Physiology - Theses

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    The structure and expression of the porcine relaxin gene
    Haley, John Douglas ( 1984)
    Relaxin is primarily a hormone of pregnancy, discovered in 1926 by Hisaw, which acts on connective tissue to lengthen the pubic ligaments and soften the cervix in preparation of parturition. Relaxin has a role in inhibiting myometrial contractility in pregnancy and less well defined functions in ovulation, mammary gland growth in pregnancy, implantation and maintainance of the fetus, in the timing of parturition, an in the male reproductive tract. It is produced primarily in the corpus luteum of pregnancy in most species studied, though relaxin activity has been detected in the non-pregnant ovary of both follicular and luteal origin, in the uterus, in the placentae and associated membranes, in male reproductive organs, and interestingly in several non-mammalian species. Relaxin is a 6,300 dalton polypeptide hormone sharing considerable structural homology with insulin and the insulin-like growth factors and they are likely derived from a single ancestral gene.
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    Studies of purification and biological action of synergistic factor from WEHI-3B conditioned medium
    McNiece, Ian K. ( 1986)
    Media conditioned by a murine myelomonocytic leukemic cell line (WEHI-3B) have been found to produce a haemopoietic growth factor, synergistic factor (SF), which in combination with a source of macrophage-colony stimulating factor (M-CSF), acts on a primitive macrophage progenitor cell to promote the growth of large colonies in agar. These progenitor cells are referred to as high proliferative potential colony forming cells (HPP-CFC) and have been detected in bone marrow from normal mice and mice which have been treated with 5-fluorouracil. The production of SF from WEHI-3B (S SFW) by a variety of methods has been investigated and the highest concentrations of SFW have been obtained by conditioning serum free medium with WEHI-3B cells which have been maintained by transplantation as an ascites tumour in BALB/c mice. The SFW activity was unaffected by storage at 4°C and -20°C, was stable between pH 2.0 and 10.6 and to heat up to 50°C but not above. However, the SFW was unstable to trypsin digestion, 8 M urea at pH 8.0 with and without Mercaptoethanol (ME, 4 mM), and to ME alone at pH 8.0. The SFW has an apparent molecular weight of 25,000 (determined by gel filtration chromatography), it has an isoelectric point in the range of pH 4.0 to 6.5 and does not bind to Con-A Sepharose. A number of separative techniques were investigated in an attempt to purify the SFW from the WEHI-3B CM. A four step purification schedule was devised consisting of anion exchange chromatography, phenyl sepharose chromatography, gel filtration on Sephadex G-100 and mono-0 anion exchange on high pressure liquid chromatography. This schedule resulted in approximately a 128 fold increase in the specific activity with a recovery of approximately 4% of the initial SFW activity. (For full summary open document)