Cryohemolysis, erythrocyte osmotic fragility, and supplementary hematimetric indices in the diagnosis of hereditary spherocytosis

BACKGROUND: Hereditary spherocytosis (HS) is a chronic hemolytic anemia characterized by microspherocytes in the peripheral blood and increased erythrocyte osmotic fragility (EOF). This study evaluated the cryohemolysis test (CHT); initial hemolysis (IH); immediate and incubated hemolysis percentage in 5.5 g/L NaCl (H5.5); mean corpuscular hemoglobin concentration (MCHC); red blood cell distribution width (RDW); and Hb/MCHC, Hb/RDW, and MCHC/RDW ratios for the diagnosis of HS. METHODS: Data from 13 patients with HS were evaluated at the Instituto de Bioquímica Aplicada and compared with data from 14 unaffected individuals and 11 patients with anemia due to another etiology. Total blood and reticulocyte counts, CHT, and immediate and incubated EOF were performed in all subjects; sensitivity, specificity, efficiency, and Youden index (YI) were calculated. RESULTS: Eight patients with HS had MCHC ≥345 g/L, 10 had RDW ≥14.5%, 12 had IH >5.0 g/L, 11 had immediate H5.5 ≥5%, and 13 had incubated H5.5 ≥50% (the cut-off value to consider HS). The efficiency and YI were: immediate H5.5 (0.94–0.85), incubated H5.5 (0.89–0.82), IH (0.89–0.78), MCHC (0.87–0.62), CHT (0.84–0.54), and Hb/MCHC (0.71–0.56), respectively. The calculated ratios could distinguish subjects with HS from unaffected individuals (P 0.05). CONCLUSION: Although the CHT and supplementary hematimetric indexes were useful to differentiate individuals with SH from healthy controls, they cannot distinguish from anemias of other etiology. CHT and MCHC, in addition to EOF, are recommended for diagnosing HS patients because of their low cost and efficiency.

Foxo3 gene expression and oxidative status in beta-thalassemia minor subjects

ABSTRACT Background: Oxidative stress may aggravate symptoms of hemolytic anemias such as beta-thalassemia. FoxO3 activation results in resistance to oxidative stress in fibroblasts and neuronal cell cultures. Objective: The purpose of this research was to study FoxO3 gene expression and oxidative status in beta-thalassemia minor individuals. Methods: Sixty-three subjects (42 apparently healthy individuals and 21 with beta-thalassemia minor) were analyzed at the Universidad Nacional de Tucumán, Argentina, between September 2013 and June 2014. A complete blood count, hemoglobin electrophoresis in alkaline pH and hemoglobin A2 levels were quantified. Moreover, thiobarbituric acid reactive species, erythrocyte catalase activity and iron status were evaluated. Beta-thalassemia mutations were determined by real-time polymerase chain reaction. FoxO3 gene expression was investigated by real-time reverse transcription-polymerase chain reaction using mononuclear cells from peripheral blood. Results: Subjects were grouped as children (≤12 years), and adult women and men. The analysis of erythrocyte catalase activity/hemoglobin ratio revealed a significant difference (p-value <0.05) between healthy and beta-thalassemia minor adults, but no significant difference was observed in the thiobarbituric acid reactive species levels and FoxO3 gene expression (p-value >0.05). Thiobarbituric acid reactive species and the erythrocyte catalase activity/hemoglobin ratio were not significantly different on comparing the type of beta-thalassemia mutation (β0 or β+) present in carriers. Conclusions: The lack of systemic oxidative imbalance demonstrated by thiobarbituric acid reactive species is correlated to the observation of normal FoxO3 gene expression in mononuclear cells of peripheral blood. However, an imbalanced antioxidant state was shown by the erythrocyte catalase activity/hemoglobin ratio in beta-thalassemia minor carriers. It would be necessary to study FoxO3 gene expression in reticulocytes to elucidate the role of FoxO3 in this pathology.

Prevalencia de anemia y deficiencia de hierro, ácido fólico y vitamina b12 en una comunidad wichí del noroeste argentino / Prevalence of anemia and iron, folic acid and vitamin b12 deficiencies in a wichí community from the argentine northwest region

La anemia y la malnutrición son prevalentes en comunidades de bajo nivel socioeconómico y con higiene inadecuada, como las aborígenes wichí que habitan en el noroeste argentino, región con alta prevalencia de enfermedad de Chagas. Objetivo: determinar la prevalencia de anemia y deficiencia de hierro (Fe), ácido fólico (AF) y vitamina B12 (B12) en una comunidad wichí de Salta, y evaluar la presencia de Chagas y talasemias como etiología de anemia. Material y métodos: se realizó un estudio observacional descriptivo en mayo de 2008. Se estudiaron 35 adultos wichí (21 mujeres, 14 hombres) de la comunidad La Unión. El grupo control consistió en 36 criollos adultos (21 mujeres, 15 hombres). Se realizó hemograma, Fe, transferrina, ferritina, AF, B12, electroforesis de hemoglobina (EHb) y serología para Chagas. Resultados: la prevalencia de anemia fue 76% (16/21; Intervalo de Confianza [IC] 95%=57-95%) en las mujeres wichí; en las criollas, 29% (6/21; IC95%=10-48%); en el grupo masculino criollo, 7% (1/15; IC95%=0-20%) y en los wichís, 29% (4/14; IC95%=16-42%). En las mujeres wichí, el 75% (12/16) de los casos fue por deficiencia de Fe. B12 y EHb fueron normales. El 15% (5/33) del grupo wichí y el 31% (11/35) del criollo presentó deficiencia de AF. La prevalencia de Chagas fue 31% (11/35; IC95%=16-46%) en wichís y 28% (10/36; IC95%=13-43%) en criollos, y no estuvo asociada a anemia. Conclusiones: la alta prevalencia de anemia y deficiencia de hierro indican un problema de salud y de nutrición importante en esta comunidad.

Anemia and malnutrition are prevalent in communities of low socioeconomic status and poor hygiene, including Aboriginal Wichí inhabiting the Argentine Northwest, a region with high prevalence of Chagas disease. Objetive: to investigate the prevalence of anemia and deficiencies of iron, folic acid (FA) and vitamin B12 in community wichí of Salta, and assess the presence of Chagas and thalas semia as etiology of anemia. Material and methods: a descriptive study was performed in May 2008. 35 wichí adults (21 women, 14 men) of La Union community were studied. The control group consisted of 36 adult creoles (21 women, 15 men), who shared environmental conditions. Complete blood count, iron, transferrin, ferritin, FA, vitamin B12, hemoglobin electrophoresis (HbE) and Chagas disease serology were performed. Results: the prevalence of anemia was 76% (16/21 Confidence Interval [CI] 95%=57-95%) in wichí women; in creole women, 29% (6/21, 95% CI=10-48%); in creole male group, 7% (1/15, 95% CI=0-20%) and wichí men group, 29% (4/14, 95% CI=16-42%). In anemic wichí women, 75% (12/16) of cases were produced by iron deficiency. Vitamin B12 levels and HbE were normal. FA deficiency was presented in 15% (5/33) of wichí group and 31% (11/35) of creole people. Chagasic infection prevalence was 31% (11/35, 95% CI=16-46%) in wichís and 28% (10/36, 95% CI=13-43%) in creoles, and was not associated with anemia. Conclusions: the high prevalence of anemia and iron deficiency indicates an important health and nutrition problem in this community.

Resistencia osmótica eritrocitaria en el diagnóstico de anemias hereditarias en Tucumán, Argentina / Red cell osmotic resistance in the hereditary anemias diagnosis in Tucumán, Argentina / Resistência osmótica eritrocitária no diagnóstico de anemias hereditárias em Tucumán, Argentina

Las anemias hereditarias más frecuentes en Tucumán (Argentina) son el rasgo beta talasémico (RBT), las hemoglobinopatías estructurales (HBP) y la esferocitosis hereditaria (EH). La resistencia osmótica eritrocitaria inmediata y 24 horas post-incubación constituye el método diagnóstico de la EH, y como tubo único (ROETU) es usada para cribado de RBT. El propósito del trabajo fue determinar el comportamiento de ROETU (4,0 y 5,5 g/L de NaCl) en el diagnóstico de anemias hereditarias. Se estudiaron 125 pacientes: 34 normales (GN), 59 con RBT (GRBT), 21 con HBP (GHBP) y 11 con EH (GEH), que fueron agrupados en niños (≤12 años), mujeres y hombres (>12 años). Se realizaron hemograma (Coulter AcT10 y Sysmex KX-21N), índices de Mentzer y de Shine&Lal, ROETU, hierro, transferrina y saturación de transferrina (Wiener Lab), reticulocitos (azul brillante de cresilo), prueba de falciformación y electroforesis de hemoglobina a pH alcalino y ácido. GRBT presentó anemia microcítica hipocrómica, y GEH y GHBP, anemia normocítica normocrómica. El hierro fue normal. GRBT y GHBP fueron resistentes en ROETU 4,0 g/L, aunque GRBT mostró mayor resistencia (p<0,05). GEH fue menos resistente que GN en ROETU 5,5 g/L (p<0,05). ROETU 4,0 y 5,5 g/L serían recomendables en el diagnóstico presuntivo de RBT y EH, respectivamente.

Beta thalassaemia trait (BTT), structural hemoglobinopathies (SHB) and hereditary spherocytosis (HS) are the most frequent hereditary anaemias in Tucumán (Argentina). Immediately and 24 hours post-incubation red cell osmotic resistance is the diagnosis method of HS, and as a single tube (RORST), it is used for clínicamenBTT screening. The purpose of this study was to determine the RORST (NaCl 4.0 and 5.5 g/L) behaviour in the diagnosis of hereditary anemia. The study encompassed 125 patients : 34 normal patients (NG), 59 with BTT (BTTG), 21 with SHB (SHBG) and 11 with HS (HSG), who were divided into children (≤12 years), women and men (> 12 years). Blood count (Coulter AcT10 and Sysmex KX-21N), Mentzer and Shine&Lal indexes, RORST, iron, transferrin and transferrin saturation (Wiener Lab), reticulocytes (brilliant cresyl blue), sickling and hemoglobin electrophoresis at alkaline and acid pH were performed. BTTG showed hypochromic microcytic anemia, and SHBG and HSG, normochromic normocytic anemia. Iron was normal. BTTG and SHBG were resistant in RORST 4.0 g/L, but BTTG showed more resistance (p<0.05). SHG was less resistant than NG at RORST 5.5 g/L (p<0.05). RORST at values of 4.0 and 5.5 g/L would be recommended for the presumptive diagnosis of BTT and SH, respectively.

As anemias hereditárias mais comuns em Tucumán (Argentina) são o traço beta talassemia minor (BTM), as hemoglobinopatias estruturais (HBP) e esferocitose hereditária (EH). A resistência osmótica dos eritrócitos imediata e 24 horas pós-incubação é o método de diagnóstico da EH, e como um único tubo (ROETU) é usado para a detecção de BTM. O objectivo deste estudo foi determinar o comportamento de ROETU (4,0 e 5,5 g/L de NaCl) para o diagnóstico de anemias hereditárias. Foram estudados 125 pacientes: 34 normais (GN), 59 com BTM (GBTM), 21 com HBP (GHBP) e 11 com EH (GEH), que foram reunidos em crianças (≤12 anos), mulheres e homens (>12 anos). Foi realizado hemograma (Coulter AcT10 e Sysmex KX-21N), índices de Mentzer e Shine&Lal, ROETU, ferro, transferrina e saturação de transferrina (Wiener Lab), reticulócitos (azul de cresil brilhante), teste de falcização e eletroforese de hemoglobina em pH alcalino e ácido. GBTM mostrou anemia microcítica hipocrômica, e GEH e GHBP, anemia normocítica normocrômica. O ferro foi normal. GRBT e GHBP foram resistentes em ROETU 4,0 g/L, mas GBTM mostrou maior resistência (p<0,05). GEH foi menos resistente que GN em ROETU 5,5 g/L (p<0,05). ROETU 4,0 e 5,5 g/L seria recomendado para o diagnóstico presuntivo da BTM e EH, respectivamente.

Prevalencia y etiología de anemia en el embarazo: estudio observacional descriptivo en el Instituto de Maternidad de Tucumán / Prevalence and etiology of anemia in pregnancy: a descriptive observational study in the Maternity Institute of Tucumán

INTRODUCCIÓN: En Argentina, la anemia por deficiencia de hierro (ADH) constituye la patología de mayor prevalencia en el grupo materno-infantil. OBJETIVO: Determinar la prevalencia y etiología de anemia en embarazadas que efectuaron su control prenatal en el primer o segundo trimestre del embarazo en la Maternidad de Tucumán, entre octubre de 2009 y mayo de 2010. MÉTODOS: Se efectuó un estudio observacional descriptivo. Se realizó hemograma, análisis de ferremia,transferrina, ferritina, vitamina B12, ácido fólico y electroforesis de hemoglobina a 122 embarazadas. RESULTADOS: La prevalencia de anemia fue del 7,4% (9/122; intervalo de confianza [IC]95% = 2,8%-12%). De las gestantes, 4 tenían ADH y 5, anemia no ferropénica. El 29,5 % (IC 95% = 21-37%) de la población estaba en alguna etapa de ADH (manifiesta, latente o silente);un 4,4% tenía deficiencia de vitamina B12, mientras que el ácido fólico era normal. No se detectaron anemias hereditarias. CONCLUSIONES: El diagnóstico de ADH debería comprender la determinación de hemoglobina y ferritina para poder brindar a la gestante el tratamiento adecuado.

INTRODUCTION: In Argentina, iron deficiency anemia (IDA) is the most prevalent disease in the mother childgroup. OBJECTIVE: To determine the prevalence and etiology of anemia in pregnant women who made their prenatal control in the 1st or 2nd trimester of pregnancy at the Maternity Institute of Tucumán, between October 2009 and May 2010. METHODS: A descriptive observational study was conducted. The analysis included blood count, serum iron, transferrin, ferritin, vitamin B12, folic acid and hemoglobin electrophoresis in 122 pregnant women. RESULTS: The prevalence of anemia was 7,4% (9/122 ; confidence interval[CI] 95% = 2,8-12,0%). 4 pregnant women had IDA, and 5, non-IDA. 29,5% (CI 95% = 21-37%) of the population was in some stage of IDA (evident, latent, silent), 4,4% had vitaminB12 deficiency, and folic acid level was normal. Hereditary anemia was not found. CONCLUSIONS: The diagnosis of IDA should include the determination of hemoglobin and ferritin, in order to provide an appropriate treatment for pregnant women.

Perfil hematológico de la beta-talasemia menor en Tucumán / Hematological profile of beta-thalassemia minor in Tucuman

La b-talasemia menor es uno de los desórdenes genéticos más comunes y constituye la principal causa de anemia hereditaria. Si se exceptúan las provincias de Buenos Aires y Santa Fe, es escasa la información bibliográfica acerca de la distribución de la talasemia en la Argentina. Dado que no existen registros sobre el perfil hematológico de la b-talasemia en la región noroeste de la Argentina, el propósito del presente trabajo fue realizar un estudio exploratorio descriptivo de las características hematológicas y electroforéticas de una población de la provincia de Tucumán portadora de b-talasemia. Se estudiaron 52 pacientes derivados para investigación de síndrome talasémico. Se realizó hemograma, reticulocitos, ferremia, electroforesis de hemoglobina, dosaje de hemoglobinas F y A2. En el 46% de los pacientes se confirmó el diagnóstico de rasgo b-talasémico, detectándose leve anemia con microcitosis y Hb A2 aumentada. El estudio del perfil hematológico no demostró diferencias significativas con respecto a edad y sexo y fue similar a lo ya publicado por otros autores. Según el origen étnico, la población estudiada estuvo constituida por un 58% de individuos de origen italiano, 34% de españoles y 8% de árabes, con predominio de la población italiana, similar a trabajos previos en la Argentina.

The b-thalassemia minor is one of the most common genetic blood disorder and it represents the main cause of hereditary anemia. There is scant information in the scientific literature about b-thalassemia minor distribution in Argentina, except for the provinces of Buenos Aires and Santa Fe. There is no published study of this disorder in the northwest of Argentina. The objective of this descriptive and explorative study is to determine the hematological and electrophoretic characteristics of a b-thalassemia minor population in the province of Tucumán. A total of 52 patients with suspected thalassemia syndrome were studied; haemogram, reticulocytes, serum iron, hemoglobin electrophoresis, hemoglobin F and hemoglobin A2 were performed. Forty-six percent of the patients presented a b-thalassemia minor diagnosis, with the following findings: mild anemia with microcytosis and elevated Hb A2. The hematological profile showed no significative differences with respect to age and sex, and it was similar to previous studies, published by different authors. The ethnic origins were as follow: Italians 58%, Spaniards 34% and Arabians 8%, with preponderance of the Italian population, similar to previous studies in Argentina.