Physicochemical and Biochemical Characterization of Enzymes Immobilized on lnorganic Matrices

Biotechnology is currently considered as a useful altemative to conventional process technology in industrial and catalytic fields. The increasing awareness of the need to create green and sustainable production processes in all fields of chemistry has stimulated materials scientists to search for innovative catalysts supports. lmmobilization of enzymes in inorganic matrices is very useful in practical applications due to the preserved stability and catalytic activity of the immobilized enzymes under extreme conditions. Nanostructured inorganic, organic or hybrid organic-inorganic nanocomposites present paramount advantages to facilitate integration and miniaturization of the devices (nanotechnologies), thus affording a direct connection between the inorganic, organic and biological worlds. These properties, combined with good chemical stability, make them competent candidates for designed biocatalysts, protein-separation devices, drug delivery systems, and biosensors Aluininosilicate clays and layered double hydroxides, displaying, respectively, cation and anion exchange properties, were found to be attractive materials for immobilization because of their hydrophilic, swelling and porosity properties, as well as their mechanical and thermal stability.The aim of this study is the replacement of inorganic catalysts by immobilized lipases to obtain purer and healthier products.Mesocellular silica foams were synthesized by oil-in-water microemulsion templating route and were functionalized with silane and glutaraldehyde. " The experimental results from IR spectroscopy and elemental analysis demonstrated the presence of immobilized lipase and also functionalisation with silane and glutaraldehyde on the supports.The present work is a comprehensive study on enzymatic synthesis of butyl isobutyrate through esterification reaction using lipase immobilized onto mesocellular siliceous foams and montmorillonite K-10 via adsorption and covalent binding. Moreover, the irnrnobil-ization does not modify the nature of the kinetic mechanism proposed which is of the Bi-Bi Ping—Pong type with inhibition by n-butanol. The immobilized biocatalyst can be commercially exploited for the synthesis of other short chain flavor esters. Mesocellular silica foams (MCF) were synthesized by microemusion templating method via two different routes (hydrothermal and room temperature). and were functionalized with silane and glutaraldehyde. Candida rugosa lipase was adsorbed onto MCF silica and clay using heptane as the coupling medium for reactions in non-aqueous media. I From XRD results, a slight broadening and lowering of d spacing values after immobilization and modification was observed in the case of MCF 160 and MCF35 but there was no change in the d-spacing in the case of K-10 which showed that the enzymes are adsorbed only on the external surface. This was further confirmed from the nitrogen adsorption measurements

Propyl Gallate Synthesis Using Acidophilic Tannase and Simultaneous Production of Tannase and Gallic Acid by Marine Aspergillus awamori BTMFW032

Marine Aspergillus awamori BTMFW032, recently reported by us, produce acidophilic tannase as extracellular enzyme. Here, we report the application of this enzyme for synthesis of propyl gallate by direct transesterification of tannic acid and in tea cream solubilisation besides the simultaneous production of gallic acid along with tannase under submerged fermentation by this fungus. This acidophilic tannase enabled synthesis of propyl gallate by direct transesterification of tannic acid using propanol as organic reaction media under low water conditions. The identity of the product was confirmed with thin layer chromatography and Fourier transform infrared spectroscopy. It was noted that 699 U/ml of enzyme could give 60% solubilisation of tea cream within 1 h. Enzyme production medium was optimized adopting Box–Behnken design for simultaneous synthesis of tannase and gallic acid. Process variables including tannic acid, sodium chloride, ferrous sulphate, dipotassium hydrogen phosphate, incubation period and agitation were recognized as the critical factors that influenced tannase and gallic acid production. The model obtained predicted 4,824.61 U/ml of tannase and 136.206 μg/ml gallic acid after 48 h of incubation, whereas optimized medium supported 5,085 U/ml tannase and 372.6 μg/ml of gallic acid production after 36 and 84 h of incubation, respectively, with a 15-fold increase in both enzyme and gallic acid production. Results indicated scope for utilization of this acidophilic tannase for transesterification of tannic acid into propyl gallate, tea cream solubilisation and simultaneous production of gallic acid along with tannase

Foreign exchange reserves in India and China

Professor Irma Glicman Adelman, an Irish Economist working in California University at Berkely, in her research work on ‘Development Over Two Centuries’, which is published in the Journal of Evolutionary Economics, 1995, has identified that India, along with China, would be one of the largest economies in this 21st Century. She has stated that the period 1700 - 1820 is the period of Netherlands, the period 1820 - 1890 is the period of England the period 1890 - 2000 is the period of America and this 21st Century is the century of China and India. World Bank has also identified India as one of the leading players of this century after China. India will be third largest economy after USA and China. India will challenge the Global Economic Order in the next 15 years. India will overtake Italian economy in 2015, England economy in 2020, Japan economy in 2025 and USA economy in 2050 (China will overtake Japan economy in 2016 and USA economy in 2027). India has the following advantages compared with other economies. India is 4th largest GDP in the world in terms of Purchasing Power. India is third fastest growing economy in the world after China and Vietnam. Service sector contributes around 57% of GDP. The share of agriculture is around 17% and Manufacture is 16% in 2005 - 2006. This is a character of a developed country. Expected GDP growth rate is 10% shortly (It has come down from 9.2% in 2006 - 2007 to 6.2% during 2008 - 2009 due to recession. It is only a temporary phenomenon). India has $284 billion as Foreign Exchange Reserve as on today. India had just $1 billion as Foreign Exchange Reserve when it opened its economy in the year 1991. In this research paper an attempt has been made to study the two booming economies of the globe with respect to their foreign exchange reserves. This study mainly based on secondary data published by respective governments and various studies done on this area

Application of biogas slurries from energy crops to arable soils and their impact on carbon and nitrogen dynamics

The use of renewable primary products as co-substrate or single substrate for biogas production has increased consistently over the last few years. Maize silage is the preferential energy crop used for fermentation due to its high methane (CH4) yield per hectare. Equally, the by-product, namely biogas slurry (BS), is used with increasing frequency as organic fertilizer to return nutrients to the soil and to maintain or increase the organic matter stocks and soil fertility. Studies concerning the application of energy crop-derived BS on the carbon (C) and nitrogen (N) mineralization dynamics are scarce. Thus, this thesis focused on the following objectives: I) The determination of the effects caused by rainfall patterns on the C and N dynamics from two contrasting organic fertilizers, namely BS from maize silage and composted cattle manure (CM), by monitoring emissions of nitrous oxide (N2O), carbon dioxide (CO2) and CH4 as well as leaching losses of C and N. II) The investigation of the impact of differences in soil moisture content after the application of BS and temperature on gaseous emissions (CO2, N2O and CH4) and leaching of C and N compounds. III) A comparison of BS properties obtained from biogas plants with different substrate inputs and operating parameters and their effect on C and N dynamics after application to differently textured soils with varying application rates and water contents. For the objectives I) and II) two experiments (experiment I and II) using undisturbed soil cores of a Haplic Luvisol were carried out. Objective III) was studied on a third experiment (experiment III) with disturbed soil samples. During experiment I three rainfall patterns were implemented including constant irrigation, continuous irrigation with periodic heavy rainfall events, and partial drying with rewetting periods. Biogas slurry and CM were applied at a rate of 100 kg N ha-1. During experiment II constant irrigation and an irrigation pattern with partial drying with rewetting periods were carried out at 13.5°C and 23.5°C. The application of BS took place either directly before a rewetting period or one week after the rewetting period stopped. Experiment III included two soils of different texture which were mixed with ten BS’s originating from ten different biogas plants. Treatments included low, medium and high BS-N application rates and water contents ranging from 50% to 100% of water holding capacity (WHC). Experiment I and II showed that after the application of BS cumulative N2O emissions were 4 times (162 mg N2O-N m-2) higher compared to the application of CM caused by a higher content of mineral N (Nmin) in the form of ammonium (NH4+) in the BS. The cumulative emissions of CO2, however, were on the same level for both fertilizers indicating similar amounts of readily available C after composting and fermentation of organic material. Leaching losses occurred predominantly in the mineral form of nitrate (NO3-) and were higher in BS amended soils (9 mg NO3--N m-2) compared to CM amended soils (5 mg NO3--N m-2). The rainfall pattern in experiment I and II merely affected the temporal production of C and N emissions resulting in reduced CO2 and enhanced N2O emissions during stronger irrigation events, but showed no effect on the cumulative emissions. Overall, a significant increase of CH4 consumption under inconstant irrigation was found. The time of fertilization had no effect on the overall C and N dynamics. Increasing temperature from 13.5°C to 23.5°C enhanced the CO2 and N2O emissions by a factor of 1.7 and 3.7, respectively. Due to the increased microbial activity with increasing temperature soil respiration was enhanced. This led to decreasing oxygen (O2) contents which in turn promoted denitrification in soil due to the extension of anaerobic microsites. Leaching losses of NO3- were also significantly affected by increasing temperature whereas the consumption of CH4 was not affected. The third experiment showed that the input materials of biogas plants affected the properties of the resulting BS. In particular the contents of DM and NH4+ were determined by the amount of added plant biomass and excrement-based biomass, respectively. Correlations between BS properties and CO2 or N2O emissions were not detected. Solely the ammonia (NH3) emissions showed a positive correlation with NH4+ content in BS as well as a negative correlation with the total C (Ct) content. The BS-N application rates affected the relative CO2 emissions (% of C supplied with BS) when applied to silty soil as well as the relative N2O emissions (% of N supplied with BS) when applied to sandy soil. The impacts on the C and N dynamics induced by BS application were exceeded by the differences induced by soil texture. Presumably, due to the higher clay content in silty soils, organic matter was stabilized by organo-mineral interactions and NH4+ was adsorbed at the cation exchange sites. Different water contents induced highest CO2 emissions and therefore optimal conditions for microbial activity at 75% of WHC in both soils. Cumulative nitrification was also highest at 75% and 50% of WHC whereas the relative N2O emissions increased with water content and showed higher N2O losses in sandy soils. In summary it can be stated that the findings of the present thesis confirmed the high fertilizer value of BS’s, caused by high concentrations of NH4+ and labile organic compounds such as readily available carbon. These attributes of BS’s are to a great extent independent of the input materials of biogas plants. However, considerably gaseous and leaching losses of N may occur especially at high moisture contents. The emissions of N2O after field application corresponded with those of animal slurries.

Effects of different tillage treatments on labile soil organic matter pools and stabilization processes

An improved understanding of soil organic carbon (Corg) dynamics in interaction with the mechanisms of soil structure formation is important in terms of sustainable agriculture and reduction of environmental costs of agricultural ecosystems. However, information on physical and chemical processes influencing formation and stabilization of water stable aggregates in association with Corg sequestration is scarce. Long term soil experiments are important in evaluating open questions about management induced effects on soil Corg dynamics in interaction with soil structure formation. The objectives of the present thesis were: (i) to determine the long term impacts of different tillage treatments on the interaction between macro aggregation (>250 µm) and light fraction (LF) distribution and on C sequestration in plots differing in soil texture and climatic conditions. (ii) to determine the impact of different tillage treatments on temporal changes in the size distribution of water stable aggregates and on macro aggregate turnover. (iii) to evaluate the macro aggregate rebuilding in soils with varying initial Corg contents, organic matter (OM) amendments and clay contents in a short term incubation experiment. Soil samples were taken in 0-5 cm, 5-25 cm and 25-40 cm depth from up to four commercially used fields located in arable loess regions of eastern and southern Germany after 18-25 years of different tillage treatments with almost identical experimental setups per site. At each site, one large field with spatially homogenous soil properties was divided into three plots. One of the following three tillage treatments was carried in each plot: (i) Conventional tillage (CT) with annual mouldboard ploughing to 25-30 cm (ii) mulch tillage (MT) with a cultivator or disc harrow 10-15 cm deep, and (iii) no tillage (NT) with direct drilling. The crop rotation at each site consisted of sugar beet (Beta vulgaris L.) - winter wheat (Triticum aestivum L.) - winter wheat. Crop residues were left on the field and crop management was carried out following the regional standards of agricultural practice. To investigate the above mentioned research objectives, three experiments were conducted: Experiment (i) was performed with soils sampled from four sites in April 2010 (wheat stand). Experiment (ii) was conducted with soils sampled from three sites in April 2010, September 2011 (after harvest or sugar beet stand), November 2011 (after tillage) and April 2012 (bare soil or wheat stand). An incubation study (experiment (iii)) was performed with soil sampled from one site in April 2010. Based on the aforementioned research objectives and experiments the main findings were: (i) Consistent results were found between the four long term tillage fields, varying in texture and climatic conditions. Correlation analysis of the yields of macro aggregate against the yields of free LF ( ≤1.8 g cm-3) and occluded LF, respectively, suggested that the effective litter translocation in higher soil depths and higher litter input under CT and MT compensated in the long term the higher physical impact by tillage equipment than under NT. The Corg stocks (kg Corg m−2) in 522 kg soil, based on the equivalent soil mass approach (CT: 0–40 cm, MT: 0–38 cm, NT: 0–36 cm) increased in the order CT (5.2) = NT (5.2) < MT (5.7). Significantly (p ≤ 0.05) highest Corg stocks under MT were probably a result of high crop yields in combination with reduced physical tillage impact and effective litter incorporation, resulting in a Corg sequestration rate of 31 g C-2 m-2 yr-1. (ii) Significantly higher yields of macro aggregates (g kg-2 soil) under NT (732-777) and MT (680-726) than under CT (542-631) were generally restricted to the 0-5 cm sampling depth for all sampling dates. Temporal changes on aggregate size distribution were only small and no tillage induced net effect was detectable. Thus, we assume that the physical impact by tillage equipment was only small or the impact was compensated by a higher soil mixing and effective litter translocation into higher soil depths under CT, which probably resulted in a high re aggregation. (iii) The short term incubation study showed that macro aggregate yields (g kg-2 soil) were higher after 28 days in soils receiving OM (121.4-363.0) than in the control soils (22.0-52.0), accompanied by higher contents of microbial biomass carbon and ergosterol. Highest soil respiration rates after OM amendments within the first three days of incubation indicated that macro aggregate formation is a fast process. Most of the rebuilt macro aggregates were formed within the first seven days of incubation (42-75%). Nevertheless, it was ongoing throughout the entire 28 days of incubation, which was indicated by higher soil respiration rates at the end of the incubation period in OM amended soils than in the control soils. At the same time, decreasing carbon contents within macro aggregates over time indicated that newly occluded OM within the rebuilt macro aggregates served as Corg source for microbial biomass. The different clay contents played only minor role in macro aggregate formation under the particular conditions of the incubation study. Overall, no net changes on macro aggregation were identified in the short term. Furthermore, no indications for an effective Corg sequestration on the long term under NT in comparison to CT were found. The interaction of soil disturbance, litter distribution and the fast re aggregation suggested that a distinct steady state per tillage treatment in terms of soil aggregation was established. However, continuous application of MT with a combination of reduced physical tillage impact and effective litter incorporation may offer some potential in improving the soil structure and may therefore prevent incorporated LF from rapid decomposition and result in a higher C sequestration on the long term.

Spatial and harvesting influence on growth, yield, quality and economic potential of Kalmegh (Andrographis paniculata Wall Ex. Nees)

Andrographis paniculata, commonly known as Kalmegh, is used both in Ayurvedic and Unani system of medicines because of its immunological, antibacterial and hepatoprotective properties. This study was carried out to investigate the influence of four harvesting times (120,135,150 days after planting and at seed maturity) and four planting distances (30×15, 30×10, 20×15 and 20×10 cm) on growth, dry herbage biomass, seed yield and quality traits of Andrographis paniculata at CCS Haryana Agricultural University, Hisar, India in the two years 2005 and 2006. The treatments were laid out in a split plot design with three replications. The maximum values for dry herbage biomass yield (5.14 t ha^(-1)), net returns (760.00 EUR ha^(-1)), B:C ratio (2.59), andrographolide content (2.63%) and total yield (135.00 kg ha^(-1)) were detected 135 days after planting with an optimum planting distance of 30×15 cm. However, the maximum iron content was estimated 120 days after planting. The highest dry herbage (4.58 t ha^(-1)) and maximum seed yield (19.7 kg ha^(-1)) were registered at plants that were lined out with a distance of 20×10 cm.

Factors that have shaped the eEvolution of bilateral trade between Canada and Colombia in the long run (1953-2003)

Currently bilateral trade between Canada and Colombia is low. In 2003, this trade amounted to US $489 million, with Co­lombian imports from Canada totaling US $ 213 million (DOTS, 2004). Canada supplied approximately 4.0% of Colombia’s total trade (total imports and total ex­ports), in contrast Colombia supplied only around 0.1% of the Canadian total trade. In 2005, Colombia was the third largest Canadian partner in South America after Brazil and Venezuela. The objective of this paper is to analyze the trade performance between Canada and Colombia using a modified gravity equation to identify the most relevant historical factors that have shaped the evo­lution of this bilateral trade in the long run (during the period from 1953 to 2003). The analysis includes traditional economic va­riables (such as population and income of the importing and exporting countries). In order to understand the qualitative nature of the evolution of this bilateral trade the second objective is to review the evolving nature of trade including commodity com­position and trade characteristics.  

Factores asociados a complicaciones post quirúrgicas en pacientes adultos con obstrucción intestinal mecánica en Bogotá, Colombia, período 2008 - 2014

Introducción: La obstrucción intestinal es una patología de alta prevalencia e impacto en los servicios de cirugía general a nivel mundial. El manejo de esta entidad puede ser médico o quirúrgico. Cuando se requiere intervención quirúrgica, se busca evitar el desarrollo de isquemia intestinal y resecciones intestinales; durante el postoperatorio, pueden existir complicaciones. El objetivo de este estudio es identificar los factores asociados al desarrollo de complicaciones post operatorias en un grupo de pacientes con obstrucción intestinal mecánica llevados a manejo quirúrgico. Metodología: Estudio analítico tipo casos y controles en un grupo de pacientes con diagnóstico de obstrucción intestinal mecánica llevados a manejo quirúrgico de su patología. Los casos corresponden a los pacientes con complicaciones postoperatorias y los controles aquellos que no presentaron complicaciones. Se identificaron factores asociados a complicación post operatoria mediante modelos estadísticos bivariados y multivariados de regresión logística para factores como edad, sexo, antecedente quirúrgico, presentación clínica, paraclínica y diagnóstico postoperatorio de malignidad, entre otras. Resultados: Se identificaron un total de 138 pacientes (54 casos y 129 controles). Los rangos de edad entre 55-66 años y mayor de 66 años fueron asociados con complicaciones postoperatorias (OR 3,87 IC95% 1,58-9,50 y OR 3,62 IC95% 1,45-9,08 respectivamente). El déficit de base inferior a 5 mEq/litro se relaciona con complicaciones postoperatorias (OR 2,64 IC95% 1.33-5,25) Otras pruebas de laboratorio, características radiológicas, hallazgos de malignidad en el postoperatorio y la evolución de los pacientes no fueron asociados con complicaciones. Conclusiones: Las disminución de las complicaciones durante el manejo quirúrgico de obstrucción intestinal mecánica continúa siendo un reto para la cirugía general. Factores no modificables como edad avanzada y modificables como el equilibrio ácido base deben ser tenidos en cuenta dada su correlación en el desarrollo de complicaciones postoperatorias.

Estimation of age- and stage-specific Catalan breast cancer survival functions using US and Catalan survival data

During the last part of the 1990s the chance of surviving breast cancer increased. Changes in survival functions reflect a mixture of effects. Both, the introduction of adjuvant treatments and early screening with mammography played a role in the decline in mortality. Evaluating the contribution of these interventions using mathematical models requires survival functions before and after their introduction. Furthermore, required survival functions may be different by age groups and are related to disease stage at diagnosis. Sometimes detailed information is not available, as was the case for the region of Catalonia (Spain). Then one may derive the functions using information from other geographical areas. This work presents the methodology used to estimate age- and stage-specific Catalan breast cancer survival functions from scarce Catalan survival data by adapting the age- and stage-specific US functions. Methods: Cubic splines were used to smooth data and obtain continuous hazard rate functions. After, we fitted a Poisson model to derive hazard ratios. The model included time as a covariate. Then the hazard ratios were applied to US survival functions detailed by age and stage to obtain Catalan estimations. Results: We started estimating the hazard ratios for Catalonia versus the USA before and after the introduction of screening. The hazard ratios were then multiplied by the age- and stage-specific breast cancer hazard rates from the USA to obtain the Catalan hazard rates. We also compared breast cancer survival in Catalonia and the USA in two time periods, before cancer control interventions (USA 1975–79, Catalonia 1980–89) and after (USA and Catalonia 1990–2001). Survival in Catalonia in the 1980–89 period was worse than in the USA during 1975–79, but the differences disappeared in 1990–2001. Conclusion: Our results suggest that access to better treatments and quality of care contributed to large improvements in survival in Catalonia. On the other hand, we obtained detailed breast cancer survival functions that will be used for modeling the effect of screening and adjuvant treatments in Catalonia

Common black coatings - reflectance and ageing characteristics in the 0.32-14.3 mu m wavelength range

The diffuse and regular reflectances of five optically absorbing coatings frequently used in optical systems, were measured over the 0.32-14.3 mu m wavelength range, before and after exposure to heat and intense optical radiation. The measured coatings included Nextel Velvet Black, an anodised coating and NPL Super Black. The anodised coating exhibited substantial variations in its diffuse and regular reflectance values after thermal and simulated solar ageing. Solar and thermal ageing of the Nextel Velvet Black resulted in increases of its reflectance. However, thermal ageing tended to decrease the reflectance of the other paint samples examined. Thermal and solar ageing of the NPL Super Black resulted in only minor changes in its reflectance characteristics. All measurements are traceable to the UK National Standards.

Monitoring and modelling the three-dimensional flow of water under drip irrigation

The interpretation of soil water dynamics under drip irrigation systems is relevant for crop production as well as on water use and management. In this study a three-dimensional representation of the flow of water under drip irrigation is presented. The work includes analysis of the water balance at point scale as well as area-average, exploring uncertainties in water balance estimations depending on the number of locations sampled. The water flow was monitored by detailed profile water content measurements before irrigation, after irrigation and 24 h later with a dense array of soil moisture access tubes radially distributed around selected drippers. The objective was to develop a methodology that could be used on selected occasions to obtain 'snap shots' of the detailed three-dimensional patterns of soil moisture. Such patterns are likely to be very complex, as spatial variability will be induced for a number of reasons, such as strong horizontal gradients in soil moisture, variations between individual sources in the amount of water applied and spatial variability is soil hydraulic properties. Results are compared with a widely used numerical model, Hydrus-2D. The observed dynamic of the water content distribution is in good agreement with model simulations, although some discrepancies concerning the horizontal distribution of the irrigation bulb are noted due to soil heterogeneity. (c) 2006 Elsevier B.V. All rights reserved.

The impact of fallowing and green manuring on soil conditions and the growth of sugarcane

There are currently concerns within some sugar industries that long-term monoculture has led to soil degradation and consequent yield decline. An investigation was conducted in Swaziland to assess the effects of fallowing and green manuring practices, over a seven-month period, on sugarcane yields and the physical properties of a poorly draining clay soil. In the subsequent first sugarcane crop after planting, yields were improved from 129 t ha(-1) under continuous sugarcane to 141-144 t ha(-1) after fallowing and green manuring, but there were no significant responses in the first and second ratoon crops. Also, in the first crop after planting, root length index increased from 3.5 km m(-2) under continuous sugarcane to 5.2-6.8 km m(-2) after fallowing, and improved rooting was still evident in the first ratoon crop where there had been soil drying during the fallow period. Soil bulk density, total porosity and water-holding capacity were not affected by the fallowing practices. However, air-filled porosity increased from 11% under continuous sugarcane to 16% after fallowing, and steady state ponded infiltration rates were increased from 0.61 mm h(-1) to 1.34 mm h(-1), but these improvements were no longer evident after a year back under sugarcane. Levels of soil organic matter were reduced in all cases, probably as a result of the tillage operations involved. In the plant crop, root length was well correlated with air-filled porosity, indicating the importance of improving belowground air supply for crop production on poorly draining clay soils.

AtTRB1, a telomeric DNA-binding protein from Arabidopsis, is concentrated in the nucleolus and shows highly dynamic association with chromatin

AtTRB1, 2 and 3 are members of the SMH (single Myb histone) protein family, which comprises double-stranded DNA-binding proteins that are specific to higher plants. They are structurally conserved, containing a Myb domain at the N-terminus, a central H1/H5-like domain and a C-terminally located coiled-coil domain. AtTRB1, 2 and 3 interact through their Myb domain specifically with telomeric double-stranded DNA in vitro, while the central H1/H5-like domain interacts non-specifically with DNA sequences and mediates protein–protein interactions. Here we show that AtTRB1, 2 and 3 preferentially localize to the nucleus and nucleolus during interphase. Both the central H1/H5-like domain and the Myb domain from AtTRB1 can direct a GFP fusion protein to the nucleus and nucleolus. AtTRB1–GFP localization is cell cycle-regulated, as the level of nuclear-associated GFP diminishes during mitotic entry and GFP progressively re-associates with chromatin during anaphase/telophase. Using fluorescence recovery after photobleaching and fluorescence loss in photobleaching, we determined the dynamics of AtTRB1 interactions in vivo. The results reveal that AtTRB1 interaction with chromatin is regulated at two levels at least, one of which is coupled with cell-cycle progression, with the other involving rapid exchange.

In vitro evaluation of fibrolytic enzymes as additives for maize (Zea mays L.) silage - II. Effects on rate of acidification, fibre degradation during ensiling and rumen fermentation

A study was carried out to determine the influence of fibrolytic enzymes derived from mesophilic or thermophilic fungal sources, added at ensiling, on time-course fermentation characteristics and in vitro rumen degradation of maize silage. The mesophilic enzyme was a commercial product derived from Trichodenna reesei (L), whereas the thermophilic enzyme was a crude extract produced from Thermoascus aurantiacus (Ta) in this laboratory. The fungus was cultured using maize cobs as a carbon source. The resulting fermentation extract was deionised to remove sugars and characterised for its protein concentration, main and side enzymic activities, optimal pH, protein molecular mass and isoelectric point. In an additional study, both enzymes were added to maize forage (333.5 g DM/kg, 70.0, 469.8, 227.1 and 307.5 g/kg DM of CP, NDF, ADF and starch, respectively) at two levels each, normalized according to xylanase activity, and ensiled in 0.5 kg capacity laboratory minisilos. Duplicate silos were opened at 2, 4, 8, 15, and 60 days after ensiling, and analysed for chemical characteristics. Silages from 60 days were bulked and in vitro gas production (GP) and organic matter degradability (OMD) profiles evaluated using the Reading Pressure Technique (RPT), in a completely randomised design. The crude enzyme extract contained mainly xylanase and endoglucanase activities, with very low levels of exoglucanase, which probably limited hydrolysis of filter paper. The extract contained three major protein bands of between 29 and 55 kDa, with mainly acidic isoelectric points. Ensiling maize with enzymes lowered (P < 0.05) the final silage pH, with this effect being observed throughout the ensiling process. All enzyme treatments reduced (P < 0.05) ADF contents. Treatments including Ta produced more gas (P < 0.05) than the controls after 24 h incubation in vitro, whereas end point gas production at 96 h was not affected. Addition of Ta increased (P < 0.01) OMD after 12 h (410 and 416 g/kg versus 373 g/kg), whereas both L and Ta increased (P < 0.05) OMD after 24 h. Addition of enzymes from mesophilic or thermophilic sources to maize forage at ensiling increased the rate of acidification of the silages and improved in vitro degradation kinetics, suggesting an improvement in the nutritive quality. (C) 2003 Elsevier B.V All rights reserved.

Fungicide and cultivar affect post-anthesis patterns of nitrogen uptake, remobilization and utilization efficiency in wheat

Three successive field experiments (2000/01-2002/03) assessed the effect of wheat cultivar (Consort.. Hereward and Shamrock) and fungicide (epoxiconazole and azoxystrobin) applied at and after flag leaf emergence on the nitrogen in the above-ground crop (Total N) and grain (Grain N), net nitrogen remobilization from non-grain tissues (Remobilized N). grain dry matter (Grain Dill), and nitrogen utilization efficiency (NUtE(g) = Grain DM/Total N). Ordinary logistic curves were fitted to the accumulation of Grain N, Grain DM and Remobilized N against thermal time after anthesis and used to simultaneously derive fits for Total N and NUtE(g). When disease was controlled, Consort achieved the greatest Grain DM, Total N, Grain N and NUtEg; in each case due mostly to longer durations, rather than quicker rates, of accumulation. Fungicide application increased final Grain Dill.. Grant N, Total N and Remobilized N, also mostly through effects on duration rather than rate of accumulation. Completely senesced leaf laminas retained less nitrogen when fungicide had been applied compared with leaf laminas previously infected severely with brown rust (Puccinia recondita) and Septoria tritici, or with just S. tritici. Late movement of nitrogen out of fungicide-treated laminas contributed to extended duration of both nitrogen remobilization and grain N filling, and meant that increases in NUtE(g) could occur without simultaneous reductions in grain N concentration.