A novel 3-base deletion (IVS3+2_4delTGG) of the hydroxymethylbilane synthase gene in a Brazilian patient with acute intermittent porphyria

Acute intermittent porphyria (AIP, OMIM 176000) is an autosomal dominant metabolic disease caused by mutations in the gene encoding hydroxymethylbilane synthase (HMBS; EC 4.3.1.8; formely named porphobilinogen deaminase, PBGD), mapped to chromosome 11q23.3. We describe a novel mutation of the HMBS gene, a de novo 3-base deletion in the splicing donor site of intron 3 (IVS3+2_4delTGG) in a woman affected by AIP. RT-PCR analysis revealed an abnormal HMBS mRNA, compatible with exon 3 skipping.

Red cell aspartate aminotransferase saturation with oral pyridoxine intake

CONTEXTO E OBJETIVO: A enzima aspartato aminotransferase apresenta o piridoxal fosfato como coenzima, oriunda da piridoxina existente em alimentos vegetais frescos. A anemia sideroblástica responsiva à vitamina B6, mielofibrose e síndrome de Peyronie respondem a altas doses de piridoxina. O objetivo foi investigar a máxima resposta da aspartato aminotransferase à suplementação oral com piridoxina. TIPO DE ESTUDO E LOCAL: Experimento controlado, na Seção de Hematologia, Instituto Adolfo Lutz. MÉTODOS: A atividade da aspartato aminotransferase eritrocitária foi determinada (antes e após) em voluntários que receberam suplementação por 15-18 dias (30 mg, 100 mg e 200 mg diariamente). Estudo in vitro também foi realizado, com sangue de sete indivíduos. As atividades enzimáticas antes e após a incubação foram determinadas, seguindo o mesmo protocolo do estudo in vivo. RESULTADOS: O estudo in vivo revelou um aumento gradativo da saturação da aspartato aminotransferase com doses crescentes de piridoxina. 83% de saturação foi alcançada com 30 mg diariamente, 88% com 100 mg e 93% com 200 mg. O estudo in vitro não revelou saturação de 100%.CONCLUSÕES: Tanto in vivo quanto in vitro, não se revelou saturação completa da aspartato aminotransferase por sua coenzima piridoxal-5-fosfato nos eritrócitos. Entretanto, a dose de 200 mg diariamente poderia ser empregada com segurança no tratamento da anemia sideroblástica, mielofibrose e síndrome de Peyronie. Embora a saturação máxima nos eritrócitos não seja atingida, os eritroblastos e outras células nucleadas que contenham as organelas citoplasmáticas certamente atingirão a saturação completa, possivelmente à razão dos resultados obtidos nas doenças citadas.

The absolute recommendation of chamber Neubauer method for platelets counting instead of indirect methods in severe thrombocytopenic patients

Accurate and precise platelet counting is crucial for recommending platelets transfusion for thrombocytopenic patients, principally when platelet counts are bellow 30,000/µl. As most laboratories still use the indirect methods for confirming low automated platelet counts, this work compared two indirect methods used in practice (Fonio and Nosanchunk et al.) with the International Committee for Standardization in Hematology recommended direct method (Brecher and Cronkite). The obtained data show that the indirect methods present low precision and accuracy, and that the direct method should always be employed in severe thrombocytopenic samples thanks to its high precision

Is automated platelet counting still a problem in thrombocytopenic blood?

CONTEXT: Reliable platelet counting is crucial for indicating prophylactic platelet transfusion in thrombocytopenic patients. OBJECTIVE: To evaluate the precision and accuracy of platelet counting for thrombocytopenic patients, using four different automated counters in comparison with the Brecher & Cronkite reference method recommended by the International Committee for Standardization in Hematology (ICSH). TYPE OF STUDY: Automated platelet counting assessment in thrombocytopenic patients. SETTING: Hematology Laboratory, Hospital do Servidor Público Estadual de Säo Paulo, and the Hematology Division of Instituto Adolfo Lutz, Säo Paulo, SP, Brazil. MAIN MEASUREMENTS: Brecher & Cronkite reference method and four different automated platelet counters. PARTICIPANTS: 43 thrombocytopenic patients with platelet counts of less than 30,000/æl RESULTS: The ADVIA-120 (Bayer), Coulter STKS, H1 System (Technicom-Bayer) and Coulter T-890 automatic instruments presented great precision and accuracy in relation to laboratory thrombocytopenic samples obtained by diluting blood from normal donors. However, when thrombocytopenic patients were investigated, all the counters except ADVIA (which is based on volume and refraction index) showed low accuracy when compared to the Brecher & Cronkite reference method (ICSH). The ADVIA counter showed high correlation (r = 0.947). However, all counters showed flags in thrombocytopenic samples. CONCLUSION: The Brecher & Cronkite reference method should always be indicated in thrombocytopenic patients for platelet counts below 30,000 plt /µl obtained in one dimensional counters

Aminoquinolone WR6026 as a feasible substitute for gentian violet in Chagas' disease prophylaxis in preserved blood for transfusional purposes

The search for a colorless, nontoxic and efficient drug to prevent transfusion-associated Chagas' disease (TACD) has been underway unsuccessfully since 1953 when gentian violet was preconized and to date is still being used as the only in vitro trypanocidal agent. The recent findings of aminoquinolone "WR6026" as a trypanocidal agent, led the authors to study the metabolism of red cells stored with this compound, the main objective of which was to define its applicability in TACD control. Ten units of human whole blood collected in CPDA-1 were divided into two equal satellite bags. One had "WR6026" (final concentration 62.5æg/mL) added and the other was used as a control, both were stored at 4ºC. At baseline, day 7, 14, 21 and 28, samples were taken for the following measurements: adenosine triphosphate (ATP), hemoglobin, electrolytes (sodium and potassium), gases (pO2 and pCO2) and osmotic fragility. The results of tests and control were analyzed through parametric t-student test. The results were similar in both groups throughout the experiment except for the level of ATP on day 14, which presented significantly higher values in the tests when compared with the controls (p = 0.012). It was concluded that WR6026 does not interfere in the preservation and probably the viability of the erythrocytes also until day 28 of storage. Consequently the authors suggest that WR6026 could emerge as a colorless substitute for gentian violet in the control of TACD in endemic areas

Normal erythrocyte calpain I activity on membrane proteins under near-physiological conditions in patients with essential hypertension

CONTEXT: It has been reported that the equilibrium between the erythrocyte protease calpain I and its physiological inhibitor calpastatin is disrupted in patients with essential hypertension. OBJECTIVE: To investigate the activity of non-purified calpain I in hemolysates against the erythrocytic membrane proteins, rather than against other substrates. DESIGN: Evaluation of calpain I red cell activity upon its own physiological substrates in hypertensive patients, in a near-physiological environment. SETTING: LIM-23 and LIM-40 of Hospital das Clinicas of the Faculty of Medicine of USP. SAMPLE: Patients with moderate primary hypertension over 21 years of age who were given amlodipine (n:10) and captopril (n:10) for 8 weeks, plus normal controls (n:10). MAIN MEASUREMENTS: Red cell membrane proteins were incubated with and without protease inhibitors and with and without calcium chloride and underwent polyacrylamide gel electrophoresis. RESULTS: Digestion of bands 2.1 and 4.1 was observed, indicating calpain I acitivity. No statistical differences regarding bands 2.1 and 4.1 were observed before treatment, between the controls and the hypertensive patients, either in ghosts prepared without calcium or with increasing concentrations of calcium. Nor were statistical differences observed after treatment, between the controls and the patients treated with amlodipine and captopril, or between the patients before and after treatment with both drugs. CONCLUSION: The final activity of non-purified calpain I upon its own physiological substrate, which was the approach utilized in this study, may more adequately reflect what happens in red cells. Under such conditions no imbalance favoring calpain I activity increase was observed. The protective factor provided by calpastatin against calpain I activity may diminish under hypertension

Enzymes and membrane proteins of ADSOL-preserved red blood cells

CONTEXT: The preservative solution ADSOL (adenine, dextrose, sorbitol, sodium chloride and mannitol) maintains red cell viability for blood trans-fusion for 6 weeks. It would be useful to know about its preservation qualities over longer periods. OBJECTIVE: To determine some red cell biochemical parameters for peri-ods of up to 14 weeks in order to determine whether the red cell metabo-lism integrity would justify further studies aiming at increasing red cell preservation and viability. DESIGN: Biochemical evaluation designed to study red cell preservation. SETTING: Sao Paulo University erythrocyte metabolism referral center. SAMPLE: Six normal blood donors from the University Hospital of the Universidade Federal do Paranß, Curitiba, Brazil. MAIN MEASUREMENTS: Weekly assay of erythrocyte adenosine-5Ý-triphosphate (ATP), 2,3-diphosphoglycerate (2,3DPG), hexokinase (HX), phosphofructokinase (PFK), pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G-6-PD), 6-phosphogluconic dehydrogenase (6-PGD), glyceraldehyde-3-phosphate dehydrogenase (GAPD), glutathione reduc-tase (GR), glutathione peroxidase (GSHPx), plasma sodium and potas-sium, blood pH, and membrane proteins of red cells preserved in ADSOL were studied during storage for 14 weeks storage. RESULTS: During ADSOL preservation, erythrocyte ATP concentration decreased 60 percent after 5 weeks, and 90 percent after 10 weeks; the pH fell from 6.8 to 6.4 by the 14th week. 2,3-DPG concentration was stable during the first week, but fell 90 percent after 3 weeks and was exhausted after 5 weeks. By the end of the 5th week, an activity decrease of 16-30 percent for Hx, GAPD, GR, G-6-PD and 6-PGD, 35 percent for PFK and GSHPx, and 45 percent for PK were observed. Thereafter, a uniform 10 percent decay was observed for all enzymes up to the 14th week. The red blood cell membrane pro-teins did not show significant alterations in polyacrylamide gel electro-phoresis (SDS-PAGE) during the 14 weeks. CONCLUSION: Although the blood viability was shown to be poor from the 6th week up to the 14th week of storage due to ATP and 2,3-DPG depletion, the other biochemical parameters remained in fairly good condition for longer storage. As there is a gradual and uniform decay in activity throughout these 14 weeks, it seems that ADSOL-preserved red cells may be used as red cell enzyme standards and membrane proteins as well

Search of intravascular hemolysis in patients with the cutaneous form of loxoscelism

Dezenove pacientes que apresentaram a forma clinica cutanea do loxoscelismo foram investigados com o proposito de pesquisar hemolise intravascular sub-clinica. lancando mao da dosagem de haptoglobina, um metodo altamente sensivel que permite detectar discreta presenca de hemolise intravascular. Nao fois encontrada diminuicao do veneno da Loxosceles nestes pacientes.

Prevalência de deficiência de ferro em gestantes de primeira consulta em centros de saúde de área metropolitana, Brasil: etiologia da anemia / The prevalence of iron deficiency in pregnant women at their first consultation in health centers in a metropolitan area, Brazil: etiology of anemia in anemic pregnant women

No período compreendido entre abril e outubro de 1988, foram estudadas 363 gestantes de primeira consulta, que estavam inscritas no Programa de Atendimento à Gestante em oito Centros de Saúde da Secretaria da Saúde do Estado de Säo Paulo (Brasil). Na ocasiäo da coleta de material estas gestantes näo faziam uso de medicamentos que continham ferro, ácido fólico, vitamina B12 ou associaçöes destes. A idade média das gestantes foi de 25 anos; 65,9 por cento delas pertenciam a famílias com renda de até um SMPC (salário mínimo per capita) e apenas 3,1 por cento pertenciam a famílias com renda superior a 3 SMPC. Tomando-se a saturaçäo da transferrina inferior a 15 por cento como índice mínimo para definir a deficiência de ferro, a prevalência de deficiência de ferro no primeiro trimestre (4,6 por cento) foi significativamente menor do que a observada no segundo (17,3 por cento) e esta foi menor do que no terceiro trimestre (42,8 por cento). A prevalência de deficiência de ferro total agrupada nos três primeiros trimestres foi de 12,4 por cento. Näo houve diferença significativa entre as prevalências de deficiência de ferro segundo o número de partos. Esta prevalência foi maior no grupo das gestantes que pertenciam a famílias com renda de até 0,5 SMPC. Nas gestantes anêmicas, 46,7 por cento eram deficientes de ferro 44,4 por cento de ácido fólico, 20,0 por cento de ferro e ácido fólico e nenhuma delas eram deficientes de vitamina B12

Infecções por helmintos e protozoarios intestinais em gestantes de primeira consulta atendidas em centros de saude da rede estadual no subdistrito do Butanta, municipio de Sao Paulo / Infections due to helminths and intestinal protozoans in pregnant women of first consultation attended in health Centers of the State in the Butantä Subdistrict, Säo Paulo City

No periodo compreendido entre abril e outubro de 1988, foram estudadas 481 gestantes de primeira consulta, as quais estavam inscritas no Programa de Atendimento a Gestante em oito Centros de Saude da Secretaria do Estado da Saude de Sao Paulo. Do total estudado, 86 gestantes nao trouxeram amostras de fezes para analise, mesmo apos varias solicitacoes. A idade media da populacao estudada foi de 24,5 anos (idade minima de 14 e maxima de 46 anos); a renda media das familias das gestantes foi de 0,97 SMPC (salario minimo per capita) e o numero medio de pessoas que compunham suas familias foi de quatro (um a quinze pessoas). A prevalencia de verminose foi de 45,1 por cento (n=395). Os parasitas mais frequentes foram: Ascaris lumbricoides (19,0 por cento); ancilostomideos (16,7 por cento) e Trichuris trichiura (15,9 por cento). Das 248 gestantes infectadas de enteroparasitas, 70 (28,2 por cento) eram portadoras somente dos seguintes protozoarios: Endolimax nana, Entamoeba coli e Iodamoeba butschllii. A prevalencia de parasitas intestinais foi significativamente maior (p<0,05) nas gestantes pertencentes as familias com renda de ate 0,5 SMPC e compostas por 5 pessoas ou mais.

Generation of reduced nicotinamide adenine dinucleotide phosphate mediated by glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in neutrophils of thoroughbred horses

Em vinte cavalos Puro-Sangue Inglês determinou-se a atividade da glicose-6-fosfato-desidrogenase e da 6-fosfogliconato-desidrogenase de neutrófilos, encontrando-se atividades específicas de 945 ñ 288 mUI mg-1 de proteína e 375 ñ 88 mUI mg-1 de proteína, respectivamente, por minuto a 37§C

Prevalência de anemia em gestantes de primeira consulta em centros de saúde de área metropolitana, Brasil / The prevalence of anemia in first consultation pregnant women of health centers in a metropolitan area, Brazil

No período de abril a outubro de 1988, foram estudadas 363 gestantes de primeira consulta, as quais estavam inscritas no Programa de Atendimento à Gestante da Secretaria do Estado da Saúde de Säo Paulo, subsdistrito do Butantä, Município de Säo Paulo, Brasil. Na ocasiäo da coleta do material biològico estas gestantes näo faziam uso de medicamentos contendo ferro, ácido fòlico, vitamina B12 ou associaçöes destes. A idade média das gestantes foi de 25 anos, 65,9 por cento delas pertenciam a famílias com renda até 1 SMPC (salário mínimo per capita) e apenas 3,1 por cento pertenciam a famílias com renda superior a 3 SMPC. A prevalência de anemia foi de 12,4 por cento. A média da concentraçäo de hemoglobina (g/dl) no primeiro trimestre foi significativamente maior que as médias no segundo e terceiro trimestre gestacional. A prevalência de anemia no primeiro trimestre (3,6 por cento) foi significativamente menor do que aquelas encontradas no segundo (20,9 por cento) e terceiro trimestre (32,1 por cento). As gestantes que tiveram mais que três partos tiveram prevalência de anemia significativamente maior do que aquelas com até três partos. A prevalência de anemia foi maior no grupo das gestantes que pertenciam a famílias com renda até 0,5 SPMC