Recombinant human macrophage colony stimulating factor beta (rhm-CSFβ) is a glycoprotein that stimulates the proliferation, differentiation and survival of cells belonging to the monocyte-macrophage lineage. It contains nine inter-subunit and intra-subunit disulfide bonds and represents an excellent model system for studying disulfide bond formation during protein folding because the assembly of its monomeric subunits and the maturation of its biological activity depend on the progressive formation of the correct disulfide structure during in vitro folding. Knowledge obtained from these studies can be potentially useful in understanding the roles of disulfide bond formation during protein folding in general. rhm-CSF8 was modified by partial reduction of disulfide bonds, yielding CN¹⁵⁷'¹⁵⁹-modified rhm-CSFβ. The modification did not affect the biological activity, stability, or the overall conformation of the protein. However, the C-terminal regions near the modification sites were shown to exhibit faster deuterium exchange behavior as a result of the chemical modification, indicating that the C-terminal regions became more flexible. Folding kinetics of rhm-CSFβ and CN¹⁵⁷'¹⁵⁹-modified rhm-CSFβ were shown to be essentially the same, suggesting that the modification did not affect the folding kinetics of the oxidized rhm-CSFβ. The denatured and reduced rhm-CSFβ was refolded with the aid of a chemical oxidant. The data indicated that the in vitro folding rhm-CSFβ proceeded via multiple pathways involving monomeric and dimeric intermediates. Disulfide bond shuffling catalyzed by GSH/GSSG represented an important isomerization step in folding. A dimeric intermediate, D-SS8-cam2, was isolated and identified as a kinetic trap, perhaps requiring significant structural arrangement to convert to the native protein. The heterogeneous folding mixture detected by both disulfide bond quenching and H/D pulsed labeling indicate that rhm -CSFβ folding is a diffusion like process as described by the folding funnel model. / Graduation date: 2003

Thyroid hormones are well known important to be in development and growth in birds and that signaling of thyrotropin (TSH) regulates the photo induced seasonal reproduction. A mutation at the thyroid stimulating hormone receptor (TSHR) gene in domestic breeds of chicken could be involved in the release of the photoperiodic regulation. Furthermore, TSH can affect a wide range of domestication related phenotypes, such as behaviour, growth rate and pigmentation. The aim of this study was to investigate the behaviours expressed in the different genotypes on the TSHR gene in chickens. Four standard tests were conducted, aerial predator, fear of human, social dominance and tonic immobility. An advanced intercross line of chickens between red junglefowl and White leghorn was used. Male domestic type chickens explored more, showed more less fear behaviours and showed least fear behaviours in the fear of human test. Increased activity and flight response has been interpreted as a lower fear response, which is in line with this study. The wild type chickens showed more social dominance than domestic type chickens which are in line with previous results. In tonic immobility there was a difference between the wild type male and heterozygous male chickens in latency until first head movement. The conclusion of this study is that there is a difference between the wild type and domestic type chickens. This indicates that the TSHR gene is involved in behavioural changes during domestication, but whether it is due to passive or active selection is the question.

Thesis (Ph. D.)--West Virginia University, 2002. / Title from document title page. Document formatted into pages; contains viii, 71 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 61-70).

Thesis (Ph. D.)--Washington State University, December 2009. / Title from PDF title page (viewed on Dec. 16, 2009). "School of Molecular Biosciences." Includes bibliographical references.

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Factors affecting bovine ovarian responsiveness to stimulation by pregnant mare's serum gonadotropin (PMSG) were studied. Initially, the effects of plasma progesterone concentration on the response were considered using a progesterone-releasing intravaginal device (PRID) to provide an artificial source of the hormone. Due to inherent biological variation in vivo, in vitro methods were developed in which cAMP and progesterone production by granulosa cells were measured. Regardless of the size of the follicles from which the cells originated, PMSG stimulated significant cAMP accumulation. Cyclic AMP production was similar between aspirated granulosa cells and those scraped from the follicle wall, between ovaries with and without a corpus luteum from the same animal, and between follicles from animals early ( 10 days) in their estrous cycles. The PMSG failed to stimulate bovine granulosa cells to synthesize significantly more progesterone than untreated cells. Unlike porcine granulosa cells, bovine cells from antral follicles of any size appeared to luteinize spontaneously in culture. / Hormonal changes in the preovulatory follicle were measured using the PMSG/hCG-treated prepubertal gilt as a model. After hCG administration, follicular fluid levels of cAMP peaked at 4 hr followed 24 hr later by a rise in prostaglandins F and E (PGF, PGE) concentrations which peaked near the expected time of ovulation. Indomethacin injection blocked ovulation and the prostaglandin rise, although the inhibition could be reversed by the administration of PGF(,2)(alpha). Temporal changes in estrone, estradiol-17(beta), progesterone, androstenedione, testosterone and 5 (alpha)-dihydrotestosterone were also measured. / In an effort to decrease endogenous levels of inhibin, thereby increasing endogenous FSH and thence follicular development, heifers, ewes and does were actively immunized against porcine follicular fluid or proteinaceous fractions of bovine seminal plasma. In some animals, plasma FSH concentrations were elevated although ovulation rates and estrous cycle lengths were not altered. / A culdoscopy technique was developed for repeated monitoring of ovarian morphological changes in cows.

Pre-clinical studies suggest that granulocyte-colony stimulating factor (GCSF) holds promise for the treatment of ischaemic heart disease (IHD). However, its safety and efficacy in this setting, and in particular in patients with chronic refractory 'no-option' IHD, is unclear. Therefore, a clinical study was initiated in 20 such 'no-option' patients, with the aim of assessing the safety and efficacy of both G-CSF administration, and also, that of an intracoronary infusion of G-CSF mobilised CD133+ cells. The study involved initial baseline cardiac ischaemia assessment (symptom based questionnaire, exercise stress test (EST), nuclear Sestamibi (MIBI) and dobutamine stress echocardiographic (DSE) imaging). Stable 'no-option' IHD patients then received open-label G-CSF commencing at 10μg/kg s/c for five days, with an EST on days four and six (to facilitate myocardial cytokine generation and stem cell trafficking). After three months, cardiac ischaemia assessment and the same regimen of G-CSF and ESTs were repeated, but in addition, leukapheresis and then a randomised double-blinded intracoronary infusion of CD133+ or unselected cells were performed. Final cardiac ischaemia assessment was three months thereafter. Eighteen male and two female subjects (mean age 62.4) were enrolled. Eight events occurred that fulfilled pre-specified 'adverse event' criteria: four ischaemic (troponin positive) episodes, two episodes of transient thrombocytopaenia (one profound), one episode of gout and one unscheduled hospitalisation for exhaustion. Troponin was positive on 17 further occasions (all CK-MB negative), however, at these instances angina severity was identical to baseline. Importantly, no adverse event(s) resulted in any detectable long-term adverse sequelae for any subject. From baseline to final follow-up, the administration of two cycles of G-CSF was associated with statistically significant improvements in a range of subjective outcomes, including anginal symptoms, quality of life and EST performance (all p < 0.005). However, the objective MIBI and DSE scans showed only trends towards improvement (all p > 0.1). Compared to unselected cells, an intracoronary infusion of CD133+ cells did not improve either subjective or objective outcomes. In conclusion, administering G-CSF to patients with refractory 'no-option' IHD warrants careful monitoring, but may be performed with safety. A larger, randomised double-blind placebo-controlled trial of G-CSF in these patients appears warranted.

Amendments to thesis in pocket on back cover. / Copy of author's previously published article in pocket on back cover. / Bibliography: leaves 124-172. / viii, 172, [101] leaves, [22] leaves of plates : ill. (chiefly col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Studies the biological effects and leukaemic potential of h[beta]c mutants using murine models. / Thesis (Ph.D.)--University of Adelaide, Dept. of Medicine, 1999?

Errata posted inside back end-paper (leaf 250). / Bibliography: leaves 204-249. / xxiv, 250 leaves, [8] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Investigates the nature of the seminal vesicle-derived trigger(s) that stimulate the granulocyte-macrophage colony-stimulating factor release from the murine uterine epithelium. / Thesis (Ph.D.)--University of Adelaide, Dept. of Obstetrics and Gynaecology, 1999

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.