Purification Of Microbial β-galactosidase From Kluyveromyces Fragilis By Bioaffinity Partitioning

This work investigated the partitioning of β-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-β-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme β-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where β-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of β-galactosidase (K = 2 x 10-5) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase.304324331Aguiñaga-Díaz, P.A., Guzmán, R.Z., Affinity partitioning of metal ions in aqueous polyethylene glycol/salt two-phase systems with PEG-modified dictators (1996) Sep. Sci. Technol., 31 (10), pp. 1483-1499Albertsson, P.Å., (1971) Partition of Cell Particles and Macromolecules. 2nd Ed., p. 323. , New York: InterscienceAlbertsson, P.Å., Tjemeld, F., Phase Diagrams (1994) Meth. Enzumol., 228, pp. 3-13Baskir, J.N., Hatton, T.A., Suter, U.W., Protein partitioning in two-phase aqueous polymer systems (1989) Biotechol. Bioeng., 34 (4), pp. 541-558Birkenmeier, G., Partitioning of blood proteins using immobilized dyes (1994) Meth. Enzymol., 228, pp. 154-167Brena, B.M., Rydén, L.G., Porath, J., Immobilization of β-galactosidase on metal-chelate-substituted gels (1994) Biotechnol. Appl. Biochem., 19 (2), pp. 217-231Chung, B.H., Bailey, D., Arnold, F.H., Metal affinity partitioning (1994) Meth. Enymol., 228, pp. 167-179Delgado, C., Patel, J.N., Francis, G.E., Fisher, D., Coupling of poly (ethyleneglycol) to albumin under very mild conditions by activation with chloride: Characterization of the conjugate by partitioning in aqueous two-phase systems (1990) Biotechnol. Appl. Biochem., 12, pp. 110-128Diamond, A.D., Hsu, J.T., Aqueous two-phase systems for biomolecule separation (1992) Adv. Biochem. Eng./Biolechnol., 47, pp. 89-135Flores, S.H., Alegre, R.M., β-galactosidase by Erwinia aroideae grown in cheese whey (1996) Arq. Biol. Tecnol., 39 (4), pp. 879-886Franco, T.T., Andrews, A.T., Asenjo, J.A., Use of chemically modified proteins to study the effect of a single protein characteristics in aqueous two-phase systems. Effect of surface hydrophobicity (1996) Biotechnol. Bioeng., 49, pp. 300-308Godfrey, T., Reichelt, J., (1983) Industrial Enzymology, pp. 514-548. , Hampshire: MacMillian Publishers LtdGuy, E.J., Bingham, E.W., Properties of β-galactosidase of Saccharomyces lactis in milk and milk products (1978) J. Dairy Sci., 61 (2), pp. 147-151Kitahata, S., Fujita, K., Hara, K., Hashimoto, H., Enzymatic synthesis of 4-O-β-galactosyl-maltopentocose by Bacillus circulans β-galactosidase (1991) Agricult. Biolog. Chem. J., 55 (9), pp. 2433-2434Kopperschaläger, G., Affinity extraction with dye ligands (1994) Meth. Enzymol., 228, pp. 121-136Kula, M.-R., Kroner, K.H., Hustedt, H., Purification of enzymes by liquid-liquid extraction (1982) Adv. Biochem. Engin., 24, pp. 73-118Kula, M.R., Trends and future of aqueous two-phase extraction (1990) Bioseparation, 1, pp. 181-189Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685Landman, O., Neurospora lactose. II. Enzyme formation in the standard strain (1984) Arch. Biochem. Biophys., 52, pp. 93-109Mahoney, R.R., Whitaker, J.R., Purification and physicochemical properties of β-galactosidase from Kluyveromyces fragilis (1978) J. Food Sci., 43, pp. 584-591Nilsson, K., Mosbach, K., Immobilization of ligands with organic sulphonyl chlorides (1984) Meth. Enzymol., 104, pp. 56-69Pastore, G.M., Park, Y.K., Purification and characterization of β-galactosidase from Scopulariopsis sp (1980) J. Ferm. Technol., 58 (1), pp. 79-81Prenosil, J.E., Stuker, E., Bourne, J.R., Formation of oligosaccharides during enzymatic lactose hydrolysis and their importance in a whey hydrolysis process: Part II: Experimental (1987) Biotechnol. Bioeng., 30, pp. 1026-1031Price, N.C., Stevens, L., (1989) Fundamentals of Enzymology, 2nd Ed., , Oxford: Science PublicationsSedmak, J.J., Grossberg, S.E., A rapid, sensitive, and versatile assay for protein using coomassic brilliant blue G250 (1977) Anal. Biochem., 79 (1), pp. 544-552Steers Jr., E., Cuatrecasas, P., Pollard, H.B., The purification of β-galactosidase from Escherichia coli by affinity chromatography (1971) J. Biolog. Chem., 246 (1), pp. 196-200Silva, M.E., Pellogia, C., Piza, F.A.T., Franco, T.T., Purification of three different microbial β-galactosidases by partitioning in aqueous two-phase systems (1997) Cienc. Tecnol. Alim., 17 (3), pp. 219-223Veide, A., Strandberg, L., Enfors, S.O., Extraction of β-galactosidase fused protein A in ATPS (1957) Enz. Microb. Technol., 9, pp. 730-738Wallenfels, K., Lehman, J., Malhotra, O.P., Die spezifität der β-galactosidase von Escherichia coli ML309 (1960) Biochemische Zeitschrift, 33

Removal Of Cyanobacteria Toxins From Drinking Water By Adsorption On Activated Carbon Fibers

Natural fibers from macadamia nut shell, dried coconut shell endocarp, unripe coconut mesocarp, sugarcane bagasse and pine wood residue were used to prepare activated carbon fibers (ACF) with potential application for removing microcystins. The ACF from pine wood and sugar cane bagasse were used to remove [D-Leucine1 MCYST-LR from water. After 10 minutes of contact time, more than 98% of toxin was removed by the ACF. The microcystin adsorption monolayer, qm, in the ACF recovered 200 and 161 μg.mg-1, with the Langmuir adsorption constant, KL, of 2.33 and 1.23 L.mg-1. Adsorption of [D-Leucine1]MCYST-LR in continuous process was studied for a fixed-bed ACF prepared from coconut shell and sugar cane bagasse and for two commercial activated carbon samples from treatment water plants of two Brazilian hemodialysis centers. Saturation of the beds occurred after 80 to 320 minutes, and the adsorption capacity for that toxin varied from 4.11 to 12.82 μg.mg-1.113371380Honda RY, Mercante CTJ, Vieira JMS, Esteves KE, Cabianca MAA, Azevedo MTP. Cianotoxinas em pesqueiros da região metropolitana de São Paulo. In: Esteves KE, Sant'Anna CL. (Org.). Pesqueiros sob uma visão integrada de meio ambiente, saúde pública e manejo. São Carlos: Rima2006. p. 105-120Sant'anna, C.L., Azevedo, M.T.D., Contribution to the knowledge of potentially toxic Cyanobacteria from Brazil (2000) Nova Hedwigia, 71 (3-4), pp. 359-385Landsberg, J.H., The effects of harmful algal blooms on aquatic organisms (2002) Reviews in Fisheries Science, 10 (2), pp. 113-390Hoffman, J.R.H., Removal of Microcystis toxins in water purification process (1976) Water S.A, 2 (2), pp. 58-60Keijola, A.M., Himberg, K., Esala, A.L., Sivonen, K., Hiisvirta, L., Removal of cyanobacterial toxins in water-treatment processes - laboratory and pilot-scale experiments (1988) Toxicity Assessment, 3 (5), pp. 643-656Falconer, I.R., Runnegar, M.T.C., Buckley, T., Huyn, V.L., Bradshaw, P., Use of powdered and granular activated carbon to remove toxicity from drinking water containing cyanobacterial toxins (1989) Journal American Water Works Association, 18 (2), pp. 102-105Himberg, K., Keijola, A.M., Hiisvirta, L., Pyysalo, H., Sivonen, K., The effect of water treatment processes on the removal of hepatotoxins from Microcystis and Oscillatoria cyanobacteria: A laboratory study (1988) Water Research, 23 (8), pp. 979-984Ministério do Desenvolvimento, Indústria e Comércio Exterior do Brasil (2005) Importation and exportation of activated carbon from Brazil, , http://aliceweb.desenvolvimento.gov.br, BRASIL, Available from:, Accessed on: JanuaryMatthiensen, A., Beattie, K.A., Yunes, J.S., Kaya, K., Codd, G.A., D-Leu(1) microcystin-LR, from the cyanobacterium Microcystis RST 9501 and from a Microcystis bloom in the Patos Lagoon estuary, Brazil (2000) Phytochemistry, 55 (5), pp. 383-387Oliveira, A.C.R., Magalhães, S.V.F., Soares, R.M., Azevedo, S.M.F.O., Influence of drinking water composition on quantitation and biological activity of dissolved microcystin (Cyanotoxin) (2005) Environmental toxicology, 20 (2), pp. 126-130Albuquerque-Jr, E.C., Mendez, M.O., Coutinho, A.R., Franco, T.T., Production and characterization of activated carbon from Brazilian agricultural residues (2005) Proceedings of the third Brazilian Carbono Congress, pp. 401-410. , Castro AT ed, November 7-11Rio de Janeiro, Brazil. Rio de Janeiro: Department of Science and Thecnology and Thecnological Center of the Brazilian Army;Webb, P.A., Orr, C., Surface area and pore structure by gas adsorption (1997) Analytical methods in fine particle technology, , Atlanta: Micromeritics Instrument Corp. Atlanta;, 301 pKuroda EK, Albuquerque-Jr EC, Di Bernardo L, Trofino JC. Caracteriza&ão e escolha do tipo de carvão ativado a ser empregado no tratamento de aguas contendo microcistinas. In: Brazilian Association of Sanitation and Environmental Engineering (ed.). Brazilian Environmental Sanitation: utopia or reality? Proceedings of the twentieth third Brazilian Congrees of Sanitary and Environmental Engineering2005 18-23Mato Grosso, Brazil. Campo Grande: Brazilian Association of Sanitary and Environmental Engineering2005. p. 1-10Albuquerque-Jr EC, Melo LFC, Franco TT. Use of solid-phase extraction, high-performance liquid chromatography, and MALDI-TOF identification for [D-Leu1]MCYST-LR analysis in treated water: Validation of the analytical methodology. Canadian Journal of Analytical Sciences & Spectroscopy. 200752(1). (in press)Ruthven, D.M., Goddard, M., Sorption and diffusion of C-8 aromatic-hydrocarbons in faujasite type zeolites. 1. equilibrium isotherms and separation factors (1986) Zeolites, 6 (4), pp. 275-282Ho, Y.S., McKay, G., Pseudo-second order model for sorption processes (1999) Process. Biochem, 34 (5), pp. 451-465Ho, Y.S., McKay, G., Kinetic models for the sorption of dye from aqueous solution by wood (1998) J. Environ. Sci. Health Part B: Process Safety Environ. Protect, 76 (B2), pp. 183-191Low, M.J.D., Kinetics of chemisorption of gases on solids (1960) Chem. Rev, 60 (3), pp. 267-312Chien, S.H., Clayton, W.R., Application of Elovich equation to the kinetics of phosphate release and sorption in soils (1980) Soil Sci. Soc. Am. J, 44 (2), pp. 265-268Santos, E.S., Guirardello, R., Franco, T.T., Preparative chromatography of xylanase using expanded bed adsorption (2002) Journal of chromatography A, 944 (1-2), pp. 217-224Barros, M.A.S.D., Zola, A.S., Arroyo, P.A., Aguiar, E.F.S., Tavares, C.R.G., Equilibrium and dynamic ion exchange studies of Cr3+ on zeolites NaA and NaX (2002) Acta Scientiarum, 24 (6), pp. 1619-1625Geankoplis, C.J., (2003) Transport Processes and Separation Process Principles, , 4 ed. USA: PTR Prentice Hall;Quinlivan, P.A., Li, L., Knappe, D.R.U., Effects of activated carbon characteristics on the simultaneous adsorption of aqueous organic micropollutants and natural organic matter (2005) Water Research, 39 (8), pp. 1663-1673Donati, C., Drikas, M., Hayes, R., Newcombe, G., Microcystin-LR adsorption by powdered activated carbon (1994) Water Research, 28 (8), pp. 1735-1742Pendleton, P., Schumann, R., Wong, S.H., Microcystin-LR adsorption by activated carbon (2001) Journal of Colloid and Interface Science, 240 (1), pp. 1-8Toles, C.A., Marshall, W.E., Johns, M.M., Surface functional groups on acid-activated nutshell carbons (1999) Carbon, 37 (8), pp. 1207-1214Mcdermott, C.M., Feola, R., Plude, J., Detection of cyanobacterial toxins (microcystins) in waters of northeastern Wisconsin by a new immunoassay technique (1995) Toxicon, 33 (11), pp. 1433-1442Ueno, Y., Nagata, S., Tsutsumi, T., Hasegawa, A., Yoshida, F., Suttajit, M., Mebs, D., Vasconcelos, V., Survey of microcystins in environmental water by a highly sensitive immunoassay based on monoclonal antibody (1996) Natural toxins, 4 (6), pp. 271-276Sivonen, K., Jones, G., Cyanobacterial toxins (1999) Toxic Cyanobacteria in Water, pp. 55-124. , A guide to their public health consequences, monitoring and management, and, ed, Routledge, UK: E&FN Spon;Robillot, C., Vinh, J., Puiseux-Dao, S., Hennion, M.C., Hepatotoxin Production Kinetics of the Cyanobacterium Microcystis aeruginosa PCC 7820, as Determined by HPLC-Mass Spectrometry and Protein Phosphatase Bioassay (2000) Environmental Science & Technology, 34 (16), pp. 3372-3378Pyo, D., Moon, D., Adsorption of microcystin LR by activated carbon fibers (2005) Bulletin of the Korean Chemical Society, 26 (12), pp. 2089-2092Yan, H., Gong, A., He, H., Zhou, J., Wei, Y., Lv, L., Adsorption of microcystins by carbon nanotubes (2006) Chemosphere, 62 (1), pp. 142-148Mohamed, Z.A., Carmichael, W.W., An, J., El-Sharouny, H.M., Activated carbon removal efficiency of microcystins in an aqueous cell extract of Microcystis aeruginosa and Oscillatoria tenuis strains isolated from Egyptian freshwaters (1999) Environmental toxicology, 14 (1), pp. 197-20

Moduli Stabilization in Stringy ISS Models

We present a stringy realization of the ISS metastable SUSY breaking model with moduli stabilization. The mass moduli of the ISS model is stabilized by gauging of a U(1) symmetry and its D-term potential. The SUSY is broken both by F-terms and D-terms. It is possible to obtain de-Sitter vacua with a vanishingly small cosmological constant by an appropriate fine-tuning of flux parameters.Comment: 14 pages, v2: minor corrections, refereces added, v3: better parameters and more figures, published versio

η\eta Production in Peripheral Heavy-Ion Collisions

We estimate the impact parameter dependence of the production cross section for $\eta_c$ and $\eta_b$ mesons in peripheral heavy-ion collisions collisions. Total and elastic $\gamma\gamma$ cross sections are calculated in an equivalent photon approximation.Comment: 9 pages, uuencoded postscrip

Scaling violations: Connections between elastic and inelastic hadron scattering in a geometrical approach

Starting from a short range expansion of the inelastic overlap function, capable of describing quite well the elastic pp and $\bar{p}p$ scattering data, we obtain extensions to the inelastic channel, through unitarity and an impact parameter approach. Based on geometrical arguments we infer some characteristics of the elementary hadronic process and this allows an excellent description of the inclusive multiplicity distributions in $pp$ and $\bar{p}p$ collisions. With this approach we quantitatively correlate the violations of both geometrical and KNO scaling in an analytical way. The physical picture from both channels is that the geometrical evolution of the hadronic constituents is principally reponsible for the energy dependence of the physical quantities rather than the dynamical (elementary) interaction itself.Comment: 16 pages, aps-revtex, 11 figure

Crescimento de eucalipto sob efeito de desfolhamento artificial

O objetivo deste trabalho foi avaliar os efeitos do desfolhamento total, realizado após o plantio e ao longo do primeiro ano de cultivo, sobre o crescimento de Eucalyptus grandis, desde a implantação até ao corte do povoamento. Foram avaliados cinco tratamentos: sem desfolhamento; um desfolhamento aos 56 dias após o plantio (DAP); dois desfolhamentos, aos 56 e 143 DAP; dois desfolhamentos, aos 56 e 267 DAP; e três desfolhamentos, aos 56, 143 e 278 DAP. Foram mensurados os diâmetros do tronco a 1,3 m e a altura total de 60 árvores por tratamento, em oito avaliações, do 21º ao 92º mês de cultivo. O crescimento médio em cada tratamento foi descrito por modelos de regressão não lineares e comparados por testes de identidade para comparar as tendências entre a testemunha e os demais tratamentos. O desfolhamento causou reduções significativas nas taxas de crescimento em diâmetro e altura das plantas, e diminuição expressiva no faturamento ao final da rotação, mesmo quando realizado uma única vez, no início do plantio. Maiores danos, no entanto, foram verificados após consecutivos desfolhamentos ao longo do primeiro ano de cultivo. A manutenção de áreas que tenham sofrido desfolhamento total na fase inicial de plantio pode tornar-se uma medida economicamente inviável.The objective of this work was to evaluate the effects of total defoliation at planting initial stages, and along the first year of cultivation, on Eucalyptus grandis growth, from planting to plantation cut. Five treatments were tested: without defoliation; one defoliation, at 56th day after planting (DAP); two defoliations, at 56th and 143th DAP; two defoliations, at 56th and 267th DAP; and three defoliations, at 56th, 143th and 278th DAP. Trunk diameter at 1.30-m height and the total height of 60 trees were measured from the 21st to the 92th cultivation months. The average growth of each treatment was described by nonlinear models and compared by identity tests in order to estimate the tendencies between control and the other treatments in each variable. Defoliation significantly reduces diameter of the trunk and height growth rates, and expressively decreases the income at the plantation cut. However, greater losses were verified after consecutive defoliation, along the first cultivation year. Maintaining areas that suffered severe defoliations at initial planting stages can become economically unfeasible

Trends and outcome of neoadjuvant treatment for rectal cancer: A retrospective analysis and critical assessment of a 10-year prospective national registry on behalf of the Spanish Rectal Cancer Project

Introduction: Preoperative treatment and adequate surgery increase local control in rectal cancer. However, modalities and indications for neoadjuvant treatment may be controversial. Aim of this study was to assess the trends of preoperative treatment and outcomes in patients with rectal cancer included in the Rectal Cancer Registry of the Spanish Associations of Surgeons. Method: This is a STROBE-compliant retrospective analysis of a prospective database. All patients operated on with curative intention included in the Rectal Cancer Registry were included. Analyses were performed to compare the use of neoadjuvant/adjuvant treatment in three timeframes: I)2006–2009; II)2010–2013; III)2014–2017. Survival analyses were run for 3-year survival in timeframes I-II. Results: Out of 14, 391 patients, 8871 (61.6%) received neoadjuvant treatment. Long-course chemo/radiotherapy was the most used approach (79.9%), followed by short-course radiotherapy ± chemotherapy (7.6%). The use of neoadjuvant treatment for cancer of the upper third (15-11 cm) increased over time (31.5%vs 34.5%vs 38.6%, p = 0.0018). The complete regression rate slightly increased over time (15.6% vs 16% vs 18.5%; p = 0.0093); the proportion of patients with involved circumferential resection margins (CRM) went down from 8.2% to 7.3%and 5.5% (p = 0.0004). Neoadjuvant treatment significantly decreased positive CRM in lower third tumors (OR 0.71, 0.59–0.87, Cochrane-Mantel-Haenszel P = 0.0008). Most ypN0 patients also received adjuvant therapy. In MR-defined stage III patients, preoperative treatment was associated with significantly longer local-recurrence-free survival (p < 0.0001), and cancer-specific survival (p < 0.0001). The survival benefit was smaller in upper third cancers. Conclusion: There was an increasing trend and a potential overuse of neoadjuvant treatment in cancer of the upper rectum. Most ypN0 patients received postoperative treatment. Involvement of CRM in lower third tumors was reduced after neoadjuvant treatment. Stage III and MRcN + benefited the most

Global data on earthworm abundance, biomass, diversity and corresponding environmental properties

14 p.Earthworms are an important soil taxon as ecosystem engineers, providing a variety of crucial ecosystem functions and services. Little is known about their diversity and distribution at large spatial scales, despite the availability of considerable amounts of local-scale data. Earthworm diversity data, obtained from the primary literature or provided directly by authors, were collated with information on site locations, including coordinates, habitat cover, and soil properties. Datasets were required, at a minimum, to include abundance or biomass of earthworms at a site. Where possible, site-level species lists were included, as well as the abundance and biomass of individual species and ecological groups. This global dataset contains 10,840 sites, with 184 species, from 60 countries and all continents except Antarctica. The data were obtained from 182 published articles, published between 1973 and 2017, and 17 unpublished datasets. Amalgamating data into a single global database will assist researchers in investigating and answering a wide variety of pressing questions, for example, jointly assessing aboveground and belowground biodiversity distributions and drivers of biodiversity change