Lignocellulosic biomass is under extensive investigation as a bioethanol and bio-based materials feedstock. However, the complex structural and chemical mechanisms of lignocellulosic plant, which cause resistance to deconstruction during saccharification, require a pretreatment process. In this study, raw materials (corn bran, wheat bran and wheat straw) were selected because of their production and consumption in Turkey and also their accessibilities to be used as bioethanol source. Microfluidization pretreatment (high-pressure fluidization), which stands as a new approach for nano-cellulosic fibers production, was studied at 500 bar and 2000 bar to observe the qualitative and quantitative modifications in enzymatic hydrolysis depending on its effects on lignocellulosic structure. Optimum cellulase concentrations were determined for microfluidized samples as 4.5 U/g dry biomass for wheat bran, corn bran and 6.0 U/g dry biomass for wheat straw samples for the first 150 min interval. Effective usage of solid loads were found as 5.0 %, 2.5 %, and 7.5 % (dw/v) for wheat bran, wheat straw and corn bran, respectively. X-ray diffraction and SEM results of the microfluidized samples have indicated that the pretreatment has increased crystallinity index of all the samples and resulted in a scattered structure. Comparisons with other methods (softening, dilute-acid and lime pretreatments) have shown that microfluidization is advantageous over others by reducing the time required for enzymatic hydrolysis and thus can be a promising alternative pretreatment.

In this thesis, an intramolecular Pauson-Khand reaction of chiral enynes derived from homoallyl, allyl and homopropargyl alcohols is described. For this purpose, 2-heteroaryl substituted homoallyl, allyl and homopropargyl alcohols are easily and efficiently resolved through enzymatic resolution in a high ee (91-99%) with a known stereochemistry. Each enantiomerically enriched enyne derived from homoallyl and homopropargyl alcohols affords the conformationally most stable diastereomeric cyclopenta[c]pyran ring system as a sole product, whereas enantiomerically enriched enynes derived from allyl alcohols give the diastereomeric cis:trans mixture of cyclopenta[c]furan ring system. In addition these, an intramolecular Pauson&ndash / Khand reaction of camphor tethered enynes derived from homoallyl, homomethallyl, and homopropargyl alcohols is also described. Accordingly, homoallyl, homomethallyl, and homopropargyl moieties are easily constructed on the camphor carbonyl group with excellent diastereoselectivity due to endo-face selectivity, and with known stereochemistry. Each enantiomerically pure enyne affords the conformationally most stable diastereomeric spirocyclic cyclopenta[c]pyran ring system.

The aim of this study is to investigate the deterioration in mechanical integrity of the collagen network in bovine bone with aging, which are related to fracture toughness. Age-related changes in collagen molecular structures formed by non-enzymatic glycation were examined and indentation fracture technique was used as a method for measuring the microstructural toughness of cortical bone. Microcrack propagation characteristics of bone for fragility were also studied. Young and old group of bovine cortical bone specimens were grouped into 2 as ribosylated and non-ribosylated which were rested in solutions for four weeks. Series of indentations were made on bone specimen groups for each of five masses 10g, 25g, 50g, 100g and 200g for 10 sec to detect the effect of applied indentation load. The applied load was increased to 300g, 500g, 1000g and 2000g for 10 sec to be able to make microcracks. Series of indentations were made on bone specimen groups for each of five durations 5sec, 10sec, 20sec, 30sec for 100g to study the effect of indentation duration. Specimens were examined in the wet and dry state while studying the factors effecting microhardness measurement. Microhardness values measured by 10g of load for 10sec were indifferent between the ribosylated and non-ribosylated groups in the young and old bovine bone pointing that this load is not indicative of the structural collagen changes. Loads of 25g, 50g, 100g and 200g for 10 sec were able to differ ribosylated bone from non-ribosylated bone for the young and old bovine bones. Degree of microhardness increased with increased incubation period. Microhardness of dry specimens being either ribosylated or non-ribosylated were found to be statistically higher than wet specimens in young and old bone except for 10g for 10sec. It has been shown that the calculated fracture toughness measured by the indentation method is a function of indentation load. Additionally, effect of indentation size might have resulted in a higher toughness measurement for higher indent loads with longer cracks even if the toughness is not actually higher.Methods using indentation technique has difficulty in relating the resistance to crack growth to the Mode I fracture toughness definition.Indentation fracture toughness allows sampling only one point on the R&shy / curve methods and was not considered as successful for assessing materials with rising R&shy / curve. Toughness is ranked incorrectly among riboslated and non-ribosylated bovine bone by this technique. Presence of extrinsic toughening mechanisms including crack bridging due to uncracked ligaments and collagen fibers were directly observed by scanning electron microscope. Ribosylated bone was found to have lower number of collagen bridging compared ton on-ribosylated bovine bone.As a summary, indentation fracture method by Vickers indentation in bone is a method for measuring the fracture toughness.

Alternative fuels are becoming important due to diminishing fossil fuel reserves and the environmental hazards associated with exhaust gases. Biodiesel is an attractive alternative fuel because it is environmentally friendly and can be synthesized from edible/non-edible oils. Though several methods are available for synthesis, transesterification is the preferred route for biodiesel synthesis. The current techniques for transesterification of the oils to biodiesel are based on acid/alkali catalysis. However, these methods do not work for oils with high free fatty acid content and also require an additional downstream step for separation of catalyst from the product. In this work, we investigate the synthesis of biodiesel by two techniques: in supercritical methanol and ethanol without any catalyst; in supercritical carbon dioxide using enzyme as catalyst. In the first technique, the transesterification of refined grade edible oil like sunflower oil, palm oil and groundnut oil, and crude non-edible oils like Pongamia pinnata and Jatropha curcas was investigated in supercritical methanol and ethanol without using any catalyst from 200 oC to 400 oC at 200 bar. The variables affecting the conversion during transesterification, such as molar ratio of alcohol to oil, temperature and time were investigated in supercritical methanol and ethanol. The conversion to biodiesel increased from 30-40% at a molar ratio of 10 to 80-85% at a molar ratio of 45 to 50. Very high conversions (96-97%) were obtained in supercritical methanol and ethanol within 30 minutes at 350 oC. The kinetics of the reaction was modeled, the rate coefficients and activation energies were determined. As an enzymatic transesterification in organic solvents is mass transfer limited, supercritical fluids are found to be a better alternative because of their gas like diffusivity. Among potentially interesting solvents for enzymatic synthesis, carbon dioxide is the most obvious choice in supercritical fluids, because it is non-toxic, nonflammable and easily available. Because the products and the enzyme do not dissolve in carbon dioxide at room conditions, separation can easily be achieved by reduction of pressure Thus, the enzymatic transesterification for production of biodiesel in supercritical fluids under mild conditions is of commercial interest. Therefore, biodiesel was synthesized using immobilized enzyme (Lipase, Novozym-435) in supercritical carbon dioxide. The effect of reaction variables like temperature, molar ratio, enzyme loading and kinetics of the reaction was investigated. The conversion to biodiesel was found to be lower in supercritical carbon dioxide than that in supercritical alcohol.

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 192-197).

Increased amounts of pesticide production and application of pesticides for agriculture, plant protection and animal health has resulted in soil, water and air pollution, consequently relating a serious risk to the environment and also to human health. Pesticides include several groups of compounds, herbicides, insecticides, rodenticides and fumigants consisting of several hundred individual chemicals. Herbicides are an integral pan of modem agriculture and for industries requiring total vegetation control. Most herbicides are soil applied and more and more concern is raised that herbicides not only affect target organisms but also the microbial community present in soil. The ESKOM sub-station Zeus, in Mpumalanga (South Africa) used to apply an industrial weed control program for the eradication of vegetation, which led to the contamination of soil by several herbicides. These herbicides consisted of Bromacil, Tebuthiuron and Ethidimuron which are all photosynthesis inhibitors, more specifically, they disrupt the plastoquinone protein during electron transport at photosystem I1 (PSII). In this study the effect of biostimulation and bio-augmentation of a specific bioremediation agent (B350) as prescribed by ESKOM, on residual herbicides, Bromacil, Tebuthiuron and Ethidimuron was evaluated by monitoring the soil physical and chemical properties, microbial attributes, including potential microbial activity and community structure, as well as the physiological effect experienced by plants (Cynodoh dactylon and Zea mays). Results from soil physical and chemical analyses were correlated with results obtained for the functional and structural diversity of microbial communities. All results were investigated through statistical and multivariate analysis and the most prominent soil physical and chemical parameters that influence the biological and biochemical properties of the soil were identified. Results obtained from this study indicated that there were no significant difference (p < 0.05) between the treatments, with bioremediation agent, irradiated agent and without the agent based on results obtained from soil microbial properties and plant physiology. Before the trial started the uncontaminated soil showed an active microbial function, characterised by dehydrogenase, urease and arylsulphatase activity, but community structure was not very diverse. The contaminated soil, irradiated contaminated soil and silica sand showed less enzymatic function and was characterised by phospholipid fatty acid groups, mid-branched saturated fatty acids, terminally branched saturated fatty acids, normal saturated fatty acids and monosaturated fatty acids which are indicative of microorganisms that survive better in harsh environments. Three weeks after the addition of the specific bioremediation took place, the uncontaminated soil showed an increase in P-glucosidase activity and percentage organic carbon (%C), which could be a result of the presence of available plant material. Furthermore, an increase in major PLFA groups were seen, suggesting that an increase in diversity within the soil community occurred. The contaminated soil, irradiated contaminated soil and silica sand once again was characterised by a low microbial function and diversity, showing no improvement. Fluorescence data clearly show a decline in PS 11 function that result in the decline of the rate of photosynthesis, which was seen from COz gas exchange rates. Furthermore, the decrease in photosynthetic activity after three weeks was too severe to supply additional information about the mechanism within photosynthesis or the photoprotective mechanisms. A detailed study was conducted in which a 3: 1 dilution of contaminated soil with silica sand, was also monitored for changes within plant physiology. Results revealed that inhibition of PS I1 function already takes place within a few days time and the decline in photosynthesis is as a result of electron transport that does not supply adenosine triphosphate (ATP) and P-nicotinamide adenine dinucleotide (NADPH) to the Calvin cycle (or Reductive Pentose Phosphate pathway). It does not appear that rubulose-1,sbisphosphate carboxylase-oxygenase (Rubisco) is affected within the Calvin cycle. As a result of PS I1 function failure, reaction centres are damaged by the production of harmful singlet oxygen and photoprotective mechanisms (xanthophyll cycle) can not be activated. Thus, except for dealing with ineffective electron transport, additional damage is caused to physiological functions. After six weeks a decrease in the estimated viable biomass for all growth mediums was found. Results of the of trans- to cis- monoenoic fatty acids and cyclopropyl fatty acids to their monoenoic precursors ratios indicated that the soil microbial community for the contaminated growth mediums, all experienced nutritional stress throughout this trail. The specific bioremediation agent (B350) used, seemed to have no effect on the microbial function and community structure within soil and as agent had no effect on the residual herbicides or the plant physiology which experienced an extreme decline in major metabolic functions. / Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2

Mining activities cause severe disturbance to the soil environment in terms of soil quality and productivity and are of serious concern worldwide. Under South African legislation, developers are required to ecologically rehabilitate damaged environments. The application of agronomic approaches for the rehabilitation of coal discard sites has failed dismally in the arid areas of southern Africa. It is obvious that compliance with mitigation and rehabilitation requirements cannot be enforced without a thorough understanding of the ecological principles that ensure ecological stability and subsequent sustainability of soil ecosystems. Soil micro organisms are crucial role-players in the processes that make energy and nutrients available for recycling in the soil ecosystem. Poor management practices and other negative impacts on soil ecosystems affect both the physical and chemical properties of soil, as well as the functional and structural properties of soil microbial communities. Disturbances of soil ecosystems that impact on the normal functioning of microbial communities are potentially detrimental to soil formation, energy transfers, nutrient cycling, plant reestablishment and long-term stability. In this regard, an extensive overview of soil properties and processes indicated that the use of microbiological and biochemical soil properties, such as microbial biomass, enzymatic activity and the analysis of microbial community structure by the quantification of specific signature lipid biomarkers are useful as indicators of soil ecological stress or restoration properties because they are more responsive to small changes than physical and chemical characteristics. In this study, the relationship between the physical and chemical characteristics and different biological indicators of soil quality in the topsoil covers of seven coal discard sites under rehabilitation in South Africa, as well as three reference sites was investigated. Through the assimilation of basic quantitative data and the assessment of certain physical, chemical and biological properties of the topsoil covers obtained from the various coal discard sites as well as the reference sites, the relative success or progress of rehabilitation and the possible correlation between the biological indicators of soil quality and the establishment of self sustaining vegetation covers was determined. Results from soil physical and chemical analyses and percentage vegetation cover were correlated with the results obtained for the functional and structural diversity of microbial communities at the various sites. All results were investigated through statistical and multivariate analysis and the most prominent physical and chemical parameters that influence the biological and biochemical properties of the soil and possibly the establishment of self-sustainable vegetation cover on these mine-tailing sites were identified. Results obtained from this study indicated no significant difference (p>0.05) between the various discard sites based on conventional microbiological enumeration techniques. However, significant differences (p<0.05) could be observed between the three reference sites. All enzymatic activities assayed for the rehabilitation sites, with the exception of urease and alkaline phosphatase displayed a strong, positive association with the organic carbon content (%C). Ammonium concentration had a weak association with all the enzymes studied and pH only showed a negative association with acid phosphatase activity. A positive association was observed between the viable microbial biomass, vegetation cover and the organic carbon content, ammonium, nitrate and phosphorus concentrations of the soil. The various rehabilitation and reference sites could be differentiated based on the microbial community structure as determined by phospholipid fatty acid (PLFA) analysis. It is hypothesised that the microbial community structure of the Hendrina site is not sustainable when classified along an r-K gradient and that the high percentage of vegetation cover and high levels of estimated viable microbial biomass are an artificial reflection of the current management practices being employed at this site. Results obtained during this study, suggest that an absence or low percentage of vegetation cover and associated lower organic matter content of the soil have a significant negative impact on soil biochemical properties (enzymatic activity) as well as microbial population size. Furthermore, prevailing environmental physico-chemical and management characteristics significantly influences the vegetation cover and subsequently the microbial community structure. The results indicate that the microbial ecosystems in the coal discard sites could become more stable and ecologically self-regulating, provided effective management to enhance the organic carbon content of the soil. This could enhance nutrient cycling, resulting in changes of soil structure and eventually an improved soil quality which could facilitate the establishment of self sustaining vegetation cover. Results obtained during this study suggest that a polyphasic assessment of physical and chemical properties; microbial activities by enzymatic analysis; the characterisation of microbial community structure by analysis of phospholipid fatty acids; and the multifactorial analysis of the data obtained can be used as complementary assessment criteria for the evaluation of the trend of rehabilitation of mine tailings and discard sites. Strategic management criteria are recommended based on the soil quality environmental sustainability indices to facilitate the establishment of self sustainable vegetation covers. The contribution of this research to soil ecology is significant with regards to the intensive investigation and explanation of characteristics and processes that drive ecological rehabilitation and determine the quality of the soil environment. The multidisciplinary approach that is proposed could, furthermore, assist in the successful rehabilitation and establishment of self-sustaining vegetation covers at industrially disturbed areas, as well as assist in improving degraded soil quality associated with both intensive and informal agriculture. Additionally, this approach could negate the negative social and environmental impacts frequently associated with these activities. / Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2004.

Most vertebrates suffer permanent damage after minutes of anoxia. Many insects however, have part of their life cycle in anoxia or constant hypoxia, such as during their egg-hatching phase, by living as deep burrowers, or at high altitudes. Insects are able to survive in anoxia from hours to days, or even months by developing various strategies through evolution. For example, the locusts (Locusta migratoria) enter a reversible coma during anoxia that is associated with an arrest of ventilation, and a reinitiation of ventilation when returned to normoxia. This coma is correlated with a surge in the concentration of extracellular potassium ions ([K+]o), and recovery from this reversible coma is dependent on re-establishing the functional [K+]o. Prior exposure to a sublethal heatshock (HS)-preconditioning grants locusts a temporary resilience to anoxia; however, the molecular mechanisms of this protection are still unclear. This project investigated the effects of HS-preconditioning on locusts’ ventilation, the total enzymatic activity of the Na+/K+-ATPase, as well as its distribution within the metathoracic ganglion and tested the hypothesis that HS-preconditioning alters locusts’ ventilation and increases the totally Na+/K+-ATPase activity and its concentration within neuronal membranes. I recorded electromyograms of locusts’ ventilatory motor patterns in the presence and absence of anoxic coma by placing a copper wire electrode on ventilatory muscles 161 or 173 in control and HS-preconditioned animals. In addition, I studied the enzymatic activity of the Na+/K+-ATPase using a pyruvate kinase/lactate dehydrogenase assay and the localization of the Na+/K+-ATPase using immunohistochemistry in control and HS-preconditioned locusts at different stages of coma. I found that the ventilatory cycle period was decreased and the ventilatory muscle burst duration was increased after recovery from anoxic coma in HS-preconditioned locusts. I also found that anoxia did not affect the activity or the localization of the Na+/K+-ATPase. However, HS-preconditioning increased the total activity of the Na+/K+-ATPase and the localization of the Na+/K+-ATPase within the neuronal membranes. From this project, I concluded that HSpreconditioning affected locusts’ ventilatory motor pattern after recover from anoxia and increased the total activity and the neuronal membrane localization of the Na+/K+-ATPase. / Thesis (Master, Biology) -- Queen's University, 2011-09-26 13:14:48.472

No description available.

Food protein-derived peptides have increasingly become important sources of ingredients for the formulation of therapeutic products. The main aim of this work was to study the in vitro and in vivo bioactive properties of structurally diverse group of peptides produced through enzymatic hydrolysis of flaxseed proteins (FP). Hydrolysis of FP with seven proteases followed by fractionation into low-molecular-weight (LMW) cationic fractions yielded multifunctional peptides that inhibited angiotensin converting enzyme (ACE) and renin activities, which are molecular targets for antihypertensive agents. The LMW peptides also exhibited antioxidant properties by scavenging free radicals and inhibiting amine oxidase activity. The peptide fractions showed inhibition of calmodulin-dependent phosphodiesterase, an enzyme that has been implicated in the pathogenesis of several chronic diseases. Moreover, FP hydrolysis with thermolysin and pronase followed by mixing with activated carbon yielded branched-chain amino acids (BCAA)-enriched multifunctional peptide mixture (Fischer ratio of 23.65) with antioxidant properties and in vitro ACE inhibition; Fischer ratio of 20.0 is considered minimum for therapeutic purposes. The BCAA-enriched peptide product can be used in clinical nutrition to treat muscle wasting symptoms associated with hepatic diseases. Furthermore, an arginine-rich peptide mixture (31% arginine versus 11% in the original flaxseed protein) was produced by hydrolysis of FP with trypsin and pronase followed by separation using electrodialysis-ultrafiltration. Arginine plays important physiological roles especially as precursor to vasodilator, nitric oxide. The arginine-rich peptide mixture exhibited in vitro ACE and renin inhibition and led to decreased systolic blood pressure (–17.9 and –11.7 mmHg, respectively at 2 and 4 h) after oral administration to spontaneously hypertensive rats. For the first time in the literature, we showed that arginine peptides have superior physiological effects when compared to the amino acid form of arginine. Lastly, quantitative structure-activity relationship studies using partial least squares (PLS) regression yielded two predictive models for renin-inhibiting dipeptides with z-scales amino acid descriptors. The PLS models indicated that hydrophobic and bulky side chain-containing amino acids contribute to renin inhibition if present at the amino- and carboxyl-terminal of dipeptides, respectively. Based on this study, Ile-Trp was discovered as potent renin-inhibiting dipeptide, and may serve as a useful template for the development of potent antihypertensive peptidomimetics.