RESUMO
The aim of this study was to develop an assay to analyze the serum profile of Mannose-binding lectin (MBL) through a simple and "in-house" method (called "dot-N-man"). Furthermore, the study attempted to associate molecular masses of MBL to the profile of MBL gene polymorphisms in patients with hepatitis C. Heterogeneity in molecular masses of MBL is due to the impairment of oligomers formation, which is linked to genetic polymorphisms in the MBL gene. Individuals with AA genotype (wild-type) produce high-molecular-mass proteins, whereas AO and OO individuals produce intermediate and low-molecular-mass proteins, respectively. Sera of thirty patients carrying the hepatitis C virus (HCV) were investigated using MBL binding assay with mannan-coated nitrocellulose (dot-N-man). Purified MBL was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Dot-N-Man assay yielded MBL with molecular masses ranging between 55 and 320â¯kDa, comparable to low and high molecular mass forms of MBL. Nonreducing SDS-PAGE showed high molecular mass bands in all AA individuals while bands of 270 and 205â¯kDa were observed in sera for a number of patients with AO and OO genotypes, respectively. Immunoblotting confirmed the MBL samples obtained from the dot-N-man. These results provide new insights to understand the MBL molecular forms profile in patients infected with HCV- which could be useful in future investigations on the influence of the MBL structure/genotype on both the progression of infection and the response to hepatitis C therapy.
Assuntos
Hepacivirus/imunologia , Hepatite C , Immunoblotting/métodos , Lectina de Ligação a Manose , Polimorfismo Genético , Colódio/química , Feminino , Hepatite C/genética , Hepatite C/imunologia , Humanos , Masculino , Mananas/química , Lectina de Ligação a Manose/genética , Lectina de Ligação a Manose/imunologiaRESUMO
Bactericidal water filters were developed. For this purpose, nitrocellulose membrane filters were impregnated with different biosynthesized silver nanoparticles. Silver nanoparticles (AgNPs) from Aspergillus niger (AgNPs-Asp), Cryptococcus laurentii (AgNPs-Cry) and Rhodotorula glutinis (AgNPs-Rho) were used for impregnating nitrocellulose filters. The bactericidal properties of these nanoparticles against Escherichia coli, Enterococcus faecalis and Pseudomona aeruginosa were successfully demonstrated. The higher antimicrobial effect was observed for AgNPs-Rho. This fact would be related not only to the smallest particles, but also to polysaccharides groups that surrounding these particles. Moreover, in this study, complete inhibition of bacterial growth was observed on nitrocellulose membrane filters impregnated with 1 mg L(-1) of biosynthesized AgNPs. This concentration was able to reduce the bacteria colony count by over 5 orders of magnitude, doing suitable for a water purification device.
Assuntos
Antibacterianos/química , Colódio/química , Membranas Artificiais , Nanopartículas Metálicas/química , Prata/química , Purificação da Água/métodos , Antibacterianos/biossíntese , Antibacterianos/farmacologia , Água Potável/química , Estabilidade de Medicamentos , Fungos Mitospóricos/metabolismo , Porosidade , Prata/metabolismo , Prata/farmacologiaRESUMO
The objective of this work was to characterize the delignification effluent originating from the delignification industry and evaluate the combination of the fungus and photocatalytic process (TiO(2)/UV system) for the treatment of this effluent. The delignification effluent has proven harmful to the environment because it presents high color (3516 CU), total phenol (876 mg/L) and TOC (1599 mg/L) and is also highly toxic even in a low concentration. The results of photocatalysis were 11%, 25% and 13% higher for reductions in color, total phenol and TOC, respectively. The combined treatments presented benefits when compared to the non-combined treatments. Fungus and photocatalysis in combination proved to be the best treatment, reducing the color, total phenol, toxicity (inhibition of Escherichia coli growth) and TOC by 94.2%, 92.6%, 4.9% and 62%, respectively.
Assuntos
Colódio/química , Fungos/fisiologia , Fotoquímica/métodos , Purificação da Água/métodos , Biodegradação Ambiental , Reatores Biológicos , Carbono/química , Catálise , Resíduos Industriais , Indústrias/métodos , Fenol/química , Fenóis , Espectrofotometria Ultravioleta/métodos , Eliminação de Resíduos Líquidos/métodos , Poluentes Químicos da ÁguaRESUMO
Twelve strains of filamentous fungi, most of them belonging to the Deuteromycetes class, were isolated from activated sludge adapted to the delignification effluent from a nitrocellulose industry and screened to be used in the treatment of the effluent. The screening experiment was carried out using the effluent without co-substrate, treated for 120 h and pH 5. Aspergillus 2BNL1, Aspergillus 1AAL1 and Lentinus edodes UEC 2019 showed the highest effluent color reduction rates between 83% and 95%. The white-rot fungus L. edodes UEC 2019 was used as the control for the decolorization. In addition to color reduction, total phenol was also reduced in 56% and 79% by Aspergillus 2BNL1 and L. edodes UEC 2019, respectively. A kinetic experiment showed that Aspergillus 2BNL1 and Aspergillus 1AAL1 reduced the effluent color in the range of 81-95% at the first 24 h while L. edodes required 72 h to achieve a similar result. UV/Visible spectra revealed that all fungi treatments were able to decrease the chromophore compounds present in the effluent, except Aspergillus 1AAL1 that increased the UV absorptions. The molar weight distribution analysis showed that the three fungi were able to change the pattern of the effluent chromatogram, probably by degradation of the high molecular weight compounds.
Assuntos
Colódio/química , Fungos/metabolismo , Resíduos Industriais , Lignina/metabolismo , Eliminação de Resíduos Líquidos/métodos , Biodegradação Ambiental , Cor , Fungos/classificação , Monofenol Mono-Oxigenase/metabolismo , Fenóis/metabolismo , Esgotos/microbiologia , Fatores de TempoRESUMO
We have developed a simple method for plasma fibronectin purification based on the well-known gelatin binding property of fibronectin. In this procedure we immobilize the melted gelatin to nitrocellulose membranes; these are then used to affinity-purify the fibronectin from the plasma sample. The fibronectin is eluted from the membrane by treatment with 8 M urea. The procedure described here gives a yield of up to 60% (from presumed fibronectin concentration) and the fibronectin obtained is homogeneous in SDS-PAGE and biologically active, as assessed by a cell migration assay. The method is rapid, simple, inexpensive, does not require the use of chromatographic equipment and is suitable for tissue culture applications.
Assuntos
Colódio/química , Fibronectinas/isolamento & purificação , Técnicas de Cultura de Tecidos/instrumentação , Fibronectinas/sangue , Fibronectinas/metabolismo , Gelatina , Vidro , Humanos , Fatores de Tempo , UreiaRESUMO
It was found that the detyrosination of tyrosinated tubulin by tubulin carboxypeptidase can occur when both the enzyme and the substrate are adsorbed on nitrocellulose. This, and the use of a specific antibody that recognizes detyrosinated tubulin allowed us to localize tubulin carboxypeptidase on a nitrocellulose membrane after agarose gel electrophoresis and blotting. The method was also extended to detect pancreatic carboxypeptidase A.