Dissertations / Theses on the topic 'Human fetal testis development'
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Mitchell, Roderick T. "Germ cell development in the human and marmoset fetal testis and the origins of testicular germ cell tumours." Thesis, University of Edinburgh, 2010. http://hdl.handle.net/1842/4818.
Full textMuczynski, Vincent. "Polluants environnementaux et développement du testicule foetal humain : effets et mécanismes des phtalates." Phd thesis, Université Paris Sud - Paris XI, 2011. http://tel.archives-ouvertes.fr/tel-00631554.
Full textYuen, Ka Chun. "Epigenetics of human fetal and placental development." Thesis, University of British Columbia, 2011. http://hdl.handle.net/2429/35689.
Full textTieppo, Paola. "The development of human fetal γδ thymocytes." Doctoral thesis, Universite Libre de Bruxelles, 2020. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/303402.
Full textDoctorat en Sciences biomédicales et pharmaceutiques (Pharmacie)
info:eu-repo/semantics/nonPublished
Jobling, Matthew S. "Fetal germ cell development in the rat testis and the impact of di (n-Butyl) phthalate exposure." Thesis, University of Edinburgh, 2010. http://hdl.handle.net/1842/4803.
Full textWillerton, Louise. "Gene expression in mouse testis during development." Thesis, Connect to electronic version, 2003. http://theses.gla.ac.uk:82/theses/available/etd-07042003-142909/.
Full textPh. D. thesis submitted to the Faculty of Veterinary Medicine, University of Glasgow, 2003. Includes bibliographical references. Electronic version also available via Glasgow University e-Theses service.
Jeffery, Nathan. "Fetal development and evolution of the human cranial base." Thesis, University College London (University of London), 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.392131.
Full textConliffe, Phyllis R. (Phyllis Rowena). "Effects of maternal diabetes on fetal development in rats." Thesis, McGill University, 1992. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=39344.
Full textFeichtinger, Julia. "Development of a bioinformatic analytical approach to identify novel human cancer testis gene candidates." Thesis, Bangor University, 2012. https://research.bangor.ac.uk/portal/en/theses/development-of-bioinfirmatic-analytical-approach-to-identify-novel-human-cancer-testis-gene-candidates(09065b27-9fc0-49df-a9eb-fb7ef8aab878).html.
Full textAsk, Björnberg Karolin. "Mercury exposure during early human development /." Stockholm, 2005. http://diss.kib.ki.se/2005/91-7140-224-1/.
Full textEddie, Sharon Lynn. "Novel regulators of human gonadal development." Thesis, University of Edinburgh, 2012. http://hdl.handle.net/1842/6530.
Full textHiley, Christopher. "Immunocytochemical studies of antioxidant enzyme expression in developing human tissue." Thesis, Keele University, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.260287.
Full textHakim, Souheil. "In vitro sheep fetal lung tissue characterization with ultrasound." Thesis, McGill University, 1995. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=23264.
Full textGitau, Rachel. "The development and attenuation of human hypothalamo-pituitary-adrenal axis responses in the fetal period." Thesis, Imperial College London, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.399544.
Full textKavun, M. P. "Morphogenesis of the liver in the late fetal period of development and newborns of human." Thesis, БДМУ, 2020. http://dspace.bsmu.edu.ua:8080/xmlui/handle/123456789/17567.
Full textMarchuk, F. D. "Development of maxillary sinuses for 3-5 months of the fetal period of human ontogenesis." Thesis, БДМУ, 2021. http://dspace.bsmu.edu.ua:8080/xmlui/handle/123456789/18451.
Full textHallmark, N. "Comparative effects of Di-butylphthalate and mono-butylphthalate in vitro on testis explants from the fetal rat and human : comparison with effects in vivo in the rat." Thesis, University of Edinburgh, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.651984.
Full textGodoi, Alana Rezende. "Programação fetal por sacarina sódica impacto sobre a saúde materna e na capacidade reprodutiva da prole masculina. /." Botucatu, 2020. http://hdl.handle.net/11449/192311.
Full textResumo: Introdução: A hipótese da “programação fetal” defende que eventos ocorridos durante a vida intrauterina exerçam influência na patogênese de doenças na vida adulta. Fatores ambientais podem programar no indivíduo o surgimento precoce de doenças cardiovasculares e metabólicas. Atualmente, há um aumento no consumo de adoçantes artificiais associados a tratamentos para a perda de peso e no controle do diabetes, sendo a sacarina sódica um dos mais consumidos. Entretanto, durante a gestação e lactação, o uso de sacarina sódica é restrito, por ser permeável a placenta, interagindo com o concepto e, por compor o leite materno. Embora os efeitos do uso de adoçantes sobre o peso corpóreo e o metabolismo sejam bastante conhecidos, não há relatos de pesquisas que relacionam a programação fetal pelo uso de sacarina sódica com o desenvolvimento pós-natal do testículo. Desta forma, o presente estudo visa investigar a influência do uso da sacarina sódica e da glicose na saúde materna e reprodutiva dos descendentes machos. Material e métodos: Ratas Sprague Dawley foram alimentadas durante a prenhez e lactação com dieta padrão para roedores, água filtrada ad libitum e suplementadas com iogurte natural desnatado (Grupo Controle Iogurte, n= 9); iogurte natural desnatado adoçado com solução de glicose (Dinâmica®) a 5% (v/v) (Grupo Glicose, n= 10); iogurte natural desnatado adoçado com solução de sacarina sódica (Dinâmica®) a 0,3% (v/v) (Grupo Sacarina Sódica, n= 10). As dietas líquidas foram prep... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Introduction: The "fetal programming" hypothesis argues that events that occur during intrauterine life have an influence on the pathogenesis of diseases in adulthood. Environmental factors can program in the individual the early onset of cardiovascular and metabolic diseases. Currently, there is an increase in the consumption of artificial sweeteners associated with treatments for weight loss and diabetes control, with sodium saccharin being one of the most consumed. However, during pregnancy and lactation, the use of sodium saccharin is restricted, since it is permeable to the placenta, interacting with the conceptus, and for composing breast milk. Although the effects of the use of sweeteners on the body weight and metabolism are well known, there are no reports of researches which relate fetal programming through the use of sodium saccharin to the postnatal development of the testis. Thus, the present study aims to investigate the influence of the use of sodium saccharin and glucose on the maternal and reproductive health of male offspring. Material and methods: Dams Sprague-Dawley rats were fed during pregnancy and lactation with a standard chow for rodents, filtered water ad libitum and supplemented with low-fat plain yogurt (Yogurt Control Group, n = 9); low-fat plain yogurt sweetened with 5% (v/v) glucose solution (Dinâmica®) (Glucose Group, n = 10); low-fat plain yogurt sweetened with 0.3% (v/v) sodium saccharin solution (Dinâmica®) (Sodium Saccharin Group, n = 10). ... (Complete abstract click electronic access below)
Mestre
Gruhl, Amanda Natalie. "Metakaryotic biology : novel genomic organization in human stem-like cells of fetal-juvenile development and carcinogenesis." Thesis, Massachusetts Institute of Technology, 2008. http://hdl.handle.net/1721.1/44864.
Full textIncludes bibliographical references (leaves 66-75).
Eight distinct nuclear shapes, or morphologies, have been discovered in human proto-organs and tumors, including bell-shaped nuclei with stem-like properties. These bell-shaped, or "metakaryotic," nuclei are abundant in fetal tissues and neoplasias, but rare in normal adult somatic tissues. Metakaryotic nuclei employ an unusual process for division in which DNA synthesis, partial genomic condensation, and separation of the two nuclei in a cup-from-cup fashion occur concurrently, as shown by Feulgen densitometry and single-stranded DNA assays by Dr. Elena Gostjeva. This is clearly different from the sequential steps of S-phase DNA synthesis, chromatin condensation, chromosomal separation, and genomic segregation that occur in mitotic eukaryotic cells. In order to discover how a genome apparently devoid of chromosomes might be organized, this thesis focused on recognizable DNA sequences common to all chromosomes: centromeres and telomeres. Fluorescence In Situ Hybridization (FISH) with pan-centromeric and pan-telomeric probes was applied to samples of human tissue. (A collaborating lab used centromeric and telomeric antibodies to confirm results.) An optimized FISH protocol was developed specifically for metakaryotic nuclei and tested in both human cell lines and eukaryotic cells as experimental controls. Staining of metakaryotic nuclei resulted in approximately 23 centromeric regions in each, unlike the expected number of 46 regions seen in eukaryotic nuclei. Many of these staining regions contained paired centromere signals, or doublets. This suggested a genomic organization of homologous chromosomes paired at their centromere regions. If this were the case, one would expect 46 telomeric signals per nuclei, if telomeres were also homologously paired.
(cont.) Unexpectedly, an average of 23 telomeric regions were found in many, if not all, bell-shaped metakaryotic nuclei. This, along with the observation of a condensed double ring around the mouth of the bell-shaped nuclei, suggested the possibility of a genome organized as paired, continuous genomic circles. Studies of telomere joining in metakaryotic nuclei by Dr. Per Olaf Ekstrom have provided further evidence for the paired genomic circle model. The results in this thesis are an original contribution to the field of stem cell physiology, a starting point for further investigation of DNA organization, synthesis, and repair in these metakaryotic cells, and hopefully will lead to a greater understanding of human development, growth, and cancer.
by Amanda Natalie Gruhl.
Ph.D.
Biriuk, I. G. "Рeculiarities of development of the colon topography at the end of the fetal period of human ontogenesis." Thesis, БДМУ, 2022. http://dspace.bsmu.edu.ua:8080/xmlui/handle/123456789/19606.
Full textMorgan, Leah. "Development of a 3D radial MR Imaging sequence to be used for (self) navigation during the scanning of the fetal brain in utero." Master's thesis, University of Cape Town, 2016. http://hdl.handle.net/11427/22735.
Full textTang, Wanjin. "Hormonal Regulation of the Human CYP27A1 and CYP7B1 Genes." Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis Acta Universitatis Upsaliensis, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7837.
Full textMiddlebrook, Aaron J. "Nicotine and TNF alpha, modulators of T cell signaling-effects on T cell development in fetal thymus organ culture." Diss., The University of Arizona, 2004. http://hdl.handle.net/10150/280628.
Full textRussell, Dana J. "Human Cranial Growth and Shape Change: Are Fetal Rates and Morphologies Extended Throughout the First Year of Life?" Digital Archive @ GSU, 2010. http://digitalarchive.gsu.edu/anthro_theses/43.
Full textSalisbury, Rachel. "Gene expression and cell cycle regulation in human pancreas development and congenital hyperinsulinism." Thesis, University of Manchester, 2015. https://www.research.manchester.ac.uk/portal/en/theses/gene-expression-and-cell-cycle-regulation-in-human-pancreas-development-and-congenital-hyperinsulinism(e9f58d6a-aba4-4179-a683-6d2380a55397).html.
Full textRutkowski, Paul, and Christian Albrecht May. "Nutrition and Vascular Supply of Retinal Ganglion Cells during Human Development." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2016. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-215952.
Full textGerginov, Marja [Verfasser], and Dirk [Akademischer Betreuer] Hellhammer. "Investigation and validation of animal models for the development of the human fetal and neonatal hypothalamic-pituitary-adrenal axis / Marja Gerginov ; Betreuer: Dirk Hellhammer." Trier : Universität Trier, 2011. http://d-nb.info/1197697411/34.
Full textRautavuoma, K. (Kati). "Human lysyl hydroxylases:characterization of a novel isoenzyme and its gene, determination of the domain structure of the lysyl hydroxylase polypeptides and generation of knock-out mice for the novel isoenzyme." Doctoral thesis, University of Oulu, 2003. http://urn.fi/urn:isbn:951427136X.
Full textPEACH, ELIZABETH ELAINE. "MATERNAL PSYCHOLOGICAL BENEFIT OF PRENATAL REPAIR FOR SPINA BIFIDA." University of Cincinnati / OhioLINK, 2001. http://rave.ohiolink.edu/etdc/view?acc_num=ucin996076806.
Full textBonnin, Edith. "Elucidating the Functional Role of Human Nucleoporin Nup88 in Health and Disease." Doctoral thesis, Universite Libre de Bruxelles, 2018. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/268017.
Full textDoctorat en Sciences
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Arroyo, Juan Pablo. "Exploring Potential Risk Factors of Fetal Origins of Diabetes| Maternal Stressors during Pregnancy and Birth Outcomes among Women in a Hospital in the Municipality of Caguas, Puerto Rico." Thesis, University of South Florida, 2013. http://pqdtopen.proquest.com/#viewpdf?dispub=1543402.
Full textPuerto Rico has the highest prevalence of type 2 diabetes, low birth-weight, and the second highest prevalence of preterm-birth in all the U.S. and its non-incorporated territories. These conditions are related. Birth-weight at both ends of the spectrum and preterm-birth are associated with an increased risk for developing type 2 diabetes and immune-inflammatory dysregulations. Maternal psychosocial stressors during pregnancy have also been recognized as potential risk factors for type 2 diabetes, and have been consistently associated with preterm-birth and low birth-weight across populations. Current evidence points toward epigenetic fetal metabolic-programming as the mechanism that underlies the increased risk for the previously mentioned morbidities. However, the particular psychosocial stressors that may contribute to the high prevalence of low birth-weight and preterm-birth in the population of Puerto Rico have not been well studied.
The present study assesses the relationships between particular psychosocial stressors, socioeconomic status, food insecurity, and birth outcomes. The results of this study show that low-risk pregnancy women were more likely to have babies with a higher ponderal index if they were exposed to stressors during gestation months 5, 6, and 7, or if exposed to "relationship stress" at any time during pregnancy. Women exposed to "financial difficulties" at any time during pregnancy were more likely to deliver babies at an earlier gestational age. Differences in birth outcomes between the exposed and non-exposed women were independent of maternal anthropometric measurements, maternal age at birth, number of previous births, and sex of the baby. Significant differences in birth outcomes were found between categories of father's self-identified and identified by others ethnicity, but sample size within categories was small. Although mothers with children at home had higher levels of food insecurity, and the level of food insecurity was correlated with higher levels of stress, no birth outcome measure was associated with food insecurity.
Some results are atypical in comparison with other populations, and therefore these findings may contribute to the understanding of population differences in the relationship between maternal stress during pregnancy and birth outcomes. The relatively small sample size and strict exclusion criteria of this study may limit the generalizability of the findings. Epidemiological similarities between Puerto Rico and other populations, and the possibility of a higher ponderal index increasing the risk for type 2 diabetes in the population of Puerto Rico need to be examined in future research.
Poulain, Marine. "Développement de la lignée germinale femelle humaine." Thesis, Paris 11, 2014. http://www.theses.fr/2014PA11T056/document.
Full textWoman fertility is partially dictated by the set up of the human female germ line. During the last ten years, which saw an increased number of couples consulting for assisted reproductive cares, the hypothesis of an early alteration in reproduction functions has emerged.In the fetal ovary, germ cells enter the path of oogenesis differentiation characterized by meiotic initiation. On this subject, vast majority of the scientific data are obtained from the mouse model, even if differences with human ovarian physiology are widely acknowledged. Therefore it is necessary to extend our knowledge on human ovarian development and identify its perturbations. The objective of my work was to assess a new model to study ovarian growth, studying regulation of meiotic entry and perturbation of germ line differentiation.We sat up a new xenograft model of early human fetal ovaries, when very early meiotic germ cells appear. Organ growth and germ cells differentiation were comparable with in vivo observations. Using this model with an RNA-interference strategy, we inhibited the expression of an oogonia germ cell gene, DMRTA2. This inhibition conducted to a significantly reduced number of germ cells gene that initiated meiosis and DMRTA2 seemed to be required for mitotic-meiotic transition. In another hand, we identified, in the ovary, the expression of germ cells markers described as specifically male in rodent (PLZF, DNMT3L, FGF9, NANOS2 ou CYP26B1). The expression of these markers in the human ovary could explain the observation of mitotic germ cells in late fetal ovaries (30 wpf).In parallel, we tested germ cells sensibility to a synthetic glucocorticoid, dexamethasone, administrated during pregnancy in some justified pathologies. We observed an increased expression of PLZF that could explain the decreased number of germ cells observed in treated ovaries.In conclusion, we identified a new gene expressed in human fetal ovaries, potentially involved in the meiotic entry, and we extended our knowledge to characterized human germ line development. However, many points have to be clarified, as the possible competence of late mitotic germ cells to form oocytes
Cavallin, Mara. "Physiopathologie moléculaire et cellulaire des anomalies du développement du cortex cérébral : le syndrome d'Aicardi WDR81 mutations cause extreme microcephaly and impair mitotic progression in human fibroblasts and Drosophila neural stem cells TLE1, a key player in neurogenesis, a new candidate gene for autosomal recessive postnatal microcephaly Mutations in TBR1 gene leads to cortical malformations and intellectual disability Aicardi syndrome: Exome, genome and RNA-sequencing of a large cohort of 19 patients failed to detect the genetic cause Recurrent RTTN mutation leading to severe microcephaly, polymicrogyria and growth restriction Recurrent KIF2A mutations are responsible for classic lissencephaly Recurrent KIF5C mutation leading to frontal pachygyria without microcephaly Rare ACTG1 variants in fetal microlissencephaly De novo TUBB2B mutation causes fetal akinesia deformation sequence with microlissencephaly: An unusual presentation of tubulinopathy A novel recurrent LIS1 splice site mutation in classic lissencephaly Further refinement of COL4A1 and COL4A2 related cortical malformations Prenatal and postnatal presentations of corpus callosum agenesis with polymicrogyria caused By EGP5 mutation Delineating FOXG1 syndrome from congenital microcephaly to hyperkinetic encephalopathy Delineating FOXG1 syndrome: From congenital microcephaly to hyperkinetic encephalopathy." Thesis, Sorbonne Paris Cité, 2019. https://wo.app.u-paris.fr/cgi-bin/WebObjects/TheseWeb.woa/wa/show?t=2213&f=18201.
Full textMalformations of cortical development (MCD) are a major cause of intellectual disability and drug-resistant epilepsy. Next Generation Sequencing (NGS) has considerably improved the identification of the molecular basis of non-syndromic MCD. However, certain forms, including complex MCD, remain unexplained. My PhD project aimed to improve the understanding of complex MCD using two disorders: Microlissencephaly (MLIS) and Aicardi Syndrome (AIC), the latter associating brain and eye malformations and only reported in girls. Trio Whole Exome Sequencing (WES) performed in 16 MLIS families allowed me to identify and functionally characterize a new MLIS gene, WDR81, in which mutations lead to cell cycle alteration. Moreover, using the same strategy, I was able to identify a pathogenic homozygous variant in TLE1 in a patient from consanguineous family with a postnatal microcephaly, suggestive of a FOXG1-like presentation. Interestingly, TLE1 is a major partner of FOXG1, a gene involved in maintaining the balance between progenitor proliferation and differentiation. In parallel, my work allowed me to redefine the phenotypic spectrum associated with RTTN, EPG5, COL4A1 and COL4A2, TBR1, KIF5C, KIF2A and FOXG1. The second part of my PhD program was aimed at identifying the genetic basis of AIC in an international cohort of 19 patients. After excluding a skewed X chromosome inactivation and the presence of chromosomal rearrangements, I performed WES in trios. The analysis of the data from WES did not allow me to identify any recurrent variants. I therefore tested a new approach combining Whole Genome Sequencing (WGS) and RNA-Sequencing (RNA-Seq) on fibroblast cells. I identified a number of deregulated transcripts implicated in brain and eye development. I compared the results of this analysis with the WGS analysis in order to find variants in these candidate genes. In conclusion, these studies have improved the knowledge of the molecular basis of complex MCD, such as TLE1 in postnatal microcephaly, and revealed the pathogenic mechanisms such as WDR81 in cell cycle progression and EPG5 in endosomes and autophagy. My work has also generated a collection of NGS data (WES, WGS and RNA-Seq) that will be shared in an international consortium to develop new analytical strategies, in particular for the non-coding DNA regions. This novel strategy provides opportunities to improve understanding of the cellular mechanisms involved in brain and eye development
Calarrão, Ana Isabel Alves Gil Barrera 1989. "Environmental chemicals and fetal testis development of human and rat." Master's thesis, 2012. http://hdl.handle.net/10451/7032.
Full textAo longo das últimas cinco décadas tem vindo a registar-se um decréscimo da taxa de fecundidade humana. Embora existam vários factores que contribuam para tal (por exemplo, alterações nos comportamentos sociais), são cada vez mais comuns problemas fisiológicos masculinos, como a diminuição da concentração e qualidade espermática, que devem ser também considerados importantes. Adicionalmente, doenças como hipospádias, criptorquidismo e cancro testicular têm também vindo a aumentar de incidência. Devido ao curto período de tempo em que se registam estas grandes diferenças, parece pouco provável que haja apenas influência genética a actuar. Como em tantos outros problemas recentes, a combinação das variações do estilo de vida e da exposição ambiental a moléculas perigosas pode prejudicar o desenvolvimento do sistema reprodutor masculino, diminuindo assim a fertilidade. Foi proposta uma origem comum na vida fetal para as doenças acima referidas, em que uma função deficiente das células somáticas do testículo fetal resultaria numa ruptura da organização e desenvolvimento normal, traduzindo-se em problemas reprodutivos masculinos. Esta teoria dá pelo nome de Síndrome da Disgénese Testicular (Testicular Dysgenesis Syndrome, TDS) e mostra o quão importante é o desenvolvimento fetal das estruturas reprodutivas masculinas para uma saúde e função reprodutiva normal posterior. Uma das doenças incluídas nesta TDS é o cancro testicular com origem na linha germinal (testicular germ cell cancer, TGCC), ao qual é atribuído a maioria das mortes relacionadas com problemas de saúde em homens entre os 15 e os 35 anos e cuja incidência duplicou nos últimos 40 anos. Durante o desenvolvimento fetal no Homem, as células germinais masculinas indiferenciadas mantêm a sua proliferação em níveis baixos após a perda de pluripotência e passagem para um estado diferenciado. Esta perda de pluripotência parece conter a hipótese de como a TGCC surge nos humanos, visto que se o processo for perturbado por factores externos, as células podem entrar num estado intermediário em que mantêm a taxa de proliferação alta, adquirindo características cancerígenas. Um tipo de molécula ambiental ao qual os fetos estão expostos, e que já demonstrou afectar negativamente o sistema reprodutor masculino noutras espécies, são os ftalatos, componentes industriais cuja exposição em roedores levou ao aparecimento de condições semelhantes às da TDS. No entanto, não há informações sobre o seu efeito nos humanos. Outros químicos ambientais cujo efeito no desenvolvimento reprodutivo pode ser negativo são os anti-inflamatórios não esteróides, tais como o paracetamol e a indometacina. Embora o mecanismo exacto ainda não seja conhecido, ambos actuam através da inibição da produção ou acção de prostaglandinas, que são lípidos libertados pelas células quando ocorre uma inflamação, embora a indometacina seja considerada um supressor mais inequívoco do que o paracetamol. A administração de paracetamol e outros analgésicos demostrou ter efeitos negativos na saúde reprodutiva em roedores. Esta tese centrou-se no estudo do efeito da exposição a químicos ambientais como o ftalato di(n-butyl) (di(n-butyl) phthalate, DBP), paracetamol e indometacina no desenvolvimento e diferenciação das células germinais fetais, baseando-se em estudos preliminares e não publicados desenvolvidos pelo grupo de investigação em que me inseri. Para estudar o efeito do DBP, foram recolhidos testículos de fetos humanos (n = 8), com idades compreendidas entre as 14 e 20 semanas de gestação, para serem xeno-enxertados debaixo da pele dorsal de ratos imunodeficientes adultos (machos castrados) em porções pequenas. Sete dias após esta operação, os quais permitem o estabelecimento de irrigação sanguínea nos enxertos, os hospedeiros recebem doses diárias de controlo, DBP ou MBP (ftalato monobutyl, o metabolito activo do DBP) dissolvidos em óleo (500mg/kg/dia), durante 21 dias. Os hospedeiros ainda são sujeitos a três injecções semanais de gonadotropina coriónica humana (20 IU), para replicar as condições normais de gravidez e manter a produção de testosterona. Após o período de tratamento, os enxertos são recolhidos, fixados, cortados em secções e sujeitos a uma imunofluorescência tripla. No total, analisaram-se pelo menos dois enxertos controlos e dois expostos a DBP/MBP para cada feto, resultando, no mínimo, em 32 amostras. O protocolo de imunofluorescência permite visualizar células germinais com antigénios para OCT3/4 (células germinais indiferenciadas), MAGE-A4 (células germinais diferenciadas) e Ki67 (células em proliferação). As imagens foram obtidas por microscopia confocal e as células incluídas num túbulo seminífero definido, que demonstrassem fluorescência para estes antigénios, foram assumidas como pertencendo à linha germinal e contadas manualmente. Os valores registados para os enxertos-controlo e expostos a DBP, do mesmo feto, foram condensados até se obter um valor médio respectivo. A média dos enxertos controlo e dos expostos ao tratamento, de cada um dos fetos, foi utilizada então para análise estatística, usando-se um Teste t para amostras emparelhadas (significância: P<0.05). Os resultados mostraram que os enxertos expostos ao DBP(MBP) registam uma diminuição significativa de mais de 10% na percentagem de células germinais positivas para OCT3/4 (indiferenciadas). Por outro lado, a percentagem de células germinais positivas para MAGE-A4 (diferenciada) em enxertos tratados com DBP(MBP) foi significativamente maior do que os controlos. Porém, a proliferação das células germinais indiferenciadas e diferenciadas não registou alterações significativas em comparação com as amostras controlo. As células germinais reduziram a expressão de OCT3/4 para aumentarem a expressão de MAGE-A4 quando se diferenciaram, o que coincidiu com a redução, e eventualmente a perda, da capacidade proliferativa, demonstrando que o modelo dos xeno-enxertos consegue recapitular o desenvolvimento fetal normal das células germinais masculinas nos humanos. Estes resultados parecem dever-se a uma apoptose selectiva das células indiferenciadas. Se a apoptose fosse geral, a proporção de células germinais indiferenciadas e diferenciadas não mudava após o tratamento com o DBP(MBP), o que não está de acordo com os resultados obtidos, já que há um aumento de células germinais diferenciadas. Devido à falta de dados sobre o efeito do DBP nas gónadas masculinas humanadas, esta hipótese estaria de acordo com dados anteriores de estudos in vitro e in vivo, em humanos e roedores respectivamente, em que a exposição a DBP reduz o número de células germinais fetais positivas para OCT3/4. O segundo projecto desenvolvido e abordado nesta tese foi analisar os efeitos da exposição ao paracetamol e indometacina no número de células germinais nos testículos fetais de ratos. Fêmeas prenhas de ratos Wistar foram tratadas diariamente com paracetamol (350mg/kg/dia), indometacina (1mg/kg/dia ou 0.8mg/kg/dia) ou tratamento controlo. O tratamento decorreu entre o dia embrionário (e) 15.5 até ao e20.5, sendo os testículos dos descendestes recolhidos ao e21.5. As amostras foram fixadas, seccionadas e processadas através de um protocolo imunohistoquímico para o antigénio VASA (marcador típico de células germinais). O número de células germinais foi obtido por estereologia, que é um método uniforme e sistemático que permite analisar a composição celular de um tecido tridimensional, neste caso o testículo, através da contagem de núcleos de células germinais em secções bidimensionais desse mesmo tecido. Para as amostras fetais e21.5, foram contadas todas as células que expressassem VASA e que estivessem contidas num túbulo seminífero definido. A comparação entre amostras controlo e as expostas a paracetamol/indometacina foi analisada estatisticamente através de uma análise de variância ANOVA (significância: p<0.05). Os resultados mostraram uma redução significativa do número de células germinais em amostras expostas à indometacina, quando comparadas com as de controlo. Embora as gónadas masculinas expostas ao paracetamol não tenham apresentado uma diferença significativa, existe uma tendência para a redução do número de células germinais. O desenvolvimento normal das células germinais no rato passa por perderem totalmente a capacidade proliferativa quando se diferenciam, num processo síncrono e uniforme, ao contrário dos humanos. Não se sabe se o decréscimo do número de células pode ser atribuído a apoptose devido à falta de dados nesta área, mas a indometacina pode induzir um aumento da taxa de diferenciação das células germinais fetais, levando à paragem da divisão celular numa idade anormalmente precoce. Outro ensaio experimental foi desenvolvido para investigar o efeito da exposição in utero à indometacina no número de células germinais em testículos de ratos obtidos no dia 25 após o nascimento (postnatal day, pnd25). O protocolo para investigar este efeito foi em tudo idêntico ao seguido para os testículos e21.5, excepto que não houve amostras expostas ao parecetamol e os testículos foram recolhidos após o nascimento (embora o período de exposição à indometacina tenha sido de e15.5 a e18.5). Visto que a gónada pnd25 já produz espermatozóides, as células germinais foram divididas em categorias que correspondem à progressão temporal da espermatogénese: espermatogónia, espermatócito I e espermatócito II. O número de células germinais de cada categoria, mais o número total de células germinais, foi contado também através de esterologia e a comparação estatística entre amostras controlo e tratadas com indometacina foi feita com um Teste t de Student (significância: p<0.05). Os resultados mostraram que o número de espermatogónias e o número total de células germinais aumentaram significativamente em amostras tratadas com indometacina, enquanto os valores para espermatócitos I e II não sofreram alterações significativas. Isto sugere que a exposição in utero à indometacina possa induzir apoptose nas células germinais e aquelas que resistem conseguem recuperar, embora faseadamente, os seus números normais após o nascimento e cessação do período de tratamento. Além disso, ambos os ensaios em ratos mostram que, pelo menos em mamíferos, o desenvolvimento fetal normal das células germinais, e posterior fertilidade dos indivíduos, é dependente de prostaglandinas. Este conjunto de estudos mostrou que a exposição fetal a químicos ambientais comuns, tais como os ftalatos e inibidores de prostaglandinas, pode afectar o desenvolvimento fetal das células germinais nos humanos e no rato, obtendo-se dados sobre este tema pela primeira vez.
Lifestyle and environmental exposure to hazardous molecules have been indicated as a cause for the decrease in human fertility. An increased incidence of hypospadias, cryptorchidism and testicular germ cell cancer has been reported and it has been hypothesized that these may form a Testicular Dysgenesis Syndrome (TDS) with a common origin in fetal life. Although testicular germ cell cancer (TGCC) occurs in young men, it appears that it arises from a failure of fetal germ cells to lose their pluripotency and which then transform into carcinoma-in-situ (CIS) cells, which are germ cells (GC) with pluripotency and proliferative features that probably lead to TGCC in young adulthood. It has been hypothesized that exposure in utero to some environmental factors could play a role in disturbing the testicular endocrine function in the fetus, possibly leading to TGCC. Di(n-butyl) phthalate (DBP), a common environmental chemical, has been shown to affect fetal testis development and endocrine function in the rat. Nonsteroidal anti-inflammatory drugs (paracetamol, indomethacin), which are believed to work through inhibition of prostaglandins, and other inflammation modulators can also affect fetal testis endocrine function in animal models. This study therefore aimed to assess the effect of DBP, paracetamol and indomethacin exposure on fetal GC development and differentiation. To study DBP effects, second-trimester human testis pieces from eight fetuses were xenografted under the backskin of nude castrated male mice, which were then treated with vehicle (control) or DBP for 21 days. Immunofluorescence for OCT3/4 (undifferentiated GC marker), MAGE-A4 (differentiated GC marker) and Ki67 (proliferative cell marker) was used to analyse GC differentiation and proliferation. Exposure to DBP led to a reduction in proportion of undifferentiated GC, although proliferative features were maintained, possibly due to selective apoptosis. To analyse the effect of indomethacin on GC numbers, e21.5 and pnd25 rat testes were evaluated in males exposed in utero to vehicle (control) or indomethacin (1mg/kg or 0.8mg/kg) treatment. Immunohistochemistry for VASA (GC marker) and stereology was used to assess GC number in e21.5 testes and spermatogenic temporal progression in pnd25 testes. Indomethacin exposed e21.5 rat testes showed a significant decrease in GC number, which is in agreement with preliminary results. Pnd25 rat testes from males exposed in utero to indomethacin showed an increase in spermatogonial numbers and total GC number, although the numbers for early and pachytene spermatocytes were not different. This shows that indomethacin affects germ cell development during the exposure period and that there might be a staggered recovery after birth toward normal GC number. Overall, these results show for the first time that exposure to common environmental chemicals affect human and rat male fetal GC development.
Jaleco, Ana Cristina Monteiro Primo. "Lymphoid development in human fetal liver." Doctoral thesis, 1998. http://hdl.handle.net/10316/10181.
Full textDisciplina afim: Imunologia Por volta da sétima semana de gestação, o timo humano é colonizado por progenitores hematopoiéticos provenientes do fígado fetal, os quais se diferenciam predominantemente em linfócitos T no seio do microambiente tímico. No Capítulo 2, é identificada e caracterizada uma população de células estromais que se encontram localizadas na região subcapsular do timo humano, e suportam a diferenciação de precursores tímicos em timócitos imaturos, levando a crer que possam ter um papel importante nos estadios mais precoces do desenvolvimento de células T. As células hematopoiéticas do fígado fetal que colonizam o timo, constituem precursores multipotentes com a capacidade de se diferenciarem em tipos celulares distinctos. No Capítulo 3, nós propusémo-nos investigar se o cometimento à linhagem T teria lugar no seio do fígado fetal, anteriormente à colonização do timo. Foi demonstrado que a capacidade de os precursores de fígado fetal se diferenciarem em linfócitos T, é exclusiva dos progenitores mais imaturos. Pelo contrário, tanto os precursores mais imaturos como os mais diferenciados possuem capacidade de diferenciação em células NK, o que implica o fígado fetal como um possível local de desenvolvimento de células NK. Apesar da medula óssea ser o orgão mais importante na formação de linfócitos B, uma série de observações sugere que a diferenciação B possa ter lugar no fígado fetal. No Capítulo 4, encontram-se descritos dois modelos experimentais de suporte estromal que induzem e suportam a diferenciação de progenitores de fígado fetal em células B, através da aquisição progressiva de marcadores associados a esta linhagem. A via de diferenciação de células B, tal como outros processos biológicos, é muito provavelmente regulada por um conjunto de factores de transcrição. Neste contexto, as proteínas E12 and E47 foram recentemente identificadas como factores cruciais para a diferenciação B no ratinho, e são reguladas negativamente por proteínas Id. No Capítulo 5, é demonstrado que a expressão forçada da proteína Id3 em precursores de fígado fetal através de transferência genética mediada por retrovírus, inibe marcadamente o desenvolvimento de linfócitos B a partir de células estaminais de fígado fetal quando estas são cultivadas nos modelos estromais descritos no Capítulo 4.
Lebel, Catherine. "Diffusion tensor imaging of human brain development." Phd thesis, 2010. http://hdl.handle.net/10048/1259.
Full text"Regulation of human oviductin mRNA expression." 2002. http://library.cuhk.edu.hk/record=b6073443.
Full text"May 2002."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (p. 149-171).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.
Jen, Kuan-Hua, and 任冠樺. "Comparing Fetal Heart Development between Human and Mouse based on Time-Series Gene Expression Profiles." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/01903043658180627167.
Full text國立交通大學
生物資訊研究所
95
Heart development is a complex process involving many genes which control cell behavior in the embryo and determine its pattern, its form, and much of its behavior. Microarray experiments can generate an enormous amount of data at one time, so we use this technology to obtain gene expression profiles in heart embryonic development. But it is usually very difficult to obtain human heart fetus sample because of the issues of ethical, legal, and social consideration. In order to help us get more understanding of human heart development, we can use the mouse model system that is most often used. Therefore, we must establish a mapping system to make a cross bridge between these two species on developmental stages. To date, the vast majority of researches have focused their study on one species. Specially, we utilize orthologous genes and incorporate the dynamic time warping algorithm in order to map the time points that human and mouse gene expression profiles having highly correlated pattern. Firstly, we apply the algorithm to select the best time-warped orthologous genes having similar pattern. Then, these genes are clustered into groups. Each group has its unique mapping pattern and different biological meaning. The following task is to find relationship and pattern in distinct groups of genes, and to get close understanding into molecular process and gene function, mechanisms of embryogenesis of the heart, and comparative genomics. Ultimately, our aim is to achieve new insights into the heart developmental biology.
"Testicular angiogenesis in rats: developmental changes and hormonal stimulation by human chorionic gonadotrophin." 1998. http://library.cuhk.edu.hk/record=b5889785.
Full textThesis (M.Phil.)--Chinese University of Hong Kong, 1998.
Includes bibliographical references (leaves 92-106).
Abstract also in Chinese.
ABSTRACT --- p.i
摘要 --- p.iii
ACKNOWLEDGMENT --- p.v
Chapter 1. --- Introduction
Chapter 1.1 --- Angiogenesis in general --- p.1
Chapter 1.1.1 --- The concept of angiogenesis --- p.1
Chapter 1.1.2 --- The process of angiogenesis --- p.1
Chapter 1.2 --- Measurement of angiogenesis --- p.3
Chapter 1.2.1 --- In vivo assays --- p.3
Chapter 1.2.2 --- In vitro assays --- p.5
Chapter 1.3 --- Angiogenic factors --- p.6
Chapter 1.4 --- Angiogenesis in the female reproductive system --- p.7
Chapter 1.5 --- Evidence of hormonally-regulated angiogenesis in endocrine tissues --- p.10
Chapter 1.5.1 --- Ovary --- p.10
Chapter 1.5.2 --- Thyroid --- p.11
Chapter 1.6 --- Angiogenesis in the testis --- p.12
Chapter 1.6.1 --- Structure of testicular vasculature --- p.12
Chapter 1.6.2 --- Angiogenic factors in the testis --- p.13
Chapter 1.6.3 --- Vascular effects of hCG/LH in the testis --- p.17
Chapter 1.6.4 --- Postnatal development of testicular vasculature --- p.17
Chapter 1.7 --- Aims of the present study --- p.19
Chapter 2. --- Materials and methods
Chapter 2.1 --- Animals --- p.20
Chapter 2.2 --- Experimental design --- p.20
Chapter 2.2.1 --- Testicular angiogenesis in adult rats - hormonal stimulation by hCG --- p.20
Chapter 2.2.1.1 --- Changes with time after hCG treatment --- p.20
Chapter 2.2.1.2 --- Effect of Leydig cell depletion --- p.22
Chapter 2.2.1.3 --- Effect of Leydig cell suppression by subcutaneous testosterone-filled silastic implants --- p.22
Chapter 2.2.1.4 --- Effect of testicular macrophage activation --- p.24
Chapter 2.2.1.5 --- Effect of testicular macrophage depletion --- p.26
Chapter 2.2.2 --- Developmental changes in testicular angiogenesis --- p.29
Chapter 2.3 --- Perfusion of testes with fixative or Indian Ink --- p.29
Chapter 2.4 --- Processing of the testes for histological sections --- p.30
Chapter 2.5 --- Immunohistochemical staining for proliferating cell nuclear antigen (PCNA) --- p.31
Chapter 2.6 --- Immunohistochemical staining for vascular endothelial growth factor --- p.32
Chapter 2.7 --- Quantification of PCNA-positive endothelial cells --- p.33
Chapter 2.8 --- Quantification of blood vessel density --- p.34
Chapter 2.9 --- Estimation of intertubular area in testis section --- p.35
Chapter 2.10 --- Preparation of liposome-entrapped dichloromethylene diphosphonate (Cl2MDP-lp) --- p.38
Chapter 2.11 --- Radioimmunoassay of serum tsetosterone --- p.38
Chapter 2.12 --- Statistical analyses --- p.40
Chapter 3. --- Results
Chapter 3.1 --- hCG-induced increase in endothelial cell proliferation in adult rat testes --- p.41
Chapter 3.1.1 --- Testicular histology --- p.41
Chapter 3.1.2 --- Changes in the number of PCNA-positive endothelial cells --- p.41
Chapter 3.1.3 --- Changes in blood vessel density --- p.44
Chapter 3.1.4 --- Changes in testis weight and serum testosterone concentration --- p.44
Chapter 3.2 --- Effect of Leydig cell depletion by ethane dimethane sulphonate (EDS) on hCG-induced endothelial cell proliferation in adult rat testes --- p.48
Chapter 3.2.1 --- Testicular histology --- p.48
Chapter 3.2.2 --- Changes in the number of PCNA-positive endothelial cells --- p.48
Chapter 3.2.3 --- Changes in serum testosterone concentration and testis weight --- p.52
Chapter 3.3 --- Effect ofLeydig cell suppression by testosterone-filled subcutaneous silastic implants on hCG-induced endothelial cell proliferation in adult rat testes --- p.54
Chapter 3.3.1 --- "Changes in serum testosterone concentration, testis weight, and testicular intertubular area" --- p.54
Chapter 3.3.2 --- Changes in the number of PCNA-positive endothelial cells --- p.58
Chapter 3.3.3 --- Changes in the level of vascular endothelial growth factor (VEGF) immunoreactivity in the testis --- p.60
Chapter 3.4 --- Effect of testicular macrophage activation by polystyrene latex beads on hCG-induced endothelial cell proliferation in adult rat testes --- p.60
Chapter 3.4.1 --- Testicular histology --- p.60
Chapter 3.4.2 --- Changes in the number of PCNA-positive endothelial cells --- p.63
Chapter 3.4.3 --- Changes in testis weight and serum testosterone concentration --- p.65
Chapter 3.5 --- Effect of testicular macrophage depletion by liposome-entrapped C12MDP treatment on hCG-induced endothelial cell proliferation in adult rat testes --- p.67
Chapter 3.5.1 --- Testicular histology --- p.68
Chapter 3.5.2 --- Changes in the number of PCNA-positive endothelial cells --- p.68
Chapter 3.5.3 --- Changes in testis weight and serum testosterone --- p.72
Chapter 3.6 --- Endothelial cell proliferation in rat testes during postnatal development --- p.74
Chapter 3.6.1 --- Changes in the number of PCNA-positive endothelial cells --- p.74
Chapter 3.6.2 --- Changes in blood vessel density --- p.74
Chapter 3.6.3 --- Changes in testis weight and intertubular area of the testes --- p.77
Chapter 4. --- Discussion
Chapter 4.1 --- hCG-induced endothelial cell proliferation and changes in blood vessel density --- p.79
Chapter 4.2 --- Role of Leydig cells in hCG-induced endothelial cell proliferation in adult rat testes --- p.82
Chapter 4.3 --- Role of testicular macrophages in hCG-induced endothelial cell proliferation in adult rat testes --- p.86
Chapter 4.4 --- Testicular angiogenesis during postnatal development --- p.88
Chapter 5. --- References --- p.92
"Studies on some factors critical for the development of pancreatic progenitor cells derived from human fetal pancreas." 2011. http://library.cuhk.edu.hk/record=b5896938.
Full textThesis (M.Phil.)--Chinese University of Hong Kong, 2011.
Includes bibliographical references (leaves 179-204).
Abstracts in English and Chinese.
Abstract --- p.I
摘要 --- p.IV
Publications --- p.VII
Acknowledgements --- p.VIII
Table of contents --- p.IX
List of figures --- p.XV
List of tables --- p.XVII
List of abbreviations --- p.XVIII
Chapter Chapter 1 --- General Introduction
Chapter 1.1 --- The Pancreas --- p.2
Chapter 1.1.1 --- Anatomy of Pancreas --- p.2
Chapter 1.1.2 --- The Exocrine Pancreas --- p.4
Chapter 1.1.3 --- The Endocrine Pancreas --- p.5
Chapter 1.1.3.1 --- Structure of Islets --- p.5
Chapter 1.1.3.2 --- "Functions of α-, β-, y-, ð-, Σ-and PP-cells in Islets" --- p.7
Chapter 1.1.4 --- Overview of Pancreas Development --- p.9
Chapter 1.1.4.1 --- Organ Morphology --- p.10
Chapter 1.1.4.2 --- Cyto-differentiation --- p.12
Chapter 1.1.4.3 --- Control by Transcriptional Factors --- p.14
Chapter 1.1.5 --- Postnatal Pancreas Development and Regeneration --- p.18
Chapter 1.1.5.1 --- Proliferation of Pre-existing β-cells --- p.19
Chapter 1.1.5.2 --- Neogenesis from Precursor Cells --- p.20
Chapter 1.1.5.3 --- Transdifferentiation of other Cells --- p.20
Chapter 1.2 --- Diabetes Mellitus --- p.22
Chapter 1.2.1 --- Pathophysiology of Diabetes Mellitus and Current Treatments --- p.24
Chapter 1.2.1.1 --- Type I Diabetes Mellitus --- p.24
Chapter 1.2.1.2 --- Type II Diabetes Mellitus --- p.25
Chapter 1.2.1.3 --- Gestational Diabetes --- p.27
Chapter 1.2.1.4 --- Secondary Diabetes --- p.28
Chapter 1.3 --- Stem Cell therapy --- p.29
Chapter 1.3.1 --- Stem Cell --- p.29
Chapter 1.3.1.1 --- Mesenchymal Stem Sell --- p.31
Chapter 1.3.1.2 --- Embryonic Stem Cell --- p.35
Chapter 1.3.1.3 --- Induced Pluripotent Stem Cell --- p.36
Chapter 1.3.2 --- Islets Engineering --- p.37
Chapter 1.3.2.1 --- Genetic Modification --- p.37
Chapter 1.3.2.2 --- Directed Differentiation --- p.38
Chapter 1.3.2.3 --- Microenvironment --- p.38
Chapter 1.3.2.4 --- In vivo Regeneration --- p.39
Chapter 1.3.2.5 --- Cell Fusions --- p.40
Chapter 1.3.2.6 --- Combinatory Treatments --- p.40
Chapter 1.4 --- The Vitamin A & Vitamin D System --- p.42
Chapter 1.4.1 --- The Vitamin A --- p.42
Chapter 1.4.2 --- Vitamin A Metabolism --- p.44
Chapter 1.4.3 --- Roles of vitamin A in Pancreatic Development --- p.46
Chapter 1.4.4 --- The Vitamin D --- p.48
Chapter 1.4.5 --- Vitamin D Metabolism --- p.49
Chapter 1.4.6 --- Metabolic Functions of Vitamin D in Islets --- p.51
Chapter 1.4.7 --- Cod Liver Oil --- p.53
Chapter 1.4.8 --- Interactions between Vitamin A and Vitamin D --- p.53
Chapter 1.5 --- The Relations of Liver and Pancreas Development --- p.55
Chapter 1.5.1 --- Endoderm Induction for Hepatic and Pancreatic Differentiation of ESCs --- p.55
Chapter 1.5.2 --- Bipotential Precursor Population within Embryonic Endoderm --- p.56
Chapter 1.5.3 --- Pancreatic Islets Promote Mature Liver Hepatocytes Proliferation --- p.57
Chapter 1.5.4 --- Transdifferentiation --- p.57
Chapter 1.5.5 --- Transplantation in Liver Niche Promotes Maturation of Insulin-Producing Cells --- p.60
Chapter 1.5.6 --- Neuronal Relay from the Liver to Pancreatic --- p.61
Chapter 1.5.7 --- Development of Islets in the Nile Tilapia --- p.62
Chapter 1.6 --- The Insulin-like Growth Factor-I (IGF1) --- p.64
Chapter 1.6.1 --- IGF1 System --- p.64
Chapter 1.6.2 --- IGF 1 Regulation --- p.65
Chapter 1.6.3 --- Roles of IGF 1 in Pancreatic Development and Regeneration --- p.68
Chapter 1.7 --- Aims and Objectives of Study --- p.70
Chapter Chapter 2 --- General Materials and Methods
Chapter 2.1 --- Pancreatic progenitor cells (PPCs) and liver stromal cells (LSCs) isolation and cell culture --- p.72
Chapter 2.1.1 --- Tissue procurement --- p.72
Chapter 2.1.2 --- PPC and LSC culture --- p.72
Chapter 2.1.3 --- "Treatments of vitamin A, vitamin D and IGF 1" --- p.76
Chapter 2.1.4 --- "Cell culture of Caco-2, HepG2 and DU-145" --- p.76
Chapter 2.2 --- Induction of Islet-like Cell Clusters (ICCs) Differentiation --- p.77
Chapter 2.2.1 --- In vitro Directed Differentiation --- p.77
Chapter 2.2.2 --- In vitro LSC Microenvironment --- p.77
Chapter 2.3 --- RNA Expression Detection --- p.79
Chapter 2.3.1 --- RNA isolation --- p.79
Chapter 2.3.2 --- Reverse Transcription --- p.79
Chapter 2.3.3 --- Polymerase Chain Reaction (PCR) --- p.80
Chapter 2.3.4 --- Realtime PCR --- p.81
Chapter 2.4 --- Immunocytochemistry --- p.83
Chapter 2.5 --- Western Blotting --- p.85
Chapter 2.5.1 --- Protein extraction and quantification --- p.85
Chapter 2.5.2 --- Western Blotting --- p.85
Chapter 2.6 --- Enzyme-linked Immunosorbent Assay (ELISA) --- p.87
Chapter 2.6.1 --- Detection of cell viability --- p.87
Chapter 2.6.2 --- Detection of cell proliferation --- p.87
Chapter 2.6.3 --- Measurement of Cell death --- p.88
Chapter 2.6.4 --- Measurement of IGF 1 level in condition medium --- p.89
Chapter 2.6.5 --- Measurement of glucose induced insulin secretion --- p.90
Chapter 2.7 --- Regeneration model --- p.92
Chapter 2.7.1 --- Regeneration model in neonatal-STZ rat --- p.92
Chapter 2.7.2 --- Change in IGF1 expression in pancreas and liver --- p.92
Chapter 2.8 --- Statistical Data Analysis --- p.93
Chapter Chapter 3 --- Vitamin D and vitamin A receptor expression and the proliferative effects of ligand activation of these receptors on the development of pancreatic progenitor cells derived from human fetal pancreas. (Stem Cell Rev. 2011;7:53-63)
Chapter 3.1 --- Abstract --- p.95
Chapter 3.2 --- Introduction --- p.97
Chapter 3.3 --- Materials and Methods --- p.101
Chapter 3.3.1 --- Fetal Tissue Procurement --- p.101
Chapter 3.3.2 --- Culture of Pancreatic Progenitor Cells --- p.101
Chapter 3.3.3 --- RNA Expression Analysis by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.102
Chapter 3.3.4 --- Western Blot Analysis --- p.103
Chapter 3.3.5 --- Immunocytochemstry --- p.105
Chapter 3.3.6 --- PPC Proliferation Assays --- p.106
Chapter 3.3.7 --- PPC Cell Death Assays --- p.107
Chapter 3.3.8 --- Statistical Data Analysis --- p.108
Chapter 3.4 --- Results --- p.110
Chapter 3.4.1 --- "Expression and Localization of RAR, VDR and RXR, CYP26 and CYP24 in PPCs" --- p.110
Chapter 3.4.2 --- Incubation of PPC with atRA Enhances PPC Viability due to Increased Proliferation and Anti-apoptosis --- p.111
Chapter 3.4.3 --- Incubation of PPCs with Calcitriol Enhances Viability due to Increased Proliferation --- p.111
Chapter 3.4.4 --- Both atRA and Calcitriol Induce Up-regulation of both the RAR and the VDR but not the RXR --- p.112
Chapter 3.4.5 --- Combination Treatment with atRA and Calcitriol on Cell Viability and NGN3 Expression --- p.112
Chapter 3.5 --- Discussion --- p.114
Chapter Chapter 4 --- Human fetal liver stromal cell co-culture enhances the growth and differentiation of pancreatic progenitor cells into islet-like cell clusters (In submission to Gastroenterology)
Chapter 4.1 --- Abstract --- p.128
Chapter 4.2 --- Introduction --- p.129
Chapter 4.3 --- Materials and Methods --- p.133
Chapter 4.3.1 --- Use of human and animal tissues --- p.133
Chapter 4.3.2 --- "Cell preparation, characterizations and Differentiation" --- p.133
Chapter 4.3.3 --- Examination of PPC growth and ICC differentiation and functions with LSC co-culture --- p.133
Chapter 4.3.3 --- Identification of growth factors and investigation of their effects --- p.134
Chapter 4.3.4 --- Statistical Analysis --- p.135
Chapter 4.4 --- Results --- p.136
Chapter 4.4.1 --- "Isolation, Culture and Characterizations of LSCs" --- p.136
Chapter 4.4.2 --- Establishment of LSC co-culture system --- p.136
Chapter 4.4.3 --- LSC co-culture enhances PPC-derived ICC differentiation --- p.137
Chapter 4.4.4 --- Differential expression of mRNA for cytokines and growth factors between 1st and 2nd trimester LSCs --- p.138
Chapter 4.4.5 --- Characterization of IGF 1 receptors in PPCs and the effects of exogenous IGF1 on PPC growth and ICC differentiation --- p.139
Chapter 4.4.6 --- Neutralizing antibodies against IGF1R inhibit ICC differentiation --- p.140
Chapter 4.5 --- Discussion --- p.142
Chapter 4.6 --- Supplementary Materials and Methods --- p.147
Chapter 4.6.1 --- Cell Preparation and culture --- p.147
Chapter 4.6.2 --- In Vitro ICC differentiation --- p.148
Chapter 4.6.3 --- RNA expression analysis --- p.149
Chapter 4.6.4 --- Immunocytochemistry --- p.149
Chapter 4.6.5 --- PPC viability and cell count assays --- p.150
Chapter 4.6.6 --- IGF1 and insulin ELISA --- p.151
Chapter 4.6.7 --- Western blotting analysis --- p.152
Chapter 4.6.8 --- Neonatal streptozotocin regeneration model --- p.153
Chapter Chapter 5 --- General Discussion and Future Studies
Chapter 5.1 --- General Discussion --- p.165
Chapter 5.1.1 --- Proliferative effects and enhance expression of NGN3 by vitamin A and vitamin D on PPC --- p.166
Chapter 5.1.2 --- Induction of PPC derived ICCs by LSCs --- p.169
Chapter 5.1.3 --- Potential effects of liver stroma derived IGF1 on PPC derived ICCs differentiation --- p.172
Chapter 5.1.4 --- Significance of islet engineering in the management of diabetes --- p.174
Chapter 5.1.5 --- Conclusions --- p.176
Chapter 5.2 --- Future Studies --- p.177
Chapter Chapter 6 --- Reference
Reference --- p.180
Andrä, Paul. "Analysis and functional characterization in embryonic mouse neocortex of a set of human-specific genes expressed in neural progenitor cells of fetal human neocortex." 2020. https://tud.qucosa.de/id/qucosa%3A73384.
Full textIntroduction: The higher cognitive functions that characterize modern humans can be attributed to the cerebral neocortex and its remarkable expansion in size during the last 5 – 7 million years of human evolution. The identification of the underlying genomic changes will be not only of importance to better understand the unique complexity of the human brain, but also its susceptibility to neurological and psychiatric diseases. Recently, 15 human-specific genes preferentially expressed in neural progenitor cells (NPCs) of the human fetal neocortex were identified (Florio et al., 2018). Three of them, FAM72B, C and D belong to the Family of sequence similarity 72 (FAM72) and occurred in the human genome by gene duplication 3.4 – 1 mya. Aims & Approaches: Specifically, it was asked whether FAM72D plays a diverse role compared to the ancestral FAM72A (Results II, III, IV) due to the specific sets of amino acid substitutions it acquired (Results I). Effects of FAM72A and FAM72D on the proliferative capacity and gene expressions of embryonic mouse NPCs were analyzed upon ectopic expression either of FAM72A or FAM72D during embryonic mouse neocortical development. Methods: In utero electroporation (IUE) of embryonic mouse brains was performed to drive the expression of a red or green fluorescent protein (RFP or GFP) either plus empty DNA vector (pCAGGS; control), pCAGGS-FAM72A or pCAGGS-FAM72D plasmids in the dorsolateral neocortex at mid-neurogenesis (embryonic day 13.5, E13.5; Results II) or in the medial neocortex at late-neurogenesis (E15.5; Results III). NPC proliferation was evaluated by immunofluorescence of Ki67 (immunohistochemistry, IHC), a cell proliferation marker, and phosphorylated Histone H3 (PH3), a marker of cell mitosis. Moreover, the abundance of NPCs using immunofluorescence of basal intermediate progenitor (Tbr2) and apical and basal radial glia (Sox2) markers, and the gliogenesis by Olig2 immunofluorescence was analyzed. Additional experiments were carried out to study the capacity of NPCs to reenter the cell cycle upon IUE of FAM72D. To this end, pregnant mice were intraperitoneally injected with the thymidine analog 5-Ethynyl-2´-deoxyuridine (EdU) 24 h post-IUE, to label all cells undergoing S-phase of the cell cycle (i.e., all cells that reentered the cell cycle after IUE) in the developing mouse brains. Embryonic brains were collected 24 h after EdU injection and co-stained with Ki67. Ki67 and EdU double positive cells were considered as cells that reentered the cell cycle. To execute the transcriptome analysis E13.5 mice were electroporated with pCAGGS-GFP either plus an empty DNA vector (pCAGGS, control), a vector driving expression of FAM72A (pCAGGS-FAM72A) or FAM72D (pCAGGS-FAM72D). Subsequently, the electroporated dorsolateral neocortical areas were microdissected at E14.5 and dissociated into single cells. The electroporated (GFP+) cells were isolated from the single cell suspensions by the fluorescence-activated cell sorting (FACS). The isolated cells were processed for RNA sequencing. Data analysis was performed as previously reported (Florio et al., 2015). Results: By immunohistochemistry, no significant changes in any of the proliferative parameters or in the abundance of progenitors in the ventricular zone (VZ) and subventricular zone (SVZ) of the developing mouse neocortex upon ectopic expression of FAM72D compared to FAM72A and control samples were detected (Results II, III). However, the transcriptome analysis (Results IV) showed 88 significantly up- and 52 down-regulated genes upon FAM72A and 91 significantly up- and 67 downregulated genes upon FAM72D expression compared to the control. Only two of these differentially expressed genes were found to be upregulated upon FAM72A and FAM72D with an expression >1 fpkm: Syde1 and Shisa5. Besides, six genes specifically upregulated upon ectopic expression of FAM72D exhibiting fpkm > 1 were identified and characterized using the existing literature: Tapbp, Mtfp1, Slitrk5, Parp9, Cnp, Rbm43. Beyond that, gene ontology analysis showed significant enrichment of angiogenesis-related genes (e.g., Vegfc) upon FAM72A expression. Interestingly, there were more genes found to be enriched in NPCs that were upregulated compared to control upon FAM72D than FAM72A expression, but more NPC enriched genes downregulated upon FAM72A compared to FAM72D expression. In the case of differentially expressed neuron-enriched genes, the data was were inverse, which slightly supports the idea that FAM72D rather than FAM72A could positively affect the maintenance of NPC characteristics. Conclusions: In a previous study knockdown of Fam72a in adult mouse NPCs increased neurogenesis (Benayoun et al., 2014). This suggests, in conjunction with the present results, that FAM72A and FAM72D are not sufficient, but may be required, to promote NPC maintenance (Results II, III). This is why the gain of function experiments conducted in this study should be complemented by a loss of function approach in the developing mouse neocortex, in chimpanzee or human-derived brain organoids. Because of their expression in the NPCs of the developing human neocortex, it might be productive to analyze the potential synergistic effect on NPC proliferation of the FAM72s and the 12 other human-specific genes such as ARHGAP11B. Among other mechanisms discussed based on the gene expression analysis in this thesis (Results IV and Discussion), the upregulation of Slitrk5 upon ectopic expression of the human-specific FAM72D could be particularly remarkable. Slitrk5 is known to be involved in the recycling of the TrKB receptor (Song et al., 2015), which affects fundamental aspects of brain development. While FAM72A was found to inhibit the TrKB receptor (Nehar et al., 2009), the occurrence of FAM72D could indirectly rescue the TrKB receptor function via Slitrk5 and thereby prolonging or enhancing essential features such as precursor cell survival and neurogenesis in humans. Therefore, this study provides the first functional characterization of the evolutionary highly interesting region in the human genome comprising the FAM72 genes during embryonic neocortical development in vivo and offers numerous starting points for further investigations, that will collectively facilitate a comprehensive understanding of the genomic adaptations underlying the astonishing evolution of the human brain.:1 INTRODUCTION 11 1.1 WHAT MADE US HUMAN? 11 1.2 THE NEOCORTEX 12 1.2.1 Origin and structure 12 1.2.2 Neurogenesis in the developing neocortex 14 1.2.3 How to increase the neuronal output 18 1.3 EVOLUTION AND GENE DUPLICATION 19 1.3.1 Gene duplication and evolutionary novelty 19 1.3.2 Mechanisms of replication 21 1.3.3 Fates of duplicated genes 22 1.3.4 Which genes tend to duplicate? 24 1.3.5 Human adaptation and gene duplication 24 1.4 HUMAN-SPECIFIC SIGNATURES OF NEOCORTICAL EXPANSION 25 1.5 IDENTIFICATION OF HUMAN-SPECIFIC GENES EXPRESSED IN THE DEVELOPING NEOCORTEX.. 25 1.6 FAMILY WITH SEQUENCE SIMILARITY 72 (FAM72) 26 1.6.1 Evolutionary origin 26 1.6.2 Subcellular localization 27 1.6.3 Cell cycle regulation 28 1.6.4 NPC maintenance 28 2 AIMS & APPROACHES 30 3 RESULTS I 31 3.1 FROM GENES TO PROTEINS: 1 FAMILY – 4 PARALOGUES 31 3.2 FAM72 MRNA EXPRESSION LEVELS IN THE DEVELOPING MOUSE AND HUMAN NEOCORTEX 32 3.3 COMPUTATIONAL ANALYSES 34 3.3.1 Proportion of cysteines 34 3.3.2 Transmembrane domain 34 3.4 AMPLIFICATION, SUBCLONING AND MUTAGENESIS 36 3.4.1 Amplification from human cDNA 36 3.4.2 Verification of the pCAGGs vectors 36 4 RESULTS II 38 4.1 ECTOPIC EXPRESSION OF FAM72A AND FAM72D IN THE MOUSE DORSOLATERAL NEOCORTEX AT MID-NEUROGENESIS 38 4.2 NPC PROLIFERATION 39 4.2.1 Assessment of NPC proliferation using Ki67 immunofluorescence 39 4.2.2 Cell cycle reentry 41 4.2.3 Assessment of mitosis using PH3 immunofluorescence 43 4.2.4 Conclusion 45 4.3 NPC ABUNDANCE 46 4.3.1 Assessment of NPC abundance using Tbr2 and Sox2 immunofluorescence 46 4.3.2 Conclusion 49 5 RESULTS III 50 5.1 ECTOPIC EXPRESSION OF FAM72A and FAM72D IN THE MOUSE MEDIAL CORTEX AT LATE-NEUROGENESIS 50 5.2 NPC PROLIFERATION 51 5.2.1 Assessment of the NPC proliferation using Ki67 immunofluorescence 51 5.3 NPC ABUNDANCE 52 5.3.1 Assessment of NPC abundance using Tbr2 and Sox2 immunofluorescence 52 5.4 GLIOGENESIS 53 5.4.1 Assessment of gliogenesis using Olig2 immunofluorescence 53 5.5 CONCLUSION 54 6 RESULTS IV 55 6.1 DIFFERENCES IN GENE EXPRESSION UPON ANCESTRAL FAM72A AND HUMAN-SPECIFIC FAM72D EXPRESSION AT MID-NEUROGENESIS 55 6.1.1 Rationale and experimental setup 55 6.2 DIFFERENTIALLY EXPRESSED GENES UPON ECTOPIC FAM72A AND FAM72D EXPRESSION IN THE DEVELOPING MOUSE DORSOLATERAL NEOCORTEX 56 6.3 UPREGULATED GENES UPON THE ECTOPIC FAM72A OR FAM72D EXPRESSION 57 6.3.1 Upregulated genes upon the ectopic FAM72A and FAM72D expression 57 6.3.2 Upregulated genes upon the ectopic FAM72A or D expression – cut off: fpkm >1 58 6.3.3 Upregulated genes upon the ectopic FAM72A and D expression – cut off: fpkm >1 59 6.4 UPREGULATED GENES SPECIFICALLY UPON THE ECTOPIC FAM72D EXPRESSION – CUT OFF: FPKM >1 60 6.4.1 Tapbp (TAP binding protein, Tapasin) 60 6.4.2 Mtfp1 (mitochondrial fission protein 1, Mtp18) 61 6.4.3 Slitrk5 (Slit and Ntrk-like protein 5) 61 6.4.4 Parp9 (Poly(ADP-ribose) polymerase 9) 63 6.4.5 Cnp (2',3'-Cyclic-nucleotide 3'-phosphodiesterase) 63 6.4.6 Rbm43 (RNA binding motif protein 43) 65 6.5 DOWNREGULATED GENES UPON THE ECTOPIC FAM72A OR FAM72D EXPRESSION 66 6.5.1 Downregulated genes upon the ectopic FAM72A or D expression – cut off: fpkm >1 66 6.5.2 Downregulated genes upon ectopic FAM72A expression – cut off: fpkm >1 66 6.5.3 Downregulated genes upon ectopic FAM72D expression – cut off: fpkm >1 67 6.6 GENES PREVIOUSLY SHOWN TO BE DIFFERENTIALLY EXPRESSED UPON FORCED FAM72A EXPRESSION 69 6.6.1 Cell cycle regulators 69 6.6.2 Tumor suppressor genes 69 6.6.3 PROTEINS PREVIOUSLY OBSERVED TO INTERACT WITH FAM72A 70 6.7 EFFECT OF ECTOPIC FAM72A AND FAM72D EXPRESSION ON GENES IMPLICATED IN NEURAL LINEAGE FATE DECISION 70 6.7.1 Upregulated and NPC-enriched genes 71 6.7.2 Downregulated and NPC-enriched genes 73 6.7.3 Upregulated and neuron-enriched genes 74 6.7.4 Downregulated and neuron-enriched genes 75 6.8 GO ENRICHMENT ANALYSIS 75 6.9 CONCLUSION 75 7 DISCUSSION 78 7.1 WHAT MAKES US HUMAN? 78 7.2 IN UTERO ELECTROPORATION OF A HUMAN-SPECIFIC GENE IN THE DEVELOPING MOUSE NEOCORTEX 81 7.2.1 Opportunities and limitations of the approach 81 7.3 THE FAMILY OF SEQUENCE SIMILARITY 72 AND HUMAN UNIQUENESS 83 7.3.1 Cell cycle regulation and NPC maintenance 83 7.3.2 Cell death 84 7.3.3 Neurogenic period 85 7.3.4 TrkB signaling 85 7.3.5 Mitochondria 86 7.3.6 Angiogenesis 88 7.3.7 An evolutionary immunological adaptation in the brain? 89 7.3.8 FAM72 and SRGAP2 90 7.3.9 FAM72, Neanderthals, and lncRNAs 91 7.4 FUTURE DIRECTIONS 92 7.4.1 Loss of function 92 7.4.2 Gain of function 92 8 SUMMARY / ZUSAMMENFASSUNG 95 8.1 SUMMARY 95 8.2 ZUSAMMENFASSUNG 98 9 MATERIALS AND METHODS 101 9.1 CHART OF ALL EXPERIMENTS 101 9.2 COMPUTATIONAL ANALYSIS 101 9.2.1 Reference sequences and multiple sequence alignments 101 9.2.2 Transmembrane domain prediction 102 9.3 AMPLIFICATION, SUBCLONING, MUTAGENESIS 102 9.3.1 Amplification from human brain cDNA 102 9.3.2 Subcloning 103 9.2.3 Mutagenesis 103 9.4 PLASMID VERIFICATION 104 9.4.1 Transfection of Cos7 cells 104 9.4.2 Immunoblots 104 9.4.3 In situ hybridization (ISH) 105 9.5 MICE 105 9.6 IN UTERO ELECTROPORATION 105 9.7 FIXATION AND CRYOSECTIONS 106 9.8 IMMUNOFLUORESCENCE AND ANTIBODIES 106 9.9 EDU DETECTION 107 9.10 IMAGE ACQUISITION 108 9.11 STATISTICS 108 9.12 MICRODISSECTION AND SINGLE CELL SUSPENSION 108 9.13 FACS 109 9.14 RNA SEQUENCING 109 9.15 TRANSCRIPTOME ANALYSIS 110 10 REFERENCES 111 11 APPENDIX 145 11.1 CONFERENCE PRESENTATION 145 V. ACKNOWLEDGMENTS 146
Kruger, Adéle. "Development and implementation of ontology-based systems for mammalian gene expression profiling." Thesis, 2009. http://hdl.handle.net/11394/3289.
Full textThe use of ontologies in the mapping of gene expression events provides an effective and comparable method to determine the expression profile of an entire genome across a large collection of experiments derived from different expression sources. In this dissertation I describe the development of the developmental human and mouse eVOC ontologies and demonstrate the ontologies by identifying genes showing a bias for developmental brain expression in human and mouse, identifying transcription factor complexes, and exploring the mouse orthologs of human cancer/testis genes.Model organisms represent an important resource for understanding the fundamental aspects of mammalian biology. Mapping of biological phenomena between model organisms is complex and if it is to be meaningful, a simplified representation can be a powerful means for comparison. The implementation of the ontologies has been illustrated here in two ways.Firstly, the ontologies have been used to illustrate methods to determine clusters of genes showing tissue-restricted expression in humans. The identification of tissue restricted genes within an organism serves as an indication of the finetuning in the regulation of gene expression in a given tissue. Secondly, due to the differences in human and mouse gene expression on a temporal and spatial level, the ontologies were used to identify mouse orthologs of human cancer/testis genes showing cancer/testis characteristics. With the use of model systems such as mouse in the development of gene-targeted drugs in the treatment of disease, it is important to establish that the expression characteristics and profiles of a drug target in the model system is representative of the characteristics of the target in the system for which it is intended.
Chang, Chia-Huang, and 張嘉晃. "Health effects of maternal nonylphenol exposure on fetal development and neonatal health-Coupling a model of human amniotic fluid-derived stem cells." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/42098384229721482070.
Full text國立陽明大學
環境與職業衛生研究所
102
Nonylphenol (NP) is an environmental hormone with proven estrogenic effects. Although its adverse effects on animals are well documented, the effects of NP exposure on humans remain unclear, and those on the human fetus are completely unknown. Additionally, human amniotic fluid-derived mesenchymal stem cells (AFMSCs) containing heterogeneous population of stem cells from various fetal organs have the capacity to differentiate into multiple lineages. AFMSCs may provide a promising cells source to identify the effects of NP on human embryo. The aim of this study was to establish a pregnant cohort to explore the association between maternal NP exposure level and birth outcomes. Furthermore, AFMSCs were treated with NP in 3 concentration levels by different time periods to assess possible effects on characteristics of AFMSCs and their Oct4, Nanog, and Sox2 expressions. A pregnant cohort was followed-up. Maternal urine samples were collected at the first, second, and third pregnancy trimester. The umbilical cord blood at delivery and amniotic fluid for those undergoing amniocentesis were collected as well. NP levels were determined for every specimen. Fetal development were determined by ultrasonic scan and birth outcomes were assessed by a pediatrician. AFMSCs were isolated and cultured by a two-stage culture protocol and then treated with NP (10, 50, 100μM) for 24, 48, and 72 hours, respectively. The effect of NP on the proliferation of AFMSCs was determined by the trypan blue dye exclusion assay. The total number of viable cells was calculated in the microscopy. Reverse transcription and quantitative PCR (polymerase chain reaction) were used to assess the Oct4, Nanog, and Sox2 expressions of AFMSCs. A total number of 201 pregnant women consented to participate. But complete data were available from 162 singletons for data analyzed. After adjusting for the urinary creatinine concentration, NP concentrations during the three trimesters were 4.27, 4.21, and 4.10 μg/g cre. respectively. The NP concentrations were 8.22 ng/ml and 5.91 ng/ml in amniotic fluid and cord blood respectively. No statistically significant correlations between urinary NP concentrations and gestational ages or maternal body weights were observed in a mixed-effects model using a generalised estimating equation. Maternal NP concentrations in each trimester were not associated with birth sex, preterm status, or low birth weight. Data analysed further stratified women by the median urinary NP concentrations in the first, second, and third trimester. Pregnant women with above-median concentration during the second trimester gave birth to the neonatal body with shorten length in the multivariable regression model (β=-0.47 cm, p value=0.04). Additionally, maternal weight gain was also low for women in the group with NP above-median concentration during the second trimester (β=-1.55 kg, p value=0.02). High NP level in the second trimester had a significant association with neonatal body weight especially in the primiparas (β=-182.49 g, p value=0.02). The odds ratios (ORs) of low infant birth weight, comparing pregnant women with different NP levels, was increased by decreasing the cutoff percentile for birth weight in the logistic regression model (ORs=1.18 for the 50th percentile, 2.12 for the 25th percentile, and 7.81 for the 10th percentile). When treating AFMSCs with NP, the growth rate of AFMSCs was dose- and time-dependently decreased. The higher level of NP as well as the longer of NP exposure, the stronger Oct4, Nanog, and Sox2 gene expressions were found (Oct4: 1.5 fold, Nanog: 2.6 fold, Sox: 3.2 fold). These results indicated that NP might influence the process of cellular differentiation or organgenesis during fetal development. This study demonstrates that maternal high NP exposure is associated with small for gestational age (SGA), decreased fetal body length at birth, and low maternal weight gain. Additionally, NP might influence the process of cellular differentiation or organgenesis during fetal development. The effects of this endocrine-disrupting substance on pregnant women and fetuses should be a concern during gestation.
Möbius, Marius Alexander. "On the isolation, functional characterization and oxygen- induced impairment of resident mesenchymal stromal cells from the human fetal lung." 2017. https://tud.qucosa.de/id/qucosa%3A73094.
Full textBackground: Despite great achievements in neonatal and perinatal medicine over the past decades, the immature lung remains the most critical organ to care for after premature birth. As a consequence, impairment of of postnatal lung development – bronchopulmonary dysplasia or BPD – remains the most common complication of extreme prematurity and a major healthcare burden. There is no therapy for BPD, except prevention of premature birth. Recently, exogenous mesenchymal stromal cells (MSC) have been shown to prevent and rescue impaired lung development in animal models. Understanding the mechanisms behind the beneficial action of these cells is crucial for a successful, safe, and effective clinical translation of these promising MSC-based cell therapies in neonates. Hypothesis: Endogenous lung-resident MSC contribute to normal lung development and become impaired in conditions resembling premature birth, thus playing a part in the pathogenesis of BPD. Exogenous MSC act by supporting and/or preserving the endogenous mesenchymal cell’s function. Methods and Results: Using lung tissue from aborted fetuses, a novel enzyme/density gradient technique was employed to obtain endogenous human fetal lung mesenchymal cells (FLMSC). The vast majority of the so-isolated cells fulfilled all criteria of MSC, making the herein presented work the first complete description of MSC from human fetal lung tissue. Human umbilical cord-derived (UC)MSC were isolated by enzymatic digestion of the Wharton’s jelly. When cultured in hypoxic atmospheres resembling intrauterine conditions, resident FLMSC exerted a gene expression- and cytokine profile supporting epithelial and endothelial lung development, and secreted extracellular matrix components crucial for normal lung growth. After exposure to hyperoxia – thus mimicking premature birth and subsequent treatment on a neonatal intensive care unit – FLMSC showed signs of transdifferentiation, acquired a pro-inflammatory / anti-angiogeneitic secretory profile, diminished production of crucial extracellular matrix components and send out danger signals to other cells. Conversely, UCMSC secreted various paracrine factors protecting lung cells, and proteins contributing to lung growth and alveolarization. Discussion and Conclusions: The human fetal lung’s mesenchyme at the late canalicular stage of development mainly consists of MSC rather than fibroblasts, thus implying a complex mesenchymal stem-/progenitor cell hierarchy and previously undescribed cellular transdifferentiation processes of human endogenous lung mesenchymal progenitors during late pregnancy. Evidence for a contribution of FLMSC to normal lung development was generated in vitro, suggesting a co-ordination of endothelial and epithelial cell fate by human endogenous lung MSC. When challenged with hyperoxia, FLMSC cells acquire a phenotype contributing to the pathogenesis of the BPD. Conversely, UCMSC harbor the potential to provide the factors that these damaged resident MSC lack to produce. The endogenous MSC may therefore represent a potential target of cell-based therapies of BPD. However, in vivo data obtained from premature animals is inevitable to gain further insights into the contribution of endogenous lung MSC to normal and disrupted lung development and to clinically translate potent and safe MSC-based therapeutics for our most vulnerable patient population - premature infants.
Somsen, Elisabeth. "Mortalidade fetal tardia versus mortalidade neonatal precoce : estudo comparativo Portugal / União Europeia (2004-2013)." Master's thesis, 2015. http://hdl.handle.net/10400.2/4700.
Full textCom o objetivo de verificar a existência de relação entre a diminuição da mortalidade fetal tardia e o aumento da mortalidade natal e natal precoce, foi consultada bibliografia relevante sobre o assunto e bases de dados disponíveis, em Portugal e na União Europeia. Apesar da limitação de informação, principalmente a nível dos dados europeus, verificaram-se vários indicadores relevantes, como reprodução medicamente assistida, doenças na grávida, como diabetes e hipertensão arterial, fatores de risco como obesidade e tabagismo, bem como no feto, o baixo peso à nascença e a gemelaridade. Foi também considerado o Índice de Desenvolvimento Humano como fator relevante na evolução. Apurados os números e em função dos resultados foi efetuada uma comparação entre Portugal e a União Europeia durante a década de 2004-2013. Esta comparação foi efetuada utilizando gráficos elaborados a partir das tabelas apuradas e permitiu concluir que até à data não existe evidência de tal relação, mas que a mesma não é de excluir, pelo que mais investigação sobre o tema deverá ser efetuada. Para tal deverão ser considerados alguns novos indicadores para futuros estudos.
In order to verify the existence of a relationship between the decrease of late foetal mortality and the increase of neonatal and early neonatal mortality, relevant literature on the subject and available databases, in Portugal and the European Union, were consulted. Despite limited information on European Union data, there were several relevant indicators, such as assisted reproduction, diseases in pregnant women as diabetes and hypertension, risk factors such as obesity and smoking, as well as in the foetus, low birth weight and gemelarity, The Human Development Index was also considered as relevant factor on this evolution. Upon the results, a comparison between Portugal and the European Union during the decade of 2004-2013 was made. This comparison was established using graphics from elaborated tables and came to the conclusion that up until now there is no evidence of such a relationship, but that it cannot be excluded, so more research on this subject should be made. Some new indicators are also mentioned for future studies.
Ahmed, Asra. "Apoptosis and caspase-3 activity in isolated fetal rat lung cells, human A549 cells and rat periodontal ligament fibroblasts following exposure to cigarette smoke extract." 2012. http://hdl.handle.net/1993/5205.
Full text"The inhibitory effect of trans fatty acids on maternal and neonatal essential fatty acid metabolism." 1997. http://library.cuhk.edu.hk/record=b5889120.
Full textThesis (M.Phil.)--Chinese University of Hong Kong, 1997.
Includes bibliographical references (leaves 145-155).
Acknowledgment --- p.i
Abstract --- p.ii
List of Tables --- p.vii
List of Figures --- p.x
List of Abbreviations --- p.xii
Chapter Chapter 1 --- Literature review
Chapter 1.1 --- Historical background --- p.1
Chapter 1.2 --- Chemistry of trans and cis fatty acids --- p.3
Chapter 1.3 --- Dietary source of trans fatty acids --- p.6
Chapter 1.4 --- Consumption of trans fatty acids among Western countries --- p.9
Chapter 1.5 --- Current health concern for excessive intake of trans fatty acids --- p.10
Chapter 1.6 --- Metabolism of trans fatty acids --- p.13
Chapter 1.6.1 --- Absorption --- p.15
Chapter 1.6.2 --- Oxidation --- p.15
Chapter 1.6.3 --- Incorporation --- p.16
Chapter 1.6.4 --- Selectivity --- p.17
Chapter 1.7 --- Impact of trans fatty acids on essential fatty acid metabolism --- p.19
Chapter 1.8 --- Desaturation and elongation of trans fatty acids --- p.21
Chapter 1.9 --- Trans fatty acids and neonatal growth --- p.23
Chapter Chapter 2 --- Amount of trans fatty acids in Hong Kong fast foods
Chapter 2.1 --- Introduction --- p.25
Chapter 2.2 --- Objective --- p.25
Chapter 2.3 --- Materials and methods --- p.26
Chapter 2.4 --- Results --- p.27
Chapter 2.5 --- Discussion --- p.31
Chapter Chapter 3 --- Cross-cultural study of trans fatty acids in human milk
Chapter 3.1 --- Introduction --- p.35
Chapter 3.2 --- Objective --- p.35
Chapter 3.3 --- Materials and methods --- p.36
Chapter 3.4 --- Results
Chapter 3.4.1 --- Dietary information --- p.38
Chapter 3.4.2 --- Fatty acid composition of Chinese and Canadian human milk --- p.40
Chapter 3.4.3 --- Difference between Chinese and Canadian human milk --- p.40
Chapter 3.4.4 --- Difference between Hong Kong and Chongqing Chinese human milk --- p.43
Chapter 3.4.5 --- The change in milk fat and LCPUFA as lactation progresses --- p.43
Chapter 3.5 --- Discussion
Chapter 3.5.1 --- Trans fatty acids in human milk --- p.46
Chapter 3.5.2 --- Content of LCPUFA in human milk --- p.47
Chapter 3.5.3 --- Content of 18:2n-6 in human milk --- p.48
Chapter 3.5.4 --- Fat content in Hong Kong and Chongqing Chinese human milk --- p.49
Chapter 3.6 --- Conclusion --- p.50
Chapter Chapter 4 --- Trans fatty acids and maternal and neonatal essential fatty acid metabolism
Chapter 4.1 --- Introduction --- p.51
Chapter 4.2 --- Objectives --- p.53
Chapter 4.3 --- Materials and methods --- p.53
Chapter 4.4 --- Results
Chapter 4.4.1 --- Experiment1
Chapter 4.4.1.1 --- Relationship between the trans fatty acids in maternal diet and those in milk --- p.64
Chapter 4.4.1.2 --- Relationship between the trans fatty acids in maternal diet and those in neonatal liver --- p.64
Chapter 4.4.1.3 --- Content of 20:4n-6 in milk and in neonatal liver relative to that in maternal diet --- p.72
Chapter 4.4.2 --- Experiment2
Chapter 4.4.2.1 --- Amount of trans fatty acids in rat milk --- p.75
Chapter 4.4.2.2 --- Trans fatty acids in rat liver phospholipids --- p.75
Chapter 4.4.2.3 --- Linoleic acid (18:2n-6) content in rat and its relation to maternal diets --- p.86
Chapter 4.4.2.4 --- Content of 20:4n-6 in rat milk --- p.86
Chapter 4.4.2.5 --- Content of20:4n-6 in rat liver --- p.89
Chapter 4.4.2.6 --- Suppression of the synthesis of 20:4t isomers in maternal and neonatal liver --- p.89
Chapter 4.5 --- Discussion
Chapter 4.5.1 --- Relationship between fatty acid composition of diet and that of milk --- p.93
Chapter 4.5.2 --- 20:4n-6 in rat milk --- p.95
Chapter 4.5.3 --- Transfer of trans fatty acids from maternal diet to neonatal liver phospholipids --- p.98
Chapter 4.5.4 --- The inhibitory effect of trans fatty acids on synthesis of 20:4n-6 in neonatal liver --- p.99
Chapter 4.5.5 --- Effect of 18:2n-6 supplement on 20:4n-6 level of neonatal liver --- p.101
Chapter 4.5.6 --- Suppression of 18:2n-6 supplement on synthesis of 20:4t isomers --- p.101
Chapter 4.6 --- Conclusion --- p.104
Chapter Chapter 5 --- Accumulation and turnover of trans fatty acids
Chapter 5.1 --- Introduction --- p.105
Chapter 5.2 --- Objective --- p.105
Chapter 5.3 --- Materials and methods --- p.106
Chapter 5.4 --- Results
Chapter 5.4.1 --- Accumulation of trans fatty acids in liver and adipose tissue --- p.108
Chapter 5.4.2 --- Selectivity of individual 18:2 trans isomersin liver and adipose tissue --- p.112
Chapter 5.4.3 --- Turnover of trans fatty acids --- p.112
Chapter 5.4.4 --- Accumulation and turnover of 18:lt in brain --- p.115
Chapter 5.5 --- Discussion
Chapter 5.5.1 --- Accumulation of trans fatty acids in liver and adipose tissue --- p.120
Chapter 5.5.2 --- Turnover of trans fatty acids --- p.122
Chapter 5.5.3 --- Accumulation and turnover of trans fatty acidsin brain --- p.124
Chapter 5.6 --- Conclusion --- p.125
Chapter Chapter 6 --- In vivo Oxidation of trans fatty acids in rat
Chapter 6.1 --- Introduction --- p.126
Chapter 6.2 --- Objective --- p.127
Chapter 6.3 --- Materials and methods --- p.127
Chapter 6.4 --- Results --- p.129
Chapter 6.4.1 --- Apparent oxidation of saturated fatty acids --- p.136
Chapter 6.4.2 --- Apparent oxidation of 18:lt relative to 18:ln-9 --- p.136
Chapter 6.4.3 --- Oxidation of 18:2t isomers relative to 18:2n-6 --- p.137
Chapter 6.4.4 --- Effect of 18:2n-6 supplement in PHCO diet on oxidation per se --- p.137
Chapter 6.5 --- Discussion --- p.138
Chapter 6.5.1 --- Oxidation of 18:lt and 18:2t isomers --- p.139
Chapter 6.5.2 --- Effect of 18:2n-6 supplement on oxidation per se --- p.140
Chapter 6.6 --- Conclusion --- p.141
General conclusion --- p.142
References --- p.145
Monfils, Sarah. "Métabolisme énergétique cardiaque fœtal dans un modèle de restriction de croissance intra-utérine chez le rat." Thèse, 2011. http://hdl.handle.net/1866/5210.
Full textA low sodium diet was given to pregnant rats during the last week of gestation. This diet diminished the maternal expansion of blood volume, the uterine arteries diameter, and placental weight, when compared to their controls. Together, these results suggest a lower placenta perfusion and a decreased output of nutrients to the fetus. The offspring of these pregnant rats were born with an intra-uterine growth retardation (IUGR). The fetal heart utilizes glucose through glycolysis as the major cardiac energy substrate. At birth, the principal source of energy switches to the oxidation of fatty acids, through β-oxydation. We hypothesized that within our IUGR model, the fetal heart could respond to a diminished nutritional intake due to the maternal input when a decreased cardiac energy metabolism was present. The pregnant rats of both groups (controls and on the low sodium diet) were sacrificed on day 22 of a 23 day gestation. The fetal hearts were then analyzed looking for signs of the limiting proteins glycolysis and β-oxidation. The GLUT1, GLUT4, HK1, HK2, CPT2, CPT1β, cytochrome c, PFK1, PKM1/2 proteins obtained through a Western immunoblot method were similar between the hearts of the IUGR and their control fetuses, whether they were male or female. The protein expression of CPT1α was lower only in female IUGR fetal hearts. There was no significant difference between the groups with respect to the enzymatic activity of PKM1/2. Our results suggest that the metabolic profile changes with regards to the fetus gender and could affect the fetal cardiac metabolism, due to a lower maternal blood flow caused by a sodium controlled diet, by diminishing its lipid metabolism and sparing glucose metabolism. To characterize the energy metabolism, the enzymatic activity of the other principal limiting enzymes glycolysis (HK1, HK2, PFK1), the intra-mitochondrial flux of acyl CoA through the CPTs and the total quantity of acetyl CoA must also be analyzed.