Evaluation of epithelial cell proliferating activity and fibroblast nuclear kariometry in recurrent pterygium treated with mitomycin C

OBJETIVO: Avaliar a eficácia da mitomicina C (MMC) na prevenção da recorrência quando previamente utilizada no transplante autólogo de conjuntiva (TAC). A avaliação da proliferação celular epitelial pelo antígeno Ki-67 e a cariometria do núcleo dos fibroblastos foram usados como auxiliares na avaliação do tratamento. MÉTODOS: Vinte e nove pacientes com pterígio recidivado foram divididos em três grupos: Grupo (G) 1-TAC e colírio placebo (PLA); G2-TAC, MMC 0,015% subconjuntival e PLA; G3-TAC e colírio de MMC 0,02%. A imuno-histoquímica foi realizada no tecido excisado para o antígeno Ki-67, como a cariometria dos núcleos dos fibroblastos (divididos em lado nasal e temporal). A cariometria dos núcleos dos fibroblastos foi avaliada de acordo com os seguintes parâmetros: volume (Vl) e área (Ar) em pelos menos 50 células por paciente. RESULTADOS: A porcentagem das células epiteliais positivas para o antígeno Ki-67 no lado nasal e temporal após o tratamento dos três grupos estudados foi: nasal (3,30% G1, 4,49% G2 e 3,38% G3) e temporal (3,30% G1, 4,46% G2 e 4,14% G3) não mostrando diferença significativa. A cariometria do núcleo dos fibroblastos foi: Vl nasal (792,1 µ3 G1, 605,1 µ3 G2, e 549,9 µ3 G3) e a Ar (100,58 µ2 G1, 83,13 µ2 G2, e 78,41 µ2 G3). Os três grupos mostraram uma diferença significativa p=0,039 e p=0,035, respectivamente do Vl e da Ar no lado nasal. Após seis meses de tratamento, os três grupos apresentaram a seguinte taxa de recidiva: 22,22% G1, 18,18%, G2 e 33,33% G3 respectivamente. CONCLUSÃO: O uso da MMC não interferiu nas células epiteliais positivas para o antígeno Ki-67 no pterígio recidivado, mas acarretou diminuição do volume e área dos núcleos dos fibroblastos no lado nasal do pterígio. As células epiteliais positivas para o antígeno Ki-67 parecem não ter relação com a recidiva do pterígio após seis meses da cirurgia. Outros estudos devem ser realizados para avaliar o papel da proliferação das células epiteliais na recorrência do pterígio.

Glioma cordoide do terceiro ventrículo: descrição de um novo caso

O glioma cordoide é um tumor cerebral raro, recentemente descrito, localizado na região do terceiro ventrículo e com características histológicas, imuno-histoquímicas e ultraestruturais peculiares. Este estudo ilustra um caso de glioma cordoide do terceiro ventrículo em uma paciente de 59 anos de idade.

Influência da terapêutica hormonal prévia sobre os indicadores de prognóstico do câncer de mama em mulheres na pós-menopausa

OBJETIVO: avaliar a influência da terapêutica hormonal (TH) prévia sobre alguns indicadores de prognóstico do câncer de mama, em pacientes na pós-menopausa. MÉTODOS: estudo transversal por meio da aplicação de questionários e levantamento de prontuários. Foram entrevistadas 157 pacientes com diagnóstico de câncer de mama na pós-menopausa, registrando-se dados clínicos, antecedentes pessoais e familiares, uso de TH e mamografias. Nos prontuários foram obtidas informações sobre o câncer de mama quanto ao diâmetro do tumor, tipo de cirurgia e estudo imuno-histoquímico. Para a estatística empregou-se ANOVA e teste do chi2. RESULTADOS: 38,2% das pacientes eram ex-usuárias de TH e 61,8% não usuárias. O tempo médio de uso da TH foi de 3,7±3,6 anos. As ex-usuárias eram de menor faixa etária e com menor tempo de menopausa quando comparadas às não usuárias (p<0,05). Constatou-se que 26,8% das pacientes apresentavam antecedentes familiares de câncer de mama, em ambos os grupos. Entre as ex-usuárias de TH, 43,3% foram submetidas a mamografias prévias, ao passo que entre as não usuárias, apenas 11,3% (p<0,001). O diâmetro médio do tumor foi menor entre as ex-usuárias de TH (2,3±1,1 cm), com predomínio de quadrantectomias (60%), quando comparadas as não usuárias (3,3±1,5 cm e 32%, respectivamente) (p<0,001). No estudo imuno-histoquímico, observou-se correlação positiva entre a presença de receptores de estrogênio e progesterona positivos e o uso de TH (p<0,001). Não houve correlação entre TH e c-erbB-2 e p53. CONCLUSÃO: nesta casuística, as mulheres na pós-menopausa que usaram TH prévia ao diagnóstico de câncer de mama apresentaram indicadores de prognóstico mais favoráveis quando comparadas às não usuárias.

Histopathological findings and detection of parasites in the eyes of dogs infected naturally with Leishmania chagasi

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

Genetic variability in natural populations of Zeyheria montana mart. from the Brazilian Cerrado

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

Interferentes eritrocitários e ambientais na anemia falciforme

Erythrocytes and environmental interferences on sickle cell anaemia Sickle cell anaemia runs na extremely variable clinical course At one end of spectrum, it is characterized by a crippling haemolitic anaemia, interspersed with severe exacerbations, or crises, yet it may be an extremely mild disorder, which is found only by chance on rotine haematological examination. The reasons are only partly understood for these remarkable differences in phenotypic expression of what appears to be the same genetic defect: they include the level of Hb Fetal, coinheritance of the alpha thalassaemia and of other genetic variantsthat has influence as genetic modulation in sickle cell anaemia. However, other genetics abnormalities of erythrocytes: G-6PD deficiency, spherocytosis and deficiencies of anti-oxidant enzymes(SOD, GPx and Catalase) probably interfereon the clinical course of sickle cell anaemia. The haplotypes of the chromosome (Bantu, Benin, Camaroon and Arab-Indian) bearing the sickle gene is associated with assorted haematological and clinical features that are likely, at least in part, to be mediated throgh effects on Hb Fetal concentration. Beyond these factors characterizes as erythrocytes interferents, there are the environmental interferents. Between environmental interferents become detached the socio-economic and cultural situation of each patient. These aspects have influence on the life of affected individuals including social interactions, family relations, peer interaction, intimate relationships, education, enployment, violence, spiritual attitudes and navigating complexities of the health care system, providers and their ancillary functions. As a result of this article it is proposed a protocol of laboratorial management of sickle cell syndrome with detach to sickle cell anaemia.

Diagnóstico laboratorial de hemoglobinopatias em populações diferenciadas

The inherited haemoglobinopathies are a heterogeneous group of recessive disorders that include the thalassaemias and sickle cell disease. Nearly a thousand mutant alleles have now been characterized. The mutations are regionally specific and in most cases the geographical and ethnic distribution shave been determined providing the foundation for a program of control through screening, genetic counseling and prenatal diagnosis. The diagnosis of hemoglobinopathies requires care for the methodologies applied and the population group which will be evaluated. The information about the abnormal hemoglobin, the medical and psychological aspects and genetic counseling of the carriers and their families are goals of great importance for the success of preventive programs in this area. Aiming to evaluate the laboratory methods for hemoglobinopathy screening and their use in clinical laboratories, we have compared abnormal hemoglobins incidence in the different population groups: blood donors, anemia carriers, newborn and students. The laboratory methods applied involved eletrophoretic proceedings, cytological and biochemical analysis. Within the period from September 1999 through January 2000, we analyzed 524 individuals with varied types of abnormal hemoglobins. Among blood donors, we diagnosed two sickle cell carriers, which suggest the necessity for better care in the process of selection of blood donor candidates. The current interest in the medical and social aspects of sickle cell anemia has resulted in a great increase in methodology research leading to the development of sickle cell screening techniques.

Hemoglobin as AS/alfa talassemia - Importância diagnóstica

Sickle Cell disease is a generic term for a group of genetic disorders characterized by the predominance of hemoglobin S. These disorders include Sickle Cell anemia, the Sickle Cell beta Thalassemia syndromes and Hemoglobinopathies in which hemoglobin S is in association with another abnormal hemoglobin, such as hemoglobin S/C. The Sickle Cell trait (hemoglobin AS) associated with Alpha Thalassemia presents alterations in the red blood cells morphology, usually absent in the heterozygous for this hemoglobin variant. The interaction between hemoglobin Sand alpha Thalassemia has been described as one of the factors responsible for the improvement in the clinical picture of homozygous of hemoglobin S (Sickle Cell Anemia), decreasing the number of episodes of pain. The genetic mechanisms of this influence are evaluated using molecular analyses of the human globin genes. With the objective of verifying the presence of alpha Thalassemia in heterozygous of hemoglobin S, with anemia, sent to the Laboratory of Hemoglobins, Department of Biology, UNESP, São José do Rio Preto, SP, we analyzed 1002 blood samples with Sickle Cell trait, in the period from 1990 to 1998. The samples were picked with EDTA 5% as anticoagulant, after previous authorization of the carriers. Appropriated counseling and management requires definitive diagnosis. For the laboratorial diagnosis the blood samples were submitted to electrophoretic procedures in alkaline and acid pH and cytological evaluation of hemoglobin H. The electrophoretic procedures confirmed the presence of hemoglobin AS. The cytological evaluation evidenced the presence of alpha Thalassemia. Of this total analyzed, 16(1,59%) blood samples presented the association between hemoglobin AS and alpha Thalassemia and two individuals belonged of the same family. Our results addressed us to suggest to the routine laboratories, that is important to accomplish the research of alpha Thalassemia among the Sickle Cell trait, with anemia, to verify the interaction with alpha Thalassemia, supplying to the carriers a important information on its hematological profile, genetic pattern of hemoglobinopathies and the appropriated counseling. Rev.bras.hematol.hemoter.,2000,22(3):388-394.

Hemoglobinas anormais e dificuldade diagnóstica

The human hemoglobins, with genetically defined inheritance patterns, have shown characteristic polymorphic variation within the Brazilian population, depending on the racial groups of each region. They have appeared under the form of hemoglobin variants or thalassemias, the variant types S and C and the alpha and beta thalassemias being more common, all of them in heterozygote form. During the year of 1999, blood samples from 506 individuals, with suspected anemia or that had already passed through hemoglobinopathies screening, were sent to the Hemoglobin Reference Center - UNESP for diagnostic confirmation and submitted to electrophoresis proceedings, biochemical and cytological analyses in order to characterize the type of abnormal hemoglobins. The goal of the present study was to verify which abnormal hemoglobin types show greater diagnostic difficulty. The samples came from 24 cities in twelve states. The results showed that 354 (69.96%) individuals presented abnormal hemoglobins, 30 (5.93%) being Hb AS, 5 (0.98%) being Hb AC, 76 (15.02%) suggestive of heterozygote alpha thalassemia, 134 (26.48%) suggestive of heterozygote beta thalassemia and 109 (21.54%) with other forms of abnormal hemoglobin, including rare variants and different forms of thalassemias and variant hemoglobin interactions. It has been concluded that, despite the improved techniques currently available and a constant influx of capacitated personnel, the heterozygote form of thalassemias (210 individuals -41.50%) is challenging to diagnose, followed in difficulty by rare variant characterization and interactive forms of hemoglobinopathies (109 individuals-21,54%), suggesting that the capacity for production of qualified professionals and information about these genetic changes in our population should be increased.

Genotipagem do locus ABO (9q34.1) em doadores de sangue da região noroeste do estado de São Paulo

The advances of molecular genetics enabled us to understand the molecular basis of the ABO locus. Considering Us importance as a genetic marker and its applications, the aim of this study was to verify the distribution of the ABO genotypes in a Brazilian population from the Northwest region of the Sào Paulo Stale, Brazil. The genomic DNA was extracted from three hundred and twenty four healthy Brazilian blood donors (O ] 50; A 118; B 32 and AB 24) and analyzed by PCR amplification followed by restriction enzyme digestion. Fourteen genotypes were identified and the relative frequencies of the O , O , O , A and B genes ivere estimated at 44.6%, 16.9%, 4.1%, 25.3% and 9.1%, respectively. Tloese results demonstrate that the ABO locus presents a high polymorphism as revealed by molecular analysis.

Prevenção de hemoglobinopatias a partir do estudo em gestantes

The Brazilian population, presents genes for abnormal hemoglobins with variable frequencies, which are influenced by the founding racial groups. Thus, the detection of carriers of the genetic alterations is important for public health, since they represent sources of new beterozygotes and possible homozygotes. The control of the hemoglobin pathologies has been possible by means of genetic counseling and early diagnosis. The clinical follow-up of the homozygotes and the orientation of the beterozygotes and especially the couples at risk represent a more effective mode of acting to avoid the birth of children who are carriers of a genetic disease, that is frequently lethal. For these reasons this work had as its objectives: to evaluate the importance of testing in pregnant women for the detection of hemoglobin pathologies with the purpose of investigating the prevalence, attaining prevention, a familial study and awareness; for the positive cases such as couples at risk, orient as to appropriate medical attendance; and to evaluate the response to the program. Of the total of 696 pregnant women analysed, 10.7% revealed hemoglobin pathologies with the following rates: alpha Thalassemia 6.75%; Hb AS 2.01%; beta minor Thalassemia 1.29%; Hb AC 0.28%; Hb AJ 0.14%; Hb AS/Alpha Thalassemia 0.14% and P.H.H.F. 0.14%. The high rates of hemoglobin pathologies encountered in the population of pregnant women studied shows the necessity of the implantation of tests for these abnormalities in the pre-natal routine, since in this period the mothers are more apt to be preoccupied with their own health and that of their babies and, however earlier diagnosed the alterations in the hemoglobins, better and more adequate will be the orientations given the couple.

Avaliação dos produtos de degradação oxidativa da Hb S em eritrócitos de doentes falcêmicos

Analysis of the products of oxidative degradation of Hb S was made by methahemoglobin measurement and a count of red blood cells with Heinz bodies. Free radicals originating from oxidation cause extensive injury to erythrocytes, decreasing their useful survival period especially in Hb S carriers. The Superoxide ion (O 2) is the most responsible for the oxidation process of Hb forming membrane-bound haemachromes which afterwards evolve to Heinz bodies, damaging the membrane and provoking erythrocytes hemolysis. The results from this work showed that the SS genotype is more susceptible to the action of the free radicals than the S/Tal genotype. The β genotype has a lower oxidative susceptibility than the SS because it has only one β s mutation. The results allowed us to conclude that: a) the simple presence of Hb S, independent of its genotype and its concentration, is sufficient to produce methaemoglobin from this Hb; b) there is not a direct relationship between methaetnoglobin concentration and the Heinz bodies count; c) the intensity of Heinz bodies in the sickled erythrocytes seems to be independent of the Hb Fetal concentration; d) The genotype SS is more susceptible to Hb oxidation with the release of products of oxidative degradation; e) methaemoglobin formation in blood of people with Hb AA and Hb AS, assessed over 24, 48, 72 and 163 hourly-periods, showed greater oxidative intensity in the Hb AS compared with Hb AA.

Estudo das alterações cariotípicas, do rearranjo gênico BCR/ABL e do cromossomo 20 em leucemias

Leukemia is a genetic disease from a noncontrolled abnormal process of the hematopoietic cells' differentiation and proliferation. Some alterations of structure and number of chromosomes have been well and specifically observed in leukemia. The detection of these alterations is highly significant in providing the patients' diagnosis, prognosis and treatment as well as the understanding of the genetic bases of this disease. The purpose of this work is to study some chromosomal alterations in peripheral blood and/or bone marrow in patients with different leukemia types by means of conventional cytogenetic techniques, and also to investigate the presence of BCR/ABL gene rearrangement and some alterations in chromosome 20 by the FISH technique. Samples of peripheral blood and/or bone marrow of 28 patients, who were not under chemoor radio-therapeutic treatment, were studied: 15 with CML, 11 with AML and 2 with ALL. The alteration most frequent was t(9;22) in the CML, whose presence or absence was related to a good or bad prognosis, respectively. A case of AMI showed inv(16)(p13q22), related to a good prognosis. Some alterations not reported previously in the literature were found, such as the trisomy in chromosome 2 associated to chromosome Ph showing some disease progress in one of the CML cases and t(5;16)(q13;q22) in an AML patient. One of the cases was submitted to an allogeneic hone marrow transplant. The monitoring after the 23 rd day of transplant, detected 95% of the donor cells suggesting the procedure had succeeded. Two patients, an AMI and the other ALL, showed trisomy of chromosome 20 in the neoplastic cells. The results showed the importance of the cytogenetic analysis in relation to leukemia, its direct benefits to the patients and the biological mechanisms involved in this disease. They also allowed the introduction in the Genetic Service of FAMERP techniques to obtain the bone marrow metaphases and the FISH technique.

Aspectos estruturais da membrana eritrocitária

This article describes the structures and functions of the erythrocyte membrane and its importance in transfusional medicine. The erythrocyte membrane is one of the best known membranes in terms of structure, function and genetic disorders. As any other plasma membrane, it mediates transport functions. It also provides the erythrocytes with their resilience and deformability. According to the International Society of Blood Transfusion (ISBT), more than 500 antigens are expressed in the erythrocyte membrane, and around 270 are involved in transfusion reaction cases and hemolytic diseases of the fetus and newborn. In the ISBT classification, the high frequency series is represented by antigens in more than 99% of population (high prevalence antigen). In transfusion, the absence of these antigens determines severe problems as for example, one woman without the P antigen suffered 6 repetitive miscarriages due to placental insufficiency, which was caused by an antibody formed against the absent P antigen. Some important erythrocyte membrane proteins are described here including Band 3, Glycophorins and spectrin. The most abundant integral membrane protein is Band 3 and its main function is to mediate exchange of chloride and bicarbonate anions across the plasma membrane. The second most abundant integral membrane protein in the human erythrocyte is sialoglycoprotein glycophorin A (GPA). With its high sialic acid content, GPA is the main contributor to the net negative cell-surface charge and is thus critical for minimizing cell-cell interactions and preventing red cell aggregation. Glycophorin C (GPC) is the receptor for PfEBP-2 (baebl, EBA-140), the newly identified erythrocyte binding ligand of Plasmodium falciparum. The ternary complex of spectrin, actin and 4.1R defines the nodes of the erythrocyte membrane skeletal network, and is inseparable from membrane stability when under mechanical stress. This erythrocyte membrane review is important for a better understanding of transfusion reactions, where the antibody formation against high prevalence antigens makes compatible transfusions difficult. The study of antigen diversity and biochemical characterization of different proteins will contribute to healthcare, as well as diagnosis, development of technology such as monoclonal antibody production and the therapeutic conduct of many diseases.