubin ≥425 umol/L) in Region Örebro County during 2014-2019, and the reason behind this is unknown. Aim: This study aimed to review cases of extreme neonatal hyperbilirubinemia regarding the compliance to the local guidelines, and to explore potential benefits of an alternative method considering bilirubin’s rate of rise, the ruler method. Method: In this case series, a retrospective medical record review was performed on 63 newborns who were delivered at ≥35 gestational weeks and developed extreme hyperbilirubinemia before or during an admission to a hospital in Region Örebro County within the first 14 days of life (2014-2020). Results: The incidence was 2.7 cases per 1000 live births during 2014-2020. Forty-three (68.3%) cases were related to failed detection/treatment initiation and 20 (31.7%) to failed treatment. Out of the newborns classified as failed detection/treatment initiation, 27 individual newborns (62.8%) could potentially have been prevented from developing extreme hyperbilirubinemia if there were no cases of non-compliance (30.2%), if a pre-discharge screening had been performed (14.0%) and if the ruler method had been applied (19/31 investigated). Conclusion: The local guidelines used in Region Örebro County might not be sufficient in preventing the development of extreme neonatal hyperbilirubinemia. However, mandatory pre-discharge screening and a consideration of bilirubin’s current rate of rise when scheduling follow-ups could potentially lower the incidence further.

Introduction: Prolonged neonatal jaundice is defined as persistent jaundice at two-three weeksof age. Prolonged neonatal jaundice is usually harmless but one in 2500 newborns have jaundicedue to cholestasis, why further investigation must be made. Region Örebro County introduced anew referral routine for prolonged neonatal jaundice 2021-02-12 that allows for follow up inLindesberg or Karlskoga instead of Örebro alone, and only three variables need to be mentionedfor the referrals to be considered complete, contrary to previous six. Aim: To compare Region Örebro County’s current and previous referral routine for prolongedneonatal jaundice in regard to compliance and complete referrals. Methods: A chart review was performed of all children born in Region Örebro County between2021-02-12 and 2022-02-01 with either sampled bilirubin and/or diagnosis code p.55, p.57-59. Results: A statistically significant difference was observed between the routines regarding stoolcolour (p=0.004), general condition (p&lt;0.001), complete referrals (p&lt;0.001) and length ofinvestigation (p&lt;0.001). Significantly fewer patients were lost during investigation (p&lt;0.002) orhad no feedback on their test results (p&lt;0.011). Two cases of cholestasis were found. The meanvalue of conjugated bilirubin was higher in patients who saw a doctor. Few children werereferred from Lindesberg or Karlskoga. Conclusion: The current routine had more complete referrals, shorter investigation times andless absence of feedback as well as fewer patients lost during investigation. Sick patients wereidentified before getting critically ill. Shifting the entire investigation to primary care andimplementing stool charts could possibly improve the routine further.

Present work has been focused on the importance of the products of the heme catabolic pathway, in particular under conditions of unconjugated hyperbilirubinemias (neonatal jaundice and Crigler-Najjar syndrome (CNS)). The second part of the project was focused on the improvement of some pharmacological approaches used in the treatment of these diseases, as well as on studies of bilirubin products that are formed during the treatment by phototherapy (PT). Neonatal jaundice is one of the most common complications in neonates. Currently, there is no efficient pharmacotherapy and the treatment with blue light is used as a gold standard for severe neonatal jaundice. However, the absolute safety of PT has still not been confirmed. In this context, it is important to note that some neonatologists start the PT before serum bilirubin levels reach the recommended values and that patients with CNS type I (CNSI) are forced to be on life-long PT (unless undergoing liver transplantation). The focus of the present project was to study biological effects of bilirubin photoisomers (PI) in an in vitro model of the human neuroblastoma SH-SY5Y cells that are used for studies of the neuronal metabolism. In further studies performed on animal model of hyperbilirubinemic rats and mice, we investigated a suitable gene...

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. / 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.

Present work has been focused on the importance of the products of the heme catabolic pathway, in particular under conditions of unconjugated hyperbilirubinemias (neonatal jaundice and Crigler-Najjar syndrome (CNS)). The second part of the project was focused on the improvement of some pharmacological approaches used in the treatment of these diseases, as well as on studies of bilirubin products that are formed during the treatment by phototherapy (PT). Neonatal jaundice is one of the most common complications in neonates. Currently, there is no efficient pharmacotherapy and the treatment with blue light is used as a gold standard for severe neonatal jaundice. However, the absolute safety of PT has still not been confirmed. In this context, it is important to note that some neonatologists start the PT before serum bilirubin levels reach the recommended values and that patients with CNS type I (CNSI) are forced to be on life-long PT (unless undergoing liver transplantation). The focus of the present project was to study biological effects of bilirubin photoisomers (PI) in an in vitro model of the human neuroblastoma SH-SY5Y cells that are used for studies of the neuronal metabolism. In further studies performed on animal model of hyperbilirubinemic rats and mice, we investigated a suitable gene...