La dysplasie broncho-pulmonaire (DBP), complication fréquente de la prématurité, atteint 30% des nouveau-nés de faible poids de naissance. L’hypertension artérielle pulmonaire persistante du nouveau-né (HTAPP), associé ou non à la DBP, résulte d’une mauvaise adaptation à la vie extra-utérine et survient dans diverses situations pathologiques (prématurité, sepsis, inhalation de méconium, hernie diaphragmatique congénitale…). Ces 2 pathologies sont grevées d’une morbidité et d’une mortalité importante en période périnatale. En effet, certaines situations d’HTAPP ou de DBP sévères restent réfractaires aux thérapeutiques actuelles.Les acides gras polyinsaturés oméga 3 (AGPI ω-3) sont des nutriments aux propriétés bénéfiques sur le système circulatoire et pulmonaire, mais également sur le développement fœtal, démontrés par de nombreuses études expérimentales et cliniques. La déhydroépiandrostérone (DHEA) est une hormone stéroïdienne dont le taux de sécrétion chez l’homme diminue avec l’âge. Des études récentes ont démontré un effet cardio-protecteur mais également un effet vasodilatateur pulmonaire et préventif de lésions de DBP dans des modèles expérimentaux.Les buts de notre travail étaient 1) d’étudier l’effet d’une supplémentation en AGPI ω-3 dans un modèle expérimental de DBP induite par hyperoxie chez le raton, 2) d’étudier l’effet circulatoire d’injection d’AGPI ω-3 (in vivo) dans un modèle d’étude de la circulation pulmonaire chez le fœtus de brebis chroniquement instrumenté, et d’étudier les mécanismes d’action AGPI ω-3 (anneaux vasculaires isolés) , enfin 3) d’étudier l’effet circulatoire de la DHEA (in vivo) dans le modèle de fœtus de brebis et d’étudier les mécanismes d’actions de la DHEA sur la circulation pulmonaire fœtale (in vivo)._x000D_Nous avons démontré que la supplémentation par voie orale en AGPI ω-3 de rates gestantes à la fin de la gestation et après la naissance permettait de prévenir, chez les ratons nouveau-nés, les lésions de DBP induites par une exposition chronique à l’hyperoxie. Ces lésions étaient retrouvées dans les groupes contrôles (eau et AGPI ω-6). Cette étude n’avait pas retrouvée d’effet bénéfique des AGPI ω-3 sur le remodelage vasculaire induit.L’injection d’acide eicosapentaènoique (EPA) chez le fœtus de brebis a révélé un effet vasodilatateur pulmonaire puissant avec une baisse significative et prolongée des résistances vasculaires pulmonaires (RVP), en comparaison à l’injection d’acide docosahéxaènoique (DHA) ou de l’excipient (faible dose d’éthanol). L’effet vasorelaxant de l’EPA sur des anneaux isolés pré-contractés était plus important que celui du DHA à dose équivalente, et il était dose- et endothélium-dépendent. Enfin, cet effet impliquait la voie de production du NO puisqu’il était diminué lors du traitement des anneaux par le L-Nitro-Arginine (LNA), inhibant la NO synthase.L’étude de perfusion en bolus de DHEA dans le lit pulmonaire vasculaire chez le fœtus de brebis instrumenté mettait en évidence un effet vasodilatateur bref. Cet effet était dose-dépendant avec une baisse plus prononcée des RVP et une durée plus importante pour des doses de DHEA plus importantes. Enfin l’étude des mécanismes d’action retrouvait une inhibition de l’effet de la DHEA par le LNA, démontrant une action vasodilatatrice par activation de production du NO.L’ensemble de ces travaux permet de suggérer que les AGPI ω-3 représentent des nutriments intéressants en période périnatale (grossesse, allaitement et per os), notamment en traitement préventif dans les situations à risque de DBP, ou curatif en cas d’HTAPP. La DHEA reste une piste dans le traitement de l’HTAP, mais semble pour l’instant plus difficile à instaurer en clinique humaine. / Bronchopulmonary dysplasia (BPD), a common complication of prematurity, reached in 30% of newborns with very low birth weight. Persistent pulmonary hypertension of the newborn (PPHN), with or without BPD, results in poor adaptation to extrauterine life and occurs in various pathological conditions such as prematurity, sepsis, inhaled meconium, or diaphragmatic hernia Congenital. The mortality and morbidities of these two diseases are high in the perinatal period. Severe PPHN or BPD are refractory to current treatment.Polyunsaturated fatty acids omega-3 (ω-3 PUFA) are nutrients with beneficial properties on the circulatory and pulmonary system, but also on fetal development, demonstrated by many experimental and clinical studies. Dehydroepiandrosterone (DHEA) is a steroid hormone whose secretion levels in humans decreases with age. Recent studies have demonstrated a cardio-protective effect of diet DHEA supplementation but also a pulmonary vasodilator and preventive effect of DBP injury in experimental models.The aims of our study were : 1) to study the effect of PUFA ω-3 supplementation in an experimental model of hyperoxia-induced DBP in pups; 2) to study effect on pulmonary circulation of infusion of ω-3 PUFAs (in vivo) in model of chronically instrumented fetal sheep, and to analyze the mechanisms of action of ω-3 PUFA (isolated vascular rings); and finally 3) to study the in vivo effect of DHEA in fetal pulmonary circulation in the same model of fetal sheep and to understand the mechanisms of action of DHEA._x000D_We have demonstrated that supplementation with diet PUFA ω-3 on pregnant rats at the end of gestation and after birth prevent BPD injuries induced by chronic exposure to hyperoxia in pups. These lesions were found in the control groups (water and ω-6 PUFA). ω-3 PUFA supplementation did not prevent vascular remodeling.Infusion of eicosapentaenoic acid (EPA) in sheep fetus showed a potent pulmonary vasodilator effect as compared to docosahexaenoic acid (DHA) or excipient (low dose of ethanol). Vasorelaxant effect of EPA on pre-contracted isolated rings was more important than DHA at equivalent dose, and was dose- and endothelium-dependent. This effect involves NO production.Bolus DHEA perfusion in the pulmonary vascular bed study on instrumented fetal sheep highlighted an acute vasodilator effect. This effect was dose-dependent with a more pronounced and sustained decrease in PVR at highest doses of DHEA. Finally, mechanisms of action study found an inhibition of the effect of DHEA by the LNA, indicating that DHEA-induced vasodilation is NO dependant.Taken together, our results suggest that supplementation with ω-3 PUFAs and DHEA within the perinatal period may prevent BPD and PPHN in high risk conditions including preterm birth, premature rupture of the membrane or intrauterine growth restriction.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-04-15T12:35:53Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Trabalho e Artigo. BIBLIOTECA.pdf: 1144641 bytes, checksum: 49381fb9cd2af1b7f9dccd4c30011139 (MD5) / Made available in DSpace on 2016-04-15T12:35:53Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Trabalho e Artigo. BIBLIOTECA.pdf: 1144641 bytes, checksum: 49381fb9cd2af1b7f9dccd4c30011139 (MD5) Previous issue date: 2015-09-11 / CAPEs / O câncer de colo uterino é o terceiro câncer mais comum em mulheres. A infecção e persistência do papilomavirus humano (HPV) tem papel fundamental no surgimento e evolução das lesões cervicais, promovendo alterações no ciclo celular e proliferação celular descontrolada através das oncoproteínas E6 e E7. Entretanto, fazem-se necessários diversos outros fatores para o desenvolvimento de neoplasias. Entre estes, encontram-se as metaloproteinases de matriz (MMP), endopeptidases capazes de digerir matriz extracelular, membrana basal e induzir fatores de crescimento, que participam dos processos de invasão, metástase, angiogênese e recidiva tumorais. Em lesões neoplásicas, a síntese de MMPs encontra-se aumentada. Sua atividade normalmente é contrarregulada por inibidores endógenos, sendo muito comum que haja desequilíbrio nesta relação em lesões tumorais. A despeito de muito estudo sobre sua relação com o câncer cervical, sabe-se pouco sobre o papel das MMPs e seus inibidores na progressão de lesões cervicais causadas por HPV. Este estudo procura correlacionar a expressão de TIMP2 e RECK às lesões cervicais causadas por HPV. Foram utilizadas 115 amostras teciduais, obtidas por conização de lesões cervicais uterinas entre 2011 e 2015 no Hospital das Clínicas de Recife, nas quais foi realizado estudo histopatológico e imunohistoquímico. Foi observada reatividade fraca no citoplasma de células do tecido escamoso de 28,5% dos controles, sem nenhuma lâmina demonstrar reatividade moderada ou forte. Quanto ao núcleo, a quase totalidade das amostras não apresentou TIMP-2, enquanto 28,5% dos controles o fez. No epitélio glandular nenhuma amostra do Grupo Controle, NIC I ou NIC II foi positiva para TIMP2 no citoplasma e núcleo. Das amostras de NIC III, 6% demonstraram positividade no citoplasma. Os resultados de RECK no citoplasma do epitélio escamoso mostraram que a expressão de RECK no citoplasma de células epiteliais escamosas é significativamente maior quanto maior o grau de lesão do tecido, exceto no CC, onde a expressão é menor que a das lesões NIC III (p = 0,019). Os resultados demonstraram que a expressão nuclear de RECK em células epiteliais escamosas é significativamente menor nos tecidos displásicos (p < 0,001). Nas análises de citoplasma do epitélio glandular , nenhuma amostra do Grupo Controle, NIC I e NIC II foi positiva, havendo 3,6% de positividade nas lesões de NIC III, todas com reatividade fraca ou moderada, e 10% de positividade moderada nas amostras de CC. Nenhuma das amostras apresentou positividade nuclear. Os resultados obtidos demonstram menor expressão nuclear de TIMP2 e RECK na presença de displasia e maior expressão citoplasmática de RECK nas células escamosas. Esta foi maior quanto mais alto o grau de displasia, mas foi menor nas amostras de carcinoma do que nas de NIC III. Conclui-se que os inibidores de MMPs podem ter utilidade como marcadores imunohistológicos nas lesões cervicais, sendo necessários mais estudos para sua validação prática. / Cervical cancer is the third most common cancer in women. The infection and persistence of human papillomavirus (HPV) plays a key role in the emergence and evolution of cervical lesions, promoting changes in the cell cycle and uncontrolled cell proliferation through the oncoproteins E6 and E7. However, make up several other factors required for the development of malignancies. Among these are the matrix metalloproteinases (MMPs), endopeptidases capable of digesting the extracellular matrix, basement membrane and induce growth factors, that participate in the processes of invasion, metastasis, angiogenesis and tumor recurrence. In neoplastic lesions, MMPs synthesis is increased. Its activity is usually contrarregulada by endogenous inhibitors, being very common there is imbalance in this relationship in tumor lesions. Despite much study on its relationship with cervical cancer, little is known about the role of MMPs and their inhibitors in the progression of cervical lesions caused by HPV. This study tries to correlate the expression of RECK and TIMP2 the cervical lesions caused by HPV. 115 tissue samples were used, obtained by conization to uterine cervical lesions between 2011 and 2015 at the Hospital das Clinicas of Recife, in which was conducted histopathological and immunohistochemical study. Weak reactivity was observed in the cytoplasm of the squamous tissue cells of 28.5% of controls, with no slide show moderate or strong reactivity. As for the core, almost all of the samples showed no TIMP-2, while 28.5% of controls did. Glandular epithelium in any sample of the control group, CIN I or CIN II was positive for TIMP2 in the cytoplasm and nucleus. Samples of CIN III, 6% showed positivity in the cytoplasm. The results of RECK in the cytoplasm of squamous epithelium showed that RECK expression in the cytoplasm of squamous epithelial cells is significantly higher the higher the degree of tissue injury, except DC, where expression is lower than that of CIN III lesions (p = 0.019). The results showed that nuclear RECK expression in squamous epithelial cells is significantly lower in dysplastic tissues (p <0.001). In the cytoplasm analysis of glandular epithelium, no sample of the control group, CIN I and CIN II was positive, with 3.6% of positivity in CIN III lesions, all with low or moderate reactivity, and 10% moderate positivity in the samples DC. None of the samples showed nuclear positivity. The results showed lower nuclear expression TIMP2 and RECK in the presence of dysplasia and increased cytoplasmic expression of RECK in squamous cells. This was greater the higher the degree of dysplasia, but was lower in carcinoma samples than in CIN III. We conclude that MMP inhibitors may have utility as immunohistological markers in cervical lesions, more research is needed to validate your practice.

Introduction: Chronic inflammation plays an important role on the transformation and tumor progression during oral carcinogenesis. There is a great number of chronic inflammatory lesions (CIL) in the oral cavity which are related to dysplastic processes of the epithelium, immune response and changes on the collagen deposition. Objectives: To investigate the presence of dysplasia and to histologically grade them in the CIL of traumatic cause, as well as toaccessthe density of mast cells and different types of collagen fibers in cases of epithelial dysplasias and oral squamous cell carcinomas (OSCC). Material and Method: Initially, 183 CIL cases were evaluated as to the presence of dysplasia and also classified according to its degree of epithelial dysplasia. Among those lesions, 45 CIL cases were selected and divided into two groups: group 1 (15 cases of mild dysplasia), group 2 (15 cases of moderate/severe dysplasia). The control group was composed by 15 cases of OSCC.They were stained with toluidine blue in order to quantify the mast cells and picrosirius red to semi-quantify the collagen type fibers. Results: The mast cells were detected in all groups presenting a mean of 6,76 cells/mm2, 10.82 cells/mm2 and 19.18 cells/mm2 in the control, group 1and 2 respectively. Regarding the collagen fibers, type III was more prevalent on groups 2 and control while type I fibers were more abundant on group 1. Conclusion: Oral chronic inflammatory lesions showed dysplastic changes in most analyzed cases. The results suggests an active participation of mast cells in the stage of tumor transformation, since it was detected a higher density onthe dysplasia cases when compared to the OSCC cases. Nevertheless, the gradual change of collagen type fibers indicates that collagenproducing cells become altered during the stages of dysplasia (tumor transformation). / Introdução: A inflamação crônica tem um papel importante na transformação e progressão tumoral durante a carcinogênese oral. Muitas lesões inflamatórias crônicas (LIC) da cavidade oral estão relacionadas a processos displásicos do epitélio, à resposta imune e à mudança na deposição do colágeno. Objetivos: Investigar a presença de displasia e graduá-las histologicamente nas LIC de origem traumática, como também, avaliar a densidade de mastócitos e de diferentes tipos de fibras colágenas nas LIC com displasias epiteliais e comprar aos casos de carcinomas de células escamosas (CCE). Material e Métodos: Inicialmente 183 LIC foram avaliadas quanto à presença de displasia e classificadas em relação ao grau. Em seguida, 45 casos foram divididos em: Grupo controle (CCE), Grupo 1 (displasia leve- DL), Grupo 2 (displasia moderada/severa- DM/S). Foram corados com Azul de Toluidina para quantificar os mastócitos e Picrosirius Red para avaliação dos tipos de fibras colágenas I e III. Resultados: As LIC foram mais frequentes em mulheres (n=107) com idade de 36,6 anos. O sítio mais afetado foi a mucosa do lábio inferior (29,7%), já a lesão mais frequente foi o fibroma traumático (39,2%). A displasia leve esteve presente em 56,3% da amostra. Os mastócitos foram evidenciados nos três grupos: grupo controle (6,76 mastócitos/mm), grupo 1 (10,82 mastócitos/mm2) e grupo 2 (19,18 mastócitos/mm2).Quando analisadas as fibras colágenas, observouse no grupo controle e no grupo 2 que as fibras tipo III foram mais prevalentes, já no grupo 1 prevaleceu-se as fibras tipo I. Conclusão: Lesões inflamatórias crônicas orais apresentaram alterações displásicas na maior parte dos casos. O estudo sugere uma participação dos mastócitos na fase de transformação tumoral. E a alteração gradativa dos colágenos tipo I e III indica alteração das células produtoras de colágeno, durante transformação tumoral.

A displasia frontonasal (DFN) compreende quadros de aparência facial variável, sendo clinicamente caracterizada por dois ou mais dos seguintes sinais: hipertelorismo ocular com consequente alargamento da base nasal; fissura facial mediana afetando o nariz ou o nariz e lábio superior e, por vezes, o palato; fissura alar (uni ou bilateral); ponta nasal ausente; crânio anterior bífido oculto, e implantação em &#39;V&#39; dos cabelos na fronte. A DFN pode ser vista como um defeito de desenvolvimento que pode ocorrer por si só ou como parte do quadro clínico de várias síndromes. A maioria dos casos de DFN é esporádica, e em raras circunstâncias foram observadas alterações cromossômicas em alguns indivíduos. Até o momento, quatro genes foram relacionados à patogênese molecular de algumas das síndromes com DFN, EFNB1, associado a uma forma de DFN ligada ao X e os genes ALX1, ALX3 e ALX4, todos associados a formas de DFN com herança autossômica recessiva. Embora esteja claro haver heterogeneidade etiológica, na maioria dos casos de DFN a causa não é conhecida, dificultando o adequado aconselhamento genético aos pacientes e seus familiares. Sendo assim, realizamos estudos com diferentes estratégias metodológicas buscando melhor compreender as possíveis causas genéticas da DFN. Ao todo foram analisados 10 pacientes: um caso familial de DFN leve com herança aparentemente autossômica dominante, um caso clinicamente sugestivo de mutação em ALX1, e oito casos de DFN associada a atraso de desenvolvimento com ou sem outras anomalias, dos quais um apresentava um rearranjo de novo aparentemente balanceado entre os cromossomos 4 e 12. Optamos por realizar sequenciamento dos genes previamente relacionados a fenótipos com DFN em todos os casos; para aqueles em que não foram detectadas mutações patogênicas, realizamos análise de variações de número de cópias (CNV) por microarray de polimorfismos de base única e, para o paciente com rearranjo cromossômico, realizamos o mapeamento do ponto de quebra por hibridação in situ fluorescente. Constatamos uma mutação em heterozigose no gene ALX4 co-segregando com o fenótipo do caso familial, sendo esta a primeira descrição de alteração em tal gene causando uma forma de DFN com herança dominante, e sugerimos pela primeira vez um mecanismo de dominância negativa. No caso sugestivo de mutação em ALX1, o diagnóstico foi confirmado através da identificação de uma mutação em homozigose neste gene do paciente; este caso consiste no 3o da literatura mundial e evidencia pela primeira vez que mutações em ALX1 não necessariamente levam a atraso de desenvolvimento ou deficiência intelectual. Os estudos citogenéticos e moleculares dos pontos de quebra do paciente com rearranjo cromossômico sugeriram os genes ARAP2 e CAND1 como possíveis responsáveis por seu quadro clínico, enquanto o estudo de CNVs nos indivíduos com DFN associada a atraso de desenvolvimento apontou os genes DNAJB12 e ENOX2 como possíveis candidatos para explicar o fenótipo de dois dos pacientes. É preciso que novos estudos sejam realizados a fim de melhor compreender o significado de tais achados e a real contribuição de cada gene para o desenvolvimento craniofacial humano e para a etiologia da DFN. Para os casos em que não foram identificadas alterações conclusivas no presente estudo, embora causas ambientais não possam ser descartadas, é preciso que seja investigada também a existência de fatores genéticos e epigenéticos não detectáveis pelas metodologias utilizadas, bem como a hipótese de mosaicismo somático. Nossos resultados, além de corroborarem o envolvimento dos genes ALX1 e ALX4 em fenótipos com DFN, sugerem também novos genes candidatos: ARAP2, CAND1, DNAJB12 e ENOX2 / Frontonasal dysplasia (FND) is a rare group of disorders that comprises cases with a variety of facial appearances, and is clinically characterized by two or more of the following signs: ocular hypertelorism with consequent broadening of the nasal root; median facial cleft affecting the nose and/or upper lip and palate; clefting of the alae nasi (uni or bilateral); lack of formation of the nasal tip; anterior cranium bifidum occultum; and a V-shaped frontal hairline. FND is a developmental defect that can occur alone or as part of several syndromes. Most cases of FND are sporadic, and in rare circumstances chromosomal alterations were observed in affected individuals. To date, four genes have been related to the molecular pathogenesis of some syndromes with DFN, one (EFNB1) is associated with an X-linked form while the 3 others (ALX1, ALX3 and ALX4) are associated with autosomal recessive forms. Although it is clear that FND is etiologic heterogeneous, the causative mechanism is unknown in most cases which makes it hard to give proper genetic counseling to patients and their families. In order to get new insights into the genetic mechanisms leading to FND, we performed studies with different methodologies. Altogether, 10 patients were analyzed: a familial case of a mild form of FND with an apparently autosomal dominant inheritance pattern, a case clinically suggestive of mutation in ALX1, and eight cases of FND associated with developmental delay with or without other anomalies, one of which with an apparently balanced de novo rearrangement between chromosomes 4 and 12. We chose to sequence the genes previously associated with FND phenotypes in all cases; for those in which pathogenic mutations were not detected, we conducted an analysis of copy number variations (CNV) by single nucleotide polymorphisms microarrays; for the patient with chromosomal rearrangement, we also mapped the breakpoints by using fluorescence in situ hybridization. We found a heterozygous mutation in ALX4 co-segregating with the phenotype of the familial case; this is the first description of mutation in this gene causing a form of FND with dominant inheritance pattern, and we suggested for the first time a dominant negative mechanism. In the case suggestive of mutation in ALX1, the diagnosis was confirmed by the identification of a homozygous mutation in this gene; this is the third case of the literature and shows for the first time that mutations in ALX1 are not necessarily related to developmental delay or intellectual disability. Breakpoints cytogenetic and molecular studies done with the patient with chromosomal rearrangement suggested ARAP2 and CAND1 genes as causative candidates for his condition, while the study of CNVs in individuals with FND associated with developmental delay pointed DNAJB12 and ENOX2 genes as possible candidates to explain the phenotypes of two of the patients. Further studies are necessary to better understand the significance of such findings and the actual contribution of each of these genes to human craniofacial development and the etiology of FND. Although environmental causes cannot be ruled out, it should also be investigated the existence of genetic and epigenetic factors as well as the possibility of somatic mosaicism, among the cases negative for the molecular approaches used in our study. Our results corroborate the involvement of ALX1 and ALX4 in FND phenotypes, and suggest new candidate genes: ARAP2, CAND1, DNAJB12 and ENOX2.

Andreas Starke Quantifizierung und Klassifizierung der kaninen Ellbogeninkongruenz auf Grundlage einer standardisierten Röntgen- und Messmethode Klinik für Kleintiere der Veterinärmedizinischen Fakultät der Universität Leipzig (79 Seiten, 27 Abbildungen, 8 Tabellen, 132 Literaturangaben) Zielstellung: Ziel der vorliegenden Arbeit war die objektive Quantifizierung der knöchernen Konformation des Ellbogengelenkes gesunder und erkrankter Hunde unter standardisierten Bedingungen. Dazu sollte eine Methode entwickelt werden mit der Röntgenaufnahmen von Ellbogen unter Belastung angefertigt und anhand von Messungen an markanten Knochenstrukturen evaluiert werden können. Diese standardisierte Röntgen- und Messmethode sollte an einer Kontrollgruppe getestet werden, um die Eignung der Methodik zu untersuchen und Referenzwerte zu erstellen. Besonderer Schwerpunkt war die Überprüfung der Reliabilität der Messungen in Abhängigkeit von Lagerungsartefakten und Messwiederholungen. Danach sollten Ellbogengelenke klinisch erkrankter Hunde untersucht und mit den Ergebnissen der Kontrollgruppe verglichen werden, um herauszufinden, ob sich die Messergebnisse signifikant von gesunden Gelenken unterscheiden. Zudem wurde die Hypothese aufgestellt, dass sich die dysplastischen Ellbogengelenke anhand der röntgenologischen Messparameter in unterschiedliche Formen der humeroulnaren und humeroradialen Inkongruenz unterteilen lassen. Material und Methode: Bei 27 lahmheitsfreien Hunden wurden von 47 Ellbogengelenken Röntgenaufnahmen mit und ohne Belastung (mediolateral, kraniokaudal; Aufnahmen am liegenden, narkotisierten Tier) sowie Aufnahmen im Stehen (kraniokaudal) untersucht. Digital gemessen wurden subchondrale Gelenkspaltenabstände, subchondrale Knochenabstände und ein Winkel. Anschließend wurden 149 Hunde mit ED der Röntgen- und Messmethode unterzogen und mit den Ergebnissen der Kontrollgruppe verglichen. Anhand von gehäuft auftretenden Messwertabweichungen wurden die 149 Hunde in Untergruppen eingeteilt und erneut mit der Kontrollgruppe verglichen. Danach wurden 4 Gruppen mit jeweils unterschiedlicher Osteotomie an Radius bzw. Ulna zur zwei- und dreidimensionalen Gelenkumstellung erstellt (euthanasierte Hunde ohne Ellbogenerkrankung), der gleichen Röntgen- und Messprozedur unterzogen und mit der Kontrollgruppe und den Gruppen der kranken Hunde verglichen. Ergebnisse: Eine standardisierte und reliable Lagerung sowie Längen- und Winkelmessungen an definierten Knochenpunkten konnten mit und ohne Belastung bei gesunden und kranken Hunden etabliert werden. Mittels Normalisierung der Parameter der Längenmessungen gelang eine Vergleichbarkeit zwischen unterschiedlichen Gelenken und Projektionsebenen herzustellen. Messwerte von Aufnahmen im Stehen zeigten viele Lagerungsartefakte und eine geringe Reliabilität. Von den einzelnen Messparametern der Aufnahmen mit und ohne Belastung konnten aufgrund ihrer geringen Streuung Normwerte von der Kontrollgruppe abgeleitet werden. Eine deutliche Abgrenzung auf Grundlage signifikanter Unterschiede führte zur Bildung der Untergruppen Typ I (n=60), Typ II (n=40) und indiff (n=49) innerhalb der 149 Hunde mit ED. Unterscheidungskriterien für die Einteilung eines kranken Ellbogengelenkes in Typ I, Typ II und indiff waren 3 subchondrale Gelenkspaltenabstände (mp3, mp4, mp6), 3 aus Gelenkspaltenabständen errechnete Indizes (LI 3, LI 4, LI 6) und ein Winkel. Diese Parameter wiesen in der Kontrollgruppe eine hohe Reliabilität auf. Durch die Osteotomiemodelle konnten Parallelen zwischen Typ I und einer short ulna sowie zwischen Typ II und einem short radius ermittelt werden. Schlussfolgerungen: Röntgenaufnahmen am stehenden Hund sind nicht praktikabel, nur bedingt auswertbar und daher nicht geeignet. Anhand von Röntgenbildern mit und ohne Belastung lassen sich dagegen die Knochenverhältnisse im Gelenk beschreiben und Normwerte erstellen. Die untersuchte Methodik ist geeignet 67% erkrankter Tiere einem Inkongruenztyp (Typ I oder Typ II) zuzuordnen. Die Ergebnisse der dreidimensionalen Ulnaverkürzung und dreidimensionalen Radiusverkürzung weisen auf dreidimensionale Bewegungen der Knochenoberflächen bei Typ I bzw. Typ II im und am Gelenk hin. Ob Ellbogengelenke des Typ I (40%) von einer dreidimensionalen Osteotomie der proximalen Ulna und Ellbogengelenke des Typ II (27%) von einer Radiuselongation profitieren würden, müssen weitere Untersuchungen zeigen. Die Inkongruenz selbst konnte nicht näher quantifiziert werden.:1 EINLEITUNG 1 2 LITERATURÜBERSICHT 3 2.1 Anatomie des kaninen Ellbogengelenkes 3 2.2 Biomechanik des Ellbogengelenkes 5 2.3 Ellbogendysplasie 7 2.3.1 Inkongruenzen 8 2.3.1.1 Radioulnare Längendisparität 9 2.3.1.2 Dynamische radioulnare Längendisparitäten 10 2.3.1.3 Ellipsoide Malformation der Incisura trochlearis 10 2.3.1.4 Primäre Rotationsinstabilität von Radius und Ulna relativ zum Humerus 11 2.3.2 Angular vector model 12 2.3.3 Bildgebende Verfahren zur Darstellung einer Inkongruenz 13 2.3.3.1 Röntgenuntersuchung als planare Projektion 13 2.3.3.2 Computertomographie 16 2.3.3.3 Magnetresonanztomographie 17 2.3.3.4 Arthroskopie 17 2.3.3.5 Röntgen, CT, MRT und Arthroskopie im Vergleich 18 3 EIGENE PUBLIKATIONEN 19 3.1 Röntgenologische Quantifizierung der Ellbogengelenks- konformation mit einer neuen standardisierten Röntgentechnik unter Last: Referenzwerte für mittelgroße und große Hunde ohne Ellbogengelenksdysplasie 19 3.2 Vergleichende röntgenologische Untersuchung von kaninen Ellbogengelenken mit ED und ohne ED unter standardisierten belasteten Bedingungen 49 4 DISKUSSION 72 5 ZUSAMMENFASSUNG 76 6 SUMMARY 78 7 LITERATURVERZEICHNIS 80 8 DANKSAGUNG 91 / Andreas Starke Quantification and classification of canine elbow incongruency using a standardized method for radiography and measurement Department of Small Animal Medicine, Faculty of Veterinary Medicine, Universitiy of Leipzig (79 pages, 27 Figures, Tables, 132 references) Objective: Goal of this paper was the objective quantification of elbowjoint conformation for healthy and diseased dogs under standardized conditions. Therefor we developed a method for taking x-rays of the elbow joint under load to facilitate measurements using bone-landmarks. For this we aimed to test a standardized X-ray- and measurement methodology on a control group for evaluation of the developed methodology and to compile reference values. Emphasis has been placed on the validation of the reliability of the taken measurements especially were positioning artefacts and repeat measurements were concerned. Subsequently we examined elbow joints of clinically affected dogs and compared the results to that of the control group to determine whether signifikant differences can be found in the measurements compared to those taken in joints of healthy dogs. Additionally we formulated the hypothesis that displastic Elbow joints can be grouped into humero-ulnar and humero-radial incongruencies using the developed method of measurement. Material and methods: Plain x-rays and standardized stressed x-rays (medio-lateral, cranio-caudal taken under sedation; cranio-caudal views taken while standing) of 47 elbow-joints of 27 lameness free dogs were examined. We digitally measured subchondral jointgaps, subchondral bone gaps and an angle. Afterwards 149 dogs with ED were examined using our standardized X-ray method and we compared the results to the control group. The 149 dogs were divided into subgroups using frequent measurement deviances and the subgroups again compared to the control group. Afterwards we created 4 groups based on the kind of radial or ulnar osteotomy for two- and three- dimensional joint adjustment, took measurements based on our standardized X-ray method and compared the results to the control group and ED group. Results: A reliable standardized X-ray methodology as well as length and angle measurement on defined bone landmarks with and without load could be established on healthy dogs and those with ED. Using normalization of length-measurements we were able to establish comparability between different joints and projections. Measurements from x-rays taken in a standing position showed a lot of positioning related artefacts and low reliability. We were able to compile standard values from x-rays of the control group taken with- and without load due to the low deviation of the norm values. Due to marked delination based on significant differences we were able to establish subgroups Typ I (n=60), Typ II (n=40) and Group indiff (n=49) in the ED Group. Distinction Criterions to classify an ED elbow joint in Types I, II or indiff were 3 subchondral jointgaps (mp3, mp4, mp6), three indices calculated out of joint gaps (LI 3, LI 4 and LI 6) and an angle. In the control group these parameters showed a high reliability. Using an osteotomy model we were able to draw parallels between Typ I and a short Ulna, as well as Typ II and a short radius. Conclusions: X-rays taken in a standing position are not feasible. Using x-rays taken with and without load we were able to adequately describe bone-marker relations and could compile norm values. Using our developed method we have been able to assign 67% of the examined diseased joints to a type of incongruency (Type I or II). The results of three dimensional ulna and radius shortening indicate three dimensional movement of the bone surface in Typ I or II in the joint. Whether elbow joints of Typ I (40%) benefit of a three dimensional proximal ulna-osteotomy or joints of Typ II (27%) would benefit of a radial elongation needs to be shown in further studies.:1 EINLEITUNG 1 2 LITERATURÜBERSICHT 3 2.1 Anatomie des kaninen Ellbogengelenkes 3 2.2 Biomechanik des Ellbogengelenkes 5 2.3 Ellbogendysplasie 7 2.3.1 Inkongruenzen 8 2.3.1.1 Radioulnare Längendisparität 9 2.3.1.2 Dynamische radioulnare Längendisparitäten 10 2.3.1.3 Ellipsoide Malformation der Incisura trochlearis 10 2.3.1.4 Primäre Rotationsinstabilität von Radius und Ulna relativ zum Humerus 11 2.3.2 Angular vector model 12 2.3.3 Bildgebende Verfahren zur Darstellung einer Inkongruenz 13 2.3.3.1 Röntgenuntersuchung als planare Projektion 13 2.3.3.2 Computertomographie 16 2.3.3.3 Magnetresonanztomographie 17 2.3.3.4 Arthroskopie 17 2.3.3.5 Röntgen, CT, MRT und Arthroskopie im Vergleich 18 3 EIGENE PUBLIKATIONEN 19 3.1 Röntgenologische Quantifizierung der Ellbogengelenks- konformation mit einer neuen standardisierten Röntgentechnik unter Last: Referenzwerte für mittelgroße und große Hunde ohne Ellbogengelenksdysplasie 19 3.2 Vergleichende röntgenologische Untersuchung von kaninen Ellbogengelenken mit ED und ohne ED unter standardisierten belasteten Bedingungen 49 4 DISKUSSION 72 5 ZUSAMMENFASSUNG 76 6 SUMMARY 78 7 LITERATURVERZEICHNIS 80 8 DANKSAGUNG 91

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19401号 / 医博第4052号 / 新制||医||1012(附属図書館) / 32426 / 京都大学大学院医学研究科医学専攻 / (主査)教授 髙田 穣, 教授 平岡 眞寛, 教授 松本 智裕 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Bronchopulmonary dysplasia (BPD) is a chronic lung disease and one of the most severe complications that develop in premature infants following mechanical ventilation, exposure to supplemental oxygen, and inflammation. The hallmarks of the lung pathology are arrested lung development, including fewer and larger alveoli with less septation, thickening of alveolar septa, and impaired development of the capillary network. BPD is associated with increased mortality, respiratory morbidity, neurodevelopmental impairment, and increased healthcare costs. Significant advancements in neonatology in the last several decades, including antenatal steroids and exogenous surfactant replacement therapy, more gentle ventilation methods, and judicious oxygen use, have allowed for the survival of more preterm infants. However, the incidence of BPD still remains high and currently, there is no cure for the disease. Novel effective interventions at this stage of life are of exceptional value. Considering their great potential in promoting tissue regeneration and modulating inflammation, mesenchymal stromal cells (MSCs) represent a promising avenue for treating several disorders, including BPD. Umbilical cord-derived MSCs (UC-MSCs) offer biological advantages over other MSC sources (easily available, high proliferative capacity, and better repair efficacy). Pioneering work in our lab showed that MSCs prevent injury to the developing lung in a rat model mimicking BPD. However, there are still considerable challenges that must be overcome before MSCs can be effectively implemented in clinical trials. As such, UC-MSC heterogeneity is poorly understood, with concerns regarding variations from donors and batches. Thus, to improve the reproducibility of basic research and clinical applications, and to identify the optimal therapeutic cell product, better molecular characterization of UC-MSCs and the development of standardized BPD models will be essential in the clinical translation of MSC therapy for BPD. Moreover, considering that BPD is a disease of prematurity, the therapeutic potential of UC-MSCs isolated from preterm birth is of major interest. In the study presented here, using single-cell RNA sequencing (scRNA-seq), we characterized MSCs isolated from the UC of term and preterm pregnancies at delivery (term and preterm donors), as well as non-progenitor control cell line, human neonatal dermal fibroblasts (HNDFs). Moreover, we associated UC-MSC transcriptomic profiles with their therapeutic potential in hyperoxia-induced lung injury in neonatal rats. Finally, we developed and characterized a novel two-hit (2HIT) BPD model in neonatal mice, assessed UC-MSCs' optimal route of injection, timing, and dose, and evaluated their therapeutic effects in that model. We showed that UC-MSCs isolated from the majority of term and preterm donors, including preterm donors with pregnancy-related complications, have limited heterogeneity and possessed a transcriptome enriched in genes related to cell cycle and cell proliferation activity (termed "progenitor-like" cells). In contrast, UC-MSCs isolated from one term and two preterm donors with preeclampsia displayed a unique transcriptome comprised of many genes related to fibroblast activity, including extracellular matrix (ECM) organization (termed "fibroblast-like" cells). In addition, treatment with progenitor-like UC-MSCs, but not with fibroblast-like cells nor HNDFs, significantly improved lung structure, function, and pulmonary hypertension (PH) in hyperoxia-induced lung injury in neonatal rats. We identified marker genes for the therapeutic UC-MSCs (progenitor-like cells) and non-therapeutic cells (fibroblast-like cells and HNDFs). Among them, the high expression of major histocompatibility complex class I (MHCI) is associated with a reduced therapeutic effect. Furthermore, we developed a novel 2HIT BPD mice model with in-depth characterization of the innate immune response and lung injury. 2HIT injury caused a transient type 1 proinflammatory cytokine response and a significant decrease in type 2 anti-inflammatory cytokine lung expression and number of anti-inflammatory M2 type alveolar macrophages. Moreover, 2HIT mice showed impaired lung compliance and growth. Repeated intravenous (i.v.) injections of UC-MSCs at a dose of 20×10⁶ cells/kg body weight (BW) on postnatal day (PD) one and two improved survival, BW, lung compliance, and growth of 2HIT animals. In conclusion, scRNA-seq experimentation provided evidence that UC-MSCs isolated from different donors harbor different transcriptomes with progenitor-like or fibroblast-like characteristics. Only progenitor-like cells provided a therapeutic effect in hyperoxia-induced lung injury in neonatal rats. The development of a novel murine 2HIT BPD model allowed us to characterize the innate immune response and lung pathology and confirm the optimal dose of UCMSCs to provide therapeutic potential in that model. These results will enable better therapeutic selection of UC-MSCs and help improve treatment regimen prior to ultimate clinical translation.

Introduction Developmental dysplasia of the hip has since 1953 in Sweden most commonly been treated with the von Rosen splint. Over time, different types of splints have been developed, one of which is the Orebro splint. In some countries it is advised to have long-term follow-ups with radiological exams of patients treated with a splint. In Orebro, there is currently no mandatory follow-up after treatment with the Orebro splint. Aim The aims were to investigate how many children, despite treatment with the Orebro splint, developed dysplasia in the adolescence period, to study the hip function in children treated with the Orebro splint and investigate if there were any differences in treatment outcomes between males and females. Methods All children born between 2000 and 2012, treated with the Orebro splint, were eligible for inclusion. The outcome measures for quality of life and hip function were EQ-5D-Y-VAS and CHOHES score. The outcome measures for the pelvic radiological exams were Acetabular Index and Center Edge angle. Results Data from 46 patients were collected, 7 males and 39 females. 5,3% of the patients showed residual dysplasia. Both males and females showed overall good results in the radiological images, the surveys, and the clinical exams. The results did not show significant differences between the genders. Conclusion Even though our participants reported an overall good quality of life, our results showed some cases of residual dysplasia. However, the small sample size makes it difficult to assess whether the Orebro splint is equal to other splints regarding treatment outcomes.

Thesis (MScEng)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The shape of the knee’s trochlear groove is a very important factor in the overall stability of the knee. However, a quantitative description of the normal three-dimensional geometry of the trochlea is not available in the literature. This is also reflected in the poor outcomes of patellofemoral arthroplasty (PFA). In this study, a standardised method for femoral parameter measurements on three-dimensional femur models was established. Using software tools, virtual femur models were aligned with the mechanical and the posterior condylar planes and this framework was used to measure the femoral parameters in a repeatable way. An artificial neural network (ANN), incorporating the femoral parameter measurements and classifications done by experienced surgeons, was used to classify knees into normal and abnormal categories. As a result, 15 knees in the database were classified by the ANN as being normal. Furthermore, the geometry of the normal knees was analysed by fitting B-spline curves and circular arcs on their sagittal surface curves to prove and reconfirm that the groove has a circular shape on a sagittal plane. Self-organising maps (SOM), which is a type of ANN, was trained with the acquired data of the normal knees and in this way the normal trochlear geometry could be predicted. The prediction of the anterior-posterior (AP) distance and the trochlear heights showed an average agreement of 97 % between the actual and the predicted normal geometries. A case study was conducted on four types of trochlear dysplasia to determine a normal geometry for these knees, and a virtual surface reconstruction was performed on them. The study showed that the trochlea was deepened after the surface reconstruction, having an average trochlea depth of 5.5 mm compared to the original average value of 2.9 mm. In summary, this research proposed a quantitative method for describing and predicting the normal geometry of a knee by making use of ANN and the femoral parameters that are unaffected by trochlear dysplasia. / AFRIKAANSE OPSOMMING: Die vorm van die trogleêre keep is ’n belangrike faktor in patella-stabiliteit. Tog is ’n kwantitatiewe beskrywing van die normale driedimensionele geometrie van die troglea nog nie beskikbaar nie, wat duidelik blyk uit die swak uitkomste van patellofemorale artroplastie (PFA). In hierdie studie is ’n gestandaardiseerde metode vir die meting van femorale parameters op grond van driedimensionele femurmodelle ontwikkel. Die femurmodel is in lyn gebring met die meganiese en posterior kondilêre vlak, welke raamwerk gebruik is om die femorale parameters op ’n herhaalbare wyse te meet. Die normale knieë is geklassifiseer met ’n kunsmatige neurale netwerk (ANN), wat die femorale parameter-mate sowel as die chirurgiese klassifikasie ingesluit het, en 15 knieë is gevolglik as normaal aangewys. Die normaleknie-geometrie is ontleed deur B-latkrommes en sirkelboë op die sagittale oppervlak-kurwes aan te bring om te bewys en te herbevestig dat die keep uit ’n sirkelvorm op ’n sagittale vlak bestaan. Die ingesamelde data van die normale knieë is ingevoer by selfreëlende kaarte (SOM), synde ’n soort ANN, wat die navorser in staat gestel het om die normale trogleêre geometrie te voorspel. Die voorspelling van die anterior-posterior (AP) afstand en die trogleêre hoogtes toon ’n gemiddelde ooreenkoms van meer as 97 % tussen die werklike en voorspelde normale geometrie. ’n Gevallestudie is op vier soorte trogleêre displasie uitgevoer om die normale geometrie te voorspel en ’n oppervlakrekonstruksie daarop uit te voer. Hierdie studie het getoon dat die troglea ná oppervlakrekonstruksie verdiep was, met ’n gemiddelde trogleadiepte van 5.5 mm in vergelyking met die aanvanklike gemiddelde waarde van 2.9 mm. Hierdie navorsing het dus ’n metode aan die hand gedoen vir die kwantitatiewe beskrywing en voorspelling van normale geometrie met behulp van ANN sowel as met die femorale parameters wat nie deur die trogleêre displasie geraak word nie.

<p>Uvod: Bronhopulmonalna displazija (BPD) je najče&scaron;ća i najteža respiratorna posledica prematuriteta. Utvrđivanje najznačajnijih faktora rizika za nastanak BPD kod novorođenčadi porođajne mase (PM) ispod 1500g može omogućiti procenu rizika za&nbsp; nastanak bolesti i identifikaciju novorođenčadi u visokom riziku, &scaron;to je važno za pružanje informacija roditeljima o prognozi,&nbsp; planiranje preventivnih i terapijskih mera i stratifikovanje novorođenčadi koja su u riziku za sprovođenje budućih istraživanja. Cilj: Utvrđivanje incidencije, stepena težine BPD, smrtnosti, identifikacija najznačajnijih prenatalnih i postnatalnih faktora rizika za nastanak BPD, konstrukcije modela predikcije za nastanak BPD. Materijal i metode: Istraživanje je sprovedeno na 504&nbsp; prevremeno rođene novorođenčadi PM&lt;1500g koja su rođena u porodili&scaron;tima u AP Vojvodini i lečena u tercijarnom Centru za neonatologiju i intenzivnu negu i terapiju, na Institutu za zdravstvenu za&scaron;titu dece i omladine Vojvodine u periodu od&nbsp; 2006.-2011. godine. Retrospektivno je analizirano prisustvo BPD, prema stepenima težine, smrtnost. Podaci su izdvojeni iz&nbsp; istorija bolesti za svako novorođenče, 30 potencijalnih prenatalnih i postnatalnih faktora je opisano deskriptivnom i univarijantnom statistikom. Statstički najznačajniji faktori su uneti u multifaktorsku logističku regresionu analizu u cilju&nbsp; konstrukcije prediktivnih modela za nastanak BPD u 1.,14. i 21. danu neonatalnog života. Podaci su obrađeni u StatSoft-ovom&nbsp; programskom paketu Statistica 10.0.&nbsp; Validacija modela predikcije je sprovedena u prospektivnom delu istraživanja, na 100&nbsp;&nbsp;&nbsp; prevremeno rođene novorođenčadi&lt;1500g, u periodu od 2012-2013. godine. Rezultati: U retrospektivnom delu&nbsp; istraživanja,&nbsp; od 504&nbsp; novorođenčeta PM&lt;1500 grama, umrlo je 17.65%, BPD je imalo 45.43% (blagu BPD 19.44%, srednje te&scaron;ku 19.84%,&nbsp; te&scaron;ku&nbsp; 6.15%), srednje te&scaron;ku i&nbsp; te&scaron;ku 25.99%.Antenatalna primena kortikosteroida je zastupljena u 47.02%, surfaktant&nbsp;&nbsp; je&nbsp;&nbsp; primenjen kod 69.78% novorođenčadi. Najznačajniji prenatalni prediktivni faktor rizika za nastanak BPD/smrtnog ishoda je horioamnionitis (OR 5.72; 95% CI 3.42-9.62), dok su protektivni faktori: prenatalna primene kortikosteroida (OR&nbsp; 0.41;&nbsp; 95%CI&nbsp; 0.29-0.60), porođaj carskim rezom (OR&nbsp; 0.24; 95% CI 0.16-0.36). Najznačajniji&nbsp; postnatalni prediktivni faktori rizika su: GS&nbsp; (p&asymp;0.00), PM (p&asymp;0.00), reanimacija u porođajnoj sali (OR 7.01; 95% CI 4.12-12.01), rana&nbsp; neonatalna&nbsp; sepsa&nbsp; (OR&nbsp; 7.35;&nbsp; 95%CI&nbsp; 3.79-14.58), RDS&nbsp; (p&asymp;0.00), primena surfaktanta (OR13,3;95%CI 8,2 - 21,67), DAP (OR 4.12; 95%CI&nbsp; 2.47-6.89),&nbsp; dok&nbsp; je&nbsp; ženski&nbsp; pol&nbsp; protektivan (OR&nbsp; 0.61; 95% CI 0.42-0.89). FiO2 i IPPV su u svim posmatranim danima značajni faktori rizika. Primena IPPV u 1. danu (OR 10.71;&nbsp; 95% CI 6.67-17.26); u ostalim danima rizik od BPD raste prema rastućoj invazivnosti respiratorne&nbsp; potpore.&nbsp; Konstruisani su modeli&nbsp; predikcije za 1, 14 i 21. dan života, modeli imaju visoku prediktivnu vrednost: ukupan procenat uspe&scaron;nosti&nbsp; modela je 84.26%-90.80%, modeli sa ne&scaron;to većim uspehom predviđaju&nbsp;&nbsp; prisustvo (85.36%-94.12%), nego odusustvo BPD (81.72-86.56%). OR modela je 28.07-103.04. Modeli su uspe&scaron;no validirani&nbsp; na 102 pacijenta sa ukupnim procentom uspe&scaron;nosti (82-90%), PPV (0.86-0.94) i NPV (0.76-0.87). Zaključak:&nbsp; Kori&scaron;ćenjem&nbsp; prenatalnih i postnatalnih kliničkih podataka moguće je predvideti nastanak BPD ili smrtnog ishoda.</p> / <p>Introduction: Bronchopulmonary dysplasia (BPD) is the most common serious pulmonary morbidity in very low birth weight (VLBW) infants. It is of clinical importance to determine clinical variables that are associated with BPD in order to identify infants who are at risk of developing BPD; it contributes to BPD prevention, may enable prognostic information for parents and future studies design. Objective: The aim of this study was to determine the incidence and severity of BPD, mortality rate in VLBW infants, to identify prenatal and postnatal predictive risk factors for bronchopulmonary dysplasia and competing outcome of death and to develop predictive models. Materials and Methods: Study was conducted in 504 VLBW infants born in the maternity hospitals in Vojvodina and admitted to tertiary Center for newborn and neonatal intensive care at the Institute for Child and Youth Health Care of Vojvodina, from January 2006. to December 2011. Data were retrospectively collected from clinical records for outcomes BPD or death; prenatal and postnatal factors associated with BPD were collected at three postnatal ages and examined by descriptive and univariate statistics; factors that were significantly associated with BPD and/or death were entered into a multivariate logistic regression analysis for develop predictive models. Data were analyzed using StatSoft&#39;s software package Statistica 10.0. Validation of the models were conducted in a prospective study in 102 VLBW infants born from January 2012. to December 2013. Results: There were 504 very low birth weight infants who were eligible for this study, 17.65% died, 45.43% developed BPD (mild BPD 19.44%, moderate 19.84%, severe 6.15%), moderate and severe 25.99%. The mean birth weight for the cohort was 1125.6&plusmn;280.9g, the mean gestation age was GS 28,78&plusmn;3,01, 49.21% were male. Surfactant received 69.78%, antenatal steroids 47.02% newborns. Key risk factors for BPD and/or death were: chorioamnionitis and maternal infections at delivery (OR 5.72; 95% CI 3.42-9.62); protective prenatal factors were: antenatal corticosteroid therapy (OR 0.41; 95%CI 0.29-0.60), cesarean delivery (OR 0.24; 95% CI 0.16-0.36). Postnatal rick factors were: GS (p&asymp;0.00), birth weight (p&asymp;0.00), delivery room resuscitation (OR 7.01; 95% CI 4.12-12.01), early neonatal sepsis (OR 7.35; 95%CI 3.79-14.58), RDS (p&asymp;0.00), surfactant (OR13,3;95%CI 8,2 - 21,67), DAP (OR4.12; 95% CI 2.47-6.89), while female gender was protective (OR 0.61; 95% CI 0.42-0.89). At each time point studied, FiO2 was significantly higher in BPD/death, as well as respiratory support; on the first day invasive respiratory support was significantly associated with BPD/death (IPPV and HFOV) (OR 10.71; 95% CI 6.67-17.26), in other days BPD was associated with increasing invasiveness of respiratory support. In multifactorial logistic regression analysis separately predictive models were developed at three postnatal ages, at 1st, 14th and 21st day. Models had high predictive performance: total success of the models were 84.26% - 90.80%, models successfully predicted the presence of BPD in 85.36% -94.12%, absence of the BPD in 81.72 - 86.56% cases. OR of models were 28.07-103.04. The models were successfully validated on 102 patients with a total percentage of success 82 - 90%, with PPV 0.86-0.94 and NPV 0.76-0.87. Conclusion: Using prenatal and postnatal clinical data it is possible to predict the development of BPD and/or death in very low birth weight infants. It is very important to identify risk factors for BPD development in order to decrease the incidence of BPD and mortality rate.</p>