The production of therapeutic recombinant proteins in heterologous systems has gained significance since the last decade. For recombinant proteins that require post-translational modifications (PTMs), mammalian systems are preferred. Chinese hamster ovary (CHO) cells are the mammalian cells of choice for production of recombinant proteins. This is because of their ability to provide correct protein-folding and post-translational modifications, displaying high productivity at large scale, ability to grow in suspension mode at high densities in a serum-free media, incapable of infection by most viruses and their history of regulatory approvals. There is an established state of the art technology for development of CHO cells for recombinant protein production. This technology relies on random integration of the gene of interest and gene amplification process for obtaining high expressing clones. There is a high degree of clonal heterogeneity and instability observed in the screened clones. To overcome the process of random integration, this report describes a lentivirus based screening for search of stable and high expressing integration sites in CHO cells. The integration sites are identified by using nrLAM-PCR (non-restrictive linear amplification mediated PCR) coupled with high throughput sequencing. Lentivirus are chosen as they preferentially integrate within the coding regions rendering the possibility of obtaining stable and high expressing clones. In addition, lentivirus vector is designed to possess landing pad for recombinase mediated cassette exchange of viral sequence with foreign DNA. The report describes a successful cassette exchange reaction but with low efficiency. Genome engineering technologies such as CRISPR/Cas, TALENs can used for targeted gene insertion at integration sites and thus establishing stable and efficient production of recombinant proteins in CHO cells. Additionally, an approach for designing synthetic promoters based on Ef1α promoter architecture has been shown. Synthetic promoters are useful for expression of multi-gene cassettes as they are short in length and provide comparable expression levels to the native mammalian promoter.

Thesis (M. Phil.)--University of Hong Kong, 1999. / Includes bibliographical references (leaves 81-104).

Thesis (doctoral)--Freie Universität Berlin, 1979.

Thesis (Ph.D.)--Boston University / The role of specific streptococcal products in the production of infection by these organisms has been the subject of many studies by previous investigators. To a great extent these workers have attempted to correlate the formation of these substances by certain streptococcal strains with their virulence and disease manifestations in man as well as with the lethal effects produced by these organisms in mice. The purpose of this study was to investigate further the role of individual streptococcal products in the pathogenicity of group A streptococci, mainly by observing the effects of these products on local and systemic infections in the hamster and the mouse, respectively. Specifically, the M protein, streptokinase, streptodornase, streptolysin 0, the erythrogenic toxin, the hyaluronic acid capsule, and hyaluronidase were studied. [TRUNCATED]

This study was designed to examine the immunological privilege of the hamster cheek pouch. A comparison of the cheek pouch with the non-immunologically privileged flank was carried out by investigating responses to Glaxo BCG at the tissue, cellular and humoral levels using Histological methods, Mantoux Tests, Lymphocyte Transformation Tests, Passive Haemagglutination Tests and Immunoelectrophoresis Tests. These studies showed no differences in the responses to BCG in the cheek pouch or flank but do not preclude the possibility that the responses elicited in the cheek pouch differed qualitatively or quantitatively from those in the flank. In response to a single injection of BCG into the cheek pouch or flank compact and non-caseating epithelioid cell granulomas developed locally, with no evidence of the infection extending into the surrounding tissue or progressing to peripheral sites in the body. These findings could be related to the low virulence of Glaxo BCG for the hamster. Animals that had received a single injection of BCG or a rechallenge injection of BCG in Incomplete Freund's Adjuvant (IFA), were Mantoux negative and may also be related to the low virulence of Glaxo BCG for the hamster. The levels of specific and non-specific lymphopro-liferation to BCG and PHA, respectively were biologically insignificant in animals that had received a single injection of BCG and in those that had received a rechallenge injection of BCG in IFA. These findings can probably be attributed to the conditions of culture in the Lymphocyte Transformation Test. No specific agglutinating antibody was detected in the sera of animals that had received a single injection of BCG or a rechallenge injection of BCG in IFA. These findings could be related to the unreliability or lack of sensitivity of the Passive Haemagglutination Test.

A proteção induzida por uma vacina comercial anti-leptospirose importada, bivalente produzida com estirpes de referência dos sorovares Icterohaemorrhagiae e Canicola, foi comparada a promovida por uma bacterina experimental, bivalente produzida com as estirpes autóctones M64/06 e LO-4, respectivamente dos sorovares Copenhageni e Canicola, isoladas no Brasil e tipificadas com anticorpos monoclonais produzidos pelo Royal Tropical Institut, Laboratório de Referência da Organização Mundial de Saúde. O ensaio foi conduzido pelo teste de potência com desafio em hamsters. A vacina experimental, bivalente foi inativada com fenol a 37%, ajustada para concentração de 108 leptospiras por mL, por sorovar e produzida nas versões com e sem adjuvante hidróxido de alumínio a 0,15%. Os hamsters foram imunizados com 0,25 mL de vacina, via subcutânea em duas aplicações com 15 dias de intervalo. Os desafios foram efetuados com as estirpes LO4 (Canicola) ou L1-130 (Copenhageni) isoladas no Brasil, respectivamente nas diluições de 10-3 e 10-1 no volume de 0,5 mL via intraperitoneal por animal, 15 dias após a última dose da vacina. A estirpe de desafio do sorovar Copenhageni foi caracterizada, por anticorpos monoclonais, como idêntica a M64/06 e foi escolhida devido a apresentar melhor regularidade em termos de patogenicidade para os hamsters. Decorridos 20 dias do desafio os hamsters sobreviventes foram eutanasiados em câmara de CO2, necropsiados e nos seus rins foi efetuada a pesquisa do estado de portador de leptospiras por cultivo em meio de Fletcher. Nos desafios efetuados com a estirpe autóctone, L1-130 do sorovar Copenhageni foi obtida a proteção contra a doença clínica em oito de dez animais, para as duas vacinas experimentais, no entanto, entre os sobreviventes, houve portadores renais de leptospiras, em maior número para a vacina sem adjuvante (n=5), quando comparado a vacina acrescida de Al(OH)3, (n=3); para a vacina comercial a proteção foi total, tanto contra a doença como quanto a infecção não havendo qualquer morte por leptospirose e todos os cultivos renais dos sobreviventes foram negativos para leptospiras. Nos desafios efetuados com a estirpe autóctone, LO4 do sorovar Canicola, a proteção conferida pelas três vacinas ensaiadas foi total, não havendo mortes por leptospirose e todos os sobreviventes apresentaram resultados negativos para leptospiras nos cultivos de tecido renal. Os números de DL50 empregadas nos inóculos de desafio foram superiores a 10.000, para os dois sorovares ensaiados, pois na titulação dos inóculos morreram mais do que 50% dos animais até a última diluição testada. Foi, portanto, demonstrado a existência de proteção cruzada entre os sorovares Copenhageni e Icterohaemorrhagiae e que a bacterina importada foi capaz de induzir proteção contra a doença e contra a infecção para estirpes de leptospiras autóctones dos sorovares Canicola e Copenhageni isoladas no Brasil. Nos desafios efetuados com o sorovar Canicola a vacina experimental induziu proteção contra a doença e contra a infecção, mas nos que empregaram o sorovar Copenhageni houve proteção apenas contra a doença, constatando-se menor número de portadores renais de leptospiras entre os animais imunizados com a bacterina experimental acrescida do adjuvante de hidróxido de alumínio. / The protection induced by an imported commercial bivalent anti-leptospirosis vaccine produced with the reference strain of the serovars Icterohaemorrhagiae and Canicola was compared to one promoted by an experimental, bivalent bacterial vaccine produced with the indigenous strains M64/06 and LO-4, respectively of the serovars Copenhageni and Canicola isolated in Brazil and typified with monoclonal antibodies produced by the Royal Tropical Institut, Reference Laboratory of the World Health Organization. The assay was conducted by using the potency test with challenge in hamsters. The experimental bivalent vaccine was inactivated with 37% phenol and adjusted at a concentration of 108 leptospires/mL, for each serovar and produced in the versions with and without adjuvant (0.15% aluminium hydroxide). The hamsters were immunized by 0.25 mL of vaccine, by subcutaneous route in two applications with 15 days of interval. Fifteen days after receiving the last dose of the vaccine, the challenges were performed by employing the strains LO4 (Canicola) or L1-130 (Copenhageni), which were isolated in Brazil, respectively at the dilution of 10-3 and 10-1, in the volume of 0.5 mL/each animal by the intraperitoneal route. The challenge strain serovar Copenhageni has been characterized as identical to M64/06 strain by means of monoclonal antibodies and it was chosen due to its better regularity in terms of pathogenicity determined in hamsters. After 20 days of the challenge, the surviving hamsters were sacrificed in CO2 chamber, submitted to necropsy and the kidneys were examined to test the carrier state of leptospires by cultivation in Fletcher\'s medium. In the challenges effectuated with the indigenous strain, L1-130 of the serovar Copenhageni the protection was obtained against the clinical disease in eight of ten animals, for two experimental vaccines, however, among the survivors, there were renal carriers of leptospires, in higher number for the vaccine without containing the adjuvant (n=5), when compared to the one added with Al (OH) (n=3). For the commercial vaccine, the protection was total both against the disease and as for infection, when no death was found in consequence of leptospirosis and all the renal cultures of the survivors were negative for leptospires. In the challenges performed with the indigenous strain LO4 of the serovar Canicola, the protection conferred by the three vaccines tested was total, without observing any deaths due to leptospirosis and all the survivors presented negative culturing results for leptospires in the renal tissue. The challenge doses of the inocula were superior to 10,000 LD50, for the two serovars tested, since in the titration of the inocula have died more than 50 % of the animals up to the last tested dilution. So, the crossed protection existing between the serovars Copenhageni and Icterohaemorrhagiae was demonstrated and that the imported bacterial vaccine was able to induce protection against the disease and against the infection for the native strains of leptospires serovars Canicola and Copenhageni isolated in Brazil. In the challenges effectuated with the serovar Canicola, the experimental vaccine induced protection against the disease and against the infection, but in vaccine employing the serovar Copenhageni, protection was found only against the disease, when less number of renal carriers of leptospires was observed among the animals immunized with the experimental vaccine added with the aluminium hydroxide adjuvant.

Thesis (Ph. D.)--University of Texas at Austin, 2006. / Vita. Includes bibliographical references.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A passagem de imunoglobulinas, antÃgenos parasitÃrios solÃveis circulantes, cÃlulas imunes, citocinas ou outros fatores solÃveis atravÃs da placenta, podem induzir no feto uma resposta imunolÃgica frente a uma infecÃÃo homÃloga à materna, resultando em supressÃo ou em estimulaÃÃo da resposta imune. Estudos demonstram uma maior susceptibilidade dos machos à infecÃÃo por leishmania, porÃm a relaÃÃo entre o sexo e a susceptibilidade à leishmaniose no subgÃnero Viannia raramente à descrita. Foi avaliada a influÃncia do gÃnero e da infecÃÃo materna na resposta à Leishmania braziliensis em filhotes de hamster. Foram utilizados trinta e nove filhotes de hamster de mÃes infectadas e de mÃes nÃo-infectadas, de ambos os sexos, com 04 semanas de vida e pesos aproximados, infectados com L. braziliensis (MHOM/BR/94/H-3227). Foram estudados o tamanho da lesÃo da pata, a carga parasitÃria em patas e linfonodos de drenagem, ELISA e histopatolÃgico. ApÃs a anÃlise dos dados, concluÃmos que os filhotes machos apresentaram maior susceptibilidade à infecÃÃo por L. braziliensis, independente se nasceram de mÃe infectada ou de mÃe nÃo infectada, apresentando um padrÃo de resposta mais indolente do que as fÃmeas. A infecÃÃo materna por L. braziliensis em hamsters nÃo influenciou o curso da doenÃa na infecÃÃo homÃloga dos filhotes. A resposta inflamatÃria mais intensa nos filhotes nascidos de mÃe infectada possivelmente pode ser correlacionada a uma maior induÃÃo de molÃculas inflamatÃrias, citocinas e quimiocinas, embora isso nÃo tenha sido avaliado no presente trabalho. / Immunoglobulins, soluble parasite circulating antigens, immune cells, citokines and other cell-related products can be transferred from infected mothers to their young, leading to suppression or stimulation of the immune response to a homologous antigen. Several studies demonstrated an increased susceptibility to leishmania infection in males, however, a relation between gender and susceptibility on the subgenus Viannia is rarely described. The influence of gender and maternal infection to the Leishmania braziliensis response in hamster offspring were evaluated. Thirty-nine hamster offspring of infected mothers and non-infected mothers from both genders, with 04 weeks-old, similar weights and infected with the Leishmania braziliensis strain MHOM/BR/94/H-3227 were analyzed. Studies on footpad lesion size, parasite load on footpad and draining lymph nodes, ELISA and histopathology were conducted. After analyzing the data, we concluded that the male offspring had higher susceptibility to infection by L. braziliensis, regardless if they were born from an infected mother or a non-infected mother, and had presented a more indolent pattern of response than the females. The maternal infection by L. braziliensis in hamsters did not influence the course of the disease in the infection rate of the offspring. The more intense inflammatory response in the offspring born from an infected mother can possibly be correlated to a greater induction of inflammatory molecules, cytokines and chemokines, although this has not been evaluated in this study.

A proteção induzida por uma vacina comercial anti-leptospirose importada, bivalente produzida com estirpes de referência dos sorovares Icterohaemorrhagiae e Canicola, foi comparada a promovida por uma bacterina experimental, bivalente produzida com as estirpes autóctones M64/06 e LO-4, respectivamente dos sorovares Copenhageni e Canicola, isoladas no Brasil e tipificadas com anticorpos monoclonais produzidos pelo Royal Tropical Institut, Laboratório de Referência da Organização Mundial de Saúde. O ensaio foi conduzido pelo teste de potência com desafio em hamsters. A vacina experimental, bivalente foi inativada com fenol a 37%, ajustada para concentração de 108 leptospiras por mL, por sorovar e produzida nas versões com e sem adjuvante hidróxido de alumínio a 0,15%. Os hamsters foram imunizados com 0,25 mL de vacina, via subcutânea em duas aplicações com 15 dias de intervalo. Os desafios foram efetuados com as estirpes LO4 (Canicola) ou L1-130 (Copenhageni) isoladas no Brasil, respectivamente nas diluições de 10-3 e 10-1 no volume de 0,5 mL via intraperitoneal por animal, 15 dias após a última dose da vacina. A estirpe de desafio do sorovar Copenhageni foi caracterizada, por anticorpos monoclonais, como idêntica a M64/06 e foi escolhida devido a apresentar melhor regularidade em termos de patogenicidade para os hamsters. Decorridos 20 dias do desafio os hamsters sobreviventes foram eutanasiados em câmara de CO2, necropsiados e nos seus rins foi efetuada a pesquisa do estado de portador de leptospiras por cultivo em meio de Fletcher. Nos desafios efetuados com a estirpe autóctone, L1-130 do sorovar Copenhageni foi obtida a proteção contra a doença clínica em oito de dez animais, para as duas vacinas experimentais, no entanto, entre os sobreviventes, houve portadores renais de leptospiras, em maior número para a vacina sem adjuvante (n=5), quando comparado a vacina acrescida de Al(OH)3, (n=3); para a vacina comercial a proteção foi total, tanto contra a doença como quanto a infecção não havendo qualquer morte por leptospirose e todos os cultivos renais dos sobreviventes foram negativos para leptospiras. Nos desafios efetuados com a estirpe autóctone, LO4 do sorovar Canicola, a proteção conferida pelas três vacinas ensaiadas foi total, não havendo mortes por leptospirose e todos os sobreviventes apresentaram resultados negativos para leptospiras nos cultivos de tecido renal. Os números de DL50 empregadas nos inóculos de desafio foram superiores a 10.000, para os dois sorovares ensaiados, pois na titulação dos inóculos morreram mais do que 50% dos animais até a última diluição testada. Foi, portanto, demonstrado a existência de proteção cruzada entre os sorovares Copenhageni e Icterohaemorrhagiae e que a bacterina importada foi capaz de induzir proteção contra a doença e contra a infecção para estirpes de leptospiras autóctones dos sorovares Canicola e Copenhageni isoladas no Brasil. Nos desafios efetuados com o sorovar Canicola a vacina experimental induziu proteção contra a doença e contra a infecção, mas nos que empregaram o sorovar Copenhageni houve proteção apenas contra a doença, constatando-se menor número de portadores renais de leptospiras entre os animais imunizados com a bacterina experimental acrescida do adjuvante de hidróxido de alumínio. / The protection induced by an imported commercial bivalent anti-leptospirosis vaccine produced with the reference strain of the serovars Icterohaemorrhagiae and Canicola was compared to one promoted by an experimental, bivalent bacterial vaccine produced with the indigenous strains M64/06 and LO-4, respectively of the serovars Copenhageni and Canicola isolated in Brazil and typified with monoclonal antibodies produced by the Royal Tropical Institut, Reference Laboratory of the World Health Organization. The assay was conducted by using the potency test with challenge in hamsters. The experimental bivalent vaccine was inactivated with 37% phenol and adjusted at a concentration of 108 leptospires/mL, for each serovar and produced in the versions with and without adjuvant (0.15% aluminium hydroxide). The hamsters were immunized by 0.25 mL of vaccine, by subcutaneous route in two applications with 15 days of interval. Fifteen days after receiving the last dose of the vaccine, the challenges were performed by employing the strains LO4 (Canicola) or L1-130 (Copenhageni), which were isolated in Brazil, respectively at the dilution of 10-3 and 10-1, in the volume of 0.5 mL/each animal by the intraperitoneal route. The challenge strain serovar Copenhageni has been characterized as identical to M64/06 strain by means of monoclonal antibodies and it was chosen due to its better regularity in terms of pathogenicity determined in hamsters. After 20 days of the challenge, the surviving hamsters were sacrificed in CO2 chamber, submitted to necropsy and the kidneys were examined to test the carrier state of leptospires by cultivation in Fletcher\'s medium. In the challenges effectuated with the indigenous strain, L1-130 of the serovar Copenhageni the protection was obtained against the clinical disease in eight of ten animals, for two experimental vaccines, however, among the survivors, there were renal carriers of leptospires, in higher number for the vaccine without containing the adjuvant (n=5), when compared to the one added with Al (OH) (n=3). For the commercial vaccine, the protection was total both against the disease and as for infection, when no death was found in consequence of leptospirosis and all the renal cultures of the survivors were negative for leptospires. In the challenges performed with the indigenous strain LO4 of the serovar Canicola, the protection conferred by the three vaccines tested was total, without observing any deaths due to leptospirosis and all the survivors presented negative culturing results for leptospires in the renal tissue. The challenge doses of the inocula were superior to 10,000 LD50, for the two serovars tested, since in the titration of the inocula have died more than 50 % of the animals up to the last tested dilution. So, the crossed protection existing between the serovars Copenhageni and Icterohaemorrhagiae was demonstrated and that the imported bacterial vaccine was able to induce protection against the disease and against the infection for the native strains of leptospires serovars Canicola and Copenhageni isolated in Brazil. In the challenges effectuated with the serovar Canicola, the experimental vaccine induced protection against the disease and against the infection, but in vaccine employing the serovar Copenhageni, protection was found only against the disease, when less number of renal carriers of leptospires was observed among the animals immunized with the experimental vaccine added with the aluminium hydroxide adjuvant.

Ho Wing Ki Vicky. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 191-211). / Abstracts in English and Chinese. / Abstracts --- p.i / Abstracts in Chinese --- p.iii / Acknowledgements --- p.v / Table of Abbreviations --- p.vi / Table of Contents --- p.ix / List of Figures and Tables --- p.xiv / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1. --- The Male Accessory Sex Glands --- p.2 / Chapter 1.1.1. --- The Prostatic Complex --- p.4 / Chapter 1.1.2. --- The Coagulating Gland --- p.4 / Chapter 1.1.3. --- The Seminal Vesicles --- p.5 / Chapter 1.1.4. --- The Ampullary Gland --- p.6 / Chapter 1.1.5. --- The Cowper's gland --- p.6 / Chapter 1.1.6. --- The Littre's Gland --- p.7 / Chapter 1.2. --- The Uterus --- p.7 / Chapter 1.2.1. --- The Structure of Uterus --- p.7 / Chapter 1.2.2. --- The Endometrium --- p.8 / Chapter 1.3. --- Identifications of Proteins by Two Dimensional Gel Electrophoresis and Matrix-assisted Laser Adsorption / ionization Time-of-flight (MALDI-TOF) Mass Spectrometry --- p.9 / Chapter 1.3.1. --- Principle of Two Dimensional Gel Electrophoresis --- p.9 / Chapter 1.3.2. --- Principle of MALDI-TOF Mass Spectrometry --- p.11 / Chapter 1.4. --- Hypothesis and Aim of this Study --- p.12 / Chapter 1.4.1. --- Hypothesis --- p.13 / Chapter 1.4.2. --- Aim --- p.13 / Legends and Figures --- p.14 / Chapter Chapter 2 --- The Effect of Omission of Male Accessory Sex Gland Secretions on Post Coital Uterine Fluid --- p.17 / Chapter 2.1. --- Introduction --- p.18 / Chapter 2.1.1. --- Uterine Fluid at the Time of Mating --- p.18 / Chapter 2.1.2. --- Ventral Prostate Secretory Proteins --- p.20 / Chapter 2.1.3. --- Proteins Specifically Bind to Sperm Membrane --- p.22 / Aim of Study --- p.23 / Chapter 2.2. --- Materials and Methods --- p.24 / Chapter 2.2.1. --- Animal Model --- p.24 / Chapter 2.2.1.1. --- Female golden hamsters --- p.24 / Chapter 2.2.1.2. --- Male golden hamsters --- p.25 / Chapter 2.2.2. --- Collections of Biological Samples --- p.26 / Chapter 2.2.2.1. --- Female Hamster Serum --- p.26 / Chapter 2.2.2.2. --- Uterine Fluid --- p.26 / Chapter 2.2.2.3. --- Uterine Tissues --- p.27 / Chapter 2.2.2.4. --- Ejaculated Sperm in Uterus --- p.27 / Chapter 2.2.2.5. --- ASG Secretions --- p.27 / Chapter 2.2.2.6. --- ASG Tissues --- p.28 / Chapter 2.2.2.7. --- Epididymal Sperm --- p.28 / Chapter 2.2.3. --- Isolation and Purification of Sperm Binding Ventral Prostate Proteins (SBVPP) --- p.28 / Chapter 2.2.3.1. --- Plasma Membrane Protein from Epididymal Sperm --- p.28 / Chapter 2.2.3.2. --- Affinity Chromatography --- p.29 / Chapter 2.2.3.3. --- FPLC Gel Filtration --- p.30 / Chapter 2.2.4. --- Histology of Flushed Uterus --- p.30 / Chapter 2.2.5. --- Quantification of Protein --- p.31 / Chapter 2.2.6. --- Two dimensional Electrophoresis --- p.31 / Chapter 2.2.6.1. --- First DimensiońؤIsoelectric Focusing --- p.32 / Chapter 2.2.6.2. --- Second DimensiońؤSDS PAGE --- p.32 / Chapter 2.2.6.3. --- Visualization of Protein Spots --- p.33 / Chapter 2.2.7. --- Identification of Proteins --- p.34 / Chapter 2.2.7.1. --- Protein Spots Detection --- p.34 / Chapter 2.2.7.2. --- MALDI-TOF --- p.34 / Chapter 2.2.7.2.1. --- In-Gel Digestion for MALDI-TOF --- p.34 / Chapter 2.2.7.2.2. --- MALDI-TOF Analysis --- p.35 / Chapter 2.2.7.2.3. --- MALDI-TOF Data Analysis --- p.36 / Chapter 2.2.8. --- Confirmation of Prostate Specific Antigen (PSA) --- p.37 / Chapter 2.2.9. --- Confirmation of Prostatic Acid Phosphatase (PAP) --- p.37 / Chapter 2.2.10. --- Confirmation of Tumor Necrosis Factor Stimulated Gene-6 (TSG-6) --- p.37 / Chapter 2.2.11. --- Scanning Electron Microscopy (SEM) --- p.38 / Chapter 2.3. --- Results --- p.39 / Chapter 2.3.1. --- Morphology of Flushed Uterus --- p.39 / Chapter 2.3.2. --- Protein Concentration --- p.39 / Chapter 2.3.3. --- "Serum, Pre and Post Coital Uterine Fluid Protein Profiles" --- p.39 / Chapter 2.3.4. --- ASG Secretory Proteins and Their Fate After mating --- p.40 / Chapter 2.3.4.1. --- ASG Secretory Proteins and 1 hp.c. Uterine Fluid --- p.40 / Chapter 2.3.4.2. --- VP Secretory Proteins and SBVPP --- p.40 / Chapter 2.3.4.3. --- PAP and PSA --- p.41 / Chapter 2.3.5. --- Protein Profile at 1 h p.c --- p.41 / Chapter 2.3.6. --- Protein Profile at 4 h p. c --- p.42 / Chapter 2.3.7. --- Identification of Proteins Differentially Expressed by the Three Groups at 1 h p .c --- p.43 / Chapter 2.3.7.1. --- Confirmation of TSG-6 --- p.43 / Chapter 2.3.8. --- SEM --- p.44 / Chapter 2.4. --- Discussion --- p.45 / Chapter 2.5. --- Conclusion --- p.55 / Legends and Figures --- p.56 / Chapter Chapter 3 --- The Effect of Omission of Male Accessory Sex Glands Secretions on Periimplanation Uterine Fluid Proteins --- p.114 / Chapter 3.1. --- Introduction --- p.115 / Chapter 3.1.1. --- Cytokine & Growth Factor --- p.115 / Chapter 3.1.1.1. --- Vascular Endothelial Growth Factor (VEGF) --- p.115 / Chapter 3.1.1.2. --- Leukaemia Inhibitory Factor (LIF) --- p.116 / Chapter 3.1.1.3. --- Insulin-like Growth Factors (IGFs) --- p.116 / Chapter 3.1.1.4. --- Interleukin 1 (IL-1) --- p.117 / Chapter 3.1.1.5. --- Adhesion Molecule --- p.118 / Chapter 3.1.2. --- Uterine Secretory Proteins --- p.118 / Chapter 3.1.2.1. --- Estrogen Dependent Protein --- p.119 / Chapter 3.1.2.2. --- Progestin-Dependent Endometrial Protein --- p.120 / Chapter 3.1.2.3. --- Uteroglobin --- p.121 / Chapter 3.1.2.4. --- Uteroferrin --- p.122 / Chapter 3.1.2.5. --- Prolactin --- p.123 / Chapter 3.1.2.6. --- Glycodelin --- p.123 / Chapter 3.2. --- Aim of Study --- p.124 / Chapter 3.3. --- Method and Materials --- p.124 / Chapter 3.3.1. --- Animal and Surgery --- p.125 / Chapter 3.3.2. --- Collection of Biological Samples --- p.125 / Chapter 3.3.2.1. --- Uterine Fluid --- p.125 / Chapter 3.3.2.2. --- Uterine Tissue --- p.125 / Chapter 3.3.3. --- Quantification of Protein --- p.125 / Chapter 3.3.4. --- Two Dimensional Electrophoresis --- p.125 / Chapter 3.3.5. --- Identification of Proteins --- p.126 / Chapter 3.3.6. --- Confirmation of Growth Differentiation Factor-8 (DGF-8) by PCR --- p.126 / Chapter 3.3.7. --- Confirmation of GDF-8 by Western Blotting --- p.126 / Chapter 3.4. --- Results --- p.128 / Chapter 3.4.1. --- Protein Concentration --- p.128 / Chapter 3.4.2. --- 60 Hours Post Coitum --- p.128 / Chapter 3.4.2.1. --- Protein Profile --- p.128 / Chapter 3.4.2.2. --- Differentially Expressed Protein Spots and Proteins Identification --- p.129 / Chapter 3.4.3. --- 72 Hours Post Coitum --- p.129 / Chapter 3.4.3.1. --- Protein Profile --- p.129 / Chapter 3.4.3.2. --- Differentially Expressed Protein Spots and Proteins Identification --- p.130 / Chapter 3.4.3.3. --- Confirmation of Growth differentiation factor 8 (GDF-8) --- p.130 / Chapter 3.5. --- Discussion --- p.132 / Chapter 3.6. --- Conclusion --- p.139 / Legends and Figures --- p.140 / Chapter Chapter 4 --- General Conclusion --- p.186 / Chapter 4.1. --- Conclusion --- p.187 / Chapter 4.2. --- Further Studies --- p.190 / References --- p.191 / Appendices --- p.212