Bioreaction and separation in preparative batch chromatographic columns : the hydrology of lactose to yield glucose,galactose and oligosaccharides

The initial aim of this project was to improve the performance of a chromatographic bioreactor-separator (CBRS). In such a system, a dilute enzyme solution is pumped continuously through a preparative chromatographic column, while pulses of substrate are periodically injected on to the column. Enzymic reaction and separation are therefore performed in a single unit operation. The chromatographic columns used were jacketed glass columns ranging from 1 to 2 metres long with an internal diameter of 1.5 cm. Linking these columns allowed 1, 2, 3 and 4 metre long CBRS systems to be constructed. The hydrolysis of lactose in the presence of β~galactosidase was the reaction of study. From previous work at Aston University, there appeared to be no difficulties in achieving complete lactose hydrolysis in a CBRS. There did, however, appear to be scope for improving the separative performance, so this was adopted as an initial goal. Reducing the particle size of the stationary phase was identified as a way of achieving this improvement. A cation exchange resin was selected which had an average particle size of around half that previously used when studying this reaction. A CBRS system was developed which overcame the operational problems (such as high pressure drop development) associated with use of such a particle size. A significant improvement in separative power was achieved. This was shown by an increase in the number of theoretical plates (N) from about 500 to about 3000 for a 2 metre long CBRS, coupled with higher resolution. A simple experiment with the 1 metre column showed that combined bioreaction and separation was achievable in this system. Having improved the separative performance of the system, the factors affecting enzymic reaction in a CBRS were investigated; including pulse volume and the degree of mixing between enzyme and substrate. The progress of reaction in a CBRS was then studied. This information was related to the interaction of reaction and separation over the reaction zone. The effect of injecting a pulse over a length of time as in CBRS operation was simulated by fed batch experiments. These experiments were performed in parallel with normal batch experiments where the substrate is mixed almost instantly with the enzyme. The batch experiments enabled samples to be taken every minute and revealed that reaction is very rapid. The hydrodynamic characteristics of the two injector configurations used in CBRS construction were studied using Magnetic Resonance Imaging, combined with hydrodynamic calculations. During the optimisation studies, galactooligosaccharides (GOS) were detected as intermediates in the hydrolysis process. GOS are valuable products with potential and existing applications in food manufacture (as nutraceuticals), medicine and drug targeting. The focus of the research was therefore turned to GOS production. A means of controlling reaction to arrest break down of GOS was required. Raising temperature was identified as a possible means of achieving this within a CBRS. Studies were undertaken to optimise the yield of oligosaccharides, culminating in the design, construction and evaluation of a Dithermal Chromatographic Bioreactor-separator.

Development of a sediment yield model for South East Ghana in the regions around Lake Volta

Soil erosion is one of the most pressing issues facing developing countries. The need for soil erosion assessment is paramount as a successful and productive agricultural base is necessary for economic growth and stability. In Ghana, a country with an expanding population and high potential for economic growth, agriculture is an important resource; however, most of the crop production is restricted to low technology shifting cultivation agriculture. The high intensity seasonal rainfall coincides with the early growing period of many of the crops meaning that plots are very susceptible to erosion, especially on steep sided valleys in the region south of Lake Volta. This research investigated the processes of soil erosion by rainfall with the aim of producing a sediment yield model for a small semi-agricultural catchment in rural Ghana. Various types of modelling techniques were considered to discover those most applicable to the sub-tropical environment of Southern Ghana. Once an appropriate model had been developed and calibrated, the aim was to look at how to enable the scaling up of the model using sub-catchments to calculate sedimentation rates of Lake Volta. An experimental catchment was located in Ghana, south west of Lake Volta, where data on rainstorms and the associated streamflow, sediment loads and soil data (moisture content, classification and particle size distribution) was collected to calibrate the model. Additional data was obtained from the Soil Research Institute in Ghana to explore calibration of the Universal Soil Loss Equation (USLE, Wischmeier and Smith, 1978) for Ghanaian soils and environment. It was shown that the USLE could be successfully converted to provide meaningful soil loss estimates in the Ghanaian environment. However, due to experimental difficulties, the proposed theory and methodology of the sediment yield model could only be tested in principle. Future work may include validation of the model and subsequent scaling up to estimate sedimentation rates in Lake Volta.

Serum-free process development:improving the yield and consistency of human mesenchymal stromal cell production

Background aims: The cost-effective production of human mesenchymal stromal cells (hMSCs) for off-the-shelf and patient specific therapies will require an increasing focus on improving product yield and driving manufacturing consistency. Methods: Bone marrow-derived hMSCs (BM-hMSCs) from two donors were expanded for 36 days in monolayer with medium supplemented with either fetal bovine serum (FBS) or PRIME-XV serum-free medium (SFM). Cells were assessed throughout culture for proliferation, mean cell diameter, colony-forming potential, osteogenic potential, gene expression and metabolites. Results: Expansion of BM-hMSCs in PRIME-XV SFM resulted in a significantly higher growth rate (P < 0.001) and increased consistency between donors compared with FBS-based culture. FBS-based culture showed an inter-batch production range of 0.9 and 5 days per dose compared with 0.5 and 0.6 days in SFM for each BM-hMSC donor line. The consistency between donors was also improved by the use of PRIME-XV SFM, with a production range of 0.9 days compared with 19.4 days in FBS-based culture. Mean cell diameter has also been demonstrated as a process metric for BM-hMSC growth rate and senescence through a correlation (R² = 0.8705) across all conditions. PRIME-XV SFM has also shown increased consistency in BM-hMSC characteristics such as per cell metabolite utilization, in vitro colony-forming potential and osteogenic potential despite the higher number of population doublings. Conclusions: We have increased the yield and consistency of BM-hMSC expansion between donors, demonstrating a level of control over the product, which has the potential to increase the cost-effectiveness and reduce the risk in these manufacturing processes.

Catalytic supercritical water gasification of plastics with supported RuO2:a potential solution to hydrocarbons-water pollution problem

Here we report on a potential catalytic process for efficient clean-up of plastic pollution in waters, such as the Great Pacific Garbage Patch (CPGP). Detailed catalytic mechanisms of RuO2 during supercritical water gasification of common polyolefin plastics including low-density polyethylene (LDPE), high-density polyethylene (HDPE), polypropylene (PP) and polystyrene (PP), have been investigated in a batch reactor at 450 °C, 60 min. All four plastics gave very high carbon gasification efficiencies (CGE) and hydrogen gasification efficiencies (HGE). Methane was the highest gas component, with a yield of up to 37 mol kg−1LDPE using the 20 wt% RuO2 catalyst. Evaluation of the gas yields, CGE and HGE revealed that the conversion of PS involved thermal degradation, steam reforming and methanation; whereas hydrogenolysis was a possible additional mechanism during the conversion of aliphatic plastics. The process has the benefits of producing a clean-pressurized methane-rich fuel gas as well as cleaning up hydrocarbons-polluted waters.

Synthesis and evaluation of novel ellipticines as potential anti-cancer agents

Ellipticine is a natural product which possesses multimodal anti-cancer activity. This thesis encompasses the synthesis and biological evaluation of novel ellipticine and isoellipticine derivatives as anti-cancer agents. Expanding on previous work within the group utilising vinylmagnesium bromide, derivatisation of the C5 position of ellipticine was accomplished by reaction of a key ketolactam intermediate with Grignard reagents. Corresponding attempts to introduce diverse substitution at the C11 position were unsuccessful, although one novel C11 derivative was produced using an alkyllithium reagent. A panel of novel ellipticinium salts encompassing a range of substitutions at the N2, C9 and N6 positions were prepared. Extensive derivatisation of the N10 position of isoellipticine was undertaken for the first time. Novel substitution in the form of acid and methyl ester functionalities were introduced at the C7 position of isoellipticine while novel C7 aldehyde and alcohol derivatives were synthesised. A large panel of isoellipticinium salts were prepared with conditions adjusted for the reactivity of the alkyl halide. Novel coupling reactions to increase the yield of isoellipticine were attempted but proved unsuccessful. A panel of 54 novel derivatives was prepared and a multimodal analysis of their anti-cancer activity was conducted. The NCI 60-human tumour cell lines screen was a primary source of information on the in vitro activity of compounds with derivatives found to exert potent anticancer effects, with mean GI50 values as low as 1.01 μM across the full range of cancer types and as low as 16 nM in individual cell lines. A second in vitro screen in collaboration with researchers in the University of Nantes identified derivatives which could potently inhibit growth in a p53 mutant NSCLC cell line. The cell cycle effects of a selected panel of isoellipticines were studied in leukaemia cell lines by researchers in the Department of Biochemistry and Cell Biology, UCC. Emerging from this, the therapeutic potential of one of the derivatives in AML was then assessed in vivo in an AML xenograft mouse model, with tumour weight reduced by a factor of 7 in treated mice relative to control.

Exploring the potential of cassava in promoting agricultural growth in Nigeria

Cassava is one of the major food crops in Nigeria, with multiple uses from human consumption to industrial applications. This study explores the potential of cassava in Nigerian agriculture based on a review of cassava development policies; performs a trend analysis of the cultivation area, production, productivity, and real price of cassava and other competing crops for the period 1961–2013; identifies the sources of growth in production; and examines the production constraints at the local level based on a survey of 315 farmers/processors and 105 marketers from Delta State. The results revealed that several policies and programmes were implemented to develop the cassava sector with mixed outcomes. Although cassava productivity grew at 1.5% per annum (p.a.) during the post-structural adjustment programme period (1993–2013), its real price declined at a rate of 3.5% p.a. The effect of yield is the main source of growth in production, contributing 76.4% of the total growth followed by the area effect (28.2%). The cassava sector is constrained by inadequate market infrastructure, processing facilities, and lack of information and unstable prices at the local level. The widespread diffusion of improved tropical manioc selection technologies and investments in market and marketing infrastructure, processing technologies, irrigation/water provision and information dissemination are recommended to enhance the potential of the cassava sector to support agricultural growth in Nigeria.

Analysis of the Market Potential of Syrian Organic Fruit and Vegetables for Exports to Germany

Syria has been a major producer and exporter of fresh fruit and vegetables (FFV) in the Arabic region. Prior to 2011, Syrian FFV were mainly exported to the neighbouring countries, the Gulf States and Northern Africa as well as to Eastern European countries. Although the EU is potentially one of the most profitable markets of high quality FFV (such as organic ones) in the world, Syrian exports of FFV to Western European countries like Germany have been small. It could be a lucrative opportunity for Syrian growers and exporters of FFV to export organic products to markets such as Germany, where national production is limited to a few months due to climatic conditions. Yet, the organic sector in Syria is comparatively young and only a very small area of FFV is certified according to EU organic regulations. Up to the author’s knowledge, little was known about Syrian farmers’ attitudes towards organic FFV production. There was also no study so far that explored and analysed the determining factors for organic FFV adoption among Syrian farmers as well as the exports of these products to the EU markets. The overarching aim of the present dissertation focused on exploring and identifying the market potential of Syrian exports of organic FFV to Germany. The dissertation was therefore concerned with three main objectives: (i) to explore if German importers and wholesalers of organic FFV see market opportunities for Syrian organic products and what requirements in terms of quality and quantity they have, (ii) to determine the obstacles Syrian producers and exporters face when exporting agricultural products to Germany, and (iii) to investigate whether Syrian farmers of FFV can imagine converting their farms to organic production as well as the underlying reasons why they do so or not. A twofold methodological approach with expert interviews and a farmer survey were used in this dissertation to address the abovementioned objectives. While expert interviews were conducted with German and Syrian wholesalers of (organic) FFV in 2011 (9 interviews each), the farmer survey was administrated with 266 Syrian farmers of FFV in the main region for the production of FFV (i.e. the coastal region) from November 2012 till May 2013. For modelling farmers’ decisions to adopt organic farming, the Theory of Planned Behaviour as theoretical framework and Partial Least Squares Structural Equation Modelling as the main method for data analysis were used in this study. The findings of this dissertation yield implications for the different stakeholders (governmental institutions and NGOs, farmers, exporters, wholesalers, etc.) who are interested in prompting the Syrian export of organic products. Based on the empirical results and a literature review, an action plan to promote Syrian production and export of organic products was developed which can help in the post-war period in Syria at improving the organic sector.

Extracting DNA from ‘jaws’: high yield and quality from archived tiger shark (Galeocerdo cuvier) skeletal material

Archived specimens are highly valuable sources of DNA for retrospective genetic/genomic analysis. However, often limited effort has been made to evaluate and optimize extraction methods, which may be crucial for downstream applications. Here, we assessed and optimized the usefulness of abundant archived skeletal material from sharks as a source of DNA for temporal genomic studies. Six different methods for DNA extraction, encompassing two different commercial kits and three different protocols, were applied to material, so-called bio-swarf, from contemporary and archived jaws and vertebrae of tiger sharks (Galeocerdo cuvier). Protocols were compared for DNA yield and quality using a qPCR approach. For jaw swarf, all methods provided relatively high DNA yield and quality, while large differences in yield between protocols were observed for vertebrae. Similar results were obtained from samples of white shark (Carcharodon carcharias). Application of the optimized methods to 38 museum and private angler trophy specimens dating back to 1912 yielded sufficient DNA for downstream genomic analysis for 68% of the samples. No clear relationships between age of samples, DNA quality and quantity were observed, likely reflecting different preparation and storage methods for the trophies. Trial sequencing of DNA capture genomic libraries using 20 000 baits revealed that a significant proportion of captured sequences were derived from tiger sharks. This study demonstrates that archived shark jaws and vertebrae are potential high-yield sources of DNA for genomic-scale analysis. It also highlights that even for similar tissue types, a careful evaluation of extraction protocols can vastly improve DNA yield.

Hyperspectral sensing to detect the impact of herbicide drift on cotton growth and yield

Yield loss in crops is often associated with plant disease or external factors such as environment, water supply and nutrient availability. Improper agricultural practices can also introduce risks into the equation. Herbicide drift can be a combination of improper practices and environmental conditions which can create a potential yield loss. As traditional assessment of plant damage is often imprecise and time consuming, the ability of remote and proximal sensing techniques to monitor various bio-chemical alterations in the plant may offer a faster, non-destructive and reliable approach to predict yield loss caused by herbicide drift. This paper examines the prediction capabilities of partial least squares regression (PLS-R) models for estimating yield. Models were constructed with hyperspectral data of a cotton crop sprayed with three simulated doses of the phenoxy herbicide 2,4-D at three different growth stages. Fibre quality, photosynthesis, conductance, and two main hormones, indole acetic acid (IAA) and abscisic acid (ABA) were also analysed. Except for fibre quality and ABA, Spearman correlations have shown that these variables were highly affected by the chemical. Four PLS-R models for predicting yield were developed according to four timings of data collection: 2, 7, 14 and 28 days after the exposure (DAE). As indicated by the model performance, the analysis revealed that 7 DAE was the best time for data collection purposes (RMSEP = 2.6 and R2 = 0.88), followed by 28 DAE (RMSEP = 3.2 and R2 = 0.84). In summary, the results of this study show that it is possible to accurately predict yield after a simulated herbicide drift of 2,4-D on a cotton crop, through the analysis of hyperspectral data, thereby providing a reliable, effective and non-destructive alternative based on the internal response of the cotton leaves.

Heterosis in locally adapted sorghum genotypes and potential of hybrids for increased productivity in contrasting environments in Ethiopia

Increased productivity in sorghum has been achieved in the developed world using hybrids. Despite their yield advantage, introduced hybrids have not been adopted in Ethiopia due to the lack of adaptive traits, their short plant stature and small grain size. This study was conducted to investigate hybrid performance and the magnitude of heterosis of locally adapted genotypes in addition to introduced hybrids in three contrasting environments in Ethiopia. In total, 139 hybrids, derived from introduced seed parents crossed with locally adapted genotypes and introduced R lines, were evaluated. Overall, the hybrids matured earlier than the adapted parents, but had higher grain yield, plant height, grain number and grain weight in all environments. The lowland adapted hybrids displayed a mean better parent heterosis (BPH) of 19%, equating to 1160 kg ha− 1 and a 29% mean increase in grain yield, in addition to increased plant height and grain weight, in comparison to the hybrids derived from the introduced R lines. The mean BPH for grain yield for the highland adapted hybrids was 16% in the highland and 52% in the intermediate environment equating to 698 kg ha− 1 and 2031 kg ha− 1, respectively, in addition to increased grain weight. The magnitude of heterosis observed for each hybrid group was related to the genetic distance between the parental lines. The majority of hybrids also showed superiority over the standard check varieties. In general, hybrids from locally adapted genotypes were superior in grain yield, plant height and grain weight compared to the high parents and introduced hybrids indicating the potential for hybrids to increase productivity while addressing farmers' required traits.

Methane seep carbonates yield clumped isotope signatures out of equilibrium with formation temperatures

Methane cold seep systems typically exhibit extensive buildups of authigenic carbonate minerals, resulting from local increases in alkalinity driven by methane oxidation. Here, we demonstrate that modern seep authigenic carbonates exhibit anomalously low clumped isotope values (Delta(47)), as much as similar to 0.2 parts per thousand lower than expected values. In modern seeps, this range of disequilibrium translates into apparent temperatures that are always warmer than ambient temperatures, by up to 50 degrees C. We examine various mechanisms that may induce disequilibrium behaviour in modern seep carbonates, and suggest that the observed values result from several factors including kinetic isotopic effects during methane oxidation, mixing of inorganic carbon pools, pH effects and rapid precipitation. Ancient seep carbonates studied here also exhibit potential disequilibrium signals. Ultimately, these findings indicate the predominance of disequilibrium clumped isotope behaviour in modern cold seep carbonates that must be considered when characterizing environmental conditions in both modern and ancient cold seep settings.

Alternate furrow irrigation of four fresh-market tomato cultivars under semi-arid condition of Ethiopia – Part I: Effect on fruit yield and quality

Scarcity of freshwater due to recurrent drought threatens the sustainable crop production in semi-arid regions of Ethiopia. Deficit irrigation is thought to be one of the promising strategies to increase water use efficiency (WUE) under scarce water resources. A study was carried out to investigate the effect of alternate furrow irrigation (AFI), deficit irrigation (DI) and full irrigation (FI) on marketable fruit yield, WUE and physio-chemical quality of four fresh-market tomato cultivars (Fetan, Chali, Cochoro and ARP Tomato d2) in 2013 and 2014. The results showed that marketable yield, numbers of fruits per plant and fruit size were not significantly affected by AFI and DI irrigations. WUE under AFI and DI increased by 36.7% and 26.1%, respectively with close to 30% irrigation water savings achieved. A different response of cultivars to irrigation treatments was found for marketable yield, number of fruits and fruit size, WUE, total soluble solids (TSS) of the fruit juice, titratable acids (TA) and skin thickness. Cochoro and Fetan performed well under both deficit irrigation treatments exhibited by bigger fruit size which led to higher WUE. ARP Tomato d2 showed good yields under well-watered conditions. Chali had consistently lower marketable fruit yield and WUE. TSS and TA tended to increase under deficit irrigation; however, the overall variations were more explained by irrigation treatments than by cultivars. It was shown that AFI is a suitable deficit irrigation practice to increase fresh yield, WUE and quality of tomato in areas with low water availability. However, AFI requires suitable cultivars in order to exploit its water saving potential.

“Acid-spike” effect in spurs/tracks of the low/high linear energy transfer radiolysis of water : potential implications for radiobiology and nuclear industry

Résumé : Les ions hydronium (H3O + ) sont formés, à temps courts, dans les grappes ou le long des trajectoires de la radiolyse de l'eau par des rayonnements ionisants à faible transfert d’énergie linéaire (TEL) ou à TEL élevé. Cette formation in situ de H3O + rend la région des grappes/trajectoires du rayonnement temporairement plus acide que le milieu environnant. Bien que des preuves expérimentales de l’acidité d’une grappe aient déjà été signalées, il n'y a que des informations fragmentaires quant à son ampleur et sa dépendance en temps. Dans ce travail, nous déterminons les concentrations en H3O + et les valeurs de pH correspondantes en fonction du temps à partir des rendements de H3O + calculés à l’aide de simulations Monte Carlo de la chimie intervenant dans les trajectoires. Quatre ions incidents de différents TEL ont été sélectionnés et deux modèles de grappe/trajectoire ont été utilisés : 1) un modèle de grappe isolée "sphérique" (faible TEL) et 2) un modèle de trajectoire "cylindrique" (TEL élevé). Dans tous les cas étudiés, un effet de pH acide brusque transitoire, que nous appelons un effet de "pic acide", est observé immédiatement après l’irradiation. Cet effet ne semble pas avoir été exploré dans l'eau ou un milieu cellulaire soumis à un rayonnement ionisant, en particulier à haut TEL. À cet égard, ce travail soulève des questions sur les implications possibles de cet effet en radiobiologie, dont certaines sont évoquées brièvement. Nos calculs ont ensuite été étendus à l’étude de l'influence de la température, de 25 à 350 °C, sur la formation in situ d’ions H3O + et l’effet de pic acide qui intervient à temps courts lors de la radiolyse de l’eau à faible TEL. Les résultats montrent une augmentation marquée de la réponse de pic acide à hautes températures. Comme de nombreux processus intervenant dans le cœur d’un réacteur nucléaire refroidi à l'eau dépendent de façon critique du pH, la question ici est de savoir si ces fortes variations d’acidité, même si elles sont hautement localisées et transitoires, contribuent à la corrosion et l’endommagement des matériaux.

Assessment of the spatial variability in Tall wheatgrass forage using LANDSAT 8 satellite imagery to delineate potential management zones

Little information is available on the degree of within-field variability of potential production of Tall wheatgrass (Thinopyrum ponticum) forage under unirrigated conditions. The aim of this study was to characterize the spatial variability of the accumulated biomass (AB) without nutritional limitations through vegetation indexes, and then use this information to determine potential management zones. A 27-×-27-m grid cell size was chosen and 84 biomass sampling areas (BSA), each 2 m(2) in size, were georeferenced. Nitrogen and phosphorus fertilizers were applied after an initial cut at 3 cm height. At 500 °C day, the AB from each sampling area, was collected and evaluated. The spatial variability of AB was estimated more accurately using the Normalized Difference Vegetation Index (NDVI), calculated from LANDSAT 8 images obtained on 24 November 2014 (NDVInov) and 10 December 2014 (NDVIdec) because the potential AB was highly associated with NDVInov and NDVIdec (r (2) = 0.85 and 0.83, respectively). These models between the potential AB data and NDVI were evaluated by root mean squared error (RMSE) and relative root mean squared error (RRMSE). This last coefficient was 12 and 15 % for NDVInov and NDVIdec, respectively. Potential AB and NDVI spatial correlation were quantified with semivariograms. The spatial dependence of AB was low. Six classes of NDVI were analyzed for comparison, and two management zones (MZ) were established with them. In order to evaluate if the NDVI method allows us to delimit MZ with different attainable yields, the AB estimated for these MZ were compared through an ANOVA test. The potential AB had significant differences among MZ. Based on these findings, it can be concluded that NDVI obtained from LANDSAT 8 images can be reliably used for creating MZ in soils under permanent pastures dominated by Tall wheatgrass.

Winter cover crops on weed infestation and maize yield.

The establishment of commercial crops in succession to winter cover crops that leaves a dense straw layer provides significantly suppression of weeds. The objective of this work was to evaluate the suppressive potential of winter cover crops on weed infestation in maize and its effect on the yield of the maize sown in succession.