Alteraciones de la hemostasia en el síndrome drepanocítico / Hemostasis alterations in sickle cell syndrome

El síndrome drepanocítico (SD) comprende un grupo de anemias hemolíticas hereditarias de tipo multisistémico asociadas a la hemoglobina S. Los pacientes que padecen este síndrome tienen un mayor riesgo, en comparación con individuos sanos, de presentar accidentes cerebrovasculares, hipertensión pulmonar, necrosis avascular de articulaciones, síndrome torácico agudo y complicaciones durante el embarazo, asociados a un estado de hipercoagulabilidad inducido por alteraciones en los diferentes componentes de la hemostasia, que incluyen la activación del endotelio y de los sistemas plaquetario, de la coagulación y de la fibrinólisis. Esta revisión resume las alteraciones en la hemostasia reportadas en los pacientes con SD, en los cuales se ha demostrado: mayor interacción de células endoteliales con leucocitos, hematíes y plaquetas; aumento de la expresión de proteínas de adhesión, como el factor von Willebrand y sus multímeros de alto peso molecular; aumento de la adhesión y la agregación plaquetaria y de la expresión de proteínas en sus membranas. En el sistema de coagulación se ha detectado aumento en la expresión del factor tisular (FT) en micropartículas derivadas de diferentes células, aumento de marcadores de activación de este sistema, entre estos los fragmentos 1.2 de la protrombina y los complejos trombina-antitrombina y una disminución de las proteínas C y S que actúan como anti-coagulantes. Adicionalmente, se han encontrado aumentados los marcadores de activación del sistema fibrinolítico como los dímeros D y los complejos plasmina/antiplasmina. Todas estas manifestaciones favorecen la aparición de complicaciones trombóticas, implicadas en el deterioro de la calidad de vida de los pacientes. Se recomienda implementar en el diagnóstico y seguimiento de esta enfermedad, la determinación de variables del sistema hemostático, con el fin de identificar alteraciones en etapas tempranas y aplicar terapias que puedan prevenir complicaciones trombóticas.

Sickle cell syndrome (SCS) includes a group of congenital hemolytic anemias associated to the presence of hemoglobin S, which is characterized by acute pain episodes and progressive damage of different organs. Some patients with sickle cell syndrome have shown, when compared with healthy individuals, an increased risk of presenting stroke, pulmonary hypertension, avascular necrosis of joints, acute chest syndrome and pregnancy complications, associated to a hypercoagulable state induced by alterations in different components of hemostasis, such as changes that include activation of the endothelium, platelet activity, coagulation and fibrinolytic systems. This paper compiles hemostasis disorders, associated with thrombotic manifestations, reported until now in sickle cell syndrom. These patients have an increase in activation markers of the coagulation system, such as prothrombin fragment 1.2, thrombin-antithrombin complex, etc., depletion of natural anticoagulant proteins, abnormal activation of the fibrinolytic system and increased tissue factor expression. Similarly, abnormal expression of glycoproteins and increased adhesion and platelet aggregation have been reported. All these alterations produce a hypercoagulable state, which induces, among other things, the appearance of thrombotic complications. In view of the importance of controlling the different complications that can occur in patients with sickle cell syndrome, we recommend the implementation, in diagnosis and monitoring studies, of the evaluation of the different components of the hemostatic system, identifying alterations at an early stage and applying effective treatments to prevent thrombotic complications.

Acute toxicity of the water-soluble fraction of diesel in Prochilodus vimboides Kner (Characiformes: Prochilodontidae)

Diesel oil can be a source of contamination in aquatic environments, mainly as a result of spills. The effects of the water-soluble fraction of diesel (WSF) on Prochilodus vimboides were assessed. Fish were exposed to three different WSF dilutions for up to 96 h and were compared to a control group. Damages in the fragments of DNA were analyzed using the Comet assay. The presence of erytrocyts abnormalities was assessed by micronucleus test. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity and the accumulation of copper in gills were also analyzed. Fish exposed for 96 h had higher rates of damage than those exposed for 24 h. There was no significant difference regarding the presence of micronuclei between exposed and control fish and between 24-h and 96-h exposures. For AST, no significant difference was observed between samples collected at the two exposure times. Fish exposed to a 1:100 dilution of WSF showed higher activity of the enzyme ALT than the control fish after a 24-h exposure period. There was no bioaccumulation of copper in the gills. We conclude that the genotoxic effects of WSF in the cells are more evident in P. vimboides during an acute exposure.

O óleo diesel pode ser uma fonte de contaminação em ambientes aquáticos, principalmente como resultado de derrames acidentais. Foram avaliados os efeitos da fração solúvel do óleo diesel (FSO) em Prochilodus vimboides. Os peixes foram expostos a três diferentes diluições da FSO por até 96 horas e comparados com um grupo controle. Os danos nos fragmentos de DNA foram analisados utilizando o ensaio Cometa. A presença de anormalidades nos eritrócitos foi avaliada pelo teste do micronúcleo. A atividade da Aspartato aminotransferase (AST) e alanina aminotransferase (ALT) e da acumulação de cobre nas brânquias também foram analisados. Os peixes expostos por 96 h tiveram maiores taxas de danos do que aqueles expostos por 24 h. Não houve diferença significativa quanto à presença de micronúcleos entre os peixes expostos e controle e entre 24 e 96 h exposições. Para AST, não foi observada diferença significativa entre as amostras coletadas em dois tempos de exposição. Os peixes expostos a uma diluição de 1:100 do FSO apresentaram maior atividade da enzima ALT do que os peixes do controle após um período de exposição de 24 horas. Não houve bioacumulação de cobre nas brânquias. Os efeitos genotóxicos nas células foram os mais evidentes em P. vimboides durante a exposição aguda a FSO.

Estudio comparativo de la frecuencia de micronúcleos y anormalidades nucleares en eritrocitos de tres especies ícticas / Comparative studies of the frequency of micronuclei and nuclear abnormalities in erythrocytes of three species fishes

Los ecosistemas acuáticos cercanos a las zonas urbanas sufren un deterioro constante ocasionado por diversos factores físico-químicos relacionados con la actividad antrópica, representando un riesgo para los organismos que los habitan, en especial los peces. Estos efectos pueden ser biomonitoreados mediante técnicas de amplia utilización, como el Test de Micronúcleos (Mn) y el de Anormalidades nucleares (AN), entre otros. El objetivo de este trabajo fue verifcar y comparar la frecuencia de micronúcleos y anormalidades nucleares en tres especies ícticas colectadas en un lago urbano, Cyprinus carpio, Astyanax eigenmanniorum y Cheirodon interruptus. Luego de la captura y anestesia de los peces, se extrajo de cada individuo una muestra de sangre y se registró el peso y la longitud total. Las muestras fueron fjadas y luego teñidas con Giemsa al 10%, posteriormente se las analizó con microscopio (1000X) y se les tomaron fotografías con cámara digital. Se calculó, además, el índice de condición corporal (K) para cada uno de los ejemplares capturados. Se aplicó un análisis de correlación de rangos de Spearman (rs) entre frecuencia de Mn y AN vs K para cada especie. La prueba de Kruskal-Wallis se aplicó para evaluar diferencias de Mn y AN entre las especies. Los resultados no mostraron una correlación estadísticamente signifcativa (P> 0,05) entre las frecuencias de Mn y AN vs K. Las frecuencias de Mn entre las diferentes especies no arrojaron diferencias signifcativas (P> 0,05), mientras que las AN sí (P< 0,05). Astyanax eigenmanniorum fue la especie que mayor sensibilidad presentó. Las diferencias preliminares de AN entre las especies utilizadas, convierten a A. eigenmanniorum en una especie "centinela" y posible biomonitor de agentes xenobióticos.

Aquatic ecosystems near urban areas suffer a steady deterioration caused by physical and chemical factors related to human activity, representing a danger to organisms that inhabit them, especially the fsh. These effects may be biomonitored by widely used techniques, such as the Micronucleus Test (Mn) and Nuclear Abnormalities (NA), among others. The aim of this study was to verify and compare the frequency of micronuclei and nuclear abnormalities in three fsh species collected in an urban lake, Cyprinus carpio, Astyanax eigenmanniorum and Cheirodon interruptus. After the capture and anesthesia of the fsh, weight and length were registered, and a blood sample was obtained from each individual. The samples were fxed and stained with 10% Giemsa, subsequently they were analyzed with the microscope (1000X) and photographed with a digital camera. The body con-dition index (K) was calculated for each fsh caught. A correlation analysis Spearman rank (rs) between frequency of Mn and NA vs. K was performed for each species. The Kruskal-Wallis test was applied to assess differences in Mn and NA between species. The results showed no statistically signifcant correlation (P> 0.05) between the frequencies of Mn and NA vs K. The frequencies of Mn between different species yielded no signifcant differences (P> 0.05), whereas NA (P <0.05) did. A. eigenmanniorum was the most sensitive species. NA preliminary differences between the species used, convert A. eigenmanniorum in a sort of "sentinel specie" and a possible biomonitor of xenobiotic agents.

Análise comparativa de duas metodologias para a identificação de cilindros hemáticos urinários / Comparative analysis of two methodologies for the identification of urinary red blood cell casts

INTRODUÇÃO: A presença de hemácias dismórficas na urina é um forte indicativo da origem glomerular do sangramento, sendo uma ferramenta importante no diagnóstico de glomerulonefrites. Os cilindros hemáticos geralmente acompanham as hemácias dismórficas, sendo também fortes indicadores de hematúria glomerular, embora não sejam encontrados com frequência no exame parcial de urina. OBJETIVO: Comparar duas técnicas de concentração de amostras em uma série de exames de urina com hematúria dismórfica. MATERIAL E MÉTODOS: Foram selecionadas 249 amostras com hematúria dismórfica a partir de 4.277 amostras de urina de rotina. As amostras foram processadas utilizando-se duas técnicas: a convencional e a de concentração. O percentual de identificação dos cilindros hemáticos foi comparado de acordo com a metodologia utilizada. RESULTADOS: A presença de cilindros hemáticos pela técnica de concentração foi estatisticamente maior (52,6 por cento) em comparação com a positividade pela metodologia convencional (8,4 por cento) (p < 0,001). DISCUSSÃO E CONCLUSÃO: Sugere-se que a técnica convencional não concentrou suficientemente a amostra de urina e os cilindros hemáticos ficaram no sobrenadante, sendo descartados. A utilização da técnica de concentração aumentou a sensibilidade técnica para a pesquisa dos cilindros hemáticos. Portanto, a técnica de concentração, associada à presença de hemácias dismórficas, mostrou-se útil para aumentar a concordância dos dois parâmetros laboratoriais para a detecção da hematúria de origem glomerular como auxílio diagnóstico das glomerulopatias, importante causa de doença renal crônica.

INTRODUCTION: Dysmorphic red blood cells (RBCs) in the urine are a strong indicator of a glomerular bleeding source. RBC casts, which while generally following RBC dysmorphism are not frequently seen on routine urinalysis, are also important indicators of glomerular hematuria. OBJECTIVE: This study tested the superiority of a urine concentration technique (CT) over the standard method (SM) for RBC cast identification in a group of patients suspected of glomerular hematuria. MATERIAL AND METHODS: Of a total of 4,227 routine urinary samples, 249 with dysmorphic hematuria were selected. The samples were processed according to two techniques: standard method (SM) and concentration technique (CT). The percentages of RBC cast identification according to each method were compared. RESULTS: The CT showed a higher rate of RBC casts (52.6 percent) compared to the SM (8.4 percent) (p < 0.001). DISCUSSION AND CONCLUSION: We suggest that the SM did not sufficiently concentrate the urine sample, the RBC casts remaining in the supernatant and being discarded. The CT increased the sensitivity of the RBC cast yield. The CT, associated with the presence of RBC dysmorphism, was useful to increase the agreement of the two parameters used for identification of glomerular-based bleeding and the diagnosis of glomerular diseases, important causes of chronic kidney disease.

Clinical and gene study of three pedigrees of phytosterolemia associated with macrothrombocytopenia and hemolysis / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the clinical features and ABCG5/ABCG8 gene mutations of three pedigrees of phytosterolemia presented with macrothrombocytopenia and hemolysis.</p><p><b>METHODS</b>Erythrocyte and platelet morphology were examined under light microscope. Plasma sterol levels were measured by high pressure/performance liquid chromatography method. All of ABCG5 and ABCG8 exons and intron-exon boundaries were directly sequenced to identify mutations, the corresponding gene mutation sites of three families members and healthy individuals were detected.</p><p><b>RESULTS</b>All the patients presented macrothrombocytopenia, hemolysis, splenomegaly and xanthomas. The blood smears showed large platelets, some as large as erythrocytes, and abnormal erythrocyte shapes, such as stomatocytes. Plasma concentrations of phytosterols, especially sitosterol were markedly elevated (30 fold) in the affected patients. Four mutations were identified in these three pedigrees, ABCG5 C20896T (R446X) and A20883G, ABCG8 del43683-43724 and del1938C-1939G/ins1938T. The latter three were novel mutations reported for the first time.</p><p><b>CONCLUSIONS</b>Phytosterolemia associated with macrothrombocytopenia and hemolysis is a new subtype of this disease. Plasma phytosterols and related gene analysis should be performed when ever an unexplained macrothrombocytopenia, especially combined with haemolysis or/and stomatocytosis.</p>

Rapid Identification of Thrombocytopenia-Associated Multiple Organ Failure Using Red Blood Cell Parameters and a Volume/Hemoglobin Concentration Cytogram

Thrombocytopenia-associated multiple organ failure (TAMOF) has a high mortality rate when not treated, and early detection of TAMOF is very important diagnostically and therapeutically. We describe herein our experience of early detection of TAMOF, using an automated hematology analyzer. From 498,390 inpatients, we selected 12 patients suspected of having peripheral schistocytosis, based on the results of red blood cell (RBC) parameters and a volume/hemoglobin concentration (V/HC) cytogram. We promptly evaluated whether the individual patients had clinical manifestations and laboratory findings were consistent with TAMOF. Plasma exchanges were then performed for each patient. All 12 patients had TAMOF. The mean values of RBC parameters were significantly higher in all of the patients than with the reference range, however, 3 patients had % RBC fragments within the reference range. The mean value of ADAMTS-13 activity was slightly lower in patients compared with the reference range. Of the 12 patients, remission was obtained in 9 patients (75%) within 4 to 5 weeks using plasma exchanges. Three patients died. An increased percentage of microcytic hyperchromic cells with anisocytosis and anisochromia indicated the presence of schistocytes, making it an excellent screening marker for TAMOF. Identification of TAMOF with RBC parameters and a V/HC cytogram is a facile and rapid method along with an automated hematology analyzer already in use for routine complete blood cell counting test.

Red cell distribution width (RDW) in the diagnosis of iron deficiency with microcytic hypochromic anemia.

OBJECTIVE: To study the utility of red cell distribution width (RDW) in the diagnosis of iron deficiency among children with microcytic hypochromic anemia. METHODS: 151 children (6 months-12 years) with microcytic (MCV<75 fl) anemia were classified into iron deficient (IDA) and non-iron deficient anemia (non-IDA) on the basis of serum ferritin and total iron binding capacity (TIBC). RDW values were obtained on an automated hematology analyzer. Receiver operator curves (ROC) were constructed and the utility of RDW in diagnosis of iron deficiency was studied. RESULTS: The mean RDW value was 18.37+/-2.22% in IDA group (97 children) compared to 16.55+/-1.51 % in the non-IDA group (54 children) (p<0.0001, unpaired t test). In IDA group, the mean RDW value was 16.60+/-1.78%, 17.95+/-1.91% and 20.55+/-1.32% among mild, moderate and severely anemic children (p<0.0001, ANOVA test). The corresponding values in non-IDA group were 16.03+/-1.25%, 16.76+/-1.20% and 16.77+/-2.68% respectively (p=0.269, ANOVA test). At a cut-off value of 17.4%, as obtained from the ROC curve, the sensitivity and specificity of RDW in diagnosis of IDA were 81.0% and 53.4% and a positive and negative predictive value of 63.0% and 72.2% respectively. CONCLUSION: RDW has a limited specificity for diagnosis of IDA among children with microcytic hypochromic anemia.

On a Haemogregarine sp. [protozoa, apicomplexa, adeleina] naturally infecting the viper Bitis arientans in Saudi Arabia with reference to the host-parasite relationship

The study described the developmental stages of a haemogregarine species in the blood and tissues of the viper Bitis arientans. Two out of 9 [22.2%] snakes from the south western region of Saudi Arabia, and recorded for the first time in such locality. The erythrocytic parasites were differentiated into three forms: the youngest form [trophozoite] measuring 7.34 +/- 0.16 x 3.38 +/- 0.07 micro m; the intermediate form [developing gametocyte] measuring 13.36 +/- 0.20 x 5.11 +/- 0.11 micro m and the largest form [mature gametocyte] measuring 18.69 +/- 0.32 x 4.32 +/- 0.16 micro m. None of the leucocytes seemed to be parasitized. Also, two types of meronts were detected in lung endothelial cells of infected vipers. Small meronts [micromeronts] measured 21.86 +/- 0.28 x 16.13 +/- 0.25 micro m and yielded about 12 merozoites. The large meronts [macromeronts] measured 38.09 +/- 0.33 x 21.52 +/- 0.32 micro m and yielded 28-42 merozoites. Random distribution of nuclei was observed in early meronts of both sizes, meanwhile peripheral arrangement of nuclei characterizing the subsequent developing events of meronts [ectomerogony]. Histopatho-logical studies showed that the infected erythrocytes were hypertrophied, mechanically stretched and their cytoplasm was faintly stained due to dehaemoglobinization. The host cell nucleus was elongated and laterally displaced. Trabeculae of the infected lung exhibited marked thickening and alveoli were collapsed in various degrees. Haemorrhagic foci and spongy structures were detected in some infected lung tissues. Formation of fibrous tissues around the meronts was seen in some foci

Assessment of the mutagenic, antimutagenic and cytotoxic activities of ethanolic extract of araticum (Annona crassiflora Mart. 1841) by micronucleus test in mice

A typical Brazilian plant, araticum (Annona crassiflora Mart.), is widely used in humans as therapeutic medicine to treat several diseases such as diarrhea, rheumatism and syphilis. It contains acetogenins which present cytotoxic, antitumogenic, and antiparasitic properties. In this study, mutagenic, antimutagenic and cytotoxic effects of araticum leaves ethanolic extract were evaluated by micronucleus test in mice. To evaluate the mutagenic activity, animals were treated with ethanolic extract of araticum (EEA) using 10, 20, 50, 100 and 160 mg.kg-1. For all doses, micronucleated polychromatic erythrocytes (MNPCE) frequency was evaluated at 24, 48 and 72 hours after treatment. To evaluate the antimutagenic activity, animals were treated with 10, 20, 50 and 100 mg.kg-1 of EEA and 4 mg.kg-1 of MMC simultaneously. The frequency of MNPCE was evaluated 36 hours after exposure. Cytotoxicity was evaluated by the polychromatic and normochromatic erythrocytes ratio (PCE/NCE). In the mutagenicity assessment, all doses of EEA resulted in no significant increase of MNPCE (P > 0.05), compared to solvent- control group. Regarding administration time, no significant difference among three evaluation periods was observed (P > 0.05). Such results indicate that EEA did not exert mutagenic activity. Cytotoxicity was evident in doses of 50, 100 and 160 mg.kg-1 at 24 and 48 hours after exposure. Concerning antimutagenicity, except the 10 mg.kg-1 co-administered with 4 mg/kg of MMC, all doses reduced significantly the frequency of MNPCE compared to the positive control group (P < 0.05). These results, therefore, indicate an antimutagenic activity of the EEA. Cytotoxicity was significantly increased (P < 0.01) at 100 mg.kg-1 EEA doses co-administered with 4 mg.kg-1 of MMC.

O araticum (Annona crassiflora Mart.) é uma planta tipicamente brasileira, largamente utilizada em humanos como remédio para o tratamento de diversas doenças como diarréia, reumatismo e sífilis. Esta planta contém acetogeninas que apresentam propriedades citotóxica, antitumorigênica e antiparasitária. Neste estudo, foram avaliados os possíveis efeitos mutagênico, antimutagênico e citotóxico do extrato etanólico de folhas de araticum, pelo teste de micronúcleos em camundongos. Para a investigação da atividade mutagênica, os animais foram tratados com o extrato etanólico de araticum (EEA) utilizando 10, 20, 50, 100 e 160 mg.kg-1. Para todas as doses, as freqüências de eritrócidos policromáticos micronucleados (MNPCE) foram avaliadas em 24, 48 e 72 horas após o tratamento. Para a investigação da atividade antimutagênica, os animais foram tratados com 10, 20, 50 e 100 mg.kg-1 de EEA simultaneamente com 4 mg.kg-1 de MMC. A freqüência de MNPCE foi avaliada após 36 horas de exposição. A citotoxicidade foi avaliada pela razão de eritrócitos policromáticos e normocromáticos (PCE/NCE). Na avaliação da mutagenicidade, todas as doses de EEA não aumentaram significativamente o número de MNPCE (P > 0,05), comparativamente as do grupo solvente-controle. Em relação ao tempo de administração, não foi constatada diferença significativa entre os 3 períodos avaliados (P > 0,05). Esses resultados indicam que o EEA não exerceu atividade mutagênica.A citotoxicidade foi evidente nas doses de 50, 100 e 160 mg.kg-1 em 24 e 48 horas depois da exposição. Em relação à antimutagenicidade, exceto para a dose de 10 mg.kg-1 co-administrada com 4 mg.kg-1 de MMC, todas reduziram significativamente a freqüência de MNPCE, comparativamente as do grupo controle positivo (P < 0,05). Esses resultados, portanto, indicam uma atividade antimutagênica do EEA. A citotoxicidade foi significativamente aumentada (P < 0,01) na dose de 100 mg.kg-1 de EEA co-administrada com 4 mg.kg-1 de MMC.

[Evaluating correlation between NRBC count and fetal distress]

Nucleated red blood cells [NRBC] are occasionally observed in blood of newborns and some recent studies have reported of a relationship between NRBC count and fetal distress and hypoxia. To investigate the correlation between NRBC count and fetal distress. This was a case-control study conducted at Kosar medical centre in Qazvin, Iran. During a 6-month period in 2005, fifty women of unifetal pregnancy at third trimester of their gestation were chosen and the NRBC counts of their newborns who suffered fetal distress [case group] were evaluated. The control group composed of 100 women at their third trimester of pregnancy whose fetuses showed no sign of any distress. Data were analyzed using X2 and t test. The mean NRBC count in fetal distress group was 2406.6 +/- 2470.7 and in control group 673.43 +/- 709.9. Statistically, increased NRBC count in fetal distress group was found to be significant [p=0.000]. NRBC count in fetal distress group was significantly increased hence it could be used as a marker to evaluate the fetal distress and hypoxia in infants

Study of the genotoxic effect of cyclophosphamide on albino mice bone marrow polychromatic erythrocytes -and the protective effect of captopril

Cyclophosphamide [CYP] is widely used as an antineoplastic and an immunosuppressive drug. However, it has been found to cause DNA damage in normal tissues as well. Captopril [CAP], an angiotensin converting enzyme inhibitor, was reported to have a potential protective effect on the genotoxic effect of CYP possibly through its antioxidant effect. The aim of the present work is to experimentally detect the genotoxic effect of cyclophosphamide using in vivo micronuclei assay in albino mice bone marrow polychromatic erythrocytes and to test the protective effect of captopril on reducing the genotoxicity of CYP. In the present study thirty adult male albino mice were equally divided into six groups. Group I [control group] animals received single physiological saline, group II mice received single injection of captopril [CAP] [50mg/kg], group III animals received single injection of 25mg/kg cyclophosphamide [CYP] dissolved in physiological saline, group IV mice received single injection of 50 mg/kg CYP dissolved in physiological saline, and groups V and VI were the same as group III and IV but CYP injection was preceded by CAP [50mg/kg] injection. The number of micronucleated polychromatic erythrocytes [MNPCEs] was determined in 1000 polychromatic cells from bone marrow smears obtained after sacrificing the animals 24 hrs from exposure to CYP or the control substance. Statistical comparison of the different groups showed that the difference between group I and II was not statistically significant [P=0.106], indicating that CAP does not induce genotoxicity. Whereas, comparing Groups III, IV to group I showed that the difference was statistically significant [P=0.013, 0.00021] It was observed that CYP increased the number of MNPCEs in a dose dependent way. Comparison of groups V and Vito groups III and IV respectively showed a significantly lower number of MNPCEs confirming a protective effect of CAP when administered prior to CYP. The results of the present study confirm a protective role of CAP and support the possibility of administration of captopril prior to cyclophosphamide to ameliorate its genotoxic effect and the possibility to develop secondary cancers

Deformabilidade eritrocitária na anemia ferropriva / Erythrocyte deformability in iron deficiency

A deformabilidade é a característica que permite ao eritrócito normal de 7 a 8 micrômetros (µm) circular por capilares de até 3 µm de diâmetro. Esse fenômeno depende da geometria celular, da viscosidade interna e de propriedades visco-elásticas da membrana eritrocitária. Dentre as técnicas de estudo da deformabilidade eritrocitária (DE), como aspiração por micropipeta, filtração e reoscopia, destaca-se a ectacitometria. Esta técnica utiliza um viscosímetro de fluxo laminar no qual as modificações de forma dos eritrócitos são monitoradas continuamente por um feixe de raio laser, processadas por microcomputador, gerando o "índice de Deformabilidade" (ID), que mede a eliptocitogênese dos eritrócitos quando submetidos a uma força denominada "shear stressl". Alterações de DE foram descritas em diversas situações, como em anemias hemolíticas hereditárias ou auto-imunes. Na anemia ferropriva, os trabalhos são controversos. O presente estudo avalia a DE em 21 pacientes portadores de anemia ferropriva, utilizando a ectacitometria. Os resultados obtidos a partir do ID demonstram DE diminuída nesses doentes, quando comparada ao grupo controle (p< 0,0007). O presente estudo sugere que o fator responsável pela diminuição da DE na anemia ferropriva é a microcitose. Recentemente, relatos desta anemia associada a fenômenos trombóticos aumentaram o interesse no estudo da DE para melhor entendimento desses casos.

Deformability allows the 7 to 8 µm red cell to cirDeformability allows the 7- to 8-µm red blood cells to circulate through capillaries of 3 µm. This phenomenon depends on cellular geometry, internal viscosity and viscoelastic properties of the membrane. Among the various techniques of erythrocyte deformability analysis, such as micropipette aspiration, filtration and reoscopy, we chose ektacytometry. This technique uses a laminar flow viscometry, where red blood cell shape changes are continuously monitored by laser, processed by a computer, generating the "Deformability Index", which shows the elliptocytogenesis of the erythrocyte under "shear stressl" force. Erythrocyte deformability has been described in a number of situations like hereditary or autoimmune hemolytic anemia. In respect to iron deficiency anemia, conclusions are controversial. The present study evaluates erythrocyte deformability in 21 patients with documented iron deficiency, using ectacytometry. Results obtained from deformability Index demonstrate diminished erythrocyte deformability in individuals with iron deficiency anemia, when compared to a control group (p< 0.0007). The present study suggests that the factor responsible for diminished erythrocyte deformability in iron deficiency is microcytosis. Recently, this anemia has been associated to thrombotic phenomenon, which has raised interest in the study of erythrocyte deformability, in order to understand these cases.

Erythrocyte enzyme abnormalities in leukemias.

Red cell enzymes were assayed in a total of 67 patient including 24 patients with AML (19 relapse, 5 remission), 16 patients with ALL (10 relapse, 6 remission), 22 patients with CML and 5 patients with blastic CML. Diagnosis of leukemia was based on clinical presentation, peripheral blood smear and bone marrow examination (as per FAB classification). PK activity was significantly high in case of CML and blastic CML (p<0.01). Red cell HK was high in all leukemia subtypes. There was no alteration in red cell G6PD. Notably there was no PK deficiency in AML or G6PD deficiency in ALL. Activities of G6PD and PK could be correlated in cases of CML, AML, (p<0.05) and ALL (p<0.01) i.e. when there was increased activity of G6PD, PK activity also tended to be higher. HK activity showed a positive correlation with PK and G6PD activity in cases of CML (p<0.05), however in acute leukemia there was no such correlation. Alteration of enzyme activities among red cells in leukemia occurred only during relapse. At the time of remission there has been no significant alteration in any of the enzyme activities. It would therefore, appear that enzyme alterations seen in leukemia patients is due to abnormal pluripotent stem cell that has given to a leukemia cell. The fact that enzyme alterations have primarily occurred at the time of relapse would further substantiate that abnormalities of red cell enzymes may be the result of a derivation some circulating red cells from the abnormal pluripotent stem cell. With the recovery of normal stem cells function during remission, enzyme abnormalities tend to become normal.

Aluminium toxicity induced poikilocytosis in an air-breathing teleost, Clarias batrachus (Linn.).

The present study was undertaken to assess the toxicity of acid alone and two different sublethal concentrations of aluminium, (25% and 75% dose of 96 hr LC50 value in acidified soft water of pH 5) on red blood cells of a stenohaline catfish, C. batrachus for an acute exposure of 5 days. The scanning electron microscopic studies on all the three treated groups revealed several kinds of erythrocyte alterations and modifications with abnormal morphology. These included abnormal surface-wrinkling accompanied with excessive roughness on the membrane, erythrocytes with surface granulation in higher dose and finally the appearance of morphologically abnormal forms, the codocyte (target cell) and the stomatocyte. The results suggest that abnormality in the shape of erythrocytes could be linked to altered surface membrane area to volume ratio, decrease in cytoplasmic volume owing to reduced Hb content or increase in the amount of water content within the cell resulting from osmotic disequilibrium. In this context, the abnormal surface membrane morphology could be attributed to cytoskeleton fragility and defects in structural proteins. Further, the acid group exhibited a striking behavior of cellular adhesion and bonding to adjoining cell surfaces, culminating in several bunches which thereby reduces the surface area for gaseous exchange and could produce blocking effect while flowing through microcirculation.

Relationship between the genotype and hematologic characteristics in the fetuses with thalassemia / 中华儿科杂志

<p><b>OBJECTIVES</b>To investigate the relationship between the genotype and the hematologic characteristics in the fetuses with different types of thalassemia.</p><p><b>METHODS</b>Fetal blood samples were taken by cordocentesis, and hemograms from 572 fetuses at the gestational age of 18 to 38 weeks were examined. According to the genotypes of thalassemia, there were 117 fetuses with heterozygous alpha-thalassemia-1, and 60 with homozygous alpha-thalassemia-1. Twenty had beta-thalassemia mild, and 9 had beta-thalassemia major, respectively. The hematological parameters in these groups were compared with reference group in which 366 cases were included.</p><p><b>RESULTS</b>In alpha-thalassemia groups, hemoglobin (Hb), hematocrit (HCT), mean cell volume (MCV), mean cell hemoglobin (MCH), and mean cell hemoglobin concentration (MCHC) significantly decreased (P < 0.001), but in heterozygous alpha-thalassemia-1, red blood cell (RBC) increased. In homozygous alpha-thalassemia-1, RBC decreased significantly (P < 0.001), but white blood cell and nucleated erythrocyte increased (P < 0.001). There were no significant differences between beta-thalassemia and reference group in most hematological parameters except for decrease of MCHC.</p><p><b>CONCLUSIONS</b>In the fetal period, the hemogram of the fetuses with alpha-thalassemia changes significantly, while it does not change in beta-thalassemia. For the couple with heterozygous alpha-thalassemia, hemogram can provide some information for prenatal screening and diagnosis for those fetuses with alpha-thalassemia, especially for homozygous, but genotype detection is necessary for confirming the diagnosis.</p>

Diagnóstico diferencial de anemias microcíticas / Microcytic anemia differential diagnostic

La anemia microlítica se define comúnmente por la disminución en el volúmen corpuscular medio (VCM). Las formas más frecuentes se deben a deficiencia de hierro y a síndromes talasémicos. En este trabajo, fueron seleccionados 250 hemogramas de un total de 8.738 cuyo VCM era menor a 80 fL y que correspondían a una población formada por 76 por ciento de mujeres con una edad promedio de 37,5 años. En cada muestra se detrminó la ferremia, se realizó una electroforesis de hemoglobina (Hb) y se investigó la presencia de las mutaciones IVS1-1, IVS1-6, IVS1-110 y del codón 39 del exón 2. La frecuencia de distribución de los pacientes con microcitosis mostró dos poblaciones bien definidas, una constituida por pacientes ferropénicos (n=152) y otra, por pacientes talasémicos menores (n=81). Los pacientes con un área de distribución eritrocitaria (ADE) mayor al 20 por ciento no presentaban mutación y sí resultaron ferropénicos. Los valores hallados de HbA2 mayores a 3,5 por ciento coinciden con la presencia de algunas de las mutaciones citadas. La mutación más común fue la IVS1-110 que alcanzó un valor del 12 por ciento, que coincidió con otros estudios realizados en la Argentina

Analyse des prescriptions d'hemogramme recues au Laboratoire d'Hematologie du CHU de Yopougon

Objectifs : Analyser la prescription; les resultats de l'hemogramme au laboratoire d'Hematologie du CHU de Yopougon. Methodologie: II s'agit d'une etude transversale sur une periode de 3 mois. RESULTATS : AU PLAN EPIDEMIOLOGIQUE ET CLINIQUE : 53pour cent des sujets de notre population d'etude sont de sexe masculin ; 61;5pour cent des sujets sont des adultes et adolescents et 38;3pour cent des enfant ; 95pour cent des prescriptions proviennent du CHU de Yopougon dont 60;5pour cent relevant d'actes externes et 39;5pour cent issus des services d'hospitalisation ; Les services prescripteurs sont surtout les services d'hematologie (41;1pour cent). de pediatrie (14;7pour cent) de medecine (13;6 pour cent) ; Les motifs de prescriptions sont principalement la drepanocytose (25pour cent). le syndrome tumoral(25pour cent); l'anemie (11;1pour cent) et l'hyperthermie (11pour cent) ;L'hemogramme se prescrit sous 2 formes : NG (64;3pour cent); NFS (25;6pour cent) ; 24pour cent des prescriptions ne comportent pas de renseignements epidemiologiques necessaires a une correcte interpretation des resultats. AU PLAN HEMATOLOGIQUE : Les anomalies des GR revelent : 78pour cent d'anemie. Il s'agit d'une anemie franche 7;6(g/dl); normochrome normocytaire (56pour cent). plus frequente chez la femme (62pour cent). Au niveau des GB; nous avons note : 30pour cent d'hyper leucocytose et 10pour cent de leucopenie ; la formule leucocytaire a montre que la neutropenie (22pour cent). l'hyper eosinophilie (13pour cent) et la lymphocytose (8pour cent) sont predominantes. Au niveau des Plaquettes.; la thrombopenie (22pour cent) etait l'anomalie la plus frequente. La repartition des anomalies par tranche d'age a montre que chez les adultes; l'anemie; la neutropenie et la thrombopenie occupent les premieres places avec respectivement 45;6pour cent 15;9pour cent et 14;5pour cent tandis que chez les enfants on a l'anemie (32;4pour cent); l'hyper leucocytose (15;8pour cent) et la thrombopenie (7;5pour cent)

A medical mystery. Lead poisoning

A 25-year-old schizophrenic man presented with abdominal pain, nausea, vomiting, weight loss and anaemia. He was noted to be malnourished with generalised muscle atrophy. Laboratory investigations showed Hb 4.7 mmol/L, MCV 80fl, bilirubin 75 micromol/L and reticulocyte count 93 percent. Peripheral blood films showed anisocytosis, basophilic stippling and Cabot's rings. Electromyography confirmed typical motor nerve neuropathy. The clinical and laboratory findings were that of lead poisoning. The patient was later found to be ingesting lead-containing paint. He was treated with lead-chelating agents.

Rapid screening of PNH red cell populations using the gel test.

Paroxysmal nocturnal hemoglobinuria (PNH), an acquired clonal hematopoietic stem cell defect is underdiagnosed because of its atypical symptoms in some patients and because available methods, which are time consuming and complicated, are not widely used. The purpose of this study is to compare the results of the detection of PNH red cell populations using the PNH gel test and the Ham test. Fifty-eight blood samples obtained from 35 patients and 23 healthy blood donors were tested for PNH by the PNH gel test and the Ham test. It was found that 7 (20%) of the patients were positive for PNH by both tests. Twenty-three blood samples from healthy donors were all negative for PNH by both tests. The overall sensitivity and specificity of the gel test were 100%. This study showed that the PNH gel test was simple and could replace the Ham test as a screening test for PNH. This test would be especially easy to introduce in laboratories that are already using this system for blood grouping and antibody detection.