京都大学 / 新制・論文博士 / 博士(工学) / 乙第13495号 / 論工博第4202号 / 新制||工||1786(附属図書館) / (主査)教授 米田 稔, 教授 高野 裕久, 教授 松井 康人 / 学位規則第4条第2項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

Thesis (MSc. Medical Science (Chemical Pathology)) -- University of Limpopo, 2017 / Obesity has been associated with dyslipidaemia (increased levels of triglycerides, total cholesterol and low levels of HDL-C together with small dense lipoprotein particles) in the absence of metabolic disorders such as, type 2 diabetes mellitus and inflammation. Since community based studies in South Africa reported that obesity is more common in women, and rural Africans have a more favourable lipid profile compared to their White counterparts, the current study investigated the association of obesity in women without metabolic disorders with lipid levels and changes in proportions of small and large LDL and HDL particles. Methods The present study was part of the project “Prevention, Control and Integrated Management of Chronic Disease in a rural area, South Africa”. A total of 521 women participated in the above project. After excluding people with diabetes mellitus, insulin resistance and inflammation, 308 women were left and of these 67 were obese. Sixty seven ages matched, randomly selected non-obese women served as controls. Anthropometry variables as well as systolic and diastolic blood pressures were measured and the WHO steps questionnaire was administered to collect information on medication, lifestyle and diseases. Fasting blood levels of total cholesterol, HDL C, triglyceride, adiponectin, CRP, glucose and insulin were measured. Proportions of small and large HDL and LDL particles were determined. Results There was no significant difference in TC, TG and LDL-C levels (p=0.558, 0.087 and 0.948) between obese and non-obese women or between women with increased waist circumference (WC) and those with normal WC. The HDL-C concentration was significantly lower in obese women compared to women with non- obese (p=0.001). The lipid ratios TC/HDL-C and Apo B-100/Apo A-I were significantly higher in obese women than those with non- obese (p=0.013 and p=0.006) respectively. The same phenomenon was observed in women with xv increased waist circumference (p=0.001** and p=0.025* respectively). Adiponectin levels were significantly lower in obese women compared to non-obese women (p=0.004**) and in women with increased waist circumference compared to those with normal waist circumference (p=0.016*). The proportions of small dense HDL and LDL lipoprotein particles were similar in obese and non-obese women. Both obese and abdominally obese women had significantly higher odds ratios of low levels of HDL-C and elevated Apo B-100/Apo A-I. Adiponectin was a significant predictor of elevated TC and TG in both obese and abdominally obese women while BMI was a significant predictor of low HDL-C in obese women. Waist circumference was a significant predictor of low HDL-C in abdominally obese women. Conclusion In the current study, obesity in women was significantly associated with lipid abnormalities such as low HDL-C levels, raised lipid ratios (TC/HDL-C and Apo B 100/Apo A-I) and low levels of adiponectin, after excluding metabolic disorders / VLIR

No description available.

The purpose of this study was to determine if post-operative serum albumin and percentage of weight loss improved in patients who received formalized pre-operative nutrition counseling. Nutrition intervention was measured quantitatively. A retrospective review of records was conducted on 77 RYGB patients (68 female subjects and 9 male subjects), ages 21-64, during January 2001 through January 2006. The results indicated that patients who received pre-operative nutrition intervention had better clinical outcomes of serum albumin than those with no nutrition intervention from a registered dietitian. However, outcomes regarding percentage of weight loss varied. Both pre-operatively and at the 3 month post-operative visit the weight of subjects who received nutrition intervention seemed to be increasing by the 6 month post-operative visit the subjects with no nutrition intervention had lost more weight.

This project describes the development of a new antitumor therapeutic platform that combines the benefits of small-molecule drug conjugates (SMDCs) and antibody drug conjugates (ADCs). Valine-citrulline (VCit) dipeptide linkers are popular cathepsin B cleavable ADC linkers. Due to its instability in mouse serum, translating efficacy data from mouse to human is more difficult. It has been reported that replacing the VCit linker with glutamic acid-valine-citrulline (EVCit) improves ADC stability in mouse serum. The effect of the EVCit linker on the stability of SMDCs has not been reported so far. In a xenograft mouse model of prostate cancer, we found that incorporating the EVCit linker in PSMA-targeting SMDCs equipped with the transthyretin ligand AG10 resulted in conjugates with lower toxicity, extended half-life, and superior therapeutic efficacy compared to the standard metastatic castration-resistant prostate cancer (mCRPC) treatment option, docetaxel. This should improve the predictability of SMDC preclinical toxicity and efficacy data from mice.

Calves are born agammaglobulinemic and rely on colostrum consumption for the transfer of maternal passive immunity. Calves that fail to absorb adequate amounts of maternal antibodies from colostrum are commonly referred to as having failed transfer of passive immunity (FTPI). The overall aim of this dissertation was to explore the usefulness of FTPI testing in neonatal beef calves to predict their risk for subsequent illness or death. The objectives were to evaluate the impact of FTPI on pre-weaning disease in beef and dairy calves, quantify and compare the variance in IgG concentrations measured by radial immunodiffusion and serum total protein (STP) values measured by optical refractometry, and evaluate the correlation between herd-level prevalence of FTPI and herd-level prevalence of pre-weaning disease in beef calves. Evaluation of literature relevant to FTPI was compiled and assessed to quantify the impact of FTPI on pre-weaning disease in beef and dairy calves. A series of randomized trials were used to evaluate the variance in IgG concentrations and STP values from banked serum. Health records from multiple farms and IgG results were used to evaluate the relationship between FTPI and disease at the individual and herd-level. Failed transfer of passive immunity had a variable association with pre-weaning disease in beef and dairy calves. IgG concentrations were less precise than STP values especially when dilution was required. IgG concentrations and STP values were associated with an increased risk of disease in pre-weaned beef calves, but FTPI cut-off values poorly classified the risk for subsequent disease. The proportion of calves with FTPI was not correlated with the proportion of calves that developed pre-weaning disease. Using a single immunological factor, such as IgG concentration or STP, to predict disease results in the misclassification of disease risk and does not consider additional component causes of disease.

The serum fraction of blood is an ideal material in which to search for novel biomarkers for disease. It is easily obtained through relatively non-invasive means, routinely collected, and a rich treasure-trove of information about the health of an individual. Cells react to signal molecules, take up nutrients, and release waste products, fragments that are the result of proteolysis, and other molecules out into the bloodstream. If these components are unique to the cells in question, that part of the complex mixture that is the blood stream can potentially characterize the health of the tissue or organ those cells are a part of. Serum is dense with proteins that span over ten orders of magnitude in size and abundance. The top 22 most abundant proteins in serum account for 99% of the total protein. These abundant proteins are well-characterized and not useful in a search for novel biomarkers for disease. Removal of these large proteins is accomplished using an organic-solvent precipitation step. Analyzing the resultant mixture of low-molecular-weight serum peptides using cLC-MS produces large, data-rich, and very complex data files. We have developed a manual analysis method we have developed that is capable of performing all of the processing steps necessary to identify novel biomarkers for disease as well as a method for the sequencing of low-abundance, highly charged peptide species without additional sample preparation. These methods are applied to two serum sample sets collected to investigate two pregnancy-related diseases: preterm birth, and preeclampsia. Three novel biomarkers of preterm birth have been identified and a combination of these with 5 previously studied markers can predict women who will have preterm birth with a sensitivity of 89.9% and a specificity of 81.0%. Nineteen different molecular species have been identified that predict women at risk for preeclampsia with a p-value of <0.05. Weighted combinations of various groups of the 19 biomarkers can increase the sensitivity up to 96% and the specificity up to 100%. The use of cLC-MS in the search for novel serum biomarkers of pregnancy-related disease allows for seamless integration from potential biomarker selection to polypeptide sequence identification.

C. jejuni, a pathogenic gram-negative bacterium infecting the human gastrointestinal tract has lately been shown to cause bacteraemia to a wider extent than previously known. In some genotypes, this is thought to be related to GDP-Mannose 4,6 dehydratase encoded by the gene wcbK in the capsule polysaccharide operon and its potential phase variated regulated nature mediated by a homopolymeric guanine tract. This potential regulatory tract has been reported to be controlling the survival in serum by switching expression of wcbK “ON” or “OFF”. This master thesis report evaluates C. jejuni’s ability to survive human serum and low pH, as proxies for the conditions that bacteria meet in human blood or the stomach, respectively. By next generation sequencing, I evaluated the correlation between survival in human serum and the wcbK gene’s “ON” or “OFF” state. Furthermore, the temporal stability of the serum resistant phenotype was assessed over multiple generations. I found that a serum resistant fraction of the C. jejuni population could be enriched by selection in normal human serum. The serum resistant part of the population did not decrease during repeated subculture for 10 generations in bacterial culture medium. However, there was no correlation between the extent of serum resistance in the population and the “ON” or “OFF” state of the wcbK gene.

Serum levels of apolipoprotein ApoC1 have been described in a number of systemic tumor entities as potential biomarkers, but little is known about ApoC1 in neurosurgical patients. A total of 230 serum samples from 96 patients were analyzed using an ELISA technique. Patient diagnoses comprised 70 glioblastomas WHO IV◦ , 10 anaplastic astrocytomas III◦ , one anaplastic oligodendroglioma III◦ , one oligodendroglioma II◦ , one diffuse astrocytoma II◦ , one pilocytic astrocytoma I◦ , and a single case of a spindle cell tumor without WHO grading, as well as 11 spinal interventions. The mean ApoC1 level of the 230 samples was 132.03 µg/mL (median 86.83, SD 292.91). In the 176 glioblastoma samples, the mean ApoC1 level was 130.0 µg/mL (median 86.23, SD 314.9), which was neither different from the whole group nor from patients with spinal interventions (215.1 µg/mL, median 63.6, SD 404.9). In the postoperative samples, the mean ApoC1 level was significantly lower (85.81 µg/mL) than in the preoperative samples (129.64 µg/mL) and in samples obtained during adjuvant chemotherapy (168.44 µg/mL). While absolute ApoC1 serum levels in a patient do not allow for the distinction between neurosurgical histological entities, future analyses will examine whether the time course of ApoC1 in an individual patient can be related to certain treatment stages.

Abstract  Aggregatibacter aphrophilus, a Gram negative bacterium, found in the oral cavity, causing cerebral abscesses and infective endocarditis, has been shown to be serum resistant in previous studies. Bacterial secretion systems are important for bacteria as they transfer virulence factors into other bacteria or host cells as an attack. A. aphrophilus encodes a type VI secretion system, which is a spike-like membrane protein, mainly consisting of a hemolysin co-regulated protein (Hcp). In this work, it was tested if Hcp would contribute to serum resistance of A. aphrophilus. Firstly, to assess Hcp contribution to serum resistance, a bacterial serum killing assay-method was used and data was collected from three independent experiments. Two strains of A. aphrophilus were used in the experiments: the laboratory strain HK83 and a HK83 hcp mutant strain. The results showed that Hcp provided no significant effect on serum resistance of A. aphrophilus. Secondly, optical density measurements were made for growth curve analysis, to determine if the HK83 hcp mutant strain had an impact in growth compared to HK83. The results indicated that the HK83 hcp mutant strain had a somewhat reduced growth compared to its parental strain.