Interação entre Hb C [beta6(A3)Glu>Lys] e IVS II-654 (C>T) beta-talassemia no Brasil

Thalassemias are a heterogeneous group of inherited disorders characterized by a microcytic hypochromic anemia and an imbalance in the synthesis of the globin-chains. Hb C is the second most frequently variant of hemoglobin found in Brazil. The laboratory diagnosis of hemoglobinopathies, including thalassemias, is growing in importance, particularly because of an increasing requirement for neonatal diagnosis of abnormal hemoglobins. Screening tests were carried out using alkaline and acid electrophoresis, globin-chain analysis by cellulose acetate in alkaline pH, isoelectric focusing and HPLC. The molecular characterization was made by PCR-ASO for Hb C and beta thalassemia mutants. Large-scale screening and discriminative methodologies must provide information about the hemoglobin polymorphisms in Brazilian population. HPLC is a powerful tool in these cases. Molecular characterization is important to genetic counseling and clinical management, in particular for the Brazilian population that have an intense racial admixture, with great variability of hemoglobins. In this paper an association between Hb C and beta thalassemia (IVS-II-654) in a black family from Brazil was described.

HB D Los Angeles in a Brazilian family

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização de hemoglobina N-Baltimore em doador de sangue de São José do Rio Preto, SP

As alterações que envolvem as globinas devem-se a modificações em genes responsáveis pela seqüência e estrutura das cadeias polipeptídicas, bem como aos genes reguladores da síntese destas cadeias. Hemoglobinas variantes apresentam estrutura química diferente da hemoglobina normal correspondente, resultante de mutações em uma ou mais bases nitrogenadas, ocasionando a troca de aminoácidos nas globinas alfa, beta, delta ou gama. A hemoglobina N-Baltimore é uma variante de globina beta, com substituição da lisina, na posição 95, por ácido glutâmico, apresentando mobilidade eletroforética mais rápida que a hemoglobina A em pH alcalino. Nas análises eletroforéticas em pH alcalino realizadas em doadores de sangue do Hemocentro de São José do Rio Preto (SP) identificamos a presença de portador de hemoglobina rápida em heterozigose, posteriormente confirmada por focalização isoelétrica e cromatografia líquida de alta pressão (HPLC). Os estudos de hemoglobinas anormais em doadores de sangue permitem a identificação de variantes raras e possibilitam o aconselhamento genético adequado a cada caso com estudo familial.

Screening of bacterial strains for pectinolytic activity: characterization of the polygalacturonase produced by Bacillus sp

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

Isolado protéico de semente de goiaba (Psidium guajava): caracterização de propriedades funcionais

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of guava during different phases of the industrial processing

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

Strontium zirconate heterogeneous catalyst for biodiesel production: Synthesis, characterization and catalytic activity evaluation

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diagnóstico laboratorial de hemoglobinas semelhantes à HbS

A hemoglobina S (HbS) está presente na população brasileira com prevalência variável, dependente dos grupos raciais formadores de cada região. A migração eletroforética em pH alcalino apresenta similaridade com outras hemoglobinas, e estudos complementares para sua correta caracterização são necessários. No presente estudo objetivamos traçar um fluxograma com as metodologias disponíveis para a caracterização da hemoglobina S e das hemoglobinas que apresentam migração semelhante em pH alcalino. No período de janeiro a junho de 2000, analisamos amostras de sangue com suspeita de hemoglobina S encaminhadas ao Laboratório de Hemoglobinas da Unesp. Caracterizamos diferentes mutantes e formas interativas com hemoglobina S, por procedimentos eletroforéticos, em variados pH, análises citológicas e testes bioquímicos específicos. Os procedimentos de análise aplicados resultaram em orientação fornecida aos laboratórios de rotina sobre como proceder no diagnóstico laboratorial destas alterações de hemoglobina. Desta forma contribuímos para um melhor conhecimento sobre a variabilidade genética das hemoglobinas em nossa população, auxiliando no acompanhamento clínico e no aconselhamento genético das hemoglobinopatias com fisiopatologia relacionada à alteração.

Electrophoretic and chromatographic profile for S-like hemoglobin

Hemoglobin variants originate mainly by simple amino acid substitutions, the result of nucleotide sequence changes. Recently, the number of known abnormal hemoglobins has increased due to improvement in analysis methodologies; however, many laboratories are not prepared to correctly identify mutants. Hb S is a very well-characterized hemoglobin variant that varies in prevalence in different regions of Brazil. However, there is a type of Hb that presents electrophoretic migration in alkaline pH similar to Hb S, named S-like Hb, which can be incorrectly diagnosed; therefore, its frequency is underestimated. We obtained reference ranges for Hb S by HPLC, and we examined the electrophoretic and chromatographic profiles of S-like Hb. Hb Hasharon, Hb D-Los Angeles, Hb Korle-Bu, Hb Lepore, Hb D-Iran, Hb G-like, Hb Queens, Hb Montgomery, and Hb Q-India were found. Cases of association between two beta chain mutants were also found. Electrophoresis in alkaline and acid pH was utilized to initially screen these Hb variants, and globin chain electrophoresis at both high and low pH was performed to identify the globin chain mutant. Chromatographic analysis permitted the identification of the hemoglobin variant and also facilitated the quantification of these variants. Therefore, an association of classical laboratory diagnostic methodologies is fundamental for the correct identification of suspect Hb variants. The S and S-like hemoglobin profiles determined in this study will help in the diagnosis of these variants in health care services.

Systematics and distribution of Thorea (Thoreaceae, Rhodophyta) from central Mexico and south-eastern Brazil

Thirteen populations of Thorea were analyzed from central Mexico and south-eastern Brazil. All populations were considered as belonging to a single species [Thorea hispida (Thore) Desvaux], with wide variation of morphological features. Secondary branches varying in frequency were observed in several populations with an overlapping in the range of branch density for Thorea violacea Bory and T. hispida (0-9 and 11-41 per 30 mm, respectively). As this is the most distinguishing character and on the basis of the overlapping (within a same population or even a single plant), we regarded T. violacea as a synonym of T. hispida. 'Chantransia' stage in culture, as well as gametophyte and carposporophyte were described in detail. We confirmed the coexistence of asexual monosporangia with sexual reproductive structures (carpogonia and spermatangia) and carposporangia. Size, content, arrangement and chromosome number were the most distinctive characteristics among spermatangia, carposporangia and monosporangia. Monosporangia can be promptly differentiated from spermatangia by their granulated content and larger size but they are similar to carposporangia in shape and size; however, monosporangia are not arranged in fascicles. Structures resembling bisporangia were observed in female plants of some populations. Chromosome numbers were n = 4 for spermatangia and fascicle cells, and 2n ca8 for gonimoblast filaments, carpospores and the 'Chantransia' stage cells. The populations of Thorea from central Mexico and south-eastern Brazil corroborated the known world distribution for T. hispida, consisting dominantly of tropical to subtropical rainforests, sometimes extending into warm temperate areas. Thorea hispida occurred in warm (temperature 17.6-28.0°C), neutral to alkaline (pH 7.0-8.0), high ion content (specific conductance 59-2140 μS cm-1), moderate flowing (current velocity 17-43 cm/s) and shallow waters (depth <50 cm); these data are essentially similar to previous reports.

A structural conformation study of aquatic humic acid

In the present work, aquatic humic substances (HS) were extracted by use of adsorbent XAD 8 and the acid humic fraction (AH) was separated throught acidification. After being purified by Hyphan resin and dialyze, the aquatic AH was characterized using Fourier-transform infrared spectroscopy and elemental analysis. The influence of the aquatic HA and electrolyte concentrations, pH and aquatic AH-metal complexation time on the conformation was investigated using UV/Vis spectroscopic studies, employing the equation suggested by Doty and Steiner. The results indicated that the acid humic flexible macromolecule assumes a condensed form at acid and alkaline pH. Other factors favoring condensed conformations are longer metal complexation time (ageing) and higher aquatic AH and electrolyte concentrations. Thus considering the strong influence of the investigated parameters in the structural conformation of the humic macromolecule, we conclude that studies using UV/Vis spectroscopy to estimate the concentration, aromaticity, humification degree of the aquatic AH and so on, require rigorous control over the experimental conditions employed to provide a correct interpretation of the analytical results. ©2006 Sociedade Brasileira de Química.

Chloroquine inhibits Paracoccidioides brasiliensis survival within human monocytes by limiting the availability of intracellular iron

The mechanisms used by Paracoccidioides brasiliensis (Pb 18) to survive into monocytes are not clear. Cellular iron metabolism is of critical importance to the growth of several intracellular pathogens, including P. brasiliensis, whose capacity to multiply in mononuclear phagocytes is dependent on the availability of intracellular iron. Chloroquine, by virtue of its basic properties, has been shown to prevent release of iron from holotransferrin by raising endocytic and lysosomal pH, and thereby interfering with normal iron metabolism. Then, in view of this, we have studied the effects of CHLOR on P. brasiliensis multiplication in human monocytes and its effect on the murine paracoccidioidomycosis. CHLOR induced human monocytes to kill P. brasiliensis. The effect of CHLOR was reversed by FeNTA, an iron compound that is soluble at neutral to alkaline pH, but not by holotransferrin, which releases iron only in an acidic environment. CHLOR treatment of Pb 18-infected BALB/c mice significantly reduced the viable fungi recovery from lungs, during three different periods of evaluation, in a dose-dependent manner. This study demonstrates that iron is of critical importance to the survival of P. brasiliensis yeasts within human monocytes and the CHLOR treatment in vitro induces Pb 18 yeast-killing by monocytes by restricting the availability of intracellular iron. Besides, the CHLOR treatment in vivo significantly reduces the number of organisms in the lungs of Pb-infected mice protecting them from several infections. Thus, CHLOR was effective in the treatment of murine paracoccidioidomycosis, suggesting the potential use of this drug in patients' treatment.

Description of electrophoretic and chromatographic hemoglobin profile of Rhinoclemmys punctularia

Studies of the hemoglobin pattern in Brazilian reptiles are important for determining ecological and phylogenetic relationships, but they are scarce. Peripheral blood samples were obtained from 7 males and 18 females of Rhinoclemmys punctularia. The hematological profile was based on the total hemoglobin and hematocrit values. The hemoglobin profile was obtained using electrophoretic procedures at different pH, isoelectric focusing, globin chain electrophoresis, and HPLC. The hematocrit (31 ± 2%) and total hemoglobin (7.5 ± 0.2 g/dL) values did not indicate gender variations. Alkaline pH electrophoresis of the total blood samples treated with 1% saponin demonstrated the presence of four well-defined hemoglobin fractions, one major component (fraction I), showing cathodic migration and three others faster than fraction I with anodic migration. When the samples were precipitated with chloroform, only two hemoglobin fractions were observed, similar to fractions I and III from the first procedure. Isoelectric focusing and HPLC showed the same pattern. With acid and neutral pH electrophoresis, two fractions with anodic migration were observed. The globin chain identification at alkaline pH showed two fractions, but four fractions were observed at acidic pH, suggesting that different polypeptide chains are involved in the hemoglobin molecule. The chromatographic separation of the total blood sample demonstrated that the major fraction comprised 81.9% and the minor 18.1%. The results obtained demonstrated a similarity between these hemoglobin components and those of some Chelidae reported in the literature for both land and aquatic animals, reflecting the adaptation to environmental conditions. ©FUNPEC-RP.

Morfologia e morfometria dos tipos de fibras do músculo reto abdominal de cadelas (canis familiaris) durante a gestação

The present paper deals with the evaluation of morphological and morphometric alterations of slow-twist (I) and fast-twitch (II) fibers of rectus abdominis muscle of adult female dog during pregestational phase, at 30 and 60 days of pregnancy and at 30 days after the parturition. At the every phases, using the open biopsy technique, muscle samples were collected. The samples were plunged. Histological sections were cut in a microtome. For general morphology, some sections were stained with HE. Subsequent sections were reacted for myofibrillar ATPase (m-ATPase), after alkaline (pH 10.4) and acid (pH 4.4) pre-incubations, in order to identificate type I and II fibers. In the pre-gestational phase, muscle tissue revealed to be composed by fibers with different diameters, presenting polygonal outlines and one or more periphery nuclei. At 30 days of pregnancy, muscle fiber characteristics were similar. At 60 days, in addition to the existence of normal fibers, polymorphic and small diameter fibers were frequent. At 30 days after the parturation, the morphology of muscle fiber were similar to that observed in the pre-gestational phase. In the four phases, type II fiber diameters were lager than type I. The diameters of both fiber types showed a significant reduction in the 30 days phase and a significative increasing at 60 days. The expansion of the abdominal wall during the pregnancy represents a chronic stimulus, induced changes in the morphology and in the fiber type diameters.

Abnormal hemoglobin phenotypes in carriers of mild anemia in Latin America.

We looked for abnormal hemoglobins in blood samples sent for diagnosis of anemia. Identification of the hemoglobins was made using electrophoretic, chromatographic and molecular procedures. The 2020 blood samples were of patients from various regions of Brazil and from some other Latin American countries. Among the abnormal hemoglobins that we found, 3.5% are known to be rare, while 51% had an electrophoretic profile similar to that of Hb S at alkaline pH. Differentiation was possible only by combining electrophoretic and chromatographic methods. Hb Hasharon, an alpha globin chain mutant, was the most frequently found variant hemoglobin; it accounted for 14.3% of the abnormal DNA samples. The other abnormal hemoglobin phenotypes displayed distinct electrophoretic profiles; most of them migrated faster than Hb A. The frequencies of the different abnormal hemoglobin profiles that we found reflect the miscegenation of the Latin American population and indicate the importance of hemoglobin studies using various methods in combination for accurate diagnosis and appropriate counseling of carriers and their families.