Spelling suggestions: "subject:"croton nuclear magnetic resonance"" "subject:"kroton nuclear magnetic resonance""
1 |
Proton Nuclear Magnetic Resonance in MicaTownsend, Don H. 05 1900 (has links)
The experiments to be described here were undertaken for the purpose of determining, if possible, by NMR techniques whether or not the hydroxyl protons in mica are bound in a regular crystalline array, and, if so, whether or not the hydroxyl protons occur in reasonably isolated pairs as in waters of hydration.
|
2 |
Synthesis of the metallocenes for the production of exotic high energy ion beamsKheswa, Ntombizonke Yvonne January 2019 (has links)
Philosophiae Doctor - PhD / The Subatomic Physics Department of iThemba Laboratory for Accelerated Based Sciences (iThemba LABS) conducts experiments that require a variety of particle beams in order to study nuclear properties (reaction, structure, etc.) of various nuclides. These particle beams are accelerated using the K-200 Separated Sector Cyclotron (SSC) and delivered to different physics experimental vaults. Prior to acceleration, the particle beam is first ionised using an Electron Resonance Ion Source (ECRIS). The main goal of this study is the production of exotic metallic beams of 60Ni8+ and 62Ni8+ using ECRIS4, which are required for the Coulomb excitation experiments approved by the Programme Advisory Committee (PAC) at iThemba LABS.
In order to provide the metallic beams of nickel, a development study of organometallic materials containing 60Ni and 62Ni isotopes in a form of metallocene complexes was undertaken. The nickelocene (NiCp2) complex, a member of the organometallic family, was synthesised at the Physics Target Laboratory of iThemba LABS for the first time. Method development involved the use of natural nickel during the multi-step synthesis before the use of enriched nickel-60 (60Ni) and nickel-62 (62Ni).
Nine samples of NiCp2 were synthesised; two were isotopically enriched nickelocene (60NiCp2 and 62NiCp2). The percentage yields of the synthesised nickelocene samples ranged between 16 to 50 %, and samples were characterised by investigating their crystal structure and bonding arrangements in the complexes by X-ray diffraction (XRD) , Fourier Transform Infrared (FT-IR) spectroscopy, and Proton Nuclear Magnetic Resonance (1H NMR).
The synthesised nickelocene were further used with ECRIS4 for the production of Ni beams on the Q-line of the cyclotron. The Metal Ions from Volatile Compounds (MIVOC) technique was used for the conversion of 60Ni and 62Ni to ion species. The method used the organometallic compounds which are volatile at specific pressures at ambient temperatures.
Metallic ion beams of nickel were successfully produced after a carefully pre-sample conditioning in the MIVOC container before connecting the MIVOC set-up to the new injection system of the ECRIS4. Measured beam intensities during the experiment for both 60Ni+ and 62Ni+ were approximately 30 μA, optimum for physics measurements.
The development of the MIVOC technique opens up new beam-target combinations with the use of new exotic stable beams for new science cases at iThemba LABS. Reactions in inverse kinematics, multi-step Coulomb-excitation and other types of reactions will immensely benefit from these developments.
|
3 |
Cycling of Bioavailable Carboxyl-Rich Alicyclic Molecules and Carbohydrates in Baffin BayMcKee, Kayla 13 July 2023 (has links)
At ~662 gigatonnes of carbon (GtC), marine dissolved organic matter (DOM) is the largest reduced pool of actively cycling carbon and nitrogen in the oceans1. Operationally defined as smaller than 0.1µm in size, this carbon reservoir comprises all non-living organic matter smaller than a bacterial cell and comprises organic colloids and molecules spanning as a continuum of sizes ranging from marine viruses and large macromolecules (e.g. DNA, enzymes) to small organic molecules (e.g. polymers and monomers)2. With deep apparent 14C-ages ranging between 4900-6400 ybp 3,4, marine DOM is anomalously old given timescales of global ocean ventilation (1000-1500 years). The great age of DOM has remained one of the most elusive lines of scientific inquiry in Chemical Oceanography for decades. The size and molecular composition of DOM has been shown to be a key variable in determining its biological reactivity (e.g. cycling rate) and long-term persistence in the deep ocean5,6.
Despite the importance of DOM in the marine carbon and nitrogen cycles, we lack a detailed understanding of the molecular composition of DOM. Due to the high concentration of salts in seawater relative to DOM, it is difficult to analyze the molecular composition of seawater with conventional chemical- or size- fractionation methods without introducing bias (i.e. isolating only hydrophobic and/or high molecular weight DOM). In fact, it is commonly reported that >80% of DOM remains uncharacterized at the molecular level (e.g. not readily identifiable as an individual known biomolecule)5. Nuclear magnetic resonance (NMR) spectroscopy has been used as a tool for several decades to describe the composition of marine DOM isolates7. For example, 13C-NMR of major high molecular weight DOM functional groups at the molecular-level demonstrated that DOM is largely made up of reactive polysaccharides with low aromaticity compared to terrestrial DOM8. To date, all marine DOM NMR measurements have been made on size-fractionated DOM or chemically-fractionated (e.g. solid phase extracted) DOM isolates. In this thesis, I report the first Proton (1H) NMR composition of total seawater DOM from seawater samples collected from 10 stations in Baffin Bay aboard the CCGS Amundsen (2019). Samples were measured using 1H-NMR at uOttawa following a novel water suppression method established by Lam and Simpson9.
The use of this method has allowed for the first molecular composition assessment of total seawater DOM to be measured (e.g. without any chemical or size fractionation). I report the % relative abundance of individual biomarkers and determine molar concentrations of two compound classes of interest. These results are shown in Ocean Data View section plots, and are listed within appendix tables, to provide a comprehensive depiction of the changing concentrations of dissolved organic carbon (DOC), total carbohydrates (TCHO), and carboxyl-rich alicyclic molecules (CRAM). In this thesis, I explore changes in the abundance of these unique DOM compound classes and discuss how the composition of DOM directly determines its bioavailability and thus cycling in Baffin Bay 5. The core objective of my thesis was to measure DOM concentrations for TCHO and CRAM, as well as to calculate the production and removal of these key DOM compounds in Baffin Bay due to either physical and/or biological processes. We found that the concentration of both TCHO and CRAM decreased with depth throughout Baffin Bay. This is consistent with previous work suggesting the rapid cycling of carbohydrates, however it contradicts the current paradigm of CRAM cycling. Our results indicate between 21-43% of CRAM produced in the surface is subsequently removed at depth. Rapid cycling of a surface CRAM population suggests that not all CRAM can be considered recalcitrant DOM
We live in a time of unprecedented global change. The Arctic Ocean is warming at a rate at least four times faster than the global average10. The impact of a rapidly warming, freshening and increasingly acidified Arctic Ocean on the biogeochemistry of DOM remains unknown. It is imperative that more DOM research be conducted as early as possible in order to better understand these impacts and inform future research directions. The distribution and cycling of CRAM in Baffin Bay provide novel and fundamental knowledge of DOM cycling in a key Arctic region, but could also potentially occur throughout the global ocean. Such data will no doubt be of use in informing future iterations of Earth System Climate models seeking to forecast how the marine carbon cycle will respond to global change.
|
4 |
Análise metabolômica (1H RMN) do líquido sinovial de equinos hígidos e acometidos por osteocondrite dissecante / 1H RMN-base metabolomics for the investigation of normal and osteochondritic synovial fluid in the horseMantovani, Cristina de Faria 29 November 2013 (has links)
A osteocondrite dissecante (OCD) é uma das principais doenças que afetam os equinos jovens e uma das condições incluídas nas Doenças Ortopédicas do Desenvolvimento. Por se tratar de importante afecção articular, a OCD pode ser responsabilizada por perdas econômicas substanciais em decorrência da diminuição do desempenho atlético e reprodutivo dos animais acometidos. Sua etiologia é considerada multifatorial sendo associada a fatores nutricionais, endócrinos, genéticos e biomecânicos. A análise dos biomarcadores do líquido sinovial fornece dados fundamentais acerca das alterações representativas de inflamação e danos à cartilagem articular, por isso o monitoramento do ambiente articular caracteriza-se como uma importante ferramenta para o diagnóstico e acompanhamento da progressão da doença. A análise metabolômica por Ressonância Magnética Nuclear de prótons de hidrogênio (1H RMN) é um método de avaliação holístico, simultâneo e em tempo real, dos metabólitos de pequenas moléculas presente no líquido sinovial, capaz de fornecer dados a respeito da progressão da doença, da resposta a tratamentos e de determinar biomarcadores. No presente estudo objetivou-se estabelecer o perfil metabólico global do líquido sinovial de equinos acometidos por OCD, estudar a aplicabilidade da 1H RMN no diagnóstico da OCD e avaliar a possibilidade de distinção entre os líquidos sinoviais de articulações com OCD sintomática e assintomática. A determinação do perfil metabólico do líquido sinovial de equinos hígidos e acometidos por OCD assintomática e sintomática resultou em 32 metabólitos comuns às três condições articulares. Além destes, 2 compostos foram identificados somente nas articulações doentes, sendo eles o propilenoglicol e o composto aromático, porém, não foi possível diferenciar as articulações com OCD assintomática das articulações sintomáticas, pois ambos os grupos apresentavam metabólitos semelhantes e com intensidades similares.. Na comparação entre a intensidade dos espectros, observaram-se aumentos dos metabólitos glicose, glutamina, etanol, leucina, isoleucina, dimetilsulfona, creatina, creatinina, Nacetilglucosamina e N-acetilglutamina e -metil histidina e dos compostos propilenoglicol e composto aromático nas articulações com OCD com relação às hígidas. Dos metabólitos que apresentaram redução na intensidade dos espectros das articulações doentes têm-se o 3- hidroxibutirato e o acetato. Outros metabólitos demonstraram a mesma intensidade de sinal para os três tipos de articulação, sendo eles: piruvato, citrato, metionina, histidina, tirosina, valina, lactato, alanina, glicina, glicerol e fenilalanina. A análise estatística por PCA foi capaz de realizar o agrupamento dos espectros e atribuir a importância do composto aromático na diferenciação entre os animais hígidos e doentes. A análise metabolômica por 1H NMR mostrou-se técnica com alta reprodutibilidade entre as amostras e sensibilidade de detecção dos componentes do líquido sinovial, revelando claras diferenças entre os perfis bioquímicos de articulações hígidas e acometidas por OCD, indicativo das alterações metabólicas que ocorrem com a progressão desta doença, relacionadas principalmente com o processo de degradação cartilagínea. Este estudo salienta o potencial da análise metabolômica em fornecer uma nova perspectiva dos processos bioquímicos envolvidos na progressão da OCD. / Osteochondritis dissecans (OCD) is one of the most common diseases affecting young horses and one of the conditions classified as Developmental Orthopedic Diseases. As an important joint disorder, OCD is held responsible for substantial economic losses in consequence of the decrease in athletic and reproductive performance in affected horses. The condition presents a multifactorial ethiology, related to nutritional, endocrine, genetic and biomechanical factors, although its pathogeny is well established, associated with a disturbance of the process of endochondral ossification. Synovial fluid biomarkers analysis provide data regarding changes representative of inflammation and articular cartilage damage, which is why monitoring the joint cavity is an important tool for diagnosis and assessing disease progression. Proton Nuclear Magnetic Resonance spectroscopy is a holistic, simultaneous and real time approach, of the metabolites present in the synovial fluid, offering the potential to provide data regarding disease progression, response to treatment and to determine disease biomarkers. The aim of the present study was to establish the global metabolic profiling of normal and osteochondritic synovial fluid, to evaluate the applicability of the 1H RMN in the diagnosis of OCD, and to asses possible distinctions between symptomatic and asymptomatic horses. The metabolic profile determination resulted in 32 common metabolites to the three joint conditions. Moreover, 2 compounds were identified exclusively in the diseased joints, namely propylene glycol and aromatic compound, but we were unable to discern between asymptomatic and symptomatic individuals for both groups presented similarities in metabolites as well as their concentrations. When comparing spectral intensities, we observed marked increases in the metabolites glucose, glutamine, ethanol, leucine, isoleucine, dimethyl sulphone, creatine, creatinine, N-acetylglucosamine, N-acetylglutamine and -methyhistidine, besides the compounds propylene glycol and aromatic compound, in the affected joints compared to the healthy ones. Decrease in spectral intensities were related to the metabolites 3-hydroxybutirate and acetate in affected joints. Other metabolites showed no difference in intensities in all joint conditions, those being pyruvato, citrate, methionine, histidine, tyrosine, valine, lactate, alanine, glycine, glycerol and phenylalanine. PCA based statistical analysis was able to group spectra and to ascribe the importance of the aromatic compound in differentiating healthy from diseased joints. 1H RMN spectroscopy showed high reproductability between samples and sensitive in detecting synovial fluid compounds, unveiling clear diferences between the biochemical profiles of healthy and OCD affected joints, thus indicating metabolic alterations occuring with disease progression, related mainly with the cartilage degradation process. This study projects the potential of metabolomics analysis in providing a new perspective of the biochemical processes involved in OCD progression.
|
5 |
Análise metabolômica (1H RMN) do líquido sinovial de equinos hígidos e acometidos por osteocondrite dissecante / 1H RMN-base metabolomics for the investigation of normal and osteochondritic synovial fluid in the horseCristina de Faria Mantovani 29 November 2013 (has links)
A osteocondrite dissecante (OCD) é uma das principais doenças que afetam os equinos jovens e uma das condições incluídas nas Doenças Ortopédicas do Desenvolvimento. Por se tratar de importante afecção articular, a OCD pode ser responsabilizada por perdas econômicas substanciais em decorrência da diminuição do desempenho atlético e reprodutivo dos animais acometidos. Sua etiologia é considerada multifatorial sendo associada a fatores nutricionais, endócrinos, genéticos e biomecânicos. A análise dos biomarcadores do líquido sinovial fornece dados fundamentais acerca das alterações representativas de inflamação e danos à cartilagem articular, por isso o monitoramento do ambiente articular caracteriza-se como uma importante ferramenta para o diagnóstico e acompanhamento da progressão da doença. A análise metabolômica por Ressonância Magnética Nuclear de prótons de hidrogênio (1H RMN) é um método de avaliação holístico, simultâneo e em tempo real, dos metabólitos de pequenas moléculas presente no líquido sinovial, capaz de fornecer dados a respeito da progressão da doença, da resposta a tratamentos e de determinar biomarcadores. No presente estudo objetivou-se estabelecer o perfil metabólico global do líquido sinovial de equinos acometidos por OCD, estudar a aplicabilidade da 1H RMN no diagnóstico da OCD e avaliar a possibilidade de distinção entre os líquidos sinoviais de articulações com OCD sintomática e assintomática. A determinação do perfil metabólico do líquido sinovial de equinos hígidos e acometidos por OCD assintomática e sintomática resultou em 32 metabólitos comuns às três condições articulares. Além destes, 2 compostos foram identificados somente nas articulações doentes, sendo eles o propilenoglicol e o composto aromático, porém, não foi possível diferenciar as articulações com OCD assintomática das articulações sintomáticas, pois ambos os grupos apresentavam metabólitos semelhantes e com intensidades similares.. Na comparação entre a intensidade dos espectros, observaram-se aumentos dos metabólitos glicose, glutamina, etanol, leucina, isoleucina, dimetilsulfona, creatina, creatinina, Nacetilglucosamina e N-acetilglutamina e -metil histidina e dos compostos propilenoglicol e composto aromático nas articulações com OCD com relação às hígidas. Dos metabólitos que apresentaram redução na intensidade dos espectros das articulações doentes têm-se o 3- hidroxibutirato e o acetato. Outros metabólitos demonstraram a mesma intensidade de sinal para os três tipos de articulação, sendo eles: piruvato, citrato, metionina, histidina, tirosina, valina, lactato, alanina, glicina, glicerol e fenilalanina. A análise estatística por PCA foi capaz de realizar o agrupamento dos espectros e atribuir a importância do composto aromático na diferenciação entre os animais hígidos e doentes. A análise metabolômica por 1H NMR mostrou-se técnica com alta reprodutibilidade entre as amostras e sensibilidade de detecção dos componentes do líquido sinovial, revelando claras diferenças entre os perfis bioquímicos de articulações hígidas e acometidas por OCD, indicativo das alterações metabólicas que ocorrem com a progressão desta doença, relacionadas principalmente com o processo de degradação cartilagínea. Este estudo salienta o potencial da análise metabolômica em fornecer uma nova perspectiva dos processos bioquímicos envolvidos na progressão da OCD. / Osteochondritis dissecans (OCD) is one of the most common diseases affecting young horses and one of the conditions classified as Developmental Orthopedic Diseases. As an important joint disorder, OCD is held responsible for substantial economic losses in consequence of the decrease in athletic and reproductive performance in affected horses. The condition presents a multifactorial ethiology, related to nutritional, endocrine, genetic and biomechanical factors, although its pathogeny is well established, associated with a disturbance of the process of endochondral ossification. Synovial fluid biomarkers analysis provide data regarding changes representative of inflammation and articular cartilage damage, which is why monitoring the joint cavity is an important tool for diagnosis and assessing disease progression. Proton Nuclear Magnetic Resonance spectroscopy is a holistic, simultaneous and real time approach, of the metabolites present in the synovial fluid, offering the potential to provide data regarding disease progression, response to treatment and to determine disease biomarkers. The aim of the present study was to establish the global metabolic profiling of normal and osteochondritic synovial fluid, to evaluate the applicability of the 1H RMN in the diagnosis of OCD, and to asses possible distinctions between symptomatic and asymptomatic horses. The metabolic profile determination resulted in 32 common metabolites to the three joint conditions. Moreover, 2 compounds were identified exclusively in the diseased joints, namely propylene glycol and aromatic compound, but we were unable to discern between asymptomatic and symptomatic individuals for both groups presented similarities in metabolites as well as their concentrations. When comparing spectral intensities, we observed marked increases in the metabolites glucose, glutamine, ethanol, leucine, isoleucine, dimethyl sulphone, creatine, creatinine, N-acetylglucosamine, N-acetylglutamine and -methyhistidine, besides the compounds propylene glycol and aromatic compound, in the affected joints compared to the healthy ones. Decrease in spectral intensities were related to the metabolites 3-hydroxybutirate and acetate in affected joints. Other metabolites showed no difference in intensities in all joint conditions, those being pyruvato, citrate, methionine, histidine, tyrosine, valine, lactate, alanine, glycine, glycerol and phenylalanine. PCA based statistical analysis was able to group spectra and to ascribe the importance of the aromatic compound in differentiating healthy from diseased joints. 1H RMN spectroscopy showed high reproductability between samples and sensitive in detecting synovial fluid compounds, unveiling clear diferences between the biochemical profiles of healthy and OCD affected joints, thus indicating metabolic alterations occuring with disease progression, related mainly with the cartilage degradation process. This study projects the potential of metabolomics analysis in providing a new perspective of the biochemical processes involved in OCD progression.
|
6 |
Fermentability of dietary fibre and metabolic impacts of including high levels of fibrous feed ingedients in maize-soyabean growing pig diets supplemented with exogenous enzymesFushai, Felix 03 1900 (has links)
The objectives of the research were to examine the effects of high dietary levels of fibrous feeds, and of supplementation with Roxazyme® G2 (RX), on the digestive metabolic and physiological responses of growing pigs fed maize-soybean diets.
The nutrient and dietary fibre (DF) composition, the swelling and water-binding capacities of maize (MM), its hominy chop (HC) and cobs (MC), dehulled soybean (dSBM) and the hulls (SH), brewer’s grains (BG), lucerne hay (LH) and wheat bran (WB) were evaluated using standard procedures. Feed fibre fractions were isolated by simulating upper tract digestion in an Ankom® DaisyII Incubator, whereby each feed was digested in pepsin (porcine, 200 FIP-U/g, Merck No, 7190), followed by pancreatin (porcine, grade IV, Sigma No P-1750), with recovery of the fibrous residues. In a third step to complete the simulated pig gastro-intestinal digestion, the pepsin-pancreatin fibre extracts were digested by RX or Viscozyme L ® V2010 (VZ). Enzyme activity was measured as the coefficients of partial degradability (solubilisation) of the washed fibre extracts. The kinetics and products of fermentation of the DF were evaluated in an AnkomRF gas production system, using buffered faecal inoculum.
Among the feed ingredients, dissimilar, fibre source-dependent activities between RX (0.02 to 0.12) and VZ (0.04-0.33) were observed. The lowest RX activities were observed on the maize and soybean derived fibres, with similarly low VZ activity on MC fibre. Variation in the activity of faecal microbial enzymes was similarly indicated by the variable production of fermentation gas (51.8-299.4 mL g-1 DM) and short chain fatty acids (SCFA) (2.3-6.0 mMol g-1 DM). Soy hull, dSBH, MM and HC fibres were highly fermentable, with low fermentability of BG, MC and WB fibres. The fibres differed in the composition of fermentation SCFA, whereby SH, LH and MC shifted fermentation to Ace, and BG, dSBM, WB, MM, HC favoured Pro, while MM and HC favoured But production.
The same nutritional properties were similarly evaluated in complete diets which were formulated from the ingredients for growth, and metabolic trials. For the growth trial, a standard (STD) (control), 141 g total dietary fibre (TDF) kg-1 dry matter (DM) maize-soybean growing pig diet, and five iso-nutritive, 246 g TDF kg-1 DM nutritionally balanced diets were formulated. The high DF was achieved by partial replacement of the MM and dSBM in the STD diet with MC, SH, BG, LH or WB.
The differences in RX and VZ activities and in the fermentation characteristics which were observed on the fibre extracts from the high fibre ingredients were reflected in the DF from the respective complete diets in which they were included. However, the fibre from the basal dietary ingredients reduced the absolute values and the variation in the activities of RX (0.03-0.06) and VZ (0.16-0.22), and similarly reduced the variation in gas (126.6-187.6 mL g-1 DM) and SCFA (4.1-5.4 mMol g-1 DM) production of the DF from the fibrous diets. Enzyme activities on the STD DF were low for RX (0.03) and high for VZ (0.25). The STD DF produced 205.3 mL gas g-1 DM, which was similar to SH DF, and higher than all the other diets. The STD DF produced 5.0-mMol SCFA g-1 DM, which was quantitatively, and not statistically higher than the other fibres. The composition of SCFA was similar across all diets, except for the high percent Ace, with low Pro by the SH DF. Compared to the STD, the high DF diets increased percent Ace, with reduced Pro and But.
The STD, MC, SH, BG, LH and WB diets were each prepared in duplicate mixes, one of which was fortified with 200 mg RX kg-1 feed (as fed). Seventy-two intact Large White X Landrace, male, 32.0 ± 5.6 kg live weight (LW) pigs were allocated to the diets in two completely randomised weight blocks in a 2 (fibre source) X 2 (enzyme) factorial arrangement. The pigs were fed ad libitum for 10 weeks. Cumulative LW gain and feed intake were measured at different stages of growth, and at slaughter. Apparent total tract digestibility (ATTD) of nutrients was estimated at 65-70 kg LW, using 0.2% (as fed) chromium oxide as the indigestible marker. Ileal tissue was sampled 50 cm above the ileo-caecal valve, on which villi height and area, and crypt depth were evaluated by computerised image analysis. Blood was sampled at slaughter from the severed vena jugularis, 16 hours after feeding. Serum urea, creatinine, triglycerides, glucose, and total cholesterol were analysed chemically. The serum metabolome was further explored using Proton Nuclear Magnetic Resonance Spectroscopy (1H -NMRS).
There was fibre X RX interaction for villi height, whereby the enzyme reduced the villi height in pigs on the SH, STD and WB diets, with an opposite effect on pigs on the MC, BG, LH diets. The soluble fibre content was negatively correlated with crypt depth. Chemical analysis did not detect differences in metabolite concentration between the STD and the high fibre diets. However, more serum cholesterol was observed in pigs fed the WB compared to the LH and MC diets. 1H-NMRS indicated that feeding pigs the WB diet increased serum Cys and His, while supplementation of RX increased serum formate, glucose, and urea. There was diet X enzyme interaction for fructose, glucose, Arg, Cys, Ser, and Trp, whereby RX increased the levels in pigs on MC and WB, with an opposite effect in pigs on the other diets.
There was large DF source-dependent variation among diets in ATTD of DM (0.80-0.85), organic matter (OM) (0.81-0.87), gross energy (GE) (079-0.85) and CP (0.81-0.85), whereby, relative to the STD diet, high DF reduced the ATTD of DM (all diets except SH), organic matter (OM) and energy and CP (all diets except the MC). Positive correlation was observed between fermentability and the ATTD digestibility of DM, OM, energy, ADF, NDF, and fat. Negative correlation was observed between the swelling capacity and the ATTD of DM, OM, energy and protein, between DF solubility and DM, OM, protein, ADF and NDF, and between water binding capacity and ATTD of DM and OM, energy and NDF.
At slaughter, there was similarly large, and DF source-dependent variation among the high fibre diets in feed intake (2.31-2.71 kg as fed day-1), live weight gain (0.75-0.86 kg day-1), and feed: gain ratio (2.73-3.00). Corresponding values for the STD diet were 2.44 kg day-1, 0.83 kg day-1and 2.86 kg day-1, respectively. Relative to the STD, LH reduced feed intake and live weight gain, and MC increased the feed: gain ratio. Predictions based on the in vitro fermentability of DF and feed intake suggested that due to poor fermentability, and or restriction of feed intake, relative to a standard fibre diet, high dietary levels of MC, WB and BG may reduce fermentation in the lower gut, while similar dietary levels of SH and LH may result in substantial increases in fermentation.
At 50 kg LW, the fermentability of DF was positively correlated with feed intake and with weight gain, while water binding capacity and solubility of DF were negatively correlated with feed intake. At slaughter, the solubility of DF was negatively correlated with feed intake and feed: gain ratio. Large variation among the high fibre diets was also observed in the slaughter weight (89.2-96.8 kg), dressing % (68.6-76.4), meat colour (80.4-82.3), lean % (69.5-71.2), and fat % (10.1-12.6). In comparison, pigs on the STD diet scored 94.7 kg slaughter weight, 75.1% dressing, 81.6 cm carcass length, 82.5 meat colour, 68.4% lean, and 15.0% fat. Relative to the STD, LH reduced dressing and fat %. Lucerne hay and WB increased the lean%.
For the metabolic trial, two iso-nutritive, mixed high fibre (319 g TDF kg-1 DM), nutritionally balanced diets were formulated to contain DF of high (HF) versus low (LF) fermentability. The diets had similar content of soluble DF and similar swelling and water binding capacities. Viscozyme was more active than RX on both the HF (0.20 versus 0.04) and the LF (0.17 versus 0.07) DF. The combination of RX and VZ statistically increased the enzyme activity on the HF (0.25) and quantitatively increased enzyme activity on the LF (0.18) DF, suggesting additive or synergistic effects. More gas was produced by the HF (159.5 mL g-1 DM) compared to the LF DF (96.6 mL g-1 DM). More SCFA were produced by HF (5.0 mMol g-1 DM), compared to the LF DF (3.6 mMol g-1 DM). Compared to the STD, HF DF increased percent Ace, with reduced Pro and But. The LF DF increased percent Ace, with quantitative, and not statistical reduction of Pro and But.
In a metabolic trial, the HF and LF diets, and their duplicates containing 0.270 g RX kg-1 DM of feed (as fed) were fed ad libitum to eight ileum T-cannulised, intact Large White X Landrace male pigs weighing 65.0 ± 5.1 kg. The diets were allocated to the pigs in a duplicate 4 x 4 Latin Square design, in a 2 (enzyme) x 2 (fermentability) factorial arrangement. Each period consisted of two weeks of adaptation followed by five days of sampling. The ileal digesta was collected in each period and was similarly subjected to the fermentation test. Apparent ileal digestibility (AID) and ATTD were determined using 0.2% (as fed) chromium oxide as the indigestible marker. N excretion in faeces and urine were measured, and N retention was calculated. Blood was sampled by vena jugularis puncture on the last day of each period. Two blood samples were collected, the first 15 hours after removal from feed (15-hour serum), and the second 3 hours after re-introduction to feed (3-hour serum). Serum metabolites were evaluated by both chemical analyses and by 1H-NMRS, as described for the growth trial. Roxazyme did not affect the fermentation characteristics of the ileal digesta. In similar proportion to the fermentability of the PP digesta, the HF ileal digesta was more fermentable (65.4 mL gas g-1 DM and 6.1 mMol SCFA g-1 DM) than the LF ileal digesta (46.7 mL gas g-1 DM and 4.4 mMol SCFA g-1 DM SCFA). Prediction based on the in vitro fermentability of DF and feed intake suggested the HF diet could support one half times more fermentation in the lower gut compared to the LF diet.
The HF diet had higher AID of DM (62.5 vs. 58.6), OM (65.6 vs. 62.1), energy (64.4 vs. 61.0), fat (85.8 vs. 81.7) and ash (41.8 vs. 32.7). The AID of HO-Pro, Met and Val were higher for the LF diet. There was diet X enzyme interaction on the AID of Met, whereby the RX reduced the AID of met in the LF diet, and not that of the HF diet. The ATTD was higher for the HF diet for DM (74.2 vs. 68.4), NDF (64.7 vs. 57.4), and ADF (35.1 vs. 21.0).
There was positive correlation between the fermentability of DF and the AID DM, OM, ash, ash, fat and energy. The solubility of DF was negatively correlated with the AID of DM, OM, ash, fat, ADF and energy, and with the ATTD of DM, OM, ash, fat, energy, NDF, and ADF. Negative correlation was also observed between the swelling capacity of DF and the AID of protein, Trp and Lys. The solubility of DF was positively correlated with Ser, Ala, Val, Iso-Leu and His.
There was diet X enzyme interaction for urea in the 15- hour serum, whereby RX tended to reduce the urea in the LF diet, while it increased that of the HF diet. Fermentability negatively correlated with urea in the 15- hour serum, and positively correlated with serum glucose in the 3-hour serum. In the 3-hour sample, 1H-NMRS indicated higher fucose, Pro and cholesterol in the LF diet. 1H-NMRS also indicated fermentability x RX interaction for Ser, Tyr, Lys, creatine, and possibly, glucose or fructose, glycerol or Gly and His or Arg, whereby RX increased the levels in the LF diets, with opposite effect in the HF diet.
In conclusion, enzyme activities and fermentability were higly variable among different DF sources, and the effects were evident in the fibrous complete diets. The results of the in vitro studies supported the application of the methods to formulate fermentable insoluble fibre-rich, maize-soybean-mixed co-product diets. Correlation analyses suggested that DF fermentability, and solubility, swelling and water binding capacities explained significant proportions of the variances of the metabolic and physiological responses of the pigs to different feeds. Predictions based on the in vitro fermentability of DF and feed intake suggested that a strategy whereby pig diets are enriched in DF after the feedstuffs are screened on DF fermentability could substantially increase fermentation in the lower gut. Overall, the results suggested that productivity can be maintained in growing pigs fed diets containing up to twice the standard levels of DF, provided producers target co-product feeds that contain highly fermentable DF. The use of RX to improve nutrient digestion and to stimulate gut fermentation was not justified. / Environmental Sciences / Ph.D. (Environmental Sciences)
|
7 |
Regulation of kinases by synthetic imidazoles, nucleotides and their deuterated analoguesNkosi, Thokozani Clement 19 April 2016 (has links)
Deuteration is the replacement of a hydrogen atom by deuterium atom in a molecule. The replacement begins at the most acidic hydrogen in the molecule. In ATP, the deshielded hydrogen is C8-H which is the first replaced during deuteration. During ATP deuteration some of the ATP is hydrolysed to ADP concurrently. Using kinetic analysis, it was confirmed that the ATP hydrolysis that occurs is 1st order in ATP concentration, while the hydrogen replacement is 2nd order. The ATP and its C8 deuterated analogue were tested against three enzymes shikimate kinase (SK), acetate kinase (AK) and glutamine synthetase (GS) to determine if a kinetic isotope effect (KIE) exists in these systems. With AK and GS, the KIED increased as the KIEH decreased, while with SK the KIED decreased as the KIEH increased as the concentration of the ATP or deuterated analogue increased. Deuteration of imidazole and purine compounds reduced the specific activity of AK or SK at low concentrations in an enzyme-catalysed reaction. From a library of imidazole-containing compounds that inhibited SK, three compounds were selected and their IC50 values were determined on the SK-catalysed reaction. These compounds show a differential potency and efficiency between their protonated and deuterated analogues when compared in a 1:1 mixture. Synthesized purines incorporating three different substituents at N-9 were tested against AK or SK for their ability to lower the specific activity of the enzymes used / Physics / M. Sc. (Physics)
|
8 |
Fermentability of dietary fibre and metabolic impacts of including high levels of fibrous feed ingedients in maize-soyabean growing pig diets supplemented with exogenous enzymesFushai, Felix 03 1900 (has links)
The objectives of the research were to examine the effects of high dietary levels of fibrous feeds, and of supplementation with Roxazyme® G2 (RX), on the digestive metabolic and physiological responses of growing pigs fed maize-soybean diets.
The nutrient and dietary fibre (DF) composition, the swelling and water-binding capacities of maize (MM), its hominy chop (HC) and cobs (MC), dehulled soybean (dSBM) and the hulls (SH), brewer’s grains (BG), lucerne hay (LH) and wheat bran (WB) were evaluated using standard procedures. Feed fibre fractions were isolated by simulating upper tract digestion in an Ankom® DaisyII Incubator, whereby each feed was digested in pepsin (porcine, 200 FIP-U/g, Merck No, 7190), followed by pancreatin (porcine, grade IV, Sigma No P-1750), with recovery of the fibrous residues. In a third step to complete the simulated pig gastro-intestinal digestion, the pepsin-pancreatin fibre extracts were digested by RX or Viscozyme L ® V2010 (VZ). Enzyme activity was measured as the coefficients of partial degradability (solubilisation) of the washed fibre extracts. The kinetics and products of fermentation of the DF were evaluated in an AnkomRF gas production system, using buffered faecal inoculum.
Among the feed ingredients, dissimilar, fibre source-dependent activities between RX (0.02 to 0.12) and VZ (0.04-0.33) were observed. The lowest RX activities were observed on the maize and soybean derived fibres, with similarly low VZ activity on MC fibre. Variation in the activity of faecal microbial enzymes was similarly indicated by the variable production of fermentation gas (51.8-299.4 mL g-1 DM) and short chain fatty acids (SCFA) (2.3-6.0 mMol g-1 DM). Soy hull, dSBH, MM and HC fibres were highly fermentable, with low fermentability of BG, MC and WB fibres. The fibres differed in the composition of fermentation SCFA, whereby SH, LH and MC shifted fermentation to Ace, and BG, dSBM, WB, MM, HC favoured Pro, while MM and HC favoured But production.
The same nutritional properties were similarly evaluated in complete diets which were formulated from the ingredients for growth, and metabolic trials. For the growth trial, a standard (STD) (control), 141 g total dietary fibre (TDF) kg-1 dry matter (DM) maize-soybean growing pig diet, and five iso-nutritive, 246 g TDF kg-1 DM nutritionally balanced diets were formulated. The high DF was achieved by partial replacement of the MM and dSBM in the STD diet with MC, SH, BG, LH or WB.
The differences in RX and VZ activities and in the fermentation characteristics which were observed on the fibre extracts from the high fibre ingredients were reflected in the DF from the respective complete diets in which they were included. However, the fibre from the basal dietary ingredients reduced the absolute values and the variation in the activities of RX (0.03-0.06) and VZ (0.16-0.22), and similarly reduced the variation in gas (126.6-187.6 mL g-1 DM) and SCFA (4.1-5.4 mMol g-1 DM) production of the DF from the fibrous diets. Enzyme activities on the STD DF were low for RX (0.03) and high for VZ (0.25). The STD DF produced 205.3 mL gas g-1 DM, which was similar to SH DF, and higher than all the other diets. The STD DF produced 5.0-mMol SCFA g-1 DM, which was quantitatively, and not statistically higher than the other fibres. The composition of SCFA was similar across all diets, except for the high percent Ace, with low Pro by the SH DF. Compared to the STD, the high DF diets increased percent Ace, with reduced Pro and But.
The STD, MC, SH, BG, LH and WB diets were each prepared in duplicate mixes, one of which was fortified with 200 mg RX kg-1 feed (as fed). Seventy-two intact Large White X Landrace, male, 32.0 ± 5.6 kg live weight (LW) pigs were allocated to the diets in two completely randomised weight blocks in a 2 (fibre source) X 2 (enzyme) factorial arrangement. The pigs were fed ad libitum for 10 weeks. Cumulative LW gain and feed intake were measured at different stages of growth, and at slaughter. Apparent total tract digestibility (ATTD) of nutrients was estimated at 65-70 kg LW, using 0.2% (as fed) chromium oxide as the indigestible marker. Ileal tissue was sampled 50 cm above the ileo-caecal valve, on which villi height and area, and crypt depth were evaluated by computerised image analysis. Blood was sampled at slaughter from the severed vena jugularis, 16 hours after feeding. Serum urea, creatinine, triglycerides, glucose, and total cholesterol were analysed chemically. The serum metabolome was further explored using Proton Nuclear Magnetic Resonance Spectroscopy (1H -NMRS).
There was fibre X RX interaction for villi height, whereby the enzyme reduced the villi height in pigs on the SH, STD and WB diets, with an opposite effect on pigs on the MC, BG, LH diets. The soluble fibre content was negatively correlated with crypt depth. Chemical analysis did not detect differences in metabolite concentration between the STD and the high fibre diets. However, more serum cholesterol was observed in pigs fed the WB compared to the LH and MC diets. 1H-NMRS indicated that feeding pigs the WB diet increased serum Cys and His, while supplementation of RX increased serum formate, glucose, and urea. There was diet X enzyme interaction for fructose, glucose, Arg, Cys, Ser, and Trp, whereby RX increased the levels in pigs on MC and WB, with an opposite effect in pigs on the other diets.
There was large DF source-dependent variation among diets in ATTD of DM (0.80-0.85), organic matter (OM) (0.81-0.87), gross energy (GE) (079-0.85) and CP (0.81-0.85), whereby, relative to the STD diet, high DF reduced the ATTD of DM (all diets except SH), organic matter (OM) and energy and CP (all diets except the MC). Positive correlation was observed between fermentability and the ATTD digestibility of DM, OM, energy, ADF, NDF, and fat. Negative correlation was observed between the swelling capacity and the ATTD of DM, OM, energy and protein, between DF solubility and DM, OM, protein, ADF and NDF, and between water binding capacity and ATTD of DM and OM, energy and NDF.
At slaughter, there was similarly large, and DF source-dependent variation among the high fibre diets in feed intake (2.31-2.71 kg as fed day-1), live weight gain (0.75-0.86 kg day-1), and feed: gain ratio (2.73-3.00). Corresponding values for the STD diet were 2.44 kg day-1, 0.83 kg day-1and 2.86 kg day-1, respectively. Relative to the STD, LH reduced feed intake and live weight gain, and MC increased the feed: gain ratio. Predictions based on the in vitro fermentability of DF and feed intake suggested that due to poor fermentability, and or restriction of feed intake, relative to a standard fibre diet, high dietary levels of MC, WB and BG may reduce fermentation in the lower gut, while similar dietary levels of SH and LH may result in substantial increases in fermentation.
At 50 kg LW, the fermentability of DF was positively correlated with feed intake and with weight gain, while water binding capacity and solubility of DF were negatively correlated with feed intake. At slaughter, the solubility of DF was negatively correlated with feed intake and feed: gain ratio. Large variation among the high fibre diets was also observed in the slaughter weight (89.2-96.8 kg), dressing % (68.6-76.4), meat colour (80.4-82.3), lean % (69.5-71.2), and fat % (10.1-12.6). In comparison, pigs on the STD diet scored 94.7 kg slaughter weight, 75.1% dressing, 81.6 cm carcass length, 82.5 meat colour, 68.4% lean, and 15.0% fat. Relative to the STD, LH reduced dressing and fat %. Lucerne hay and WB increased the lean%.
For the metabolic trial, two iso-nutritive, mixed high fibre (319 g TDF kg-1 DM), nutritionally balanced diets were formulated to contain DF of high (HF) versus low (LF) fermentability. The diets had similar content of soluble DF and similar swelling and water binding capacities. Viscozyme was more active than RX on both the HF (0.20 versus 0.04) and the LF (0.17 versus 0.07) DF. The combination of RX and VZ statistically increased the enzyme activity on the HF (0.25) and quantitatively increased enzyme activity on the LF (0.18) DF, suggesting additive or synergistic effects. More gas was produced by the HF (159.5 mL g-1 DM) compared to the LF DF (96.6 mL g-1 DM). More SCFA were produced by HF (5.0 mMol g-1 DM), compared to the LF DF (3.6 mMol g-1 DM). Compared to the STD, HF DF increased percent Ace, with reduced Pro and But. The LF DF increased percent Ace, with quantitative, and not statistical reduction of Pro and But.
In a metabolic trial, the HF and LF diets, and their duplicates containing 0.270 g RX kg-1 DM of feed (as fed) were fed ad libitum to eight ileum T-cannulised, intact Large White X Landrace male pigs weighing 65.0 ± 5.1 kg. The diets were allocated to the pigs in a duplicate 4 x 4 Latin Square design, in a 2 (enzyme) x 2 (fermentability) factorial arrangement. Each period consisted of two weeks of adaptation followed by five days of sampling. The ileal digesta was collected in each period and was similarly subjected to the fermentation test. Apparent ileal digestibility (AID) and ATTD were determined using 0.2% (as fed) chromium oxide as the indigestible marker. N excretion in faeces and urine were measured, and N retention was calculated. Blood was sampled by vena jugularis puncture on the last day of each period. Two blood samples were collected, the first 15 hours after removal from feed (15-hour serum), and the second 3 hours after re-introduction to feed (3-hour serum). Serum metabolites were evaluated by both chemical analyses and by 1H-NMRS, as described for the growth trial. Roxazyme did not affect the fermentation characteristics of the ileal digesta. In similar proportion to the fermentability of the PP digesta, the HF ileal digesta was more fermentable (65.4 mL gas g-1 DM and 6.1 mMol SCFA g-1 DM) than the LF ileal digesta (46.7 mL gas g-1 DM and 4.4 mMol SCFA g-1 DM SCFA). Prediction based on the in vitro fermentability of DF and feed intake suggested the HF diet could support one half times more fermentation in the lower gut compared to the LF diet.
The HF diet had higher AID of DM (62.5 vs. 58.6), OM (65.6 vs. 62.1), energy (64.4 vs. 61.0), fat (85.8 vs. 81.7) and ash (41.8 vs. 32.7). The AID of HO-Pro, Met and Val were higher for the LF diet. There was diet X enzyme interaction on the AID of Met, whereby the RX reduced the AID of met in the LF diet, and not that of the HF diet. The ATTD was higher for the HF diet for DM (74.2 vs. 68.4), NDF (64.7 vs. 57.4), and ADF (35.1 vs. 21.0).
There was positive correlation between the fermentability of DF and the AID DM, OM, ash, ash, fat and energy. The solubility of DF was negatively correlated with the AID of DM, OM, ash, fat, ADF and energy, and with the ATTD of DM, OM, ash, fat, energy, NDF, and ADF. Negative correlation was also observed between the swelling capacity of DF and the AID of protein, Trp and Lys. The solubility of DF was positively correlated with Ser, Ala, Val, Iso-Leu and His.
There was diet X enzyme interaction for urea in the 15- hour serum, whereby RX tended to reduce the urea in the LF diet, while it increased that of the HF diet. Fermentability negatively correlated with urea in the 15- hour serum, and positively correlated with serum glucose in the 3-hour serum. In the 3-hour sample, 1H-NMRS indicated higher fucose, Pro and cholesterol in the LF diet. 1H-NMRS also indicated fermentability x RX interaction for Ser, Tyr, Lys, creatine, and possibly, glucose or fructose, glycerol or Gly and His or Arg, whereby RX increased the levels in the LF diets, with opposite effect in the HF diet.
In conclusion, enzyme activities and fermentability were higly variable among different DF sources, and the effects were evident in the fibrous complete diets. The results of the in vitro studies supported the application of the methods to formulate fermentable insoluble fibre-rich, maize-soybean-mixed co-product diets. Correlation analyses suggested that DF fermentability, and solubility, swelling and water binding capacities explained significant proportions of the variances of the metabolic and physiological responses of the pigs to different feeds. Predictions based on the in vitro fermentability of DF and feed intake suggested that a strategy whereby pig diets are enriched in DF after the feedstuffs are screened on DF fermentability could substantially increase fermentation in the lower gut. Overall, the results suggested that productivity can be maintained in growing pigs fed diets containing up to twice the standard levels of DF, provided producers target co-product feeds that contain highly fermentable DF. The use of RX to improve nutrient digestion and to stimulate gut fermentation was not justified. / Environmental Sciences / Ph.D. (Environmental Sciences)
|
9 |
Regulation of kinases by synthetic imidazoles, nucleotides and their deuterated analoguesNkosi, Thokozani Clement 19 April 2016 (has links)
Deuteration is the replacement of a hydrogen atom by deuterium atom in a molecule. The replacement begins at the most acidic hydrogen in the molecule. In ATP, the deshielded hydrogen is C8-H which is the first replaced during deuteration. During ATP deuteration some of the ATP is hydrolysed to ADP concurrently. Using kinetic analysis, it was confirmed that the ATP hydrolysis that occurs is 1st order in ATP concentration, while the hydrogen replacement is 2nd order. The ATP and its C8 deuterated analogue were tested against three enzymes shikimate kinase (SK), acetate kinase (AK) and glutamine synthetase (GS) to determine if a kinetic isotope effect (KIE) exists in these systems. With AK and GS, the KIED increased as the KIEH decreased, while with SK the KIED decreased as the KIEH increased as the concentration of the ATP or deuterated analogue increased. Deuteration of imidazole and purine compounds reduced the specific activity of AK or SK at low concentrations in an enzyme-catalysed reaction. From a library of imidazole-containing compounds that inhibited SK, three compounds were selected and their IC50 values were determined on the SK-catalysed reaction. These compounds show a differential potency and efficiency between their protonated and deuterated analogues when compared in a 1:1 mixture. Synthesized purines incorporating three different substituents at N-9 were tested against AK or SK for their ability to lower the specific activity of the enzymes used / Physics / M. Sc. (Physics)
|
Page generated in 0.1022 seconds