Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation

Endochondral calcification involves the participation of matrix vesicles (MVs), but it remains unclear whether calcification ectopically induced by implants of demineralized bone matrix also proceeds via MVs. Ectopic bone formation was induced by implanting rat demineralized diaphyseal bone matrix into the dorsal subcutaneous tissue of Wistar rats and was examined histologically and biochemically. Budding of MVs from chondrocytes was observed to serve as nucleation sites for mineralization during induced ectopic osteogenesis, presenting a diameter with Gaussian distribution with a median of 306 ± 103 nm. While the role of tissue-nonspecific alkaline phosphatase (TNAP) during mineralization involves hydrolysis of inorganic pyrophosphate (PPi), it is unclear how the microenvironment of MV may affect the ability of TNAP to hydrolyze the variety of substrates present at sites of mineralization. We show that the implants contain high levels of TNAP capable of hydrolyzing p-nitrophenylphosphate (pNPP), ATP and PPi. The catalytic properties of glycosyl phosphatidylinositol-anchored, polidocanol-solubilized and phosphatidylinositol-specific phospholipase C-released TNAP were compared using pNPP, ATP and PPi as substrates. While the enzymatic efficiency (k cat/Km) remained comparable between polidocanol-solubilized and membrane-bound TNAP for all three substrates, the k cat/Km for the phosphatidylinositol-specific phospholipase C-solubilized enzyme increased approximately 108-, 56-, and 556-fold for pNPP, ATP and PPi, respectively, compared to the membrane-bound enzyme. Our data are consistent with the involvement of MVs during ectopic calcification and also suggest that the location of TNAP on the membrane of MVs may play a role in determining substrate selectivity in this micro-compartment.

Least limiting water range in soil under crop rotations and chiseling

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Adhesion of Histoplasma capsulatum to pneumocytes and biofilm formation on an abiotic surface

The pathogenic fungus, Histoplasma capsulatum, causes the respiratory and systemic disease 'histoplasmosis'. This disease is primarily acquired via inhalation of aerosolized microconidia or hyphal fragments of H. capsulatum. Evolution of this respiratory disease depends on the ability of H. capsulatum yeasts to survive and replicate within alveolar macrophages. It is known that adhesion to host cells is the first step in colonization and biofilm formation. Some microorganisms become attached to biological and non-biological surfaces due to the formation of biofilms. Based on the importance of biofilms and their persistence on host tissues and cell surfaces, the present study was designed to investigate biofilm formation by H. capsulatum yeasts, as well as their ability to adhere to pneumocyte cells. H. capsulatum biofilm assays were performed in vitro using two different clinical strains of the fungus and biofilms were characterized using scanning electron microscopy. The biofilms were measured using a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium-hydroxide (XTT) reduction assay. The results showed that both the H. capsulatum strains tested were very efficient at adhering to host cells and forming biofilm. Therefore, this is a possible survival strategy adopted by this fungus.

A tellurium-based cathepsin B inhibitor: Molecular structure, modelling, molecular docking and biological evaluation

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biological Response to Wollastonite Doped alpha-Tricalcium Phosphate Implants in Hard and Soft Tissues in Rats

The biological response following subcutaneous and bone implantation of beta-wollastonite(beta-W)-doped alpha-tricalcium phosphate bioceramics in rats was evaluated. Tested materials were: tricalciurn phosphate (TCP), consisting of a mixture of alpha- and beta-polymorphs; TCP doped with 5 wt. % of beta-W (TCP5W), composed of alpha-TCP as only crystalline phase; and TCP doped with 15 wt. % of beta-W (TCP15), containing crystalline alpha-TCP and beta-W. Cylinders of 2x1 mm were implanted in tibiae and backs of adult male Rattus norvegicus, Holtzman rats. After 7, 30 and 120 days, animals were sacrificed and the tissue blocks containing the implants were excised, fixed and processed for histological examination. TCP, TCP5W and TCP15W implants were biocompatible but neither bioactive nor biodegradable in rat subcutaneous tissue. They were not osteoinductive in connective tissue either. However, in rat bone tissue beta-W-doped alpha-TCP implants (TCP5W and TCP15W) were bioactive, biodegradable and osteoconductive. The rates of biodegradation and new bone formation observed for TCP5W and TCP15W implants in rat bone tissue were greater than for non-doped TCP.

Structural and biological characterization of three novel mastoparan peptides from the venom of the neotropical social wasp Protopolybia exigua (Saussure)

The venom of the Neotropical social wasp Protopolybia exigua(Saussure) was fractionated by RP-HPLC resulting in the elution of 20 fractions. The homogeneity of the preparations were checked out by using ESI-MS analysis and the fractions 15, 17 and 19 (eluted at the most hydrophobic conditions) were enough pure to be sequenced by Edman degradation chemistry, resulting in the following sequences:Protopolybia MPI I-N-W-L-K-L-G-K-K-V-S-A-I-L-NH2 Protopolybia-MP II I-N-W-K-A-I-I-E-A-A-K-Q-A-L-NH2 Protopolybia-MP III I-N-W-L-K-L-G-K-A-V-I-D-A-L-NH2All the peptides were manually synthesized on-solid phase and functionally characterized. Protopolybia-MP I is a hemolytic mastoparan, probably acting on mast cells by assembling in plasma membrane, resulting in pore formation; meanwhile, the peptides Protopolybia-MP II and -MP III were characterized as a non-hemolytic mast cell degranulator toxins, which apparently act by virtue of their binding to G-protein receptor, activating the mast cell degranulation. (C) 2004 Elsevier Ltd. All rights reserved.

Vascular flora of Serra do Japi Biological Reserve, Jundiaí, southeastern Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Relationship between the nucleolar cycle and chromatoid body formation in the spermatogenesis of Phrynops geoffroanus (Reptilia Testudines)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aplicação da técnica de caminhamento elétrico em área contaminada por derivados de petróleo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biomimetic apatite formation on Ultra-High Molecular Weight Polyethylene (UHMWPE) using modified biomimetic solution

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chemical properties of soils treated with biological sludge from gelatin industry

O impacto dos resíduos orgânicos agroindustriais no ambiente pode ser reduzido com o seu uso agrícola. do ponto de vista da fertilidade do solo, o que se deseja com a aplicação dos resíduos é aumentar o teor de matéria orgânica e fornecer nutrientes para as plantas. Neste trabalho, objetivou-se avaliar o efeito do lodo biológico de indústria de gelatina em atributos químicos de dois Argissolos Vermelho-Amarelos (PVA-arenoso e PVA-textura média) e de um Latossolo Vermelho (LV-argiloso). O experimento foi conduzido por 120 dias em laboratório, em delineamento inteiramente casualizado e esquema fatorial combinando os três solos e seis doses de lodo (0, 100, 200, 300, 400 e 500 m³ ha-1), com três repetições. A aplicação de até 500 m³ ha-1 de lodo diminui a acidez do solo e aumenta a CTC efetiva e a disponibilidade de N, Ca, Mg e P, sem ultrapassar o limite de tolerância para Na. O aumento do teor de bases, maior do que o da CTC efetiva, indica que a maior parte dos cátions adicionados pelo lodo permanece em solução e pode ser perdida por lixiviação.

Spatial and linear correlations between soil and corn

O conjunto de tecnologias aplicadas ao sistema de produção agrícola tem como características principais a verticalização da produtividade, diminuição de custos, melhoria nas características físicas, químicas e biológicas do solo para proporcionarem o crescimento sustentável do meio de produção. Desta forma, o trabalho teve como objetivo determinar a variabilidade e as correlações lineares e especiais entre os atributos da planta e do solo, visando selecionar um indicador da qualidade física do solo de boa representatividade para produção de forragem. No ano agrícola de 2006, na Fazenda Bonança, município de Pereira Barreto (SP), foram analisados a produtividade de forragem do milho outonal (FDM) em sistema plantio direto irrigado e os atributos físicos do solo, num Latossolo Vermelho distrófico. O objetivo foi estudar a variabilidade e as correlações lineares e espaciais entre os atributos da planta e os do solo, visando selecionar um indicador da qualidade física do solo de boa representatividade para a produtividade da forragem. Foi instalada a malha geoestatística, para coleta de dados do solo e planta, contendo 125 pontos amostrais, numa área de 2.500 m². Os atributos estudados, além de não terem variado aleatoriamente, apresentaram variabilidade dos dados entre baixa e muito alta e seguiram padrões espaciais bem definidos, com alcances entre 7,8 e 38,0 m. Por outro lado, a correlação linear entre os atributos da planta com o do solo foi baixa e extremamente significativa. Os pares Massa Seca de forragem versus Microporosidade e Diâmetro do colmo versus Densidade do Solo foram melhor correlacionados na camada de 0-0.10m, enquanto os outros pares - Massa Seca de Forragem versus Macroporosidade - e Porosidade Total - apresentaram correlação inversa para a mesma camada. Entretanto, do ponto de vista espacial, houve uma alta correlação inversa entre Massa Seca de Forragem com Microporosidade, de modo que a microporosidade na camada de 0-0.10m pode ser considerada um bom indicador de qualidade física do solo, tendo em vista a produção de forragem de milho.

Different responses in bulk density and saturated hydraulic conductivity to soil deformation by logging machinery on a Ferralsol under native forest

Ferralsols have high structural stability, although structural degradation has been observed to result from forest to tillage or pasture conversion. An experimental series of forest skidder passes in an east Amazonian natural forest was performed for testing the effects of mechanical stress during selective logging operations on a clay-rich Ferralsol under both dry and wet soil conditions. Distinct ruts formed up to 25 cm depth only under wet conditions. After nine passes the initially very low surface bulk density of between 0.69 and 0.80 g cm(-3) increased to 1.05 g cm(-3) in the wet soil and 0.92 g cm(-3) in the dry soil. Saturated hydraulic conductivities, initially > 250 mm h(-1), declined to a minimum of around 10 mm h(-1) in the wet soil after the first pass, and in the dry soil more gradually after nine passes. The contrasting response of bulk density and saturated hydraulic conductivity is explained by exposure of subsoil material at the base of the ruts where macrostructure rapidly deteriorated under wet conditions. We attribute the resultant moderately high hydraulic conductivities to the formation of stable microaggregates with fine sand to coarse silt textures. We conclude that the topsoil macrostructure of Ferralsols is subject to similar deterioration to that of Luvisols in temperate zones. The stable microstructure prevents marked compaction and decrease in hydraulic conductivity under wetter and more plastic soil conditions. However, typical tropical storms may regularly exceed the infiltration capacity of the deformed soils. In the deeper ruts water may concentrate and cause surface run-off, even in gently sloping areas. To avoid soil erosion, logging operations in sloping areas should therefore be restricted to dry soil conditions when rut formation is minimal.

Chemical structure, surface properties and biological activities of the biosurfactant produced by Pseudomonas aeruginosa LBI from soapstock

Pseudomonas aeruginosa LBI isolated from petroleum-contaminated soil produced rhamnolipids (RLLBI) when cultivated on soapstock as the sole carbon source. HPLC-MS analysis of the purified culture supernatant identified 6 RL homologues (%): R-2 C-10 C-10 28.9; R-2 C-10 C-12:1 23.0; R-1 C-10 C-10 23.4; R-2 C-10 C-12 11.3; R-2 C-10 C-12 7.9; R-2 C-10 C-12 C-12 5.5. To assess the potential antimicrobial activity of the new rhamnolipid product, RLLBI, its physicochemical properties were studied. RLLBI had a surface tension of 24 mN m(-1) and an interfacial tension 1.31 mN m(-1); the cmc was 120 mg l(-1). RLLBI produced stable emulsions with hydrocarbons and vegetable oils. This product showed good antimicrobial behaviour against bacteria: MIC for Bacillus subtilis, Staphylococcus aureus and Proteus vulgaris was 8 mg l(-1), for Streptococcus faecalis 4 mg l(-1), and for Pseudomonas aeruginosa 32 mg l(-1). RLLBI was active against phytopathogenic fungal species, MIC values of 32 mg l(-1) being found against Penicillium, Alternaria, Gliocadium virens and Chaetonium globosum. Due to its physicochemical properties and antimicrobial behaviour, RLLBI could be used in bioremediation treatment and in the food, cosmetic and pharmaceutical industries.