HemoTypeSC point-of-care testing shows high sensitivity with alkaline cellulose acetate hemoglobin electrophoresis for screening hemoglobin SS and SC genotypes

ABSTRACT Introduction: By providing timely actionable results for prompt management, point-of-care testing (POCT) kits have revolutionised medical care for various diseases, ranging from infectious diseases like malaria to genetic disorders, such as sickle cell disease (SCD). They are, however, underutilised in the diagnosis of SCD in developing countries, where the need is greatest. Objective: The study was aimed at assessing the sensitivity of HemoTypeSC POCT among a cohort of children with SCD, previously diagnosed by Alkaline cellulose acetate hemoglobin electrophoresis (ACAE), with or without high-performance liquid chromatography (HPLC). Methods: In this descriptive cross-sectional study, HemoTypeSC test was conducted on all participants and its sensitivity was determined by comparing results with those obtained using ACAE. Discordance was verified with HPLC. Results: One hundred and forty-five children aged one to 19 years were studied. There were 84 males and 61 females (male: female ratio = 1.4:1). The HemoTypeSC was able to correctly diagnose sickle cell anemia (SCA) and hemoglobin SC in all (100%) of the children tested. Conclusion: The HemoTypeSC shows high sensitivity in detecting SCA and hemoglobin SC. Hence, it is useful for targeted screening of individuals suspected of having SCD, leading to rapid diagnosis of these hemoglobinopathies, even in resource-constrained settings.

Valor diagnóstico de la electroforesis de proteínas y cadenas ligeras libres en suero en el mieloma múltiple / Diagnostic value of serum protein electrophoresis and free light chains in the multiple myeloma's diagnosis

RESUMEN Fundamento: La electroforesis de proteínas y las cadenas ligeras libres en suero son técnicas utilizadas en el diagnóstico del mieloma múltiple. Sin embargo, la utilidad diagnóstica de ambas pruebas puede variar según el método empleado y condiciones reales del medio donde se realicen. Objetivo: Determinar el valor diagnóstico de la electroforesis de proteínas y de las cadenas ligeras libres en suero en el mieloma múltiple. Metodología: Se realizó un estudio retrospectivo de los parámetros electroforesis de proteínas en suero y cadenas ligeras libres en suero a 43 pacientes con diagnóstico de mieloma múltiple por evaluación de la médula ósea. La electroforesis de proteínas se realizó por el método convencional de separación de proteínas sobre papel de acetato de celulosa y para las cadenas ligeras libres se aplicó un ensayo inmunoturbidimétrico en el que se usó un analizador químico (Cobas 311). Se calcularon 7 parámetros que evaluaron la exactitud diagnóstica. Resultados: Todos los parámetros que evaluaron la exactitud diagnóstica estuvieron dentro de los intervalos de confianza en ambas pruebas. Conclusiones: La electroforesis de proteínas y las cadenas ligeras libres en suero son ensayos de gran utilidad en el diagnóstico del mieloma múltiple y se deben utilizar en conjunto para la mayor captación posible de casos.

ABSTRACT Background: Protein electrophoresis and serum free light chains are techniques used in the diagnosis of multiple myeloma. However, the diagnostic utility of both tests may vary according to the method used and the actual conditions of the environment where they are performed. Objective: To determine the diagnostic value of protein electrophoresis and serum free light chains in multiple myeloma. Methodology: A retrospective study of serum protein electrophoresis parameters and serum free light chains was conducted in 43 patients diagnosed with multiple myeloma by bone marrow evaluation. Protein electrophoresis was completed by the conventional method of protein separation on cellulose acetate paper and for free light chains an immunoturbidimetric assay was applied in which a chemical analyzer (Cobas 311) was used. Seven parameters were calculated to evaluate diagnostic accuracy. Results: All parameters assessing diagnostic accuracy were within confidence intervals in both tests. Conclusions: Protein electrophoresis and serum free light chains are very useful assays in the diagnosis of multiple myeloma and should be used in conjunction for the highest possible approval of cases.

Polimorfismo genético da glicose-6-fosfato desidrogenase na população da região de Araraquara, Estado de São Paulo / Genetic polymorphism of glucose-6-phosphate dehydrogenase in the population of Araraquara, State of São Paulo

The most important role played by the enzyme Glucose-6-Phosphate Dehydrogenase (G6PD) in erythrocyte metabolism is in generating energy and reducing power used to protect the cell against oxidative attack. G6PD deficiency is the erythroenzymopathy that most frequently causes hemolytic anemia, and more than 130 molecular variants have already been identified. The aim of this study was to analyze the genetic mutations in the G6PD-deficient adult males in the population of the region of Araraquara, São Paulo State. Out of 5087 male blood donors, 89 were deficient for G6PD, as confirmed by assaying the enzyme activity and electrophoresis on cellulose acetate. Thus, a frequency of 1.75% of G6PD-deficient patients was found, this value being similar to other investigations in São Paulo state. Molecular analysis was performed by amplification of genomic DNA with specific primers and digestion with restriction enzymes. In 96.6% of the patients, the G6PD A¯ variant was observed, with mutations at residues 376(A-G) and 202(G-A). Mean G6PD specific activity among the patients was 1.31 IU.g Hb-1.min-1 at 37ºC, that is 10.8% of the normal activity of the G6PD B enzyme. The variant forms G6PD A¯680(G-T) and 968(T-C) were not found. In 3.4% of the deficient individuals, the G6PD Mediterranean variant was found, with a mutation at 563(C-T). In these cases,mean enzymatic activity was 0.25 IU.g Hb-1.min-1 at 37ºC, or 2.1% of the enzymatic activity of G6PD B. Theuse of traditional techniques, allied to the identification of the different molecular variants, is important for the understanding of the structural and functional properties and hemolytic behavior of the red blood cells of the patient...

A importância da enzima Glicose-6-fosfato desidrogenase (G6PD) no metabolismo eritrocitário está na obtenção de energia calórica e redutora para a proteção celular contra agressões oxidativas. A deficiência de G6PD é a eritroenzimopatia que causa mais frequentemente anemia hemolítica, com mais de 130 variantes moleculares identificadas. O objetivo deste estudo foi realizar a análise molecular da deficiência de G6PD em uma população masculina adulta da região de Araraquara, SP, para a identificação das mutações genéticas. Nos 5087 doadores de sangue do sexo masculino pesquisados, foram encontrados 89 deficientes de G6PD, confirmados pela determinação da atividade enzimática e eletroforese em acetato de celulose, com frequência de 1,75%, valores semelhantes aos encontrados por outros pesquisadores no Estado de São Paulo. A análise molecular realizada pela amplificação do DNA genômico com iniciadores específicos e digestão com enzimas de restrição, demonstrou que 96,6% dos deficientes apresentaram a variante G6PD A¯, com as mutações 376(A-G) e 202(G-A) e atividade enzimática média de 1,31 UI.g de Hb-1.min-1 a 37ºC, correspondendo a 10,8% da atividade enzimática da enzima normal G6PD B. Não foram encontradas as formas variantes G6PD A¯ 680(G-T) e 968(T-C). Em 3,4% dos indivíduos deficientes, foi encontrada a variante G6PD Mediterrânea, mutação 563(C-T) e atividade enzimática média de 0,25 UI.g de Hb-1.min-1 a 37ºC, correspondendo a 2,1% da atividade enzimática da G6PD B. A utilização das técnicas tradicionais, aliadas à identificação da variante molecular, são importantes na compreensão das propriedades estruturais, funcionais e comportamento hemolítico dos glóbulos vermelhos do paciente...

Hemoglobin F level in different hemoglobin variants / 대한혈액학회지

BACKGROUND: Fetal hemoglobin (HbF) levels in different hemoglobin variants in Osogbo, Nigeria, were estimated using two principal methods of estimation using existing information for HbF concentration and distribution of various hemoglobin variants in the area, as well as diagnosed cases of thalassemia. Two hundred and sixty samples collected from HbSS, HbSC, HbAA, HbAS, and HbAC subjects were analyzed. HbF level and hemoglobin type were determined in this study. METHODS: The hemoglobin type was determined using cellulose acetate electrophoresis at an alka-line pH, and HbF was determined by the acid elution and alkaline denaturation methods. RESULTS: The mean+/-SD of HbF in the respective hemoglobin variants was as follows: HbSS, 2.09+/-1.94%; HbSC, 0.85+/-0.54%; HbAA, 0.69+/-0.46%; HbAS, 0.52+/-0.31%; and HbAC, 0.57+/-0.26%. The mean HbF level across the hemoglobin variants was statistically significant (P<0.05). Investigating the sex distribution of the HbF level in the studied population revealed a slightly higher mean HbF level in females than in their male counterparts. CONCLUSION: Within the study population, the HbF level was found to be highest in HbSS and lowest in HbAS. The two methods of estimating HbF are equally reliable, since there was no significant difference between the results obtained from the two methods.

Comparison of Capillary Electrophoresis with Cellulose Acetate Electrophoresis for the Screening of Hemoglobinopathies / 대한진단검사의학회지

BACKGROUND: beta-thalassemia is primarily found in individuals of Mediterranean and Southeast Asian ancestry. With rapid growth in the Southeast Asian segments of the Korean population, the geographic distribution of hemoglobinopathies is expected to become significantly different from what it is today. In this study, Hb fractions were measured in patients with hypochromic microcytosis to detect thalassemia and Hb variants. To evaluate the feasibility of replacing cellulose acetate electrophoresis (CA) with capillary electrophoresis (CE) in a clinical laboratory, both techniques were performed and the outcomes were compared. METHODS: To evaluate hemoglobinopathies, complete blood cell counts (CBC), CA, and CE were carried out on samples from healthy and microcytic hypochromic groups. The microcytic hypochromic group consisted of 103 patients whose mean corpuscular volume (MCV) was less than 75 fL and mean corpuscular hemoglobin (MCH) was less than 24 pg. Quantitative analysis of Hb fractions was performed on 143 whole blood samples. RESULTS: There was a good correlation for measurements of HbA (r=0.9370, P<0.0001), HbA2 (r=0.8973 P<0.0001), and HbF (r= 0.8010, P=0.0304) between the two methods. In the microcytic hypochromic group, there were 29 cases (28.2%) with decreased HbA2, 2 cases (1.9%) with increased HbA2, 3 cases (2.9%) with increased HbF, and 2 cases (1.9%) with increased HbA2 and HbF. CONCLUSIONS: CE is comparable to CA for reliable measurement of Hb fractions. It is suitable for screening of hemoglobinopathies in many clinical laboratories.

Estudo comparativo da eficiência da eletroforese alcalina em acetato de celulose na identificação de hemoglobinas utilizando diferentes tampões / Comparative study of the alkaline electrophoresis on cellulose acetate efficiency to identify hemoglobins by using different buffers

As hemoglobinas podem apresentar alterações moleculares denominadas hemoglobinopatias caracterizadas pela ineficiência da produção das cadeias de globinas (alfa ou beta) ou por alterações estruturais na síntese destas cadeias. Tais alterações estruturais promovem a formação de hemoglobinas variantes (anômalas) e o desbalanceamento na síntese destas cadeias tem como consequência a talassemia. O diagnóstico das hemoglobinopatias é realizado essencialmente por avaliação eletroforética. Este trabalho teve como objetivo avaliar a influência de diferentes tampões de eletroforese na caracterização de hemoglobinas variantes por meio de técnica de eletroforese alcalina em acetato de celulose. Considerando as características de separação e nitidez das bandas obtidas concluímos que o tampão TEB (Tris base 0,090 M, Ácido bórico 0,090 M, EDTA0,002 M, pH 8,2) na concentração de 0,5X foi o mais eficiente na caracterização das hemoglobinas testadas. Este tampão além de permitir uma melhor separação eletroforética entre as diferentes hemoglobinas, também permitiu uma maior estabilidade da hemoglobina H, favorecendo a identificação mais eficaz dessa hemoglobina instável.

Characterization and oxygen binding properties of des-Arg human hemoglobin

The role of chloride in the stabilization of the deoxy conformation of hemoglobin (Hb), the low oxygen affinity state, has been studied in order to identify the nature of this binding. Previous studies have shown that arginines 141α could be involved in the binding of this ion to the protein. Thus, des-Arg Hb, human hemoglobin modified by removal of the α-chain C-terminal residue Arg141α, is a possible model for studies of these interactions. The loss of Arg141α and all the salt bridges in which it participates is associated with subtle structural perturbations of the α-chains, which include an increase in the conformational flexibility and further shift to the oxy state, increasing oxygen affinity. Thus, this Hb has been the target of many studies of structural and functional behavior along with medical applications. In the present study, we describe the biochemical characterization of des-Arg Hb by electrophoresis, high-performance liquid chromatography and mass spectroscopy. The effects of chloride binding on the oxygen affinity and on the cooperativity to des-Arg Hb and to native human hemoglobin, HbA, were measured and compared. We confirm that des-Arg Hb presents high oxygen affinity and low cooperativity in the presence of bound chloride and show that the binding of chloride to des-Arg does not change its functional characteristics as observed with HbA. These results indicate that Arg141α may be involved in the chloride effect on Hb oxygenation. Moreover, they show that these residues contribute to lower Hb oxygen affinity to a level compatible with its biological function.

Serum proteins and fractions, HDL-cholesterol and total IgG and IgE levels in cases of acute and chronic paracoccidioidomycosis / Proteínas séricas e frações, HDL-colesterol e níveis de IgG e IgE totais na paracoccidioidomicose aguda e crônica

This study evaluated serum protein fractions, HDL-cholesterol, total immunoglobulin G and total immunoglobulin E levels in patients with acute and chronic paracoccidioidomycosis, by means of electrophoresis, enzymatic reaction and immunoenzymatic assay. The results demonstrated elevated levels of total immunoglobulin G, total immunoglobulin E, alpha-2 and gamma-globulins, which were more evident in acute than in chronic PCM, but no increase in HDL-cholesterol levels. There was a correlation between the levels of total immunoglobulin E and gamma-globulins and the alpha-2 and beta-globulin fractions in the acute form and between beta and gamma-globulins in both the acute and the chronic form. In conclusion, changes in total immunoglobulin G and immunoglobulin E levels and in the electrophoretic profile may be important markers for the prognosis and therapeutic follow-up of PCM cases, especially because protein electrophoresis is a simple laboratory test that can be applied when specific PCM serological tests are not available. In addition, levels of the gamma-globulin fraction greater than 2.0g/dl may suggest that the patient is developing a more severe form of PCM.

Este trabalho avaliou as frações de proteínas séricas, HDL-colesterol e imunoglobulina G e imunoglobulina E totais em pacientes com paracoccidioidomicose aguda e crônica por eletroforese, reação enzimática e ensaio imunoenzimático. Os resultados demonstraram aumento dos níveis de imunoglobulina G e imunoglobulina E totais, alfa-2 e gama-globulinas, mais evidente na forma aguda que na forma crônica, e não nos níveis de HDL-colesterol. Houve correlação entre níveis de imunoglobulina E total e gama-globulinas e fração alfa-2 e beta-globulinas na forma aguda e entre beta e gama-globulinas nas formas aguda e crônica. Concluindo, alterações nos níveis de imunoglobulina G e imunoglobulina E totais e no perfil eletroforético podem ser importantes marcadores para prognóstico e acompanhamento terapêutico da PCM, especialmente por ser a eletroforese de proteínas um exame laboratorial simples que pode ser empregado em situações onde a sorologia específica para PCM não está disponível. Adicionalmente, níveis da fração gama-globulinas acima de 2,0g/dL podem sugerir que o paciente esteja desenvolvendo uma forma mais grave de PCM.

Identification of hemoglobin AC heterozygote status in a Malay family: a decision between hemoglobin electrophoresis and high performance liquid chromotography.

Thalassemia is a common public health problem among Malays. Hemoglobin C (Hb C) is a hemoglobin beta variant resulting from a single base mutation at the 6th position of the beta-globin gene leading to the substitution of glycine for glutamic acid. Hb C is commonly detected in West Africans and in African American but has not been reported in Malaysia. It can be falsely diagnosed as HbE trait in the Malaysian Thalassemia Screening Program which utilizes cellulose acetate hemoglobin electrophoresis. This is the first reported case of Hb AC heterozygote status in a Malay family, with unusual splenomegaly in one of the family members.

Molecular characterization of human Trypanosoma cruzi isolates from endemic areas in Panama

The present work provides information on Trypanosoma cruzi genotype circulating in endemic areas of Chagas disease in Panama. A total of 26 crude stocks of T. cruzi, isolated from the blood of persons with different clinical profiles of Chagas disease were collected and crio-conserved until used. Most of the stocks had been characterized by means of isoenzyme electrophoresis on cellulose acetate membranes. The clinical profiles of infected persons included 9 (34.6 percent) asymptomatic and 17 acute (65.4 percent) including 5 (19.2 percent) fatal cases, 2 under 5 years old and 3 adults. A multiplex-PCR assay based on the amplification of the non-transcribed spacer of the mini-exon gene was performed. All stocks of T. cruzi included in the study were found to correspond to Tc I group. This result supports the predominance of T. cruzi-I in the transmission cycles affecting the human population in the Republic of Panama.

Utilization of different methodologies for the characterization of Hb Hasharon heterozygotes

Hb Hasharon has an electrophoretic mobility similar to that of Hb S in cellulose acetate and a mobility between Hb S and C at acid pH. In high-performance liquid chromatography, Hb Hasharon shows a distinct chromatographic profile and retention time. The origin of this variant is a mutation in codon 47 (GAC --> CAC) of the alpha2-globin gene, resulting in the replacement of asparagine by histidine during the translation process. Ten blood samples from individuals suspected of being Hb Hasharon carriers were analyzed. In addition to classic laboratory tests and high-performance liquid chromatography, molecular analysis by polymerase chain reaction with restriction fragment length polymorphism designed in the laboratory was performed to confirm this mutation. The study of these cases showed that a combination of classical and molecular methodologies is necessary in the diagnosis of hemoglobinopathies for a correct hemoglobin mutant identification. The accurate identification of hemoglobin variants is essential for genetic counseling and choice of therapy.

Serum electrophoresis in patients of Eale's disease

Eales disease is an obliterative vasculopathy that usually involve peripheral retina of young adults. It is thought to be a nonspecific tissue response to a number of agents and exact etiology remains unknown. Many diseases reflect some alterations in electrophoretic pattern of serum proteins. This study was carried out to investigate electrophoretic pattern of serum proteins in Eales disease so as to throw some light on possible etiological factors in these patients. Thirty-two patients of Eales disease constituted the subject material for this study. Paper electrophoresis of the sera was performed by cellulose acetate method on Whatman filter paper No.10 using barbitone buffer of pH 8.6. Both patients and controls underwent complete ocular and clinical examination and followed up for one year. This was a descriptive study. Ten patients [30%] had raised total serum proteins out of 32 patients and ten patients [30%] had raised gamma globulin. Serum albumin was decreased in seven patients [20%] out of thirty-two patients of Eales disease. In our study 30% increase in total serum proteins and gamma globulins in Eales disease patients points to a possible role of some immunological process which manifests as retinal perivasculitis in eyes. An increase in erythrocyte sedimentation rate in >85% of cases is consistent with this suggestion

Perfil proteico en líquido cefalorraquídeo: aporte al diagnóstico de esclerosis múltiple / Protein profile in cerebrospinal fluid: contribution to the diagnosis of multiple sclerosis

El presente estudio permitió determinar dos perfiles proteicos, mediante SDS-PAGE con tinción argéntica, en 104 líquidos cefalorraquídeos (LCR) de pacientes con diversas patologías. En el perfil A (PA) se observaron bandas de peso molecular (PM) 67 KDa. En el perfil B (PB) se observaron, además, bandas correspondientes a proteínas de bajo PM (20-25 KDa y 13 KDa). Se analizó la presencia de bandas oligoclonales (BO) mediante electroforesis y electroinmunofijación en acetato de celulosa con tinción argéntica, sin concentración previa. El análisis estadístico estableció una asociación entre la presencia de PB y esclerosis múltiple (EM) (p<0,0001) y la presencia de BO y EM (p<0,001) comparada con el resto de las neuropatías. Dentro del grupo de pacientes con PB se observó una asociación significativa entre la presencia de BO y el diagnóstico de EM (p<0,05). Tres pacientes sin EM presentaron BO positivas. El perfil proteico y la presencia de BO fueron útiles como herramientas diagnósticas adicionales para definir a la población de pacientes con EM. La utilización de ambas determinaciones fortalecería el aporte del Laboratorio al diagnóstico de EM, al evaluar dos eventos patológicos complementarios, la síntesis local de IgG (BO) y la destrucción axonal con liberación de proteínas de origen cerebral (PB)

Valor prognóstico da fibronectina plasmática e da classificação de Child-Pugh na cirrose hepática alcoólica: estudo comparativo / Prognostic value of plasmatic fibronectin and Child-Pugh classification in alcoholic cirrhotic patients: a comparative study

OBJETIVOS: Avaliar o valor prognóstico da fibronectina plasmática (FN), comparativamente à classificação numérica de Child-Pugh e os parâmetros bioquímicos que a compõem, no acompanhamento prospectivo de portadores de cirrose alcoólica durante 18 meses. MÉTODOS: Incluídos 50 pacientes com cirrose alcoólica, diagnosticada por biópsia ou critérios clínico-bioquímicos, excluídos aqueles com hepatocarcinoma ou hemorragia digestiva, infecção ou ingestão alcoólica continuada nos últimos 30 dias. A idade média do grupo foi 51,3±12,6 anos, 72 por cento deles do sexo masculino e classificados 17 como Child-Pugh A, 18 como B e 15 como C. Os valores das bilirrubinas foram dosados pelo método automatizado, eletroforese de proteínas em acetato de celulose e o tempo de protrombina pelo método de Quick. A FN plasmática foi dosada por imunodifusão radial, com anticorpos contra FN humana em géis de agarose a 1 por cento. RESULTADOS: Um paciente foi excluído por óbito de causa não natural e 12 foram a óbito por doença hepática. Os melhores preditores de óbito foram a pontuação de Child-Pugh [escore>10, risco relativo (RR) de 11,33) e os valores de bilirrubina (>2,5mg/dL, RR=9,47). A concentração de FN foi significantemente maior nos sobreviventes que naqueles que foram a óbito (185±66 mg/L x 131±38mg/L, p<0,01), com RR = 6,59 para FN<165mg/L. Valores de FN acima desse valor de corte, entretanto, foram os melhores indicadores de sobrevida desde que 96,5 por cento desses 29 pacientes estavam vivos ao final de 18 meses de seguimento. CONCLUSAO: Embora apresente menor acurácia em predizer o risco de óbito desses pacientes, valores de FN plasmática> 165mg/L foram melhores indicadores de sobrevida que a classificação de Child-Pugh e seus parâmetros bioquímicos isolados.

Isozyme analysis of Taenia solium isolates from Mexico and Colombia

Mexican and Colombian Taenia solium cysticerci and some species of Taenia adults were assayed using cellulose acetate electrophoresis to distinguish between isolates. Isozyme patterns for ARK, GOT, G3PD, GPI, and MPI were identical in all cysticerci suggesting homozygotic profiles. G6PD and MDH showed different patterns between Mexican and Colombian cysticerci, suggesting regional differences. ME activity was mainly detected in the adult stage suggesting that this enzyme is active in anaerobic environment, while MDH, detected in cysticerci, could be related to an environment that contains oxygen. Finally, the species of taeniid adults analyzed showed different patterns among them.

Red cell glucose-6-phosphate dehydrogenase phenotypes in Iraq

We attempted to characterize biochemically glucose-6-phosphate dehydrogenase [G6PD] variants in Iraqi individuals. Thus 758 healthy Iraqi males aged 18-60 years were randomly selected and 46 [6.1%] were G6PD deficient. Although the predominant non-deficient G6PD phenotype was G6PD B [92.6%], G6PD A+ was found in polymorphic frequency [1.3%]. In the deficient group, 31 cases were fully characterized, including 17 cases with features consistent with G6PD Mediterranean variant, while 12 had other biochemical features and were labelled as non-Mediterranean variant. The remaining two deficient cases were characterized as G6PD A- variant. The presence of a significant number of non-Mediterranean variant was unexpected and may be related to the more heterogeneous background of the Iraqi people

Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology

The aim of the method described here is to remove hemoglobin, the major contaminant in the bovine plasma obtained from slaughterhouses, by adding a mixture of 19 percent cold ethanol and 0.6 percent chloroform, followed by fibrinogen and globulin precipitation by the Cohn method and nonspecific hemagglutinin by thermocoagulation. The experimental volume of bovine plasma was 2,000 ml per batch. Final purification was performed by liquid chromatography using the ion-exchange gel DEAE-Sepharose FF. The bovine albumin thus obtained presented > or = 99 percent purity, a yield of 25.0 + or 1.2 g/l plasma and >71.5 percent recovery. N-acetyl-DL-tryptophan (0.04 mmol/g protein) and sodium caprylate (0.04 mmol/g protein) were used as stabilizers and the final concentration of albumin was adjusted to 22.0 percent (w/v), pH 7.2 to 7.3. Viral inactivation was performed by pasteurization for 10 h at 60 degres C. The bovine albumin for the hemagglutination tests used in immunohematology was submitted to chemical treatment with 0.06 percent (w/v) glutaraldehyde and 0.1 percent (w/v) formaldehyde at 37 degrees C for 12 h to obtain polymerization. A change in molecular distribution was observed after this treatment, with average contents of 56.0 percent monomers, 23.6 percent dimers, 12.2 percent trimers and 8.2 percent polymers. The tests performed demonstrated that this polymerized albumin enhances the agglutination of Rho(D)-positive red cells by anti-Rho(D) serum, permitting and improving visualization of the results

Triagem neonatal: investigação de hemoglobinoparias em recém-nascidos da cidade do Salvador - Bahia / Neonatal screening: hemoglobinopathies investigation in newborns from Salvador - Bahia

As hemoglobinopatias apresentam freqüência mundial elevada. No Brasil, tem sido registradas freqüências elevadas para a HbS, estimando-se 0,1 a 0,3% de recém-nascidos (RN) com anemia falciforme (SS). A talassemia a apresenta freqüência de 20 - 23% entre algumas populações estudadas. Um total de 600 amostras de sangue de cordão umbilical foi investigada. Os RN foram provenientes da Maternidade Pública Tsylla Balbino (SESAB) e as amostras coletadas no período de fevereiro a junho de 2000. A análise de hemoglobinas foi realizada por eletroforese em fitas de acetato de celulose pH 8,9 e em ágar - citrato pH 6,0; os dados hematológicos foram obtidos através de contador de células eletrônico (Coulter Count T - 890). O DNA foi isolado dos leucócitos e utilizado para a investigação das talassemias α2 3.7Kb e α2 4,ZKb por PCR (Reação em Cadeia da Polimerase). Os dados referentes aos RN foram obtidos através de entrevista às mães, consulta aos prontuários médicos e observação do RN. A análise estatística foi realizada através do software EPI-INFO versão 6.04. Foram encontrados 538 (90,9%) RN com perfil de hemoglobinas normal (AF); 33 (5,6%) heterozigotos para a HbS (ASF), 19 (3,2%) heterozigotos para a HbC (ACF) e 02 (0,3%) homozigotos para a HbC (CF). O peso, gênero e idade gestacional não demonstraram associação com a presença de hemoglobinas anormais. Vinte e seis RN com perfil ASF foram distribuídos de acordo com a classificação racial: 11 (42,30%) foram mulatos, 08 (30,77%) pretos e 07 (26,90%) brancos. A análise hematológica entre os RN com padrão de AF e ASF não demonstrou diferenças estatisticamente significativas. A análise hematológica entre os RN AF e ACF apresentou diferenças significativas para os valores de Ht (p=0,008), Hm (p=0,017) e CHCM (p=0,0045). Os padrões SF e SCF não foram encontrados na análise da presente amostra. A talassemia α2 3,7 Kb foi detectada em 134 (22,71%) dos RN, entre os quais 120 (20,34%) foram heterozigotos e...

case of sickle cell hemoglobin D disease-two years follow-up

Sickle cell Hb D disease is a rare disorder presenting clinically as a mild to severe sickle cell anemia. A case of a two-and-a-half-years old female child is reported here who presented with severe sickle cell disease. Patient's father carried sickle cell trait [AS] and mother an Hb D trait [AD]. She was diagnosed by Hb electrophoresis, sickling and solubility tests as well as family studies. The patient has been followed-up for two years. A two-and-a-half-years old female child presented in November, 1998, with syndrome characterized by painful swelling of hands and feet. She also gave one year history of frequent attacks of severe bone pain in legs, feet and hands. Four weeks earlier she had been admitted in a tertiary care hospital with acute respiratory tract infection, swelling and painful extremities. She was treated with antibiotics and blood transfusion as well as investigated for juvenile rheumatoid arthritis. Rheumatoid factor and anti DNA antibodies were found negative. Her parents are second degree relatives. One younger male sibling is asymptomatic. Physical examination revealed tenderness of the extremities and mild hepatosplenomegaly. Her complete blood counts showed moderate anemia [Hb 8.4 g/dl], high leucocyte count [14.0 x 109/l] and reticulocytosis of 20%. The ESR was 4 mm fall after 1 hour. Peripheral smear showed marked anisopoikilocytosis, hypochromia, microcytosis, few macrocytes, microspherocytes, irregularly contracted cells, few target cells, occasional nucleated red cell and numerous sickle cells. Hb electrophoresis showed a single band in the region of Hb S. Hb F was 4%. The sickling and Hb solubility test for hemoglobin S were positive. The differential diagnosis at this stage was between Hb SS and Hb SD as both Hb S and Hb D have similar mobility at alkaline pH and both are present in our population. Since facility for separation of Hb S from Hb D like citrate-agar gel electrophoresis at acid pH was not present, family studies were carried out to reach at a diagnosis. Cellulose-acetate electrophoresis of both the parents showed heterogenous bands of Hb A and a band in the region of Hb S/D. Sickling test was positive in the father while it was negative in mother. The patient had inherited Hb S from father and Hb D from mother. So a final diagnosis of sickle cell Hb D disease was made. Since her diagnosis the patient had been having similar attacks of bone pain and repeated respiratory tract infection. She had six hospital admissions over the past two years due to infections [thrice], vaso-occlusive crises in the form of severe bone pain [twice] and hemolytic crises [once]. Her Hb ranged from 7-10 g/dl; blood transfusions were required thrice during this period. Renal functions were normal. Spleen had regressed and was no longer palpable; liver was palpable 5 cm below right costal margin. The body growth was normal. The patient was put on prophylactic antibiotics