Thesis (M. Phil.)--Chinese University of Hong Kong. / Bibliography: l. 88-97.

Human chorionic gonadotropin (HCG), a glycoprotein hormone with two nonidentical subunits, is produced by chorionic tissue in pregnant women and by neoplastic tissue containing chorionic elements. It is used in the treatment of male hypogonadism and female sub-fertility. Quantitation of HCG is used to monitor therapy, diagnose various disease states and diagnose and monitor pregnancy. Low levels of HCG in the early and late stages of pregnancy and in various disease states has prompted the development of extremely sensitive assay procedures. Clinically, radioimmunoassay methods are most frequently used due to their precision, sensitivity and cost. However, problems with specificity have been noted. Commercial preparations of HCG must meet the standards outlined in the United States Pharmacopeia (USP). The assay procedure involves a rat uterine weight bioassay. This protocol is lengthy to perform (5 days), requires the sacrifice of a large number of animals (minimum of 60 female rats per assay) and may need to be repeated if the results do not meet the statistical requirements of the assay. Due to the use of animals and the animal care facilities required, this is an expensive assay. In addition, the bioassay is not specific for HCG. Therefore, this thesis reports the analysis of two commercial preparations of HCG, as well as USP Reference Standard HCG and commercially available purified intact HCG and purified individual subunits. Various HPLC assay procedures were evaluated to determine if HPLC would be a viable alternative to the official USP bioassay. Size exclusion HPLC, using one Protein Pak 125 sw column and two Protein Pak 300 sw columns individually and in various combinations, was used to assess all the samples of HCG. Attempts to increase resolution of HCG from interfering components found in these preparations included using both 208 nm and 278 nm for ultraviolet detection, evaluation of 32 buffers as mobile phases with the Protein Pak 300 sw column, fluorescamine derivatization of HCG followed by fluorescence detection, connection of two size exclusion columns in series, and recycling on a Protein Pak 300 sw column. Further attempts to isolate HCG from its protein contaminants involved using ion exchange HPLC with a Protein Pak DEAE 5 pw column with 20 different buffers as mobile phases as well as reversed-phase HPLC with an Ultrasphere ODS column. The greatest resolution was obtained with one Protein Pak 300 sw column with a phosphate buffer (0.15 M, pH 7.0) for the mobile phase and ultraviolet detection. Latex agglutination inhibition slide tests and electrophoresis techniques were used to evaluate commercial samples of HCG and chromatographic peak eluates. Commercial HCG samples appear to contain the individual subunits of HCG and intact HCG along with impurities. The USP Reference Standard HCG contains intact HCG but also contains other ultraviolet absorbing components that were partially separated by HPLC. Electrophoresis also indicated that this HCG sample contained impurities. In addition, the purified intact HCG and purified subunit samples contained impurities, as shown by HPLC. The size exclusion HPLC assay developed using one Protein Pak 300 sw column was unable to resolve intact HCG from the beta-subunit. This assay would be useful for a qualitative assay for purity of HCG preparations. However, at present, HPLC is not a viable alternative to the USP bioassay. / Pharmaceutical Sciences, Faculty of / Graduate

No description available.

Orientador: Albina Messias de Almeida Milani Altemani / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-06T19:27:59Z (GMT). No. of bitstreams: 1 Juliano_PriscilaBianchi_M.pdf: 2749600 bytes, checksum: a6323ddb35a6ed804b6e080a03a83c4a (MD5) Previous issue date: 2006 / Resumo: A participação da barreira trofoblástica na patogènese da vilosite placentária tem sido abordada somente em raros estudos realizados em cultura de tecidos, os quais demonstram que a expressão de ICAM-1 (intercellular adhesion molecule I) por células do trofoblasto viloso poderia permitir o influxo de células maternas para dentro da vilosidade. Para investigar a expressão de ICAM-1 pelo trofoblasto viloso, sua relação com ruptura da barreira trofoblástica e influxo de células imunes para dentro da vilosidade, foram analisadas 18 placentas com placentites (5 causada por T. gondii, 3 por T. cruzi, 2 por P. brasiliensis e 8 de etiologia desconhecida - VED) e 8 sem inflamação. Todas foram analisadas através da técnica de imunoistoquímka, em secções de parafina, utilizando-se anticorpos monoclonais anti-CD45RO, anti-CD20, MAC 387, HAM 56, anti-CD15, anti-hNK e anti-ICAM-1. Nas vilosites, o infiltrado intraviloso era composto principalmente por macrófagos HAM 56+ e linfócitos T UCHL-1+, enquanto que no espaço interviloso as células eram predominantemente monócitos MAC 387+. Na intervilosite por P. brasiliensis, não havia célula inflamatória dentro da vilosidade, enquanto que o infiltrado do espaço interviloso era composto principalmente por monócitos MAC 387+ e neutrófilos CDI5+. Todos os casos com placentite, exceto um, mostraram aumento da expressão de ICAM-1 nas vilosidades inflamadas, a qual estava localizada, quase que exclusivamente, próxima às áreas de ruptura trofoblástica. Nestas, foram observados leucócitos aderidos, apenas nos casos com vilosite. Quando a reação inflamatória ocorria somente no espaço interviloso (infecção por P. brasiliensis) apesar da ruptura trofoblástica e aumento da expressão de ICAM-I, não se observou influxo de leucócitos em direção ao estroma viloso. Nenhuma das placentas sem inflamação mostrou expressão de ICAM-1. Concluindo, o aumento da expressão de ICAM-1 pelo trofoblasto viloso ocorre durante placentites com acúmulo de leucócitos tanto no estroma viloso como no espaço interviloso e, provavelmente, desempenha um importante papel na ruptura da barreira trofoblástica. O influxo de células imunes para o vilo placentário, aparentemente, é mediado por ICAM-1, mas a localização do antígeno no interior da vilosidade é certamente um fator crucial para que isso ocorra. / Abstract: Although an in vitro study has hypothesized that expression of ICAM-1 by villous trophoblasts could be important for the influx of maternal immune cells in villitis, it remains to be shown whether the same phenomenon occurs in human villitis. To investigate the expression of ICAM-1 by villous trophoblasts, its relationship with rupture of the trophoblastic barrier and influx of immune cells into the villi, we analyzed 18 paraffin-embedded placentas with placentitis (5 by T. gondii, 3 by I cruzi, 2 by P. brasiliensis and 8 of unknown aetiology - VTJE) and 8 control placentas by immunohistochemistry using the monoclonal antibodies anti-CD45RO, anti-CD20, MAC 387, HAM 56, anti-CD15, anti-HNK and ami-ICAM-1. In all villitis cases, the inflammatory infiltrate within the villous stroma was composed mainly of HAM 56+ macrophages and UCHL-1 T lymphocytes, but in the intervillous space, the cells were mainly MAC 387+ monocytes. Otherwise, intervillosites caused by P. brasiliensis showed mainly MAC 387+ monocytes, CD15+ granulocytes and none inflammatory cell within the villi. All cases but one of placentitis showed trophoblast overexpression of ICAM-1 in the inflamed villi, located almost exclusively next to the areas of trophoblastic rupture. The villitis cases (caused by T. cruzi, T. gondii and VUE) presented leukocyte adherence in the areas of trophoblastic rupture. When the inflammatory reaction was situated in the intervillous space (placentitis by P. brasiliensis), in spite of the trophoblastic rupture and ICAM-1 overexpression there was no leukocyte influx into villi. None of the control placentas showed ICAM-1 expression. We concluded that overexpression of ICAM-1 by villous trophoblasts occurs during placentitis characterized by accumulation of leukocytes in the villous or intervillous space and probably plays an important role in the rupture of the trophoblastic barrier. The influx of immune cells into the villi appears to be mediated by ICAM-1 but the location of the antigen within villous stroma is certainly a crucial factor for its occurrence. / Mestrado / Ciencias Biomedicas / Mestre em Ciências Médicas

by Pui-Kwong Chan. / Thesis -- The Chinese University of Hong Kong. / Bibliography: leaves 139-145.

Kwok-pui Fung. / Thesis (M.Ph.)--Chinese University of Hong Kong, 1975. / Bibliography: leaves 122-131.

Thesis (M.Ph.)--Chinese University of Hong Kong. / Bibliography: leaves 87-96.

Thesis--University of Kentucky (Ph. D.), 2003. / Title from document title page. Document formatted into pages; contains vii, 65 p. : ill. Includes abstract and vita. Includes bibliographical references (p. 60-65).

Gonadotropins were used to stimulate luteal function, as determined by progesterone secretion, in both in vitro and in vivo systems. LH and hCG were capable of significantly stimulating progesterone secretion in the in vivo systems. Stimulation of progesterone secretion by hCG was greater than that for LH. PMSG failed to increase progesterone production at any level of treatment. hCG was also used to stimulate progesterone production by the corpus luteum in mares during early gestation. hCG administration resulted in a significant (p < 0.10) increase in peripheral progesterone levels in treatment mares through day 14 post-estrus. Peripheral progesterone concentrations were also higher in hCG treated mares for days 15 through 30 post-estrus in mares that conceived. hCG treatment had no influence on anterior pituitary release of LH.

The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from PDF of title page (University of Missouri--Columbia, viewed on March 25, 2010). Vita. Thesis advisor: Michael Roberts. "December 2009" Includes bibliographical references