Masters of Science / Bacterial endophytes are able to improve the growth of their hosts through a number of different mechanisms such as nutrient uptake regulation, plant hormone production and regulation, siderophore production and phosphate solubilisation. They have also been shown to be able to provide protection to plants against various abiotic stressors, through various means such as oxidative stress protection. The purpose of this study was therefore to isolate endophytic bacteria from the stems of different weeds, to characterize their ability to use some of the most important growth promoting mechanisms including the ability to produce IAA, siderophores and ACC deaminase, what effect they had on the nutrient uptake in their hosts and to determine to what extent they could promote growth in the roots, stems and leaves of Brassica napus plants. In addition to this the endophytes were tested to see to what extent they could protect Brassica napus from the negative effects of vanadium stress and how this affected the plant physiologically in terms of morphology, overall biomass, the plants nutrient profile, lipid peroxidation and levels of cell death. The effect of vanadium stress on the oxidative state of Brassica napus was also monitored by determining the levels of stress induced reactive oxygen species (ROS) and the corresponding antioxidants that are responsible for regulating these reactive oxygen species. Six different endophytes (P1, P2, P3, P4, P5, P6) were isolated from different weed samples. Each endophyte was found to be able to significantly improve germination and growth in their host plant. Each isolate was able to improve the uptake of certain macronutrients and micronutrients in their respective hosts, while all of the isolates were shown to be capable of producing siderophores and ACC deaminase. One isolate had high levels of IAA production, with the remaining isolates producing small amounts of IAA. All isolates were also unable to solubilize phosphate. The five best performing endophytes (P1, P2, P3, P5, P6) in the preliminary growth trials were used in the follow up vanadium stressed growth trials, with endophyte P4 being left out of the remaining experiments. All of the endophytes showed improvements in growth promotion in comparison to the control, with endophyte treated plants showing both increased growth and biomass in both the non-stressed and vanadium stressed treatments of the vanadium stressed growth trial; however, the leaves of the vanadium stressed plants were significantly smaller than their non-stressed counterparts. When looking at the oxidative state it was found that vanadium stress caused a significant increase in the development of O2 -, H2O2 and •OH in the control and in addition to this it was shown that treatment with endophytes was able to cause a significant decrease in the levels of stress induced H2O2 and •OH in all of the treatments and O2 - for plants treated with endophyte P5. The noted change in the oxidative state of endophyte treated plants was attributed to an increase in the antioxidant activity of these plants, as it was found that endophyte treated plants showed a combination of increased activity for Superoxide dismutase, catalase and ascorbate peroxidase. This study has shown that endophytic bacteria from plant stems can be used to improve crop growth and yield, while simultaneously producing more nutrient dense crops from the same amount of land. It has also determined that endophytes P1, P2, P3, P5 and P6 are able to successfully provide protection to crop plants from the harmful effects of exposure to vanadium stress. This has great potential for improving food security locally and around the world, by allowing those who cannot gain access to large amounts of food to take in more nutrients from the same amount of food. Furthermore, it also presents the opportunity to use endophyte treatments to grow crops on land that has been previously contaminated with certain heavy metals. / 2023-12-01

To determine if greater amounts of hydroxyl radical (·OH) are formed by dopamine (DA) denervation and treatment with L-dihydroxyphenylalanine (L-DOPA), the neostriatum was DA denervated (99% reduction in DA content) by 6-hydroxydopamine treatment (134μg icv, desipramine pretreatment) of neonatal rats. At 10 weeks the peripherally restricted dopa decarboxylase inhibitor carbidopa (12.5mg/kg i.p.) was administered 30min before vehicle, L-DOPA (60mg/kg i.p.), or the known generator of reactive oxygen species, 6-hydroxydopa (6-OHDOPA) (60mg/ kg i.p.); and this was followed 30min later (and 15 min before termination) by the spin trap, salicylic acid (8 μmoles icv). By means of a high performance liquid chromatographic method with electrochemical detection, we found a 4-fold increase in the non-enzymatically formed spin trap product, 2,3-dihydroxybenzoic acid (2,3-DHBA), with neither L-DOPA nor 6-OHDOPA having an effect on 2,3-DHBA content of the neostriatum. Basal content of 2,5-DHBA, the enzymatically formed spin trap product, was 4-fold higher vs. 2,3-DHBA in the neostriatum of untreated rats, while L-DOPA and 6-OHDOPA each reduced formation of 2,5-DHBA. We conclude that DA innervation normally suppresses ·OH formation, and that the antiparkinsonian drug L-DOPA has no effect (2,3-DHBA) or slightly reduces (2,5-DHBA) ·OH formation in the neostriatum, probably by virtue of its bathing the system of newly formed ·OH.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18899号 / 医博第4010号 / 新制||医||1009(附属図書館) / 31850 / 京都大学大学院医学研究科医学専攻 / (主査)教授 生田 宏一, 教授 三森 経世, 教授 岩井 一宏 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

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Indiana University-Purdue University Indianapolis (IUPUI) / There is no consensus on which vascular segment or what size of vessels is most important in the process of collateral growth, the degree to which these vessels can enlarge, or the mechanisms that mediate collateral vessel expansion and its impairment. Chapter I identifies the major collateral vessels that develop in response to femoral arterial occlusion in the pig, rat, and mouse hindlimbs for comparison to humans. Pre-existent small named arteries enlarged ~2-3-fold to become the major collateral vessels in each species, these major collaterals displayed characteristics similar to large arteries experiencing flow-mediated outward remodeling, and important differences in vascular wall thickness were observed between rodents and pigs. Chapter II utilized Rac2-/- and Nox2-/- mice to investigate the hypothesis that Nox2-NAD(P)H oxidase is required for major collateral growth subsequent to femoral arterial occlusion. Previous studies suggest bone marrow cell (BMC)-derived reactive oxygen species (ROS) produced by the Nox2 subunit of NAD(P)H oxidase plays an important role in neovascularization and recovery of hindlimb perfusion subsequent to femoral arterial occlusion; but did not investigate collateral growth. The hematopoietic cell restricted protein Rac2 has been shown to bind to and activate Nox2-NAD(P)H oxidase and Rac2-/- and Nox2-/- leukocytes display impaired ROS related functions. The data demonstrated that Rac2 and Nox2 are not essential for major collateral growth, but both are important for the recovery of hindlimb perfusion and preservation of distal tissue morphology. Chapter III investigated BMC and antioxidant therapy in the age-related impairment of collateral growth. Aging, like all cardiovascular disease risk factors is associated with elevated ROS and impaired collateral growth. Studies also suggest BMCs promote collateral growth by secreting paracrine factors but elevated ROS may affect the efficacy of BMCs. The data revealed that neither BMC injection nor antioxidant therapy via apocynin enhanced the process of major collateral artery growth in aged mice.

Previous studies have established that ozone-oxygen mixtures can be used to inactivate a variety of microorganisms including bacteria, fungi and viruses. Ozone is a potent reactive oxygen species (ROS) that rapidly decays into a variety of additional short half-life ROS which have been shown to cause oxidative damage to biological molecules. I hypothesize that controlled ozone exposure and the subsequent generation of additional ROS would reduce viral infectivity by lipid and/or protein peroxidation. A proprietary ozone-oxygen delivery system was used to inactivate a series of enveloped [herpes simplex virus type-1 strain McIntyre (HSV-1), vaccinia strain Elstree (VAC), vesicular stomatitis virus strain Indiana (VSV), and influenza A strain (H1N1) A/WS/33] and non-enveloped [human adenovirus type2 (Ad2)] viruses. Plaque reduction and suspension-infection viral antigen assays were used to determine inactivation kinetics. After ozonation, HSV-1 and VSV lost up to 6 log10 infectious particles in 15 min, while VAC and influenza A lost up to 5 log10 in 40 min and 30 min, respectively. In comparison, the non-enveloped Ad2 lost up to 5 log10 in 60 min. Increasing amounts of serum supplementation in the ozone treated virus suspensions slowed the rate of inactivation in both enveloped and non-enveloped viruses, suggesting the protective effect of serum against ozone. Lipid peroxidation was determined through a chromogenic assay for malondialdehyde (MDA), a byproduct of peroxidation events. MDA concentrations were inversely correlated with virus infectivity, as MDA concentrations elevated with virus exposure time to ozone. Transmission electron microscopy images of Ad2, HSV-1, VAC and VSV confirmed the drastic morphological changes that resulted from ozone treatment. The ROS-mediated attack compromised the integrity of the lipid envelopes and protein shells of the viruses. These data suggest that a wide range of viruses can be inactivated through use of an innovative ozone delivery system, thus validating my hypothesis.

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M. Tech. (Department of Biotechnology, Faculty of Applied and Computer Sciences), Vaal University of Technology. / The aim of the study is to synthesise, characterize and assess the antimicrobial activity of cobalt oxide, zinc oxide and cobalt-doped zinc oxide nanoparticles against selected waterborne pathogenic fungi (yeasts and moulds) and bacteria. Various types of oxide based nanomaterial are an attractive option for the disinfection of water due to its high chemical stability and non-toxicity towards human cells. Synthesis of Co -doped ZnO and Co3O4nanoparticles was done through mechanochemical synthesis and urea based synthesis and microwave heating was employed for the preparation of ZnO nanoparticles. The ZnO nanoparticles were produced in short reaction and it was white color. Cobalt oxide (Co3O4) nanoparticles appeared as a pink precipitate but was turned black after being calcined. The synthesis of Co- ZnO nanoparticles was successfully prepared and blue solid was obtained from pink cobalt ion solution. The nanoparticles were characterised by X- Ray Diffraction (XRD), Fourier Infrared Spectroscopy (FTIR), UV–visible spectroscopy, Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) (Yang et al. 2003). In this research project, the antibacterial activities of NPs were carried out by well diffusion method and minimum inhibitory concentration (MIC). MIC is the lowest concentration of a chemical, usually a drug, which prevents visible growth of bacterium. Bacterial strains used in the study are: Salmonella enterica, Escherichia coli, Shigella sonnei and Staphylococcus aureus, yeast and mould is: Candida albicans and Aspergillus niger. The antimicrobial results obtained showed that ZnO nanoparticles are more effective than Co- ZnO and Co3O4 nanoprticles against all the microorganisms used. The toxicity studies were performed using DAPHTOXKIT F and the 24h EC50 and 48h EC50 were calculated according to the manufactures’ instructions. The results showed that Co- ZnO nanoparticles is less toxic to Daphnia magna compared to ZnO and Co3O4 NPs.

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Diabetic kidney disease (DKD) is an emerging pandemic, paralleling the worldwide increase in obesity and diabetes mellitus. DKD is now the most frequent cause of end-stage renal disease and is associated with an excessive risk of cardiovascular morbidity and mortality. DKD is a consequence of systemic endothelial dysfunction. The endothelial-dependent cytoprotective coagulation protease activated protein C (aPC) ameliorates glomerular damage in DKD, in part by reducing mitochondrial ROS generation in glomerular cells. Whether aPC reduces mitochondrial ROS generation in the tubular compartment remains unknown. Here, we conducted expression profiling of kidneys in diabetic mice (wild-type and mice with increased plasma levels of aPC, APChigh mice). The top induced pathways were related to metabolism and in particular to oxidoreductase activity. In tubular cells, aPC maintained the expression of genes related to the electron transport chain, PGC1-α expression, and mitochondrial mass. These effects were associated with reduced mitochondrial ROS generation. Likewise, NLRP3 inflammasome activation and sterile inflammation, which are known to be linked to excess ROS generation in DKD, were reduced in diabetic APChigh mice. Thus, aPC reduces mitochondrial ROS generation in tubular cells and dampens the associated renal sterile inflammation. These studies support approaches harnessing the cytoprotective effects of aPC in DKD.