Evaluation of GFP Tag as a Screening Reporter in Directed Evolution of a Hyperthermophilic beta-Glucosidase

By applying a directed evolution methodology specific enzymatic characteristics can be enhanced, but to select mutants of interest from a large mutant bank, this approach requires high throughput screening and facile selection. To facilitate such primary screening of enhanced clones, an expression system was tested that uses a green fluorescent protein (GFP) tag from Aequorea victoria linked to the enzyme of interest. As GFP`s fluorescence is readily measured, and as there is a 1:1 molar correlation between the target protein and GFP, the concept proposed was to determine whether GFP could facilitate primary screening of error-prone PCR (EPP) clones. For this purpose a thermostable beta-glucosidase (BglA) from Fervidobacterium sp. was used as a model enzyme. A vector expressing the chimeric protein BglA-GFP-6XHis was constructed and the fusion protein purified and characterized. When compared to the native proteins, the components of the fusion displayed modified characteristics, such as enhanced GFP thermostability and a higher BglA optimum temperature. Clones carrying mutant BglA proteins obtained by EPP, were screened based on the BglA/GFP activity ratio. Purified tagged enzymes from selected clones resulted in modified substrate specificity.

Comparison of gonadorelin products in lactating dairy cows: Efficacy based on induction of ovulation of an accessory follicle and circulating luteinizing hormone profiles

This study evaluated whether the four gonadorelin products that are commercially available in the United States produce comparable ovulation responses in lactating cows. Dairy cows at 7 d after last gonadotropin-releasing hormone (GnRH) treatment of Ovsynch (Day 7), with a corpus luteum (CL) >= 15 mm and at least one follicle >= 10 mm, were evaluated for response to GnRH treatment. Selected cows were randomized to receive (100 mu g; im): (1) Cystorelin (n = 146): (2) Factrel (n = 132): (3) Fertagyl (n = 140); or (4) Ovacyst (n = 140). On Day 14, cows were examined for Ovulation by detection of an accessory CL. Circulating luteinizing hormone (LH) concentrations were also evaluated in some cows after treatment with 100 mu g (n = 10 per group) or 50 mu g (n = 5 per group) GnRH. Statistical analyses were performed with the procedures MIXED and GLIMMIX of the SAS program. Percentage of cows ovulating differed (P < 0.01) among groups, with that for Factrel being lower (55.3%) than that for Cystorelin (76.7%), Fertagyl (73.6%), or Ovacyst (85.0%), There was no effect of batch, parity, or follicle size on ovulation response. but increasing body condition score decreased Ovulation response. There was a much greater LH release in cows treated with 100 mu g than in those treated with 50 mu g, but there were no detectable differences among products in time to LH peak, peak LH concentration, or area under the LH curve and no treatment effects nor treatment by time interactions on circulating LH profile. Thus, ovulation response to Factrel on Day 7 of the cycle was lower than that for other commercial GnRH products, although a definitive mechanism for this difference between products was not demonstrated. (C) 2009 Elsevier Inc. All rights reserved.

Annotated checklist of bird and mammal species of Cocha Cashu Biological Station, Manu National Park, Peru / John W. Terborgh, John W. Fitzpatrick, Louise Emmons.

n.s. no.21(1984)

New information on the Holocystites fauna (Diploporita) of the Middle Silurian of Wisconsin, Illinois, and Indiana / Terrence J. Frest --, Donald G. Mikulic -- and Christopher R. C. Paul --.

v.35:no.6(1977)

Microbiological and biochemical properties of an agricultural Mexican soil amended with sewage sludge

The application of sewage sludge is a concern because it may affect the quality of organic matter and microbiological and biochemical soil properties. The effects of surface application of sewage sludge to an agricultural soil (at 18 and 36 t ha-1 dry basis) were assessed in one maize (Zea mays L.) growing season. The study evaluated microbial biomass, basal respiration and selected enzymatic activities (catalase, urease, acid and alkaline phosphatase, and β-glucosidase) 230 days after sewage sludge application and infrared spectroscopy was used to assess the quality of dissolved organic matter and humic acids. Sewage sludge applications increased the band intensity assigned to polysaccharides, carboxylic acids, amides and lignin groups in the soil. The organic matter from the sewage sludge had a significant influence on the soil microbial biomass; nevertheless, at the end of the experiment the equilibrium of the soil microbial biomass (defined as microbial metabolic quotient, qCO2) was recovered. Soil urease, acid and alkaline phosphatase activity were strongly influenced by sewage sludge applications.

An Evaluation of the Calderon Test, HR-80, 1963

As a result of Chang's studies, Calderon's developments, and the need for a new test procedure to determine specific physical properties of an asphalt concrete, the Iowa Highway Research Board sponsored a research project to investigate the correlation of results of the Calderon Test with the Iowa Stability Test and the Marshall and Hveem stability tests using Iowa Type A asphaltic concrete. The project was assigned to the Bituminous. Research Laboratory of Iowa State University as Project HR 80, the. Iowa Highway Research Board, and Project 442-S of the Engineering Experiment Station.

Evaluation of tractive performance of four agricultural tractors in laterally inclined terrain

The present work aimed to evaluate the tractive performance of four agricultural tractors with auxiliary front traction in function of six lateral inclinations in a lateral track of trials, which belongs to the Agronomic Sciences Faculty from the São Paulo State University, Campus of Botucatu. The lateral inclinations were 0; 5; 10; 15; 20 and 25 degrees. In all of these situations, the tractors operated with predetermined load of an imposed traction to the pulled tractor of 40 kN in the inclination of 0 degrees. Hence a delineation in random blocks was used, considering six inclinations and four tractors, and with three repetitions for each treatment. The analyzed variables were slippage, tractive force, hourly fuel consumption, and speed. It was concluded that the pneumatic tire configurations influenced in the tractive performance of the tractors, as they increased the lateral inclinations of the terrain.

Evaluation of the agricultural vocational training programmes conducted by the Krishi Vigyan Kendras (Farm Science Centres) in Indian Punjab

Krishin Vigyan Kendras-KVKs (Farm Science Centres) have been established by the Indian Council of Agricultural Research in 569 districts. The trust areas of KVKs are refinement and demonstration of technologies, and training of farmers and extension functionaries. Imparting vocational trainings in agriculture and allied fields for the rural youth is one of its mandates. The study was undertaken to do a formative and summative (outcome and impact) evaluation of the beekeeping and mushroom growing vocational training programmes in the Indian state of Punjab. One-group pre and post evaluation design was employed for conducting a formative and outcome evaluation. The knowledge tests were administered to 35 beekeeping and 25 mushroom cultivation trainees, before and after the training programmes organized in 2004. The trainees significantly gained in knowledge. A separate sample of 640 trainees, trained prior to 2004, was selected for finding the adoption status. Out of 640, a sample of 200 was selected by proportionate sampling technique out of three categories, namely: non-adopters, discontinued-adopters and continued-adopters for evaluating the long-term impact of these training programmes. Ex-post-facto one-shot case study design was applied for this impact analysis. The vocational training programmes have resulted in continued-adoption of beekeeping and mushroom cultivation enterprises by 20% and 51% trained farmers, respectively. Age and trainee occupation had significant influence on the adoption decision of beekeeping vocation, whereas education and family income significantly affected the adoption decision of mushroom cultivation. The continued adopters of beekeeping and mushroom growing had increased their family income by 49% and 24%, respectively. These training programmes are augmenting the dwindling farm income of the farmers in Indian Punjab.

Abstracts of communications: Proceedings of the thirty-nineth meeting of the Agricultural Research Modellers' Group

This group, which is concerned with the applications of mathematics to agricultural science, was formed in 1970 and has since met at approximately yearly intervals in London for one-day meetings. The thirty-ninth meeting of the group, chaired by Professor N. Crout of the University of Nottingham, was held in the Kohn Centre at the Royal Society, 6 Carlton House Terrace, London on Friday, 30 March 2007 when the following papers were read.

Agricultural tenancies: implied surrender and regrant and the operation of the Agricultural Tenancies Act 1995

Agricultural tenancies arising after 1st September 1995 are mostly governed by the Agricultural Tenancies Act 1995. As such, tenants under this Act do not benefit from the degree of protection conferred on tenancies already in existence, which remain under the Agricultural Holdings Act 1986. Section 4 of the 1995 Act seeks to protect those tenancies which subsequently inadvertently undergo a surrender and regrant and which would otherwise lose the protection of the 1986 Act. This paper seeks to investigate, by relating recent case law and statute to the situation of agricultural tenancies, the occasions where surrender and regrant might occur and whether in such instances the protection of the 1986 Act will be lost.

The magnetized steel and scintillator calorimeters of the MINOS experiment

The Main Injector Neutrino Oscillation Search (MINOS) experiment uses an accelerator-produced neutrino beam to perform precision measurements of the neutrino oscillation parameters in the ""atmospheric neutrino"" sector associated with muon neutrino disappearance. This long-baseline experiment measures neutrino interactions in Fermilab`s NuMI neutrino beam with a near detector at Fermilab and again 735 km downstream with a far detector in the Soudan Underground Laboratory in northern Minnesota. The two detectors are magnetized steel-scintillator tracking calorimeters. They are designed to be as similar as possible in order to ensure that differences in detector response have minimal impact on the comparisons of event rates, energy spectra and topologies that are essential to MINOS measurements of oscillation parameters. The design, construction, calibration and performance of the far and near detectors are described in this paper. (C) 2008 Elsevier B.V. All rights reserved.

Evaluation of tractive performance of four agricultural tractors in laterally inclined terrain

O presente trabalho teve como objetivo avaliar o desempenho operacional de quatro tratores agrícolas com tração dianteira auxiliar, em função de seis inclinações laterais, em uma pista lateral de ensaios, pertencente à Faculdade de Ciências Agronômicas da Universidade Estadual Paulista, Câmpus de Botucatu. As inclinações laterais foram 0; 5; 10; 15; 20 e 25 graus. em todas estas situações, os tratores operaram com carga predeterminada de tração imposta ao trator tracionado de 40 kN na inclinação de 0 grau. Portanto, foi utilizado um delineamento em blocos casualizados (DBC), sendo seis inclinações e quatro tratores, e com três repetições para cada tratamento. As variáveis analisadas foram patinhagem, força de tração, consumo horário de combustível e velocidade de deslocamento. Concluiu-se que a configuração dos rodados pneumáticos influenciaram no desempenho operacional dos tratores, conforme aumentaram as inclinações laterais do terreno.

A Comparative Study of the Information-Seeking Behavior of Bachelor of Science and Master of Science Agricultural Extension and Education Students

This study compares information-seeking behavior of Bachelor of Science and Master of Science students in the fields of agricultural extension and education. The authors surveyed Iranian students in departments of agricultural extension and education at four universities in Tehran, Shiraz, Mollasani, and Kermanshah. This study focused on three aspects: (1) comparison of amounts of information-seeking behavior between Bachelor of Science and Master of Science agricultural extension and education students; (2) comparison of information-seeking behavior varieties in Bachelor of Science and Master of Science agricultural extension and education students; (3) Comparison of amounts of available information resources at four universities and its effectiveness on students' information-seeking behavior; and (4) comparison of research and educational outputs in Bachelor of Science and Master of Science students. Scale free technique, division by mean method, principal components analysis technique, Delphi method, t-test, correlation and regression tools were used for data analysis. This study revealed that Bachelor of Science students' information-seeking behavior is for improving educational output, but Master of Science students' information-seeking behavior is for promoting research output. Among varieties of Internet searching skills, library searching skills, and awareness of library information-seeking methods with students' information-seeking behavior, there are not significant differences between two groups of students.