Hematological Reference Values According to Geographical Region in Jujuy, Argentina.

Armella, Flavia Noelia, and Sandra Stella Lazarte. Hematological reference values according to geographical region in Jujuy, Argentina. High Alt Med Biol. 23:232-239, 2022. Background: Hematological values vary with altitude above sea level. The purpose of this work was to determine the ranges of hematological reference, and the prevalence of anemia and erythrocytosis in adult blood donors from Jujuy, discriminated according to the geographical region, who assisted at the Centro Regional de Hemoterapia from 2016 to 2018. Materials and Methods: A cross-sectional retrospective study was carried out. Men and women donors belonging to the regions of Valles, Yungas, Quebrada, and Puna were analyzed. The percentiles 2.5 and 97.5 were obtained for the hemoglobin (Hb), hematocrit (Ht), red blood cell count (RBCC), and hematimetric indices, according to sex and region. Results: A total of 4,707 men and 3,326 women, ranging from 18 to 65 years of age, who met with the expected clinical criteria in donors, were studied. The anemia prevalence was 2.5% (95% confidence interval [CI] = 2.2%-2.8%), while the erythrocytosis was 4.9% (95% CI = 4.4%-5.4%). The RBCC, Hb, and Ht increased with altitude above sea level. Conclusions: The Hb and Ht were similar to those observed in the Andean populations living at similar altitudes. The anemia prevalence was lower than the global prevalence reported by the World Health Organization, while that of erythrocytosis increased with the altitude above the sea level. The evidence indicates the necessity for laboratories to obtain reference values for their target population. Registro Nacional de Investigaciones (Renis, Ministerio de Salud de Argentina) IS002323.

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), but not those with anemia of another etiology (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.

Intervalos hematológicos de referencia en la provincia de Jujuy según región geográfica / Hematological reference intervals in the province of Jujuy according to geographic region

INTRODUCCIÓN Numerosos estudios han detectado que existen diferencias en los valores hematológicos obtenidos a distinta altura sobre el nivel del mar, y es importante que cada laboratorio establezca sus intervalos de referencia derivados de la población general saludable que atiende. Ello evitaría que en el acto de donación, cuando se trata de poblaciones de altura, pasen como normales donantes anémicos. OBJETIVO Determinar los intervalos hematológicos de referencia y la prevalencia de anemia en una población adulta, aparentemente sana, donante de sangre de la provincia de Jujuy discriminados según región geográfica, que fueron atendidos en el Centro Regional de Hemoterapia (CRH) durante los años 2016 y 2017. MATERIALES Y MÉTODOS Se realizó un estudio observacional descriptivo retrospectivo de corte transversal. La población estuvo compuesta por donantes del CRH de ambos sexos, pertenecientes a las regiones de Valles, Yungas, Quebrada y Puna. Se obtuvieron los percentiles 2,5 y 97,5 para hemoglobina (Hb), hematocrito (Hto), recuento de glóbulos rojos (RGR) e índices hematimétricos, según sexo y región. En Valles el sexo femenino se dividió en mujeres menores y mayores de 48 años. RESULTADOS Se estudiaron 5.397 varones y 3.693 mujeres, con un rango de edades de 18 a 65 años, que cumplieron con los criterios clínicos deseables en donantes. La prevalencia de anemia fue de 2,0% (IC 95%= 1,7-2,3%). Se comprobó que RGR, Hb y Hto aumentan con la altura sobre el nivel del mar, ya que los valores más bajos pertenecían a Yungas que se encuentra a unos 500 msnm, y los superiores a Puna, situada a más de 3500 msnm. DISCUSIÓN Los valores de Hb y Hto fueron semejantes a los observados en personas que residen en altitudes similares. La prevalencia de anemia obtenida fue inferior a la prevalencia global informada por la Organización Mundial de la Salud. La evidencia de este estudio indica la necesidad de que los laboratorios obtengan los valores de referencia de su población atendida

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

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 (ß0 or ß+) 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.

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.

Expresión de genes involucrados en el estrés oxidativo e inflamación en neoplasias hematológicas / Expression of genes involved in oxidative stress and inflammation in hematological neoplasms

INTRODUCCIÓN Varios estudios han encontrado evidencia de estrés oxidativo (EOx) crónico en las neoplasias hematológicas. Sin embargo, la importancia de la asociación entre el EOx y la malignidad actualmente no está clara. OBJETIVOS Analizar la expresión de los genes de las enzimas antioxidantes catalasa (CAT), superóxido dismutasa (SOD) y peroxiredoxina- 2 (PRX-2) y de los genes de IL-6 y TNF-α, y relacionarla con los niveles de expresión del gen Nrf2; establecer la prevalencia de las neoplasias hematológicas (NH) en Tucumán, describir las características demográficas y reconocer factores de riesgo de la población bajo estudio. MÉTODOS Se realizó un estudio descriptivo, entre junio de 2017 y mayo de 2018. Se estudiaron 33 pacientes con NH y 22 controles sanos en el Instituto de Bioquímica Aplicada de UNT. Los sujetos respondieron una encuesta sobre datos personales y factores de riesgo. Se realizó hemograma, técnicas citoquímicas, e inmunofenotipo mediante citometría de flujo. Se evaluó la expresión génica de CAT, SOD, PRX-2, IL-6, TNF-α y Nrf2 por RetroTranscripción-PCR tiempo real. RESULTADOS La prevalencia obtenida fue; leucemias agudas 70%, neoplasias linfoproliferativas (NLP) 12%, síndromes mielodisplásicos (SMD) 9% y neoplasias mieloproliferativas (NMP) 9%. Se detectó asociación significativa (p< 0,05) entre padecer NH con menor nivel educativo, no poseer obra social, tener hábitos tóxicos y no realizar actividad física. El análisis comparativo de los grupos NH mostró una disminución significativa (p<0,05) en la expresión de los genes de las enzimas y de TNF-α en los grupos SMD, NLP y NMP. El nivel de expresión del gen de Nrf2 demostró influencia significativa sobre los niveles de expresión del gen de la PRX-2 (R2= 0,73). DISCUSIÓN Estos hallazgos sentarían las bases de un potencial vínculo entre la disminución de los antioxidantes y el aumento de los niveles de EOx en las NH, además de señalar a Nrf2 como posible gen regulador del sistema antioxidante

Erythrocyte Catalase Activity in More Frequent Microcytic Hypochromic Anemia: Beta-Thalassemia Trait and Iron Deficiency Anemia.

Most common microcytic hypochromic anemias are iron deficiency anemia (IDA) and ß-thalassemia trait (BTT), in which oxidative stress (OxS) has an essential role. Catalase causes detoxification of H2O2 in cells, and it is an indispensable antioxidant enzyme. The study was designed to measure erythrocyte catalase activity (ECAT) in patients with IDA (10) or BTT (21), to relate it with thalassemia mutation type (ß (0) or ß (+)) and to compare it with normal subjects (67). Ninety-eight individuals were analyzed since September 2013 to June 2014 in Tucumán, Argentina. Total blood count, hemoglobin electrophoresis at alkaline pH, HbA2, catalase, and iron status were performed. ß-thalassemic mutations were determined by real-time PCR. Normal range for ECAT was 70,0-130,0 MU/L. ECAT was increased in 14% (3/21) of BTT subjects and decreased in 40% (4/10) of those with IDA. No significant difference (p = 0,245) was shown between normal and BTT groups, while between IDA and normal groups the difference was proved to be significant (p = 0,000). In ß (0) and ß (+) groups, no significant difference (p = 0,359) was observed. An altered ECAT was detected in IDA and BTT. These results will help to clarify how the catalase activity works in these anemia types.

Resistencia osmótica eritrocitaria en el diagnóstico de anemias hereditarias en Tucumán, Argentina / Red cell osmotic resistance in the hereditary anemias diagnosis in Tucumán, Argentina / ResistÛncia 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.(AU)

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.(AU)

As anemias hereditárias mais comuns em Tucumán (Argentina) sÒo o traþo beta talassemia minor (BTM), as hemoglobinopatias estruturais (HBP) e esferocitose hereditária (EH). A resistÛncia osmótica dos eritrócitos imediata e 24 horas pós-incubaþÒo é o método de diagnóstico da EH, e como um único tubo (ROETU) é usado para a detecþÒo 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 crianþas (≤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 saturaþÒo de transferrina (Wiener Lab), reticulócitos (azul de cresil brilhante), teste de falcizaþÒo e eletroforese de hemoglobina em pH alcalino e ácido. GBTM mostrou anemia microcítica hipocr¶mica, e GEH e GHBP, anemia normocítica normocr¶mica. O ferro foi normal. GRBT e GHBP foram resistentes em ROETU 4,0 g/L, mas GBTM mostrou maior resistÛncia (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.(AU)

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.