Myelodysplastic syndrome: validation of flow cytometry multilineage score system / Síndrome mielodisplásica: validação de ficha de escore multilinhagem por citometria de fluxo

ABSTRACT Objective To validate multilineage score system correlating results of flow cytometry, cytogenetics, cytomorphology and histology from samples of patients with suspected myelodysplastic syndrome or cytopenia of unknown origin. Methods A retrospective study analyzing laboratory data of 49 patients with suspected myelodysplastic syndrome or cytopenia of unknown origin, carried out between May and September 2017. The inclusion criteria were availability of flow cytometry results, and at least one more method, such as morphology, histology or cytogenetics. Thirty-eight patients were classified as diagnosis of myelodysplastic syndromes, whereas 11 were classified as normal. Patients were evaluated based on score systems, Ogata score and flow cytometry multilineage score. Results Comparing the scores obtained in the Ogata score and the multilineage score, it was observed that in four cases the Ogata score was zero or 1 point, while the multilineage score was higher than 3 points. In addition, in 12 cases with Ogata score of 2, the multilineage score was greater than 3. Conclusion The flow cytometry multilineage score system demonstrated to be more effective in dysplasia analysis, by assessing the erythroid, monocytic, granulocytic and precursor cell lineages, apart from the parameters evaluated by the Ogata score.

RESUMO Objetivo Validar ficha de escore multilinhagem correlacionando resultados obtidos de citometria de fluxo, citogenética, citomorfologia e histologia de amostras de pacientes com suspeita de síndrome mielodisplásica ou citopenias a esclarecer. Métodos Estudo retrospectivo de análise de dados laboratoriais de 49 pacientes com suspeita clínica de síndrome mielodisplásica ou citopenias a esclarecer realizado entre maio e setembro de 2017. Os critérios de inclusão foram a disponibilidade de resultados de citometria de fluxo e de, pelo menos, outra metodologia, entre morfologia, histologia, ou citogenética. Trinta e oito pacientes foram classificados como diagnosticados com síndromes mielodisplásicas enquanto 11 foram classificados como normais. Os pacientes foram avaliados utilizando sistemas de escore, escore de Ogata e ficha multilinhagem. Resultados Comparando as pontuações obtidas no escore de Ogata e na ficha multilinhagem, observou-se que, em quatro casos, o score de Ogata foi zero ou 1 ponto, enquanto, pela ficha multilinhagem, a pontuação foi superior a 3 pontos. Além disso, em 12 casos com escore de Ogata 2, a pontuação pela ficha multilinhagem foi superior a 3. Conclusão A ficha multilinhagem demonstrou ser mais eficaz na análise de displasia por avaliar as linhagens eritroide, monocítica, granulocítica e células precursoras, além dos parâmetros avaliados no escore de Ogata.

Effect of MiR-451a on Erythroid Differentiation of K562 Cells under Hypoxia / 中国实验血液学杂志

OBJECTIVE@#To investigate the changes of GATA-1 protein expression during erythroid differentiation of K562 cells under hypoxia and how GATA-1 can regulate erythroid differentiation by up-regulating the expression of miR-451a and inhibiting the expression of 14-3-3ζ.@*METHODS@#K562 cells were divided into 2 groups: the normoxia group and the hypoxia group, after the induction of hemin for 96 h, the positive cells rate of the benzidine staining, the mRNA expression of γ-globin and the expression of CD235a were detected, and the success of the model was verified. The changes of GATA-1 and miR-451a expression in the above-mentioned 2 groups, the changes of miR-451a expression after over-expressed GATA-1 were detected by Western blot and qRT-PCR. The cells in normoxic group and hypoxia group were divided into negative control group (NC group) and miR-451a over-expression group respectively, and the degree of erythroid differentiation in the four groups was judged according to the corresponding erythroid differentiation indexes, and the expression of 14-3-3ζ was detected by Western blot after over-expressed miR-451a.@*RESULTS@#The positive cell rate of benzidine staining, mRNA expression of γ-globin and the expression of CD235a after 96 h induction by K562 cells under hypoxia were significantly higher than 0 h, suggesting that the erythroid differentiation model of K562 cells under hypoxia was replicated successfully. The expression levels of GATA-1 protein and miR-451a in the hypoxic group were significantly higher than that in the normoxic group (P<0.05). The expression level of miR-451a in hypoxia group was significantly higher than that in NC group after overexpressed GATA-1 (P<0.05). After over-expressed of miR-451a under hypoxia, the positive cell rate of benzidine staining, the mRNA expression level of γ-globin and the expression of CD235a were significantly higher than those in NC group (P<0.05). The expression level of 14-3-3ζ protein in miR-451a over-expressed group was lower than that in NC group under hypoxia (P<0.05).@*CONCLUSION@#Hypoxia can significantly increase the expression of GATA-1 protein, and the increase of GATA-1 expression can up-regulate the expression of miR-451a, thereby inhibiting the expression of 14-3-3ζ protein, which hinders the cell proliferation in erythroid differentiation model of K562 cells and plays an important role in promoting erythroid differentiation.

Inhibition of Erythroid Differentiation of Human Leukemia K562 Cells by N-acetylcysteine and Ascorbic Acid through Downregulation of ROS / 生物医学与环境科学（英文）

This study investigated the effects of N-acetylcysteine (NAC) and ascorbic acid (AA) on hemin-induced K562 cell erythroid differentiation and the role of reactive oxygen species (ROS) in this process. Hemin increased ROS levels in a concentration-dependent manner, whereas NAC and AA had opposite effects. Both NAC and AA eliminated transient increased ROS levels after hemin treatment, inhibited hemin-induced hemoglobin synthesis, and decreased mRNA expression levels of β-globin, γ-globin, and GATA-1 genes significantly. Pretreatment with 5,000 μmol/L AA for 2 h resulted in a considerably lower inhibition ratio of hemoglobin synthesis than that when pretreated for 24 h, whereas the ROS levels were the lowest when treated with 5,000 μmol/L AA for 2 h. These results show that NAC and AA might inhibit hemin-induced K562 cell erythroid differentiation by downregulating ROS levels.

Inhibitory effect of mesenchymal stem cell co-culture on erythroid differentiation of K562 cells compared to the control group

Objective: Bone marrow mesenchymal stem cells [BMMSCs] reside in the bone marrow and control the process of hematopoiesis. They are an excellent instrument for regenerative treatment and co-culture with hematopoietic stem cells [HSCs]

Materials and Methods: In this experimental study, K562 cell lines were either treated with butyric acid and co-cultured with MSCs, or cultivated in a conditioned medium from MSCs plus butyric acid for erythroid differentiation. We used the trypan blue dye exclusion assay to determine cell counts and viability in each group. For each group, we separately assessed erythroid differentiation of the K562 cell line with Giemsa stain under light microscopy, expression of specific markers of erythroid cells by flowcytometry, and erythroidspecific gene expressions by real-time polymerase chain reaction [RT-PCR]

Results: There was enhandced erythroid differentiation of K562 cells with butyric acid compared to the K562 cell line co-cultured with MSCs and butyric acid. Erythroid differentiation of the K562 cell line cultivated in conditioned medium with butyric acid was higher than the K562 cell line co-cultured with MSCs and butyric acid, but less than K562 cell line treated with butyric acid only

Conclusion: Our results showed that MSCs significantly suppressed erythropoiesis. Therefore, MSCs would not be a suitable optimal treatment strategy for patients with erythroid leukemia

Differentiation of K562 Cells Induced by Pulsatilla Saponin A into Erythroid Lineage / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the differentiation-inducing potentiality of Pulsatilla saponin A on K562 cells.</p><p><b>METHODS</b>Pulsatilla saponin A of different concentrations was used to treat K562 cells; the benzidine staining and the hemoglobinometry were applied to measure the change of hemoglobin content; the flow cytometry (FCM) was used to detect the expression of CD71 and GPA on K562 cells.</p><p><b>RESULTS</b>K562 cells treated with 4 µg/ml pulsatilla saponin A differentiated into the erythroid lineage. With the treatment of pulsatilla saponin A, the hemoglobin content in K562 cells increased significantly; CD71 and GPA expression on the K562 cell surface were up-regulated.</p><p><b>CONCLUSION</b>Pulsatilla saponin A can induce K562 cells to differentiate into erythroid lineage.</p>

Construction of Protein Phosphatase 2A Catalytic Subunit β (PPP2Cβ) Overexpression Lentiviral Vector and Its Effect on K562 Erythroid Differentiation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To construct the ovexpression lentivirus vector of PPP2Cβ, the catalytic subunit of protein phosphatase 2A, so as to obtain high-titer packaged lentivirus particles, and to examine the effect of PPP2Cβ on the erythroid differentiation Methods: The CDS of PPP2Cβ was cloned into the second generation of lentivirus vector FUGW, which should be used to co-transfect HEK 293T cells with the lentiviral expression vector and packaging vectors including pMD2G and pSPAX2. Lentiviruses were harvested at 36 and 48 hours after transfection. Titers of viral stock were determined by using flow cytometric analysis. The Western blot was performed to detect the expression level of PPP2Cβ in K562 cells transinfected with the lentiviruses. Benzidine staining and real-time PCR analysis were used to assess the erythroid differentiation of K562 cells.</p><p><b>RESULTS</b>The PPP2Cβ overexpressing lentivirus vectors were constructed, the high-titer lentiviral particles were obtained, and then the PPP2Cβ overexpression K562 cell line was established and promote erythroid differentiation of K562 cells.</p><p><b>CONCLUSION</b>This study suggests that overexpression PPP2Cβ can promote K562 cell erythroid differentiation.</p>

Correlation between methylation and expression level of P15 and P16 genes during differentiation of cord blood stem cells into erythroid lineage mediated by erythropoietin

Several influential factors such as transcription factors and intracellular signaling components are involved in differentiation of stem cells into a specific lineage. P15 and p16 proteins are among these factors. Accumulating evidences has introduced the epigenetic as a master regulator of these factors during lineage specification. The main objective of this study is to determine the correlation between the expression level and methylation pattern of P15 and P16 genes in erythroid lineage after in vitro differentiation by erythropoietin [EPO]. The purified and expanded CD34+ cord blood stem cells were differentiated into erythroid lineage in the presence of EPO. DNA was isolated from both cord blood stem cells and differentiated cells. The Real-Time PCR performed using cDNA and the isolated DNA was used in methylation Specific PCR [MSP] reaction for methylation pattern analysis in both pre and post differentiation stages. The study demonstrated that P15 and P16 genes have partial methylation after erythroid differentiation by EPO. The Expression of P15 gene was higher after differentiation and the expression of P16 gene had a slightly decreased level in post differentiation stage. Significant increase in P15 gene expression after differentiation to erythroid lineage, suggests the remarkable efficacy of this gene in erythroid function. According to upregulation of P15 gene after differentiation despite unchanged methylation status and slight down regulation of P16 gene with slight hyper-methylation of the gene it can be suggested that although the methylation can affects the expression level of P16 gene, the P15 gene is not affected by this mechanism during erythroid differentiation mediated by EPO

Differentiation potential of O Bombay human-induced pluripotent stem cells and human embryonic stem cells into fetal erythroid-like cells

There is constant difficulty in obtaining adequate supplies of blood components, as well as disappointing performance of "universal" red blood cells. Advances in somatic cell reprogramming of human-induced pluripotent stem cells [hiPSCs] have provided a valuable alternative source to differentiate into any desired cell type as a therapeutic promise to cure many human disease. In this experimental study, we examined the erythroid differentiation potential of normal Bombay hiPSCs [B-hiPSCs] and compared results to human embryonic stem cell [hESC] lines. Because of lacking ABO blood group expression in B-hiPSCs, it has been highlighted as a valuable source to produce any cell type in vitro. Similar to hESC lines, hemangioblasts derived from B-hiPSCs expressed approximately 9% KDR+CD31+ and approximately 5% CD31+CD34+. In semisolid media, iPSC and hESC-derived hemangioblast formed mixed type of hematopoietic colony. In mixed colonies, erythroid progenitors were capable to express CD71+GPA+HbF+ and accompanied by endothelial cells differentiation. Finally, iPS and ES cells have been directly induced to erythropoiesis without hemangioblast formation that produced CD71+HbF+erythroid cells. Although we observed some variations in the efficiency of hematopoietic differentiation between iPSC and ES cells, the pattern of differentiation was similar among all three tested lines

Mielolipoma suprarrenal incidental / Incidental Detection of Adrenal Myelolipoma

La implementación de nuevas tecnologías y la ampliación de la cobertura a nivel de imágenes, han facilitado encontrar casos incidentales, siendo el más común el adenoma suprarrenal no funcionante. El mielolipoma suprarrenal es una neoplasia benigna, rara, en la mayoría de casos asintomática compuesta por tejido adiposo maduro y elementos hematopoyéticos. Se presenta un caso clínico de una paciente remitida al servicio de radiología por presentar en ecografía previa, una masa en región suprarrenal derecha incidentalmente. La paciente se encontraba asintomática, por lo cual se realiza tomografía axial computarizada abdominal. A la paciente por su ausencia de síntomas se le continúo con seguimiento periódico sin detectar cambios evolutivos (MÉD.UIS. 2014;27(2):105-107) .

The implementation of new technologies and expanding coverage to level of images, have provided incidental finding cases, the most common nonfunctioning adrenal adenoma. Myelolipoma adrenal benign tumors, rarely, in most cases consisting of asymptomatic mature adipose tissue and hematopoietic elements (myeloid and erythroid cells). We present a case of a patient referred to the radiology department for filing in previous ultrasound a right adrenal mass incidentally region. The patient was asymptomatic, which is done by abdominal CT. A patient by the absence of symptoms is continued with regular monitoring undetected evolutionary changes (MÉD.UIS. 2014;27(2):105-107).

Production of mature red blood cell by using peripheral blood mononuclear cells / 中国实验血液学杂志

Most protocols for in vitro producing red blood cells (RBC) use the CD34(+) cells or embryonic stem cells from cord blood, bone marrow or peripheral blood as the start materials. This study was purposed to produce the mature RBC in vitro by using peripheral blood mononuclear cells as start material. The peripheral blood mononuclear cells (PBMNC) were isolated from buffy coat after blood leukapheresis, the mature red blood cells (RBC) were prepared by a 4-step culture protocol. The results showed that after culture by inducing with the different sets of cytokines and supporting by mouse MS-5 cell line, the expansion of PBMNC reached about 1000 folds at the end of the culture. About 90% of cultured RBC were enucleated mature cells which had the comparable morphological characteristics with normal RBC. Colony-forming assays showed that this culture system could stimulate the proliferation of progenitors in PBMNC and differentiate into erythroid cells. The structure and function analysis indicated that the mean cell volume of in vitro cultured RBC was 118 ± 4 fl, which was slight larger than that of normal RBC (80-100 fl); the mean cell hemoglobin was 36 ± 1.2 pg, which was slight higher than that of normal RBC (27-31 pg); the maximal deformation index was 0.46, which approachs level of normal RBC; the glucose-6-phosphate dehydrogenase and pyrurvate kinase levels was consistant with young RBC. It is concluded that PBMNC are feasble, convenient and low-cost source for producing cultured RBC and this culture system is suitable to generate the RBC from PBMNC.

Application of flow cytometry in detecting dysplasia of myelodysplastic syndromes / 中国实验血液学杂志

Myelodysplastic syndrome (MDS) is a heterogeneous disease characterized by dysplasia and ineffective hematopoiesis. The dysplasia is crucial in the diagnosis of MDS, but the morphologic abnormalities of bone marrow cells are not specific for MDS. When the morphological evaluation of marrow dysplasia and cytogenetics can not give enough informations, for diagnosis of MDS, the application of flow cytometry (FCM) for immunophenotyping in MDS will become particularly important. Multiparametric evaluation of myeloid, monocytic maturation and antigen expression pattern contribute to the identification of two or more aberrancies in MDS cases. FCM evaluation of erythroid dysplasia is particularly difficult, because of the limited availability of specific markers. By analyzing the proteins involved in cellular iron metabolism, MDS erythroid cells present an "iron-loaded" phenotype characterized by increased ferritin contents and reduced transferrin receptor, which reflects the degree of dysplasia assessed by morphology. The proportion of CD34(+) cells increased, abnormal expression of surface antigen is also important. The application of flow cytometry in detecting dysplasia of myelodysplastic syndrome is discussed in this article.

A case of myelodysplastic syndrome with marked eosinophilia showing favorable prognosis

Myelodysplastic syndrome (MDS) with eosinophilia is a rare condition and has yet to be classified under the 2008 World Health Organization classification. However, reports have described the prognostic significance of chronic persistent eosinophilia in MDS. Here, we report a case of a 67-year-old woman who was admitted to the hospital in July 2007 with generalized weakness, dizziness, and dyspnea on exertion persisting for 5 years. In the initial investigation, eosinophilia (22.1%) in peripheral blood and an increased proportion of eosinophils (5.6%) in normocellular bone marrow with dysplastic megakaryocytes and erythroid cells were noted. Eosinophilia was continuously detected during follow-up over 3 years. In a second bone marrow examination in August 2010, hypercellular bone marrow with similar features was observed. These findings led to the diagnosis of MDS with chronic persistent eosinophilia. To increase awareness of the prognostic significance of MDS with chronic eosinophilia, here we report a slow-progressing case of MDS with chronic persistent eosinophilia lasting over 6 years.

Reticulocyte assessment as an indirect evaluation and elaboration of erythroid marrow output in the groups of pregnant women in Maiduguri, Nigeria

Nutritional anaemia is one of Nigeria's major public health problem among pregnant women. In this locality, no large scale study has been done to assess bone marrow activity in groups of pregnant women using Reticulocyte assessment. This result will serve as indirect evaluation and elaboration of erythroid marrow output in the pregnant women. To assess bone marrow activity in groups of pregnant women using Reticulocyte assessment as an indirect evaluation. 150 women covering the three trimesters of pregnancy were recruited into the study, prospectively. 101 multiparous and 49 primiparous pregnant women, consisting of 33 in the first trimester, 74 in the second trimester and 43 in the third trimester were investigated. Red blood cell count, Reticulocyte count and assessment were conducted by standard methods. The patients in the third trimester had statistically significant [P<0.05] lower reticulocyte values than those in the first and second trimesters. The primiparous pregnant women had statistically significant [P<0.05] higher reticulocyte values than the multiparous pregnant women. Absolute reticulocyte count results were 50.0 +/- 15.2, 60.0 +/- 18.4, 34.0 +/- 12.8 and 48.0 +/- 12.0 for first, second and third trimesters and combined group, respectively. Reticulocyte index was 1.5 +/- 0.6, 1.9 +/- 0.8, 0.8 +/- 0.6 for first, second and third trimesters, respectively and 1.5 +/- 0.5 for combined group. Reticulocyte production index was 0.9 +/- 0.2, 1.1 +/- 0.4, 0.7 +/- 0.5 and 0.7 +/- 0.4 for first, second and third trimesters and combined group, respectively. In this study, bone marrow activity as assessed by reticulocyte studies is on the lower side of the normal range, more so in the third trimester of pregnancy. Severe anaemia during pregnancy therefore remains endemic despite intervention measures such as the distribution of iron and folate tablets. One of the problems yet uninvestigated is the bone marrow turnover as affected by some other nutritional differences as well as malaria, heavy loads of helminths, and other inflammatory or infectious diseases. A successful strategy to combat anaemia, therefore, should address all the casual factors after their elucidation

[ effect of adult mouse lung extracts growth factors in differentiation of umbilical cord stem cells into blood cells in vitro condition]

Stem cells [SCs] have great therapeutic indication due to their potency of self-renewal, multilineage differentiation, feasibility and safety for donors. In this study, adult mouse lung extracts containing hematopoietic growth factors were administered to umbilical cord, and evaluated the differentiation of umbilical cord stem cells into erythroid and myeloid lineages. In this basic and practical research, SCs were isolated from umbilical cord by enzyme digestion and cultured in appropriate culture medium. Subjects were divided into four groups: Experimental groups 1 and 2 [E1 and E2] which were exposed to 50% and 70% concentration of lung extract for 7 days, respectively, sham [Sh] group which did not exposed to lung extract and cultured for 7 days, and control group [C]. E1, E2 and Sh groups were incubated for 7 days. All groups were evaluated by alkaline phosphatase detection kit for stem cells. Then, blood cells count and hematopoietic growth factors were assessed. ANOVA was used for data analysis. There were significant changes in E2 groups as compared with Sh and C groups, so that E2 group cells were differentiated into erythroid and myeloid lineages. Growth factors in lung extract could have stimulatory effects on umbilical cord stem cell differentiation into blood cells

Expression of DLK1 gene in acute leukemias and its function in erythroid differentiation of K562 cell line / 中南大学学报(医学版)

OBJECTIVE@#To determine the expression of DLK1 gene in acute leukemias (AL) and its function in erythroid differentiation of K562 cells.@*METHODS@#We detected the expression of DLK1 gene in 65 different acute leukemia categories (a test group) and 34 normal bone marrow controls (a control group) with RT-PCR. DLK1 protein in 20 out of the 65 AL patients and 13 of the 34 controls was assayed by Western blot. The K562 cell line was induced to erythroid differentiation by hemin. We observed the relationship between its expression and erythroid differentiation.@*RESULTS@#Both leukemia cells and normal marrow cells expressed DLK1. The expression of DLK1 mRNA in patients in the test group was higher than that in the control group (P=0.018), while there was no significance between acute lymphoblastic leukemia and acute myelogenous leukemia (P>0.05).The expression of DLK1 mRNA in the test group at onset had no relation with the WBC and platelet count in the total peripheral blood, and the same was true for blast cell rates in bone marrow cells.The level of DLK1 protein in the test group was higher than that in the control group, which was consistent with the mRNA expression (P=0.042). The expression of DLK1 mRNA decreased gradually with K562 cells towards hemin-induced erythroid differentiation.@*CONCLUSION@#DLK1 gene may be involved in leukemia,but the mRNA level of DLK1 has no relation with some clinical characteristics of AL patients at onset. DLK1 may inhibit the erythroid differentiation of K562 cells.

Acute erythroid toxicity in visceral leishmaniasis: a rare complication of antimonial therapy.

The spectrum of side-effects of sodium stibogluconate is well described, however, little is known regarding the acute erythroid toxicity caused by this drug. We hereby present a case with this unusual complication of antimonial therapy. A 6-year-old male with leishmaniasis was started on parenteral sodium stibogluconate. During the course of treatment, his hemoglobin (Hb) dropped from 7.2 g/dl to 3.5 g/dl. Bone-marrow aspirate showed karyorrhexis in many erythroid precursors with several Leishmania donovanii bodies. Sodium stibogluconate was stopped and amphotericin-B was started. Four days after the cessation of the antimonials, the patient's Hb improved to 5 gm/dl with a corrected reticulocyte count of 10% indicating bone-marrow erythroid regeneration. The exact mechanism of this acute erythroid toxicity of sodium stibogluconate remains unexplored.

Expression of human ermap gene in umbilical cord blood mononuclear cells during differentiation and development towards erythroid lineage / 中国实验血液学杂志

The aim of study was to explore the potential of human erythroid membrane associated protein (ERMAP) gene in erythroid cell differentiation and development, mononuclear cells (MNCs) were isolated from umbilical cord blood and induced to erythroid cell differentiation by SCF, IL-3 and EPO. The cell morphology was observed by using optical microscopy, the positive rate of cells was counted by biphenylamine staining and the ratios of CD36+/CD235a-, CD36+/CD235a+, CD36-/CD235a+ cells were detected by flow cytometry, the change of human ermap gene expression level was analyzed by using fluorescent quantitative PCR (FQ-PCR). The results showed that the ermap gene expression level increased while MNCs were induced to erythroid lineage after treatment with SCF, IL-3 and EPO. It is concluded that the human ermap gene plays an important role in differentiation and development of erythroid cells.

Valores de referencia de la serie eritroide con tecnología del siglo XXI en embarazadas: Prevalencia de anemia / Reference values for the erythroid series with twenty-first century technology in pregnant women: Prevalence of anemia

La disponibilidad, en los hospitales, de contadores hematológicos con nueva tecnología hace necesario actualizar los valores de referencia para establecer el límite entre lo normal y lo patológico. Con este objetivo, se estableció cuáles son dichos valores en una población de mujeres embarazadas. Asimismo, se determinó el porcentaje de disminución de hemoglobina durante el embarazo por efecto de la dilución fisiológica, que fue del 2,2% del primero al segundo trimestre y del 1,5% del segundo al tercero, La prevalencia de anemia microcítica fue del 28% y de anemia macrocítica del 1,6%.

The availability in hospitals of blood cell counters using state-of-the-art technology has made it necessary to update the corresponding reference values. These values allow us to establish the boundary between normal and pathological. With this objective in mind, these intervals were established among pregnant women. Furthermore, the percentage of haemoglobin level decrease during pregnancy due to the effect of physiological dilution has been established as being of 2.2% between the first and the second trimester and 1.5% between the second and the third. The prevalence of microcytic anaemia was 28% and of macrocytic anaemia 1.6%.

Capsaicin promotes the development of burst-forming nits-erythroid (BFU-E) from mouse bone marrow cells

Capsaicin, the pungent component of chilli peppers, is known to induce mediators of hematopoiesis. We investigated the effect of capsaicin on hematopoiesis in mouse progenitor cells. Treatment of mouse bone marrow cells with capsaicin induced the formation of colony of burst-forming units-erythroid (BFU-E). We also found that the number of erythropoietin receptor (EpoR)-positive cells was increased by capsaicin. To clarify the effect of capsaicin on erythroid lineage, BFU-E colonies were separated from non-BFU-E colonies by colony-picking after in vitro culture of mouse bone marrow cells. Quantitative RT-PCR analysis revealed that capsaicin stimulated the expression of the erythroid-specific genes encoding EpoR, glycophorin A (GPA), beta-globin (Hbb-b1), GATA-1, PU.1, nuclear factor erythroid-derived 2 (NF-E2), and Kruppel-like factor 1 (KLF1) in the BFU-E colonies. Furthermore, capsaicin could effectively stimulate the transfected GATA-1 promoter in K562 cells. GATA-1 is known as an essential transcription factor for the development of erythroid cells. Our results show that development of the erythroid lineage from bone marrow cells can be induced by treatment with capsaicin, and that GATA-1 seems to play a role in this induced erythroid maturation.

Hematopoietic features and apoptosis in the bone marrow of severe Plasmodium falciparum-infected patients: preliminary study.

The mechanism of anemia in severe falciparum malaria is still not completely understood. The purpose of this study was to determine whether apoptosis in the erythroid lineage causes anemia in falciparum malaria. Bone marrow aspirated from 8 severe falciparum malaria patients, 3 normal volunteers and 5 retrospective normal bone marrow smears were investigated. By light microscopic study, 5 of 8 hyperparasitemic patients had hypocellular bone marrows and erythroid hypoplasia, whereas the other 3 patients had normal cellularity. The mean myeloid : erythroid ratio of these 5 patients was significantly (p < or = 0.05) higher than normal. Apoptosis of bone marrow nucleated cells (BMNC) could be determined from the exposure of phosphatidylserine (PS) on the cell membrane but not DNA fragmentation (180-250 bp) or ultrastructural morphology. The percentages of apoptotic BMNC and apoptotic erythroid cells in bone marrow from each patient and controls varied from low to high, and were not associated with parasitemia. This study suggests that destruction of erythroid lineage, particularly through apoptosis regulation, cannot solely account for anemia in falciparum malaria.