Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 4 uppsatser och 1 appendix.

No description available.

Almost a century has elapsed since the antigen-antibody interaction was first recognised as the basis of an immune response (Ehrlich, 1897). However, it was only in the 1930s, with the development of improved technologies that this concept was better understood, and led to the discovery of the amazing diversity and specificity of antibody molecules (Landsteiner, 1933). Theoretically, it is possible to make antibodies to a variety of biological substances and other chemicals, and therefore they are ideally suited as specific recognition elements to be used for analytical, cytological, functional, therapeutic and biochemical purposes. The development of the radioimmunoassay (RIA) thirty five years ago, revolutionised research in many areas of clinical and scientific investigation. This technique evolved rapidly from the discovery made by Berson et al. in 1956 that antibodies to insulin could be detected in patients treated with this hormone, by measuring the binding of radiolabeled insulin to these antibodies. Although in the past RIAs have been the most important assay system employing antibody and labelled tracer, the limitation was that reliance had to be placed on the chance development of a good polyclonal antibody. These shortcomings stimulated the search for monospecific antibodies of reproducible quality and sufficient quantity. The development and introduction of monoclonal antibody technology brought about a revolution in immune serology (Kohler and Milstein, 1975). Establishment of immortal cell lines which contained the genetic elements of antibody-producing cells was achieved by fusion between a myeloma cell line and spleen cells from an immunised donor. The resulting hybrids had the essential properties of both parents, namely, permanent growth and a high capacity for the synthesis and secretion of immunoglobulins, normally characteristics of plasmacytomas, together with the genetic elements defining a specific antibody. Gestational trophoblastic disease (GTD) is a neoplastic condition of the trophoblast and occurs as molar pregnancy in a benign or invasive form, or as choriocarcinoma in a malignant form. Effective therapy has been developed for the treatment of both choriocarcinoma and molar pregnancy, but the key to successful management of these patients lies in their prompt diagnosis and careful monitoring of response to treatment (Green-Thompson, 1986). Fortuitously, these tumours elaborate the human chorionic gonadotrophin hormone (hCG) and its free alpha (a) and beta (B) subunits and hence a ready marker for the tumour exists. Human chorionic gonadotrophin is one of a group of glycoprotein hormones, which includes luteinising hormone (LH), follicle stimulating hormone (FSH) and thyroid stimulating hormone (TSH). These hormones are composed of two dissimilar subunits designated a and B, which are bound non-covalently in the intact molecule. The B-subunit of each glycoprotein hormone is unique and is responsible for the respective biological and immunological properties of the glycoproteins. In contrast, all four hormones possess an identical a-subunit which is coded for by a single gene (Fiddes and Goodman, 1979). The measurement of hCG and its free B-subunit, as so-called BhCG, for the diagnosis and monitoring of therapy in patients with GTD is now routinely practised throughout the world (Vaitukaitis et al., 1972). However it has been demonstrated by Bagshawe (1975) that when serum BhCG can no longer be measured by current RIA methods, up to 10" tumour cells may remain undetected. In addition, there have been isolated reports of two patients with choriocarcinoma in whom BhCG was undetectable in the serum but who appeared to be secreting only the a-subunit (Dawood et al, 1977). Furthermore, it has been suggested that measurement of free a-subunit rather than intact hCG or the free B-subunit is a more effective means of detecting persistent trophoblastic disease as well as tumour recurrence following treatment (Quigley et al, 1980a and b). Radioimmunoassays which measure the free a-subunit of hCG have been developed, but in general lack the specificity and sensitivity required (Gaspard et al, 1980; Kohorn et al, 1981). These assays employ polyclonal antisera which also detect epitopes common to the pituitary gonadotrophins. Thus there is a need to produce monoclonal antibodies which recognise regions of the free a-subunit which are hidden in the intact gonadotrophins. Such antibodies would provide the required specificity for use in RIAs but are limited in their use by their inherent lack of high affinity for the antigen. Fortunately, this drawback may be overcome by using monoclonal antibodies as labelled reagents in an alternative assay system, the immunoradiometric assay (IRMA), described by Miles and Hales (1968). The IRMA, particularly the two-site sandwich version of the assay, has been shown to provide greater sensitivity in addition to allowing enhanced specificity. This is a consequence of the use of two antibodies in excess to detect the analyte, each directed at a different epitope on the target molecule. The first antibody, referred to as the capture antibody, is usually linked to a solid-phase to facilitate easy separation and is added in excess relative to the target hormone to enhance antibody-antigen interaction, thereby allowing increased sensitivity in the measurement of analyte. The second antibody, referred to as the detection antibody, is labelled with a radioactive isotope or an enzyme to detect antigen already bound to the capture antibody. The application of monoclonal antibodies specific for the free a-subunit to a highly sensitive IRMA format is an obvious need. Hence this study was undertaken firstly, to raise and characterise monoclonal antibodies to the free a-subunit, secondly to develop an IRMA using these antibodies and finally to establish whether measurement of free a-subunit has any clinical advantage. / Thesis (M.Med.Sc.)-University of Natal, Durban, 1990.

O biglicam é um proteoglicano é altamente expresso em células trofoblásticas de patologias placentárias com invasividade exacerbada. No entanto, as funções do biglicam no trofoblasto ainda não foram elucidadas. Sendo assim, verificamos a expressão e as funções de biglicam e seus receptores Toll-like (TLR)-2 e TLR-4 nas células trofoblásticas durante a gestação. As células do citotrofoblasto extraviloso (CTEV) foram positivas para todas as moléculas, menos para o biglicam em placentas a termo. Adição exógena de biglicam promoveu migração e invasão das células trofoblásticas. O biglicam estimulou a fosforilação de AKT nos sítios Thr308 e Ser473 nas células trofoblásticas. A migração e a invasão biglicam-dependentes e as fosforilações de AKT foram inibidas após a adição de anticorpos bloqueadores anti-TLR-2 e anti-TLR-4. O silenciamento gênico de AKT1 em células SGHPL-5 aboliu os efeitos do biglicam na motilidade. Em conclusão, o biglicam aumenta a motilidade de células trofoblásticas após sinalização por AKT através da ativação de TLR-2 e TLR-4. / Biglycan is a highly expressed proteoglycan in trophoblast cells from invasiveness-changed placental pathologies. However, biglycan functions in the trophoblast were not yet identified. Therefore, it was verified the expression and functions of biglycan and its receptors Toll-like (TLR)-2 and TLR-4 in trophoblast cells throughout pregnancy. The extravillous cytotrophoblast cells (EVT) were positive to all the molecules, although biglycan was negative in term placentas. Exogenous biglycan promoted migration and invasion of trophoblast cells. Biglycan stimulated AKT phosphorilation at Thr308 and Ser473 sites in trophoblast cells. The biglycan-dependent migration, invasion and AKT phosphorilation were inhibited upon addiction of anti-TLR-2 and anti-TLR-4 blocking antibodies. AKT1 genic silencing in SGHPL-5 cells abolished the motility effects. In conclusion, biglycan increases the motility of trophoblast cells after AKT signaliing throughout TLR-2 and TLR-4 activation.

A Gonadotrofina Coriônica Eqüina (eCG) é produzida pela égua prenhe e tem ação folículo estimulante e luteinizante em animais domésticos não eqüídeos. Um pool formado por plasma de 4 éguas prenhes, com média de 69 dias de gestação, foi purificado em coluna cromatográfica com resina de afinidade Blue Sepharose FF (BS). As frações que adsorveram à resina BS foram purificadas em coluna cromatográfica com resina de afinidade Concanavalina A 4B (ConA). As frações que não adsorveram à resina BS também foram purificadas em coluna cromatográfica com resina de afinidade ConA. O mesmo pool de palsma foi diafiltrado, em cartucho de hemodiálise. O diafiltrado foi aplicado em coluna cromatográfica com resina de afinidade ConA. Atividade biológica (UI/mL) do plasma, do diafiltrado e das frações purificadas foram quantificadas por ensaio biológico com ratas impúbres. As atividades biológicas encontradas no plasma e no plasma diafiltrado foram de 3,63 e 5,14UI/mL, respectivamente. A atividade biológica encontrada nas frações que adsorveram à BS foi de 3,50UI/mL. Não foi encontrarda atividade biológica nas frações que não adsorveram à BS. A atividade biológica contida nas frações que adsorveram à BS e que também adsorveram a ConA foi de 3,65UI/mL. O rendimento do processo cromatográfico onde o plasma foi adsorvido pela BS e pela ConA, foi de 69,52%. Não foi encontrada atividade biológica nas frações obtidas da aplicação do plasma diafiltrado em coluna de ConA. O processo cromatográfico com uso de BS seguido de ConA mostou-se eficaz em purificar a eCG do plasma de éguas prenhes. / The equine Chorionic Gonadotrophin (eCG) is produced by the pregnant mare and has follicle-stimulant and luteinizing actions on non-equine domestic animals. A pool formed by the plasma of 4 pregnant mares (with mean gestation of 69 days) was purified in chromatographic column with Blue-Sepharose FF affinity resin (BS resin). Fractions adsorbed by BS resin were then purified in chromatographic column with Concavalin A 4B affinity resin (ConA resin). The fractions not adsorbed by the BS resin were also purified in chromatographic column with ConA resin. The same plasma pool was dialyzed in hemodialysis cartridge. The dialyzed was applied in chromatographic column with ConA resin. Biological activities (in IU/mL) of the plasma, of the dialyzed and of the purified fractions were quantified in a biological assay with female rats that did not reach puberty. The biological activities found in the plasma and dialyzed were of 3.63 and 5.14 IU/mL, respectively. Fractions that were adsorbed by BS had a biological activity of 3.50 IU/mL. No biological activity was found in fractions that were not adsorbed by BS. Biological activity found in fractions adsorbed by both BS and ConA was of 3.65 IU/mL. When plasma was both adsorbed by BS and ConA, the chromatographic process yield had results of 69.52%. No biological activity was found in the fractions obtained from the administration of dialyzed plasma in ConA column. The BS - followed by ConA -chromatographic process showed efficacy in purifying the eCG from the plasma of pregnant mares.

Objetivo: Caracterizar as indicações das gestantes que procuram o serviço de Medicina Fetal do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo que realizaram procedimentos invasivos diagnósticos e avaliar os resultados dos cariótipos fetais e destas gestações no período de fevereiro de 2005 a dezembro de 2009. Método: Estudo observacional retrospectivo das gestantes que realizaram biópsia de vilo corial (BVC), amniocentese e cordocentese. Não foram incluídos outros procedimentos diagnósticos ou terapêuticos. O resultado da gestação foi obtido através de consulta de prontuário eletrônico e/ou físico e/ou contato telefônico. Resultados: Foram realizados 113 BVC, 340 amniocenteses e 260 cordocenteses. A principal indicação para a realização dos procedimentos invasivos foi a presença de malformações fetais (69,8%), seguido por translucência nucal aumentada (13,4%) e idade materna avançada (10,2%). A trissomia do cromossomo 18 foi a aneuploidia mais comum (8,1%), seguido pela trissomia do 21 (6,2%), 45,X0 (4,8%) e a trissomia do 13 (3,8%). Ocorreram 4,9% abortamentos, 25,7% natimortos e 13% neomortos. Oito gestantes optaram pela interrupção judicial. 99% das gestantes cujos fetos não apresentavam malformação e que apresentavam cariótipo fetal normal tiveram nativivos. CONCLUSÃO: A principal indicação para a realização dos procedimentos invasivos foi a presença de malformação fetal em 69,8% das gestantes e presença de anormalidades cromossômicas encontradas nos fetos foi de 26,23%. / Objective: The purpose of this study is to characterize the indications of pregnant women who seek the Fetal Medicine Service of Hospital das Clínicas of São Paulo University to perform invasive diagnostic procedures and evaluate the results of fetal karyotypes and their pregnancies from February 2005 to December 2009. Methods: Retrospective observational study of pregnant women who underwent CVS, amniocentesis or cordocentesis. Other diagnostic or therapeutic procedures were not included. The outcomes of pregnancies were obtained through consultation of medical records and/or telephone contact. Results: 113 CVS, 340 amniocentesis and 260 cordocentesis were performed. The main indication for performing invasive procedure was the presence of fetal anomaly (69.8%), followed by increased nuchal translucency (13.4%) and maternal age (10.2%). The trisomy of chromosome 18 was the most common aneuploidy (8.1%), followed by trisomy 21 (6.2%), 45,X0 (4.8%), and trisomy 13 (3.8%). There were 4.9 % of miscarriage, 25.7% of stillbirth and 13% of neonatal deaths. Eight women opted for legal termination of pregnancies. 99% of pregnant women whose fetus had no structural abnormalities and normal karyotype had a live child. CONCLUSION: The main indication for karyotyping was the presence of fetal malformation in 69.8% of pregnancies and chromosomal abnormalities was found in 26.23% of the fetuses.

A clonagem de animais é uma técnica em desenvolvimento e ainda requer refinamento, uma vez que as perdas embrionárias são significativas. Mesmo com animais chegando à termo, algumas deficiências verificadas ao nascimento provocam a morte destes antes do primeiro mês de vida. As alterações placentárias durante a embriogênese estão relacionadas, à formação deficiente da placenta, principalmente nas regiões de troca materno-fetal. Considerando a necessidade de estudos mais detalhados sobre a morfologia da placenta e a ausência de estudos quantitativos das estruturas placentárias em grandes mamíferos, investigamos comparativamente os vilos coriônicos da placenta de bovinos não clonados e clonados usando delineamento estereológico. Foram utilizadas placentas de quatro fêmeas de bovinos clonados e quatro de não clonados. A amostragem foi realizada de modo uniforme, sistemático e aleatório, sendo obtidas estimativas de densidade de volume, volume referência, tamanho e comprimento e número de unidades volume star. Todos os parâmetros estereológicos apresentaram valores superiores (embora não significativos) para não clonados, com exceção do número de unidades volume star. No entanto, a placenta de não clonados foi mais eficiente (p= 0.014). Apesar de não significativos, os resultados sugerem a existência de diferenças biológicas, ou seja, a placenta de bovinos não clonados apresenta vilos maiores (porém em menor número), mas distribuídos em um número maior de placentomas. Já em animais clonados, estes vilos são menores, mais numerosos e distribuídos em um número menor de placentomas. / Animal cloning is still in development it requires further improvements since embryonic losses are remarkable animals die before the first month after birth. The placental changes during embryogenesis cause a deficiency in the placenta, mostly in the maternal-fetal exchange areas. Due to the need for more detailed studies on the placenta?s morphology, especially the quantitative analysis of the placenta structures in large animals, chorionic villi were comparatively investigated in both cloned and non-cloned bovines by means of stereology designed methods. Placentas from four cloned and four non-cloned bovines were used. Specimens were collected in a random uniformand systematic procedure, and stereological designed methods were pursued to estimate volume density of villi; the volume reference of placenta (Cavilieri Principle), surface density, surface area, star volume and number of star volume units. All parameters presented larger values for non-cloned, with exception of number of star volume units, thought these results were statistically non-significant larger in non-cloned animals (p= 0.014). In conclusion, it is possible to say that there are biological differences, in the placenta of non-cloned bovines which presents larger chorionic villi, small villi number, but distributed in a larger number of placentomas. In cloned bovines, however, villi are smaller, numerous more, and they are distributed in fewer number of placentomes.

by Chung Hoi Sing. / Thesis (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

Cheung Kwok Lung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 123-136). / ABSTRACT (English) --- p.i / ACKNOWLEDGMENTS --- p.1 / LIST OF FIGURES --- p.3 / LIST OF TABLES --- p.5 / LIST OF ABBREVIATIONS --- p.7 / Chapter I. --- INTRODUCTION AND OBJECTIVES --- p.8 / Chapter II. --- LITERATURE REVIEWS --- p.11 / Chapter II.A. --- Maternal Serum Alpha-fetoprotein Screeningin Singleton Pregnancies --- p.11 / Chapter II.A.1. --- Physiology of Alpha-fetoprotein --- p.12 / Chapter II.A.2. --- Historical Background of Screening by Alpha- fetoprotein --- p.12 / Chapter II.A.3. --- Factors that Influence Maternal Serum Alpha- fetoprotein Concentration --- p.13 / Chapter ILA.4. --- Elevated Maternal Serum Alpha-fetoprotein Concentration and Adverse Pregnancy Outcomes and Complications --- p.14 / Chapter II.A.4.a. --- Low Birth Weight --- p.16 / Chapter II.A.4.b. --- Fetal Loss --- p.17 / Chapter II.A.4.c. --- Pregnancy Induced Hypertension --- p.18 / Chapter II.B. --- Maternal Serum Human Chorionic Gonadotrophin Screening in Singleton Pregnancies --- p.18 / Chapter II.B.1. --- Physiology of Human Chorionic Gonadotrophin --- p.18 / Chapter II.B.2. --- Historical Background of Screening by Human Chorionic Gonadotrophin --- p.20 / Chapter II.B.3. --- Factors that Influence Maternal Serum Human Chorionic Gonadotrophin --- p.21 / Chapter II.B.4. --- Elevated Maternal Serum Human Chorionic Gonadotrophin Concentration and Pregnancy Complications --- p.21 / Chapter II.B.5. --- Maternal Serum AFP and hCG Concentrations and Adverse Outcomes or Complications in Twin Pregnancies --- p.23 / Chapter II.C. --- Mechanism for the Association between Adverse Outcomes and Elevated Maternal Serum Alpha- fetoprotein and Human Chorionic Gonadotrophin --- p.25 / Chapter III. --- METHODS --- p.28 / Chapter III.A. --- Study Population --- p.28 / Chapter III.B. --- Sample Collection and Analysis --- p.29 / Chapter III.C. --- Clinical Information --- p.30 / Chapter III.D. --- Microparticle Enzyme Immunoassay --- p.30 / Chapter III.D.1. --- Principles --- p.30 / Chapter III.D.1.a. --- Reaction Process --- p.31 / Chapter III.D.1.b. --- MEIA Assembly --- p.33 / Chapter III.D.1.c. --- Operation --- p.34 / Chapter III.D.2. --- AFP Assay --- p.34 / Chapter III.D.2.a. --- AFP Reagents --- p.34 / Chapter III.D.2.b. --- Sample Dilution --- p.36 / Chapter III.D.3. --- Total p-hCG Assay --- p.37 / Chapter III.D.3.a. --- Total p-hCG Reagents --- p.37 / Chapter III.D.3.b. --- Sample Dilution --- p.39 / Chapter III.D.4. --- Intra- and Inter-assay Variation --- p.39 / Chapter III.E. --- Data Handling --- p.42 / Chapter III.F. --- Statistical Analysis --- p.42 / Chapter III.F.1. --- Calculations of Median Values of Maternal Serum Alpha-fetoprotein and Human Chorionic Gonadotrophin Concentrations --- p.42 / Chapter III.F.2. --- Analysis for Adverse Outcomes or Complications --- p.43 / Chapter III.F.3. --- Adjustment of Alpha-fetoprotein and Human Chorionic Gonadotrophin for Gestational Age and Maternal Weight --- p.46 / Chapter IV. --- RESULTS --- p.48 / Chapter IV.A. --- Median Values of Maternal Serum Alpha-fetoprotein Human Chorionic Gonadotrophin --- p.48 / Chapter IV.B. --- Prediction of Adverse Outcomes by Maternal Serum Alpha-fetoprotein and Human Chorionic Gonadotrophin --- p.60 / Chapter IV.B. l. --- Preterm Delivery --- p.60 / Chapter IV.B.2. --- Spontaneous Preterm Delivery --- p.64 / Chapter IV.B.3. --- Premature Delivery --- p.68 / Chapter IV.B.4. --- Spontaneous Premature Delivery --- p.68 / Chapter IV.B.5. --- Other Outcomes or Complications --- p.72 / Chapter IV.B.6. --- Single Predictor for Most Adverse Outcomes --- p.74 / Chapter IV.C. --- Adjustment of Maternal Serum Alpha-fetoprotein and Human Chorionic Gonadotrophin for Maternal Weight and Gestational Age --- p.75 / Chapter IV.C.1. --- Distribution of Alpha-fetoprotein and Human Chorionic Gonadotrophin during Mid-trimester --- p.76 / Chapter IV.C.2. --- Adjustment of Alpha-fetoprotein for Maternal Weight and Gestational Age --- p.79 / Chapter IV.C.3. --- Adjustment of Human Chorionic Gonadotrophin for Maternal Weight and Gestational Age --- p.80 / Chapter IV.D. --- Predictiveness of Alpha-fetoprotein and Human Chorionic Gonadotrophin for Adverse Outcomes after Adjusted for Maternal Weight and Gestational Age --- p.83 / Chapter IV.D.l. --- Preterm Delivery --- p.86 / Chapter IV.D.2. --- Spontaneous Preterm Delivery --- p.86 / Chapter IV.D.3. --- Premature Delivery --- p.92 / Chapter IV.D.4. --- Spontaneous Premature Delivery --- p.92 / Chapter IV.D.5. --- Other Adverse Outcomes or Complications --- p.98 / Chapter IV.D.6. --- Single Predictor for Most Adverse Outcomes --- p.98 / Chapter V. --- DISCUSSIONS --- p.100 / Chapter V.A. --- Median Values of Maternal Serum Alpha-fetoprotein and Human Chorionic Gonadtrophin --- p.100 / Chapter V.B. --- Maternal Serum Alpha-fetoprotein and Human Chorionic Gonadotrophin Screening for Adverse Outcomes --- p.103 / Chapter V.C. --- Adjustment of Alpha-fetoprotein and Human Chorionic Gonadotrophin for Maternal Weight and Gestational Age --- p.109 / Chapter V.D. --- Predictiveness of Alpha-fetoprotein and Human Chorionic Gonadotrophin for Adverse Outcomes after Maternal Weight and Gestational Age Adjustment --- p.112 / Chapter V.E. --- Conclusions --- p.113 / Chapter V.F. --- Future Directions --- p.116 / APPENDIX 1 DATA BASE OF CLINICAL INFORMATION --- p.117 / APPENDIX 2 SEVERITY AND CLASSIFICATION OF PREGNANCY INDUCED HYPERTENSION --- p.122 / REFERENCES --- p.123

A Gonadotrofina Coriônica Eqüina (eCG) é produzida pela égua prenhe e tem ação folículo estimulante e luteinizante em animais domésticos não eqüídeos. Um pool formado por plasma de 4 éguas prenhes, com média de 69 dias de gestação, foi purificado em coluna cromatográfica com resina de afinidade Blue Sepharose FF (BS). As frações que adsorveram à resina BS foram purificadas em coluna cromatográfica com resina de afinidade Concanavalina A 4B (ConA). As frações que não adsorveram à resina BS também foram purificadas em coluna cromatográfica com resina de afinidade ConA. O mesmo pool de palsma foi diafiltrado, em cartucho de hemodiálise. O diafiltrado foi aplicado em coluna cromatográfica com resina de afinidade ConA. Atividade biológica (UI/mL) do plasma, do diafiltrado e das frações purificadas foram quantificadas por ensaio biológico com ratas impúbres. As atividades biológicas encontradas no plasma e no plasma diafiltrado foram de 3,63 e 5,14UI/mL, respectivamente. A atividade biológica encontrada nas frações que adsorveram à BS foi de 3,50UI/mL. Não foi encontrarda atividade biológica nas frações que não adsorveram à BS. A atividade biológica contida nas frações que adsorveram à BS e que também adsorveram a ConA foi de 3,65UI/mL. O rendimento do processo cromatográfico onde o plasma foi adsorvido pela BS e pela ConA, foi de 69,52%. Não foi encontrada atividade biológica nas frações obtidas da aplicação do plasma diafiltrado em coluna de ConA. O processo cromatográfico com uso de BS seguido de ConA mostou-se eficaz em purificar a eCG do plasma de éguas prenhes. / The equine Chorionic Gonadotrophin (eCG) is produced by the pregnant mare and has follicle-stimulant and luteinizing actions on non-equine domestic animals. A pool formed by the plasma of 4 pregnant mares (with mean gestation of 69 days) was purified in chromatographic column with Blue-Sepharose FF affinity resin (BS resin). Fractions adsorbed by BS resin were then purified in chromatographic column with Concavalin A 4B affinity resin (ConA resin). The fractions not adsorbed by the BS resin were also purified in chromatographic column with ConA resin. The same plasma pool was dialyzed in hemodialysis cartridge. The dialyzed was applied in chromatographic column with ConA resin. Biological activities (in IU/mL) of the plasma, of the dialyzed and of the purified fractions were quantified in a biological assay with female rats that did not reach puberty. The biological activities found in the plasma and dialyzed were of 3.63 and 5.14 IU/mL, respectively. Fractions that were adsorbed by BS had a biological activity of 3.50 IU/mL. No biological activity was found in fractions that were not adsorbed by BS. Biological activity found in fractions adsorbed by both BS and ConA was of 3.65 IU/mL. When plasma was both adsorbed by BS and ConA, the chromatographic process yield had results of 69.52%. No biological activity was found in the fractions obtained from the administration of dialyzed plasma in ConA column. The BS - followed by ConA -chromatographic process showed efficacy in purifying the eCG from the plasma of pregnant mares.