Membrane adsorber for endotoxin removal

ABSTRACT The surface of flat-sheet nylon membranes was modified using bisoxirane as the spacer and polyvinyl alcohol as the coating polymer. The amino acid histidine was explored as a ligand for endotoxins, aiming at its application for endotoxin removal from aqueous solutions. Characterization of the membrane adsorber, analysis of the depyrogenation procedures and the evaluation of endotoxin removal efficiency in static mode are discussed. Ligand density of the membranes was around 7 mg/g dry membrane, allowing removal of up to 65% of the endotoxins. The performance of the membrane adsorber prepared using nylon coated with polyvinyl alcohol and containing histidine as the ligand proved superior to other membrane adsorbers reported in the literature. The lack of endotoxin adsorption on nylon membranes without histidine confirmed that endotoxin removal was due to the presence of the ligand at the membrane surface. Modified membranes were highly stable, exhibiting a lifespan of approximately thirty months

NEOTROPICAL XENARTHRANS: a data set of occurrence of xenarthran species in the Neotropics

Xenarthrans – anteaters, sloths, and armadillos – have essential functions for ecosystem maintenance, such as insect control and nutrient cycling, playing key roles as ecosystem engineers. Because of habitat loss and fragmentation, hunting pressure, and conflicts with 24 domestic dogs, these species have been threatened locally, regionally, or even across their full distribution ranges. The Neotropics harbor 21 species of armadillos, ten anteaters, and six sloths. Our dataset includes the families Chlamyphoridae (13), Dasypodidae (7), Myrmecophagidae (3), Bradypodidae (4), and Megalonychidae (2). We have no occurrence data on Dasypus pilosus (Dasypodidae). Regarding Cyclopedidae, until recently, only one species was recognized, but new genetic studies have revealed that the group is represented by seven species. In this data-paper, we compiled a total of 42,528 records of 31 species, represented by occurrence and quantitative data, totaling 24,847 unique georeferenced records. The geographic range is from the south of the USA, Mexico, and Caribbean countries at the northern portion of the Neotropics, to its austral distribution in Argentina, Paraguay, Chile, and Uruguay. Regarding anteaters, Myrmecophaga tridactyla has the most records (n=5,941), and Cyclopes sp. has the fewest (n=240). The armadillo species with the most data is Dasypus novemcinctus (n=11,588), and the least recorded for Calyptophractus retusus (n=33). With regards to sloth species, Bradypus variegatus has the most records (n=962), and Bradypus pygmaeus has the fewest (n=12). Our main objective with Neotropical Xenarthrans is to make occurrence and quantitative data available to facilitate more ecological research, particularly if we integrate the xenarthran data with other datasets of Neotropical Series which will become available very soon (i.e. Neotropical Carnivores, Neotropical Invasive Mammals, and Neotropical Hunters and Dogs). Therefore, studies on trophic cascades, hunting pressure, habitat loss, fragmentation effects, species invasion, and climate change effects will be possible with the Neotropical Xenarthrans dataset

More than 10,000 pre-Columbian earthworks are still hidden throughout Amazonia

Indigenous societies are known to have occupied the Amazon basin for more than 12,000 years, but the scale of their influence on Amazonian forests remains uncertain. We report the discovery, using LIDAR (light detection and ranging) information from across the basin, of 24 previously undetected pre-Columbian earthworks beneath the forest canopy. Modeled distribution and abundance of large-scale archaeological sites across Amazonia suggest that between 10,272 and 23,648 sites remain to be discovered and that most will be found in the southwest. We also identified 53 domesticated tree species significantly associated with earthwork occurrence probability, likely suggesting past management practices. Closed-canopy forests across Amazonia are likely to contain thousands of undiscovered archaeological sites around which pre-Columbian societies actively modified forests, a discovery that opens opportunities for better understanding the magnitude of ancient human influence on Amazonia and its current state

Production Of An Acidic And Thermostable Lipase Of The Mesophilic Fungus Penicillium Simplicissimum By Solid-state Fermentation

The production of a lipase by a wild-type Brazilian strain of Penicillium simplicissimum in solid-state fermentation of babassu cake, an abundant residue of the oil industry, was studied. The enzyme production reached about 90 U/g in 72 h, with a specific activity of 4.5 U/mg of total proteins. The crude lipase showed high activities at 35-60 °C and pH 4.0-6.0, with a maximum activity at 50 °C and pH 4.0-5.0. Enzyme stability was enhanced at pH 5.0 and 6.0, with a maximum half-life of 5.02 h at 50 °C and pH 5.0. Thus, this lipase shows a thermophilic and thermostable behavior, what is not common among lipases from mesophilic filamentous fungi. The crude enzyme catalysed the hydrolysis of triglycerides and p-nitrophenyl esters (C4:0-C18:0), preferably acting on substrates with medium-chain fatty acids. This non-purified lipase in addition to interesting properties showed a reduced production cost making feasible its applicability in many fields. © 2009 Elsevier Ltd. All rights reserved.1002152495254Aidoo, K.E., Hendry, R., Wood, B.J.B., Estimation of fungal growth in a solid-state fermentation system (1981) Appl. Microbiol. Biol., 12, pp. 6-9Boutaiba, S., Bhatnagar, T., Hacene, H., Mitchell, D.A., Baratti, J.C., Preliminary characterization of a lipolytic activity from an extremely halophilic archaeon, Natronococcus sp. (2006) J. Mol. Catal. B - Enzyme, 41, pp. 21-26Cammarota, M.C., Freire, D.M.G., A review on hydrolytic enzymes in the treatment of wastewater with high oil and grease content (2006) Bioresour. Technol., 97, pp. 2195-2210Castilho, L.R., Polato, C.M.S., Baruque, E.A., Sant'Anna Jr., G.L., Freire, D.M.G., Economic analysis of lipase production by Penicillium restrictum in solid-state and submerged fermentations (2000) Biochem. Eng. J., 4, pp. 239-247Costa, M.A.F., Peralta, R.M., Production of lipase by soil fungi and partial characterization of lipase from a selected strain (Penicillium wortmanii) (1999) J. Basic Microbiol., 39, pp. 11-15Di Luccio, M., Capra, F., Ribeiro, N.P., Vargas, G.D., Freire, D.M.G., Oliveira, D., Effect of temperature, moisture, and carbon supplementation on lipase production by solid-state fermentation of soy cake by Penicillium simplicissimum (2004) Appl. Biochem. Biotechnol., 113-116, pp. 173-180Freire, D.M.G., (1996) Seleção de Microorganismos Lipolíticos e Estudo da Produção de Lipase por Penicillium restrictum, , Tese de Doutoramento, Universidade Federal do Rio de Janeiro, Departamento de Bioquímica, Instituto de Química, Rio de Janeiro, Brasil (174 pp.)Freire, D.M.G., Gomes, P.M., Bom, E.P.S., Sant'Anna Jr., G.L., Lipase production by a new promising strain of Penicillium restrictum (1997) Rev. Microbiol., 28, pp. 6-12Gombert, A.K., Lopes, A.P., Castilho, L.R., Freire, D.M.G., Lipase, amylase and protease production by Penicillium restrictum in a solid-state fermentation using babassu oil cake (1999) Process Biochem., 35, pp. 85-90Gutarra, M.L.E., Cavalcanti, E.D.C., Castilho, L.R., Freire, D.M.G., Sant'Anna Jr., G.L., Lipase production by solid-state fermentation: cultivation conditions and operation of tray and packed-bed bioreactors (2005) Appl. Biochem. Biotechnol., 121, pp. 105-116Haki, G.D., Rahshit, S.K., Developments in industrially important thermostable enzymes: a review (2003) Bioresour. Technol., 89, pp. 17-34Hölker, U., Höfer, M., Lenz, J., Biotechnological advantages of laboratory-scale solid-state fermentation with fungi (2004) Appl. Microbiol. Biotechnol., 64, pp. 175-186Jaeger, K.E., Dijkstra, B.W., Reetz, M.T., Bacterial biocatalysts: molecular biology, three-dimensional structures, and biotechnological applications of lipases (1999) Annu. Rev. Microbiol., 53, pp. 315-351Jesus, M.F.C.O., Branco, R.N., Sant'Anna Jr., G.L., Freire, D.M.G., Silva Jr., J.G., Penicillium restrictum lipases: a comparative study and characterization of enzymes with different degrees of purity (1999) Braz. J. Chem. Eng., 16, pp. 113-118Kakugawa, K., Shobayashi, M., Suzuki, O., Miyakuwa, T., Characterization of a lipase from the glycolipid-producing yeast Kurtzmanomyces sp. I-11 (2002) Biosci. Biotechnol. Biochem., 66, pp. 978-985Kamini, N.R., Mala, J.G.S., Puvanakrishnan, R., Lipase production from A. niger by solid-state fermentation using gingelly oil cake (1998) Process Biochem., 33, pp. 505-511Karadzic, I., Masui, A., Zivkovic, L.I., Fujiwara, N., Purification and characterization of an alkaline lipase from Pseudomonas aeruginosa isolated from putrid mineral cutting oil as component of metalworking fluid (2006) J. Biosci. Bioeng., 102, pp. 82-89Lima, V.M.G., Krieger, N., Mitchell, D.A., Fontana, J.D., Activity and stability of a crude lipase from Penicillium aurantiogriseum in aqueous media and organic solvents (2004) Biochem. Eng. J., 18, pp. 65-71Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., Protein measurement with folin phenol reagent (1951) J. Biol. Chem., 193, pp. 265-275Mateos Diaz, J.C., Rodríguez, J.A., Roussos, S., Cordova, J., Abousalham, A., Carriere, F., Baratti, J., Lipase from the thermotolerant fungus Rhizopus homothallicus is more thermostable when produced using solid-state fermentation than liquid fermentation procedures (2006) Enzyme Microb. Technol., 39, pp. 1042-1050Matsumoto, Y., Saucedo-Castañeda, G., Revah, S., Shirai, K., Production of β-N-acetylhexosaminidase of Verticillium lecanii by solid-state and submerged fermentations utilizing shrimp waste silage as substrate and inducer (2004) Process Biochem., 39, pp. 665-671Nawani, N., Kaur, J., Studies on lipolytic isoenzymes from a thermophilic Bacillus sp.: production, purification and biochemical characterization (2006) Enzyme Microb. Technol., 40, pp. 881-887Ramachandran, S., Singh, S.K., Larroche, C., Soccol, C.R., Pandey, A., Oil cakes and their biotechnological applications - a review (2006) Bioresour. Technol., 93, pp. 169-174Sharma, R., Chisti, Y., Banerjee, U.C., Production, purification, characterization, and applications of lipases (2001) Biotechnol. Adv., 19, pp. 627-662Stöcklein, W., Sztajer, H., Menge, U., Schmid, R.D., Purification and properties of lipase from Penicillium expansum (1993) Biochim. Biophys. Acta, 1168, pp. 181-189Sztajer, H., Lünsdorf, H., Erdmann, H., Menge, U., Schmid, R., Purification and properties of lipase from Penicillium simplicissimum (1992) Biochim. Biophys. Acta, 1124, pp. 253-261Tan, T., Zhang, M., Xu, J., Zhang, J., Optimization of culture conditions and properties of lipase from Penicillium camenbertii Thom PG-3 (2004) Process Biochem., 39, pp. 1495-1502Viniegra-González, G., Favela-Torres, E., Aguilar, C.N., Rómero-Gomez, S.J., Díaz-Godínez, G., Augur, C., Advantages of fungal enzyme production in solid-state over liquid fermentation systems (2003) Biochem. Eng. J., 13, pp. 157-16

Successful scale-up of human embryonic stem cell production in a stirred microcarrier culture system

Future clinical applications of human embryonic stem (hES) cells will require high-yield culture protocols. Currently, hES cells are mainly cultured in static tissue plates, which offer a limited surface and require repeated sub-culturing. Here we describe a stirred system with commercial dextran-based microcarriers coated with denatured collagen to scale-up hES cell production. Maintenance of pluripotency in the microcarrier-based stirred system was shown by immunocytochemical and flow cytometry analyses for pluripotency-associated markers. The formation of cavitated embryoid bodies expressing markers of endoderm, ectoderm and mesoderm was further evidence of maintenance of differentiation capability. Cell yield per volume of medium spent was more than 2-fold higher than in static plates, resulting in a significant decrease in cultivation costs. A total of 10(8) karyotypically stable hES cells were obtained from a unitary small vessel that needed virtually no manipulation during cell proliferation, decreasing risks of contamination. Spinner flasks are available up to working volumes in the range of several liters. If desired, samples from the homogenous suspension can be withdrawn to allow process validation needed in the last expansion steps prior to transplantation. Especially when thinking about clinical trials involving from dozens to hundreds of patients, the use of a small number of larger spinners instead of hundreds of plates or flasks will be beneficial. To our knowledge, this is the first description of successful scale-up of feeder- and Matrigel™-free production of undifferentiated hES cells under continuous agitation, which makes this system a promising alternative for both therapy and research needs

Avaliação psicométrica em português do indicador de dor crônica de Helsinki em cães com sinais crônicos de osteoartrite

RESUMO Objetivou-se determinar a validade, a confiabilidade e a sensibilidade do indicador de dor crônica de Helsinki (IDCH) em português, além de estabelecer correlação entre a plataforma de pressão em cães com displasia coxofemoral (DCF). O questionário foi preenchido por tutores de 40 animais com DCF e por 16 tutores de cães saudáveis. Os animais com DCF foram tratados com carprofeno 4,4mg/kg (GT= 21) ou placebo (GP= 19), administrados uma vez ao dia, durante quatro semanas. A avaliação consistiu no preenchimento do questionário (IDCH), na escala analógica visual de dor (EAVdor) e na escala analógica visual de locomoção (EAVloc) com duas semanas (A1) e imediatamente antes do tratamento (A2), duas (S2) e quatro (S4) semanas de tratamento e após duas semanas do término do tratamento (S6). A consistência interna dos dados foi considerada excelente (alfa de Cronbach = 0,89). Houve correlação moderada entre o IDCH e a EAVdor e boa entre o IDCH e a EAVloc, porém não houve diferença entre o grupo tratado com carprofeno e placebo no IDCH. Não houve correlação entre os dados da plataforma de pressão e o IDCH. O questionário apresentou validade de constructo, critério e fidedignidade e pode-se empregá-lo em países de língua portuguesa em cães com osteoartrite