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Biomedical subjects

M Palatnik

Publications and source records attributed to M Palatnik.

At least 19 recordsLinked to original sources

Haemolytic activities of plant saponins and adjuvants. Effect of Periandra mediterranea saponin on the humoral response to the FML antigen of Leishmania donovani.

An 87.7% (P < 0.01) and 84% (P < 0.001) of protection against visceral leishmaniasis was achieved in CB hamsters and Balb/c mice, respectively, with saponin combined to the fucose-mannose ligand of Leishmania donovani (FML). However, an undesirable haemolytic effect was described for several saponins. Aiming to improve the formulation with FML/saponin, we comparatively analysed the haemolytic potential of recently characterized plant saponins and currently used adjuvants. The haemolytic activity of steroidic saponins from Agave sisalana; Smilax officinalis as well as commercial saponin (Riedel De Haën's), was higher than that of triterpenoid ones (Bredemeyera floribunda; Periandra mediterranea) and the Freund's complete adjuvant. The concentration resulting in 50% haemolysis was 500 micrograms ml-1 for aluminum hydroxide. The low haemolytic effect of P. mediterranea saponin was abolished by removal of its glycidic moiety and its sapogenin fraction as well as the Freund's Incomplete Adjuvant were non-haemolytic within this range. Furthermore, the adjuvant effect of three doses of P. mediterranea saponin injected with the FML antigen of L. donovani, was assayed in mice, either by the intraperitoneal (i.p.) or the subcutaneous (s.c.) route. The anti-FML IgG antibody levels increased and detectable levels were observed up to 3 months in the s.c. group. The response was expanded in both groups after an injection with a fourth vaccine dose. The IgG response showed increased levels of IgG2a only in the i.p. group, while IgG2b and IgG1 but not IgG3 antibodies were higher than controls in both groups. In conclusion, the results suggest that the recently described triterpenoid fractions of P. mediterranea can be safely used as adjuvant with low or non-haemolytic effect.

Adjuvants, Immunologic

Prevalence of anti-Leishmania donovani antibody among Brazilian blood donors and multiply transfused hemodialysis patients.

The prevalence of anti-Leishmania donovani antibodies was investigated in 1,500 Brazilian blood donors and multiply transfused hemodialysis patients. Sera were tested using the fucose-mannose ligand (FML) ELISA, which was shown to have 100% sensitivity and 96% specificity for kala-azar. Among 1,194 volunteer blood donors, seroreactivity was 9%, increasing to 25% in a periurban kala-azar focus. However, higher positivity (37%) was found in multiply transfused hemodialysis patients from Natal, where kala-azar is constantly present in low numbers (endemic), with sporadic outbreaks in localized regions (endemic and epidemic). Risk factors included blood transfusion, which was significantly associated with the presence of anti-Leishmania antibodies (chi2 = 8.567, P < 0.005), but did not include potential exposure to sandfly bites (chi2 = 0.033, P > 0.1). The prevalence significantly decreased to 7% in hemodialysis patients from Rio de Janeiro, where kala-azar is only occasionally seen, and was 0% in patients undergoing continuous ambulatorial peritoneal dialysis. The prospective analysis of 27 FML-seroreactive donors from Natal revealed amastigotes of Leishmania in the bone marrow of one subject while four had clinical complaints, including splenomegaly and hepatosplenomegaly. Our results point to the need for control of blood transfusion as a possible route for transmission of kala-azar in endemic areas.

Animals

Electrophoretic variation of hair proteins.

Hair follicle cells secrete a complex assortment of proteins that form the hair shaft, and can be classified into two major groups. The low-sulfur proteins are keratins that contribute to the backbone of intermediate filaments, and the high-sulfur proteins are associated with these filaments. In the present investigation we describe a comparative electrophoretic study of normal human hair proteins from 182 individuals, including some families. Hair proteins were extracted in urea buffer (pH 9.3), examined by 10% polyacrylamide gel electrophoresis (pH 8.8) in the presence of sodium dodecyl sulfate and stained with Coomassie brilliant blue. Eighteen bands appeared and were reproducible in most individuals, with apparent molecular mass ranging from 10.0 to approximately 100 kDa. Based on the most prominent bands, an electrophoretic profile defined as the "frequent profile" was observed. This profile was observed in 180 individuals and consisted of 6 prominent bands, 4 of them of apparent molecular mass in the 40-70-kDa range, which is characteristic of keratins (61.9 +/- 1.02, 58.5 +/- 1.21, 47.9 +/- 1.58, and 45.4 +/- 1.53 kDa), and 2 bands with lower molecular mass (18.9 +/- 0.75 and 13.7 +/- 0.91 kDa). In 2 samples from unrelated women, an additional band of 42.1 +/- 1.72 kDa appeared. The meaning of this variant is still under investigation.

Adult

An improved method for the isolation of erythrocyte antigen band-3.

An improved method for isolation of human and Rhesus monkey band-3 separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is described. Purified band-3 was obtained from human hemoglobin-free ghosts (Hb-free ghosts) after SDS-PAGE by chemical elution+sonication (CE+S). The section of the gel corresponding to the antigen was cut out, mechanically disrupted and incubated in 1% NaHCO3 containing 1% SDS, for 2 h, with shaking, at room temperature, followed by overnight incubation at 4 degrees C. The preparation was subsequently sonicated and clarified by centrifugation. Supernatants were dialyzed against distilled water, their protein contents were measured, and the presence of purified band-3 was demonstrated by SDS-PAGE. A calibration curve was developed for assay of CE+S material using densitometric evaluation of the protein profile on SDS-PAGE. An amount of 37.5 mg of Hb-free ghosts gave 3.15 mg of purified band-3 after CE+S, corresponding to an 8.4% yield. Rabbits were immunized with 50 micrograms CE+S antigen. Sera were collected and assayed by Western blot analysis against its proteolytic fragments, which were obtained from packed red blood cells by treatment with protease type VI from Streptomyces griseus (1 h at 37 degrees C), followed by extensive washing and hypotonic lysis. Specific antibodies recognized band-3 and its proteolytic fragments 60 and 63 kDa in human ghosts obtained from different blood donors, confirming the genetic polymorphism. Analogous serum obtained against the Rhesus monkey band-3 proteolytic fragment 63 kDa recognized the human antigen and its respective fragments. These results indicate the existence of similarities between these two species of band-3, suggesting the potential use of this technique in taxonomic and phylogenetic studies. Purification by CE+S is an efficient and rapid method for isolation of band-3 and its fragments with satisfactory yields and maintenance of both their immunogenic and antigenic properties.

Animals

The 60 and 63 kDa proteolytic peptides of the red cell membrane band-3 protein: their prevalence in human and non-human primates.

Three phenotypes based on the polymorphism of band-3 protein from human red cells are described. Limited proteolysis of intact red cells from most individuals (homozygotes) yields a peptide of 60 kDa, but in some cases (heterozygotes), there is also a 63-kDa peptide, and rarely only the single peptide of 63 kDa is found. This is the first description of the 63-kDa homozygote. The interpretation that the three phenotypes are controlled by two alleles of a single autosomal locus, with no dominance, is supported by population and family studies. The frequencies of the allele, which we designate as p63, is 0.041 +/- 0.0068 in Caucasoids and 0.125 +/- 0.0121 in Negroids. The electrophoretic profiles and molecular weights of the peptides obtained with several commercial proteases from Streptomyces griseus are similar to those obtained with chymotrypsin. Whereas band-3 protein in two New-World monkeys (Saimiri and Cebus) resisted pronase attack, an Old-World monkey (Macaca mulatta) was monomorphic for a 63-kDa fragment, and in an ape (Pan troglodytes), a doublet of 62 kDa and 64 kDa was found. Band-3 protein polymorphism appears to be a good marker for genetic differentiation in human populations.

Animals

ABO aggulutinins from Biomphalari straminea snails.

Extracts from B. straminea spawn are active against A and B normal red cells. A1 and A2 subgroups may be differentiated with trypsin-, papain- and pronase-treated cells. 0 cells treated with papain, pronase and neuraminidase react weakly to the extracts.

ABO Blood-Group System

Agglutinins and hemolysins from Biomphalaria tenagophila snails.

A saline anti-A and incomplete anti-A,B agglutinin were found in spawn extracts of Biomphalaria tenagophila snails. Significance differences after papain and pronase treatments were also found for A1 and A2 subgroups. The hemolysin titers are high, and unspecific.

ABO Blood-Group System

[Teaching by using genetic stimulation].

With the advent of the computer as an educational tool, new avenues have been opened in the teaching of biology. The development of instructional techniques using a variety of mathematical models has assisted and stimulated the study of genetics, particularly that of populations. This article refers to an instructional program by simulation of the genetic drift phenomenon, which enables the student to utilize results that would otherwise take a long time to observe. The authors feel that this teaching method is the only efficient way to teach the phenomenon, since its very complexity makes it very difficult to explain in the postgraduate classroom or through the conventional practical task. The program is being used in a graduate course in genetics; however it is felt that it can be applied to other health sciences fields and adopted to the levels of understanding of the students.

Brazil

Protectins in Argentine mollusks: immunological and immunochemical aspects.

Protectins and agglutinins in several organs, fluids and spawn from Argentine terrestrial and fresh-water gastropod species were examined. Differences or analogies with vertebrate immunoglobulin serological behaviour are summarized. Individual or group variability and the evolutionary meaning of the reproductive system-linked and the Ca++ ion-linked protectins are discussed.

ABO Blood-Group System