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Biomedical subjects

L B Carvalho

Publications and source records attributed to L B Carvalho.

14 recordsLinked to original sources

Sulfated fucan as support for antibiotic immobilization.

Xylofucoglucuronan from Spatoglossum schröederi algae was tested as a support for antibiotic immobilization. The polysaccharide (20 mg in 6 ml) was first activated using carbodiimide, 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide methiodide (20 mg in 2 ml), under stirring for 1 h at 25 masculine C and pH from 4.5 to 5.0. After adjusting the pH to 8.0, either gentamicin or amikacin (62.5 mg in 1.25 ml) was then immobilized on this chemically modified polysaccharide with shaking for 24 h in a cold room. Infrared spectra of the activated carbodiimide xylofucoglucuronan showed two bands to carbonyl (C=O at 1647.9 and 1700.7 cm(-1)) and to amide (C-NH2) groups (1662.8 and 1714.0 cm(-1)). Microbial characterization of the derivatives was carried out by the disk diffusion method using Staphylococcus aureus or Klebsiella pneumoniae incorporated in Müller Hinton medium. Inhibition halos of bacterial growth were observed for the antibiotics immobilized on this sulfated heteropolysaccharide before and after dialysis. However, the halos resulting from the samples after dialysis were much smaller, suggesting that dialysis removed either non-covalently bound antibiotic or other small molecules. In contrast, bacterial growth was not inhibited by either xylofucoglucuronan or its activated form or by gentamicin or amikacin after dialysis. An additional experiment was carried out which demonstrated that the sulfated heteropolysaccharide was hydrolyzed by the microorganism. Therefore, the antibiotic immobilized on xylofucoglucuronan can be proposed as a controlled drug delivery system. Furthermore, this sulfated heteropolysaccharide can be extracted easily from sea algae Spatoglossum schröederi.

Amikacin↗

Properties of carbodiimide treated heparin.

Heparin, a well-known anticoagulant, has been frequently used to coat surfaces for attaining blood compatibility of polymeric materials. Since carbodiimides are often used for immobilization of heparin on these biomaterials, the present study intended to evaluate some properties of carbodiimide-treated heparin. It was observed that the properties of heparin were altered after treatment with carbodiimide, mainly in the molar excess of this reagent. Thus, dye fixation and electrophoretic behaviour of heparin were modified, as well as its degradability by specific enzymes. Also, these modifications resulted in loss of anticoagulant activity. Infrared spectra of this carbodiimide-treated heparin presented evidence that can confirm its modification.

Biological Assay↗

Parkia pendula lectin as histochemistry marker for meningothelial tumour.

Lectins have been intensively used in histochemical techniques for cell surface characterization. These proteins are involved in several biological processes and their use as histochemical markers have been evaluated since they can indicate differences in cell surfaces. Parkia pendula lectin (PpeL) was evaluated as histochemical marker for meningothelial meningioma biopsies. Tissue slices were incubated with PpeL conjugated to horseradish peroxidase (PpeL-HRP) and Concanavalin A-HRP (ConA-HPR) and the binding visualized with diaminobenzidine and hydrogen peroxide. The lectin-tissue binding was inhibited with D-glucose. PpeL showed to be a useful tool for the characterization of meningothelial tumour and clinico-pathological diagnosis.

Adult↗

Polysiloxane/PVA-glutaraldehyde hybrid composite as solid phase for immunodetections by ELISA.

We developed an efficient method to prepare a hybrid inorganic-organic composite based on polyvinyl alcohol (PVA) and polysiloxane using the sol-gel disc technique. Antigen obtained from Yersinia pestis was covalently immobilized onto these discs with glutaraldehyde and used as solid phase in ELISA for antibody detection in serum of rabbits experimentally immunized with plague. Using 1.25 microg antigen per disc, a peroxidase conjugate dilution of 1:4,000 and a serum dilution of 1:200 were adequate for the establishment of the procedure. These values are similar to those used for PVA-glutaraldehyde discs, plasticized filter paper discs and the polyaniline-Dacron composite discs. This procedure is comparable to that which utilizes the adsorption of the antigen to conventional PVC plates, with the amount of antigen being one fourth that employed in conventional PVC plates (5 microg/well). In addition to the performance of the polysiloxane/PVA-glutaraldehyde disc as a matrix for immunodetection, its easy synthesis and low cost are additional advantages for commercial application.

Animals↗

Polyaniline-dacron composite as solid phase in enzyme linked immunosorbent assay for Yersinia pestis antibody detection.

Polyaniline (PANI) was chemically synthesized on a dacron disk surface and an antigen (F1 fraction) obtained from Yersinia pestis was covalently fixed onto this composite via glutaraldehyde. The enzyme linked immunosorbent assay (ELISA) or rapid ELISA procedure detected immunoglobulin G (IgG) anti-F1 fraction in human serum employing this derivative. The appropriate conditions for carrying out the test were established as an antigen concentration of 2 microg/PANI-dacron disk, peroxidase labeled goat anti-human IgG conjugate diluted 4000 times, and a serum dilution of 1:100. The PANI-dacron disks showed greater antigen retention than conventional poly(vinyl chloride) plates and less antibody unspecific adsorption.

Aniline Compounds↗

The use of filter paper plasticized with polyvinyl alcohol-glutaraldehyde in ELISA.

F1-antigen purified from Yersinia pestis was covalently linked to 5-mm diameter filter paper discs plasticized with polyvinyl alcohol-glutaraldehyde. These discs were used both for ELISA and dot-ELISA for the detection of anti-F1 IgG in rabbits. The best conditions were achieved using 1.25 microg of F1 antigen/disc, 3% w/v skim milk in PBS as blocking agent, anti-IgG peroxidase conjugate diluted 12,000 times, and serum from rabbits immunized or not against Y. pestis, diluted 6,400 times. The absorbance values obtained from the comparative study between this procedure and conventional ELISA were not significantly different but the low cost of the reagents employed in ELISA using the filter paper discs plasticized with polyvinyl alcohol-glutaraldehyde makes this method economically attractive.

Animals↗

Immobilization of xanthine oxidase on a polyaniline silicone support.

A polyaniline silicone support to immobilize xanthine oxidase is proposed as a reactor coil to monitor the action of xanthine oxidase on hypoxanthine, xanthine and 6-mercaptopurine. A purified xanthine oxidase immobilized on this support lost 80% of the initial activity after 12 min of use. Co-immobilization of superoxide dismutase and catalase increased the stability of immobilized xanthine oxidase so that the derivative maintained 79% of its initial activity after 4.6 h of continuous use in which 1.5 mumol purine bases were converted by the immobilized enzyme system. There is no evidence of either polyaniline or protein leaching from the coil during 3 h of continuous use. When solutions (10 ml) of hypoxanthine, xanthine and 6-mercaptopurine were circulated individually through the xanthine oxidase-superoxide dismutase-catalase-polyaniline coil (1 mm internal diameter and 3 m in length, 3 ml internal volume) activities of 8.12, 11.17 and 1.09 nmol min-1 coil-1, respectively, were obtained. The advantages of the reactor configuration and the redox properties of the polymer, particularly with respect to immobilized oxidoreductases, make this methodology attractive for similar enzyme systems. This immobilized enzyme system using polyaniline-silicone as support converted 6-mercaptopurine to 6-thiouric acid with equal efficiency as resins based on polyacrylamide and polyamide 11.

Enzymes, Immobilized↗

No relationship between HTLV-1 infection and filariasis--Serological study on patients with filariasis in Recife, Brazil.

The seroprevalence of antibodies against human T-cell leukemia virus was determined by ELISA in 68 patients with filarial infestation living in an endemic area. The total seropositivity was 2.9% and the HTLV-1-positive cases were detected in 2 microfilaremic patients 12 and 40 years old. This value is very close to that obtained for healthy individuals in the same region and age groups. This result suggests that there is no relationship between filariasis and HTLV-1 infection as previously proposed.

Adolescent↗

EEG and dementia indicators in AIDS patients' Rorschach test.

We studied the EEG and Rorschach test (RT) of nineteen AIDS patients and eight normal people in the same age group. Eight patients presented slow alpha rhythms (8 to 9 Hz); three, not-slow alpha rhythms (> 9 to 13 Hz); and eight, beta rhythms in background activity. Paroxystic activity, characterized by diffuse theta or delta waves, was present in eleven patients. We observed Oberholzer syndrome (organic dementia diagnosed by RT) in ten patients and Piotrowski syndrome (organic dementia diagnosed by RT) in eleven patients; six presented both. When considering only the group of AIDS patients, we did not observe a significant relation among slow alpha rhythm, not-slow alpha rhythm and the presence of paroxystic activity with the above-mentioned syndromes. AIDS patients with slow alpha rhythms showed a significantly greater number of Piotrowski syndrome dementia indicators when compared to normal individuals or those with slow alpha rhythms. We did not observe the same with Oberholzer syndrome.

AIDS Dementia Complex↗

Isolation and convulsant effect of the 'insulin-like' proteins from the pancreas of the Brazilian opossum Didelphis albiventris.

An opossum pancreatic insulin-like protein fraction was obtained at pH 5.2 which has high electrophoretic mobility. At pH 8.2, another pancreatic insulin-like protein fraction was obtained with low electrophoretic mobility. Isoelectric focusing studies showed that the high molecular weight protein has a pI value 8.35 and the low molecular weight protein has pI value of 3.9 when used at pH range between 3.5-10.0. The purified high and low molecular weight positive aldehyde-fuchsin proteins induced convulsions when administered subcutaneously to mice.

Animals↗

Action of amyloglucosidase on oxidised amylose.

The Michaelis constant and maximal velocity of alpha-amylase-free amyloglucosidase decrease with increasing periodate oxidation of amylose. These kinetic features have been explained on the basis of competitive inhibition by the oxidised non-reducing end of the (1 leads to 4)-alpha-D-glucan chain with the active centres of the enzyme. A kinetic model is proposed to demonstrate this special kind of inhibition where the concentration of inhibitor is directly proportional to the substrate concentration. The experimental data fitted this model, and the plots of 1/Km and 1/V against the ratio of oxidised/unoxidised non-reducing end-groups were straight lines.

Amylose↗

Acridinium ester conjugated to lectin as chemiluminescent histochemistry marker.

Cell differentiation/dedifferentiation includes changes in oligosaccharide composition and distribution in the cell surface glycoconjugates. Lectins have been used as auxiliary tools in histopathological diagnosis of mammary, uterus and brain pathologies. Acridinium ester (AE) conjugated to biomolecules has been employed in chemiluminescent analytical applications. This work aimed to use a lectin, concanavalin A (Con A), conjugated to AE as a chemiluminescent histochemistry tool. Biopsies of normal and infiltrating duct carcinoma (IDC) of mammary tissues were treated by a Con A-AE derivative. Photon emission, observed during the breakage of the chemical bound between Con A and AE, was quantified, expressed in relative light units (RLU) and correlated to the labelling of the normal and transformed tissues. The results demonstrated that RLU presented a linear relationship with the labelled tissue area in the range 0.125-1.0 cm2 (r=0.98). Furthermore, RLU was much higher for the IDC (1283.920x103+/-220.621x103) than the normal tissue (2.565x103+/-0.247x103), namely, about 500 times higher. The Con A-AE conjugation efficiency, differential staining of normal and IDC tissues, and quantification of results contribute to a decrease in the subjectivity in routine histopathological diagnoses and indicate that acrydinum ester can join other lectin marker to be used in histochemistry.

Acridines↗

Cellulose acetate as solid phase in ELISA for plague.

Antigen from Yersinia pestis was adsorbed on cellulose acetate discs (0.5 cm of diameter) which were obtained from dialysis membrane by using a paper punch. ELISA for human plague diagnosis was carried out employing this matrix and was capable to detect amount of 1.3 microg of antigen, 3,200 times diluted positive serum using human anti-IgG conjugate diluted 1:4,000. No relevant antigen lixiviation from the cellulose acetate was observed even after washing the discs 15 times. The discs were impregnated by the coloured products from the ELISA development allowing its use in dot-ELISA. Furthermore, cellulose acetate showed a better performance than the conventional PVC plates.

Antigens, Bacterial↗