Although the osones were first prepared by Emil Fischer sixty-five years ago and, in the intervening period, investigation of these compounds has not been neglected, there exists no modern comprehensive review of their chemistry and biochemistry. In consequence, Part I of this thesis is a full and critical survey of work published on the osones up to June 1953. Part II, a discussion of the methods and results of the author's own research on the subject, is presented under the same arrangement of sub-titles as that used in Part I. The object of this research was to investigate the preparation and properties of the osones, in particular D-glucosone, and their derivatives by the application of both modern and classical methods of carbohydrate chemistry, with a view to establishing the structural features of these compounds. An attempt has been made to correlate the author;s own results with those of other workers and, at the same time, to develop lines of attack which have hitherto received no attention. Part III is a detailed report of the author's experimental work. The material presented stands as a contribution to carbohydrate chemistry. That further investigation of the osones is justified may be realised from consideration of the following facts: no completely satisfactory methods for the preparation, characterisation, and estimation and no preparation of a crystalline derivative from which the ozone may be readily regenerated has been described; there is little definitive knowledge of ozone structure. In addition, the elucidation of the precise chemical structure of D-glucose and its behaviour in solution is essential for the complete understanding of the biological significance of the compound. 1. A critical and comprehensive review of the published work on the preparation, properties, structure, and biological significance of the osones is presented. 2. A full experimental investigation of the methods of preparation of osones is reported. By the introduction of modifications established methods have been adapted for the preparation of osones, possessing a high degree of purity, in higher yields than hitherto obtained; a number of new preparative procedures are described. 3. The physical properties of glucosone, including rotational behaviour, ultraviolet absorption spectrum, chromatographic analysis, and ionophoretic analysis of aqueous solutions, properties which have hitherto received little or no attention, have been studied; the results are discussed in relation to the structural features of the osone and to the detection and identification of glucosone when in admixture with other sugars or in biological material. 4. The effect of acids and of alkalis on glucosone has been investigated and the preparation and properties of a number of ozone derivatives with substituted hydrazines are described. Methods of estimation of glucosone in the presence of other sugars have been evolved. 5. The preparation, properties, and structures of isopropylidene derivatives of a number of osones are described. 1:2-2:3-5:6-Tri-o-isopropylidene d-and l-glucosone hydrate and 1:2-2:3-5:6-tri-o-isopropylidene l-gulosone hydrate were obtained, the first crystalline derivatives from which the corresponding osones may be readily regenerated. 6. The possible structures which may be assigned to d-glucosone on the available evidence are discussed.

Indolizine was first synthesised by Scholts1 in 1912 who proposed at that time the presently accepted ring structure (1). A number of alternative names and systems of numbering have been suggested for the heterocycle. These include pyrindole, pyrroline, pyrrosoline, 9 pyrrolopyridine and pyrrolo [1,2-a] pyridine. It is proposed to use the name indolozine and the numbering as shown in (1), in accordance with the recommendations of the I.U.P.A.C. Direct support for Scholts' formulation of the structure of indolizine followed from its catalytic reduction2 showing the presence of four double bonds, to a derivative identical with d-coniceine (a) (ostahydtoindolizine) which on degradation with ayanogen bromide yielded a1 condine (9-n-gropylpieridine). Further evidence substantiating this formulation of indolizine arises from consideration of numerous syntheses described below.

The phase relations, electrical properties and structural characteristics of doped cubic stabilised Zirconia based electroceramic materials have been investigated using a number of characterisation techniques. The phase relations of the ternary systems ZrO2 -Y2O3 -TiO2 and ZrO2 -Gd2 O3 - TiO2 at 1500°C have been investigated. Electrical characterisation in air and in low oxygen partial pressures has been carried out using 2-probe A.C. Impedance Spectroscopy and 4-probe D.C. resistivity measurements to ascertain whether compositions within these systems could be utilised as the anode materials in Solid Oxide Fuel Cells. The effect of porosity on the ionic and electronic conducting properties of the ZrO2 -Y2 O3 -TiO2 system has been investigated to provide a clearer understanding of the effect of the porosity within candidate anode materials. The effect of Al2O3 additions on the electrical properties and stability of the Solid Oxide Fuel Cell material of choice, 8 mol% Yttria stabilised Zirconia, has been investigated. Al2 O3 has been found to remain primarily as a second phase within the 8YSZ, however a small quantity of Al3+ does dissolve into the fluorite matrix. Al2 O3 has been found to have a negligible effect on the high temperature ionic conductivity of 8YSZ and improves the resistance of 8YSZ to hydrothermal degradation by stabilising the cubic structure. High temperature Time of Flight Neutron Diffraction has been used to link the change in activation energy observed in 8YSZ to a break down in local ordering of oxygen ions. Extended X-ray absorption Fine Structure Spectroscopy has been used to characterise the local structure of the cations in 8 mol% Yttria-stabilised Zirconia. Analysis of the high temperature data reveals that the local structure is quite different from the average crystallographic structure. The oxygen vacancies were determined to be associated with Zirconium ions and found to disorder at high temperatures.

In this Masters dissertation, various doses of solubulized crude F4 fimbrial protein in conjunction with the adjuvants CpG ODN and porcine â-defensin 1 (pBD-1) were used to enhance the F4-specific intestinal immune response against Enterotoxigenic Escherichia coli (ETEC) F4 in post-weaned pigs. Using the mechanically shearing method we isolated the F4 fimbrial protein of ETEC with a molecular weight of 26 kDa. We verified this using a Western blot probed with a rabbit anti-F4 fimbrial antibody. Binding of the F4 fimbrial protein to the F4 receptor (F4R), present on the brush border of the villi in the small intestine of pigs, was demonstrated using an in vitro villus adhesion assay (IVVA). To demonstrate specificity rabbit polyclonal and mouse monoclonal anti-F4 antibodies, or the F4 protein were used to inhibit the adhesion of ETEC F4ac to F4R positive (F4Rpos) villi. <p>To examine immunogenicity of the 500 micrograms (ìg) of the F4 were administered into surgically created jejunal gut-loops in pigs. Three weeks later Peyers patches (PP) from immunized and control loops as well as gut-wall tissue were analyzed for their F4-specific antibody secreting cells (ASCs) by a modified enzyme linked immunosorbent spot (ELISPOT) assay. The F4-specific immune response in the serum was analyzed by an enzyme linked immunosorbant assay (ELISA). High numbers of F4-specific ASCs were isolated from the loops of pigs that contained high levels of the F4R. Conversely nominal or low numbers of F4-specific ASCs were found in loops of pigs expressing low levels of the F4R or no F4R (F4Rneg). The IVVA was used to categorize the pigs into either F4Rpos or F4Rneg animals. <p>Next three different concentrations of the crude F4 protein 50, 250, and 500 µg in the loops of individual pigs were used to analyze if dose affected the F4-specific immune response. Interestingly dose had no effect on the magnitude of the response. Therefore we hypothesized that the F4-specific immune response in the loops could be enhanced through the use of the adjuvants CpG ODN 2007 and pBD-1. The F4 protein was co-administered with either CpG ODN 2007 or pBD-1 and immune responses were assessed after 3 weeks. However neither CpG ODN 2007 nor PBD-1 at the doses used made an improvement in the immune response. Thus, these results demonstrated that the expression level of the F4R was the most important parameter for eliciting of the local immune response against the F4 protein. Furthermore our studies revealed that both F4Rneg and F4Rpos pigs responded to F4 immunization, however the former respond only nominally to F4-immunization in the loops. Moreover, an inverse relationship existed between the level of the F4-specific IgG in the serum and the F4-specific immune response seen in the loops. Thus our findings have important implications for oral vaccination using fimbrial based antigens (Ags) that utilize a receptor for their immunogenicity. Our results indicate that only animals with high levels of enterocyte F4R will have the ability to elicit high levels of protective F4-specific anti-fimbrial antibodies in their intestine after oral immunization. Therefore unless an effective adjuvant is available, animals with low to moderate levels of the fimbrial receptor in their small intestine will mount only weak immune responses making herd immunity after vaccination currently unattainable.

In this Masters dissertation, various doses of solubulized crude F4 fimbrial protein in conjunction with the adjuvants CpG ODN and porcine â-defensin 1 (pBD-1) were used to enhance the F4-specific intestinal immune response against Enterotoxigenic Escherichia coli (ETEC) F4 in post-weaned pigs. Using the mechanically shearing method we isolated the F4 fimbrial protein of ETEC with a molecular weight of 26 kDa. We verified this using a Western blot probed with a rabbit anti-F4 fimbrial antibody. Binding of the F4 fimbrial protein to the F4 receptor (F4R), present on the brush border of the villi in the small intestine of pigs, was demonstrated using an in vitro villus adhesion assay (IVVA). To demonstrate specificity rabbit polyclonal and mouse monoclonal anti-F4 antibodies, or the F4 protein were used to inhibit the adhesion of ETEC F4ac to F4R positive (F4Rpos) villi. <p>To examine immunogenicity of the 500 micrograms (ìg) of the F4 were administered into surgically created jejunal gut-loops in pigs. Three weeks later Peyers patches (PP) from immunized and control loops as well as gut-wall tissue were analyzed for their F4-specific antibody secreting cells (ASCs) by a modified enzyme linked immunosorbent spot (ELISPOT) assay. The F4-specific immune response in the serum was analyzed by an enzyme linked immunosorbant assay (ELISA). High numbers of F4-specific ASCs were isolated from the loops of pigs that contained high levels of the F4R. Conversely nominal or low numbers of F4-specific ASCs were found in loops of pigs expressing low levels of the F4R or no F4R (F4Rneg). The IVVA was used to categorize the pigs into either F4Rpos or F4Rneg animals. <p>Next three different concentrations of the crude F4 protein 50, 250, and 500 µg in the loops of individual pigs were used to analyze if dose affected the F4-specific immune response. Interestingly dose had no effect on the magnitude of the response. Therefore we hypothesized that the F4-specific immune response in the loops could be enhanced through the use of the adjuvants CpG ODN 2007 and pBD-1. The F4 protein was co-administered with either CpG ODN 2007 or pBD-1 and immune responses were assessed after 3 weeks. However neither CpG ODN 2007 nor PBD-1 at the doses used made an improvement in the immune response. Thus, these results demonstrated that the expression level of the F4R was the most important parameter for eliciting of the local immune response against the F4 protein. Furthermore our studies revealed that both F4Rneg and F4Rpos pigs responded to F4 immunization, however the former respond only nominally to F4-immunization in the loops. Moreover, an inverse relationship existed between the level of the F4-specific IgG in the serum and the F4-specific immune response seen in the loops. Thus our findings have important implications for oral vaccination using fimbrial based antigens (Ags) that utilize a receptor for their immunogenicity. Our results indicate that only animals with high levels of enterocyte F4R will have the ability to elicit high levels of protective F4-specific anti-fimbrial antibodies in their intestine after oral immunization. Therefore unless an effective adjuvant is available, animals with low to moderate levels of the fimbrial receptor in their small intestine will mount only weak immune responses making herd immunity after vaccination currently unattainable.

No description available.

For the successful establishment of secondary tumours, blood-borne metastatic tumour cells must adhere and spread on the vascular endothelium before they can migrate through it to form secondary growths in the tissue beneath. In this study an in vitro assay was developed to study the behavourial interactions between B16F10 cells and Bovine aortic endothelial cells. It was hypothesized that molecules synthesized by the endothelial cells may be involved in the mediation of the adhesion, spreading and migration events and hence that such molecules may possibly be involved in the process of haematogenic metastasis. Endothelial derived extracts were obtained from the cell surface and from conditioned medium. The extracts were tested for their adhesion promoting abilities in a quick dot blot adhesion assay. To verify that these molecules promoted adhesion, antibodies were raised against the extracts. Partial characterisation of the molecules was achieved using SDS-PAGE and immunoprobing. The extracts were also tested for their spreading and migration promoting properties. An attempt was made to block the adhesion, spreading and migration events using antibodies directed against components of the extracts. Clearly, if endothelial-derived molecules are involved in metastasis, then preventing the mediation of adhesion, spreading and migration may ultimately have relevance in the clinical situation.

A procedure was developed for the quantitative separation of major fermentation products of uniformly labeled glucose-C¹⁴ produced by bovine rumen microorganisms in vitro. After 45 min, the fermentation mixture was fractionated into (a) one control subsample, and duplicate fractions of (b) solid matter “precipitate“, (c) ether extract, (d) “amino acid“, (e) “sugar“, (f) CO₂, and (g) CH₄. Similar fractionation of an unfermented control sample was made. A portion of the fermentation ether extract was subjected to column chromatography to resolve (a) C₁, (b) C₂, (c) C₃, (d) C₄, and (e) C₅ fatty acids, (f) succinic, and (g) lactic acids. Each fraction was analyzed in triplicate for C¹⁴ by a direct plating technique. Corrections for geometry, self absorption, and efficiency were made by direct plating additional triplicate fraction subsamples, each containing a uniformly labeled glucose-C¹⁴ internal standard. The data were expressed as per cent recovery of added C1u. The results indicated that glucose was rapidly fermented with most of the C¹⁴ found in the ether extractable fraction as acetic acid. Significant levels of C¹⁴ were found in the “precipitate“ fractions. The data were compatible with evidence that CH₄ was derived from CO₂. The results of 6 trials indicated that there was no significant difference in the distribution of products resulting from the in vitro fermentation of uniformly labeled glucose-C¹⁴ between animals, between days within animals, or between times within days. / Ph. D.

Post-weaning diarrhea is a major health challenge in the swine industry and is routinely managed by fortifying pig starter diets with antimicrobials. But there are concerns about antibiotic resistance, hence the need for identifying effective alternatives. The use of spray-dried whole egg powder containing anti-F4 antibodies (SDWE) against recombinant F4 antigens and chitosan oligosaccharide and Enterococcus fecalis probiotic combination (CPRO) was investigated in two trials using enterotoxigenic Escherichia coli K88+ (ETEC) oral challenge model in 21-d-old piglets. Pre-challenge, SDWE supported higher (P < 0.05) piglet performance whereas during the post-challenge period, SDWE and CPRO had no effect on growth performance but diarrhea incidences and severity were reduced (P > 0.05) in SDWE-fed piglets compared to the control. The results show that SDWE supported greater piglet performance pre-ETEC challenge although there was no benefit of SDWE or CPRO supplementation evident during the post-challenge period in early-weaned pigs. / October 2015

No description available.