Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 103
Filter
1.
J Clin Invest ; 130(10): 5576-5590, 2020 10 01.
Article in English | MEDLINE | ID: mdl-32663195

ABSTRACT

During hemolysis, macrophages in the liver phagocytose damaged erythrocytes to prevent the toxic effects of cell-free hemoglobin and heme. It remains unclear how this homeostatic process modulates phagocyte functions in inflammatory diseases. Using a genetic mouse model of spherocytosis and single-cell RNA sequencing, we found that erythrophagocytosis skewed liver macrophages into an antiinflammatory phenotype that we defined as MarcohiHmoxhiMHC class IIlo erythrophagocytes. This phenotype transformation profoundly mitigated disease expression in a model of an anti-CD40-induced hyperinflammatory syndrome with necrotic hepatitis and in a nonalcoholic steatohepatitis model, representing 2 macrophage-driven sterile inflammatory diseases. We reproduced the antiinflammatory erythrophagocyte transformation in vitro by heme exposure of mouse and human macrophages, yielding a distinctive transcriptional signature that segregated heme-polarized from M1- and M2-polarized cells. Mapping transposase-accessible chromatin in single cells by sequencing defined the transcription factor NFE2L2/NRF2 as a critical driver of erythrophagocytes, and Nfe2l2/Nrf2 deficiency restored heme-suppressed inflammation. Our findings point to a pathway that regulates macrophage functions to link erythrocyte homeostasis with innate immunity.


Subject(s)
Hemolysis/physiology , Liver/cytology , Liver/physiology , Macrophages/cytology , Macrophages/physiology , Phagocytes/cytology , Phagocytes/physiology , Animals , Disease Models, Animal , Female , Heme/metabolism , Humans , In Vitro Techniques , Inflammation/prevention & control , Macrophages/classification , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Mutant Strains , NF-E2-Related Factor 2/deficiency , NF-E2-Related Factor 2/genetics , NF-E2-Related Factor 2/metabolism , Non-alcoholic Fatty Liver Disease/etiology , Non-alcoholic Fatty Liver Disease/pathology , Non-alcoholic Fatty Liver Disease/physiopathology , Phagocytes/classification , Phagocytosis/physiology , Phenotype , RNA-Seq , Single-Cell Analysis , Spherocytosis, Hereditary/genetics , Spherocytosis, Hereditary/pathology , Spherocytosis, Hereditary/physiopathology
4.
Cytometry B Clin Cytom ; 94(1): 189-195, 2018 01.
Article in English | MEDLINE | ID: mdl-28103644

ABSTRACT

BACKGROUND: Osmotic fragility test (OFT) is widely considered as a sensitive indicator of red blood cells' sensitivity to the hypotonic solution. It is often used as a screening test for the diagnosis of hereditary spherocytosis (HS). Nowadays, the osmotic fragility test based on flow cytometric analysis (FCM OF) is widely used in laboratory practice. The purpose of this study was to optimize the assay sensitivity and to validate its clinical application in the diagnostic screening of childhood anemias. METHODS: The study was conducted on 175 children suffering from various types of anemia (including 30 children with proven hereditary spherocytosis, HS) and 16 healthy subjects. All children were aged between 3 months and 17 years, including 94 boys and 97 girls. FCM OF was performed on every subject according to two different analysis time patterns (hemolysis was analyzed for 214 or 300 s) using Cytomics FC500 flow cytometer. RESULTS: Significant higher sensitivity was demonstrated by the tests carried out according to the longer analysis time pattern (90.0 vs. 83.33%). The level of specificity of both the analysis patterns was similar. When an extended analysis time was used, the percentage of red cell survival levels in HS patients were significantly lowered compared to the same cases analyzed with shorter incubation times and all other non-HS anemic cases (9.31 ± 4.69 vs. 35.59 ± 15.30%, P < 0.05). During the shorter analysis time, the values obtained were 13.76 ± 7.92% for HS and 48.18 ± 19.04% for non-HS, P < 0.0001. The 300-s test is very useful in distinguishing thalassemia patients from patients with other types of anemias (94.74% sensitivity and 90.12% specificity) and provided the values of remaining red blood cells as 70.46 ± 12.29% for thalassemia and 27.16 ± 13.01% for nonthalassemia subjects, P < 0.0001. CONCLUSION: Flow cytometric osmotic fragility test with a longer (300-s) analysis time demonstrated an increased sensitivity in detecting HS in anemic children. © 2017 International Clinical Cytometry Society.


Subject(s)
Flow Cytometry/methods , Hematologic Tests/methods , Hematology/methods , Osmotic Fragility/physiology , Adolescent , Child , Child, Preschool , Erythrocytes/physiology , Female , Hemolysis/physiology , Humans , Infant , Male , Mass Screening/methods , Sensitivity and Specificity , Spherocytosis, Hereditary/diagnosis , Spherocytosis, Hereditary/physiopathology , Thalassemia/diagnosis , Thalassemia/physiopathology
5.
Neonatal Netw ; 36(5): 280-288, 2017 Sep 01.
Article in English | MEDLINE | ID: mdl-28847351

ABSTRACT

Hereditary spherocytosis (HS) is the third most common yet most frequently underrecognized, congenitally acquired hemolytic disease of the neonate. Hereditary spherocytosis is caused by a defect of one or more erythrocyte membrane proteins, which leads to an increased rate of destruction of circulating red blood cells. The HS spectrum of symptoms is varied from asymptomatic to intrauterine hydrops. Diagnostic tests range from a complete blood count (CBC) analysis to deoxyribonucleic acid (DNA) sequencing. Management in the neonatal period focuses primarily on associated comorbidities, including the prevention of severe hyperbilirubinemia and anemia. Life span implications of HS include hemolysis, jaundice, anemia, splenomegaly, and periodic gallstones. Early identification and diagnosis of HS is essential to ensure proper monitoring and medical management throughout infancy, childhood, and adulthood.


Subject(s)
Hematologic Tests/methods , Phototherapy/methods , Spherocytosis, Hereditary , Early Diagnosis , Early Medical Intervention , Humans , Infant, Newborn , Male , Spherocytosis, Hereditary/diagnosis , Spherocytosis, Hereditary/physiopathology , Spherocytosis, Hereditary/therapy , Symptom Assessment/methods , Treatment Outcome
7.
Indian Pediatr ; 54(7): 563-566, 2017 Jul 15.
Article in English | MEDLINE | ID: mdl-28368266

ABSTRACT

OBJECTIVE: To analyze growth-failure and improvement, if any, following splenectomy in children with hereditary spherocytosis. METHODS: Data collection from case-records (n=82) over 27-years (1985-2011). RESULTS: Prevalence of stunting was 26%; 32% were underweight. Stunted children were older in age (P=0.006) and presented late (P=0.003). Splenectomy (n=26) improved anemia (P<0.001). However, height-for-age did not improve at 1-year, or 4.5-years (median) following splenectomy (P=1.0). Number of underweight children did not reduce at 1- (P=0.21), or 4.5-years (P=0.21) following surgery. CONCLUSION: Growth-failure is frequent in children with hereditary spherocytosis in India. Splenectomy corrected the anemia but failed to improve the growth.


Subject(s)
Failure to Thrive/epidemiology , Spherocytosis, Hereditary , Splenectomy/statistics & numerical data , Thinness/epidemiology , Adolescent , Child , Child, Preschool , Female , Growth Disorders/epidemiology , Humans , India/epidemiology , Infant , Infant, Newborn , Male , Retrospective Studies , Spherocytosis, Hereditary/epidemiology , Spherocytosis, Hereditary/physiopathology , Spherocytosis, Hereditary/surgery
8.
Hum Pathol ; 60: 95-103, 2017 02.
Article in English | MEDLINE | ID: mdl-27771375

ABSTRACT

Hereditary spherocytosis (HS) and sickle cell disease (SCD) are associated with splenomegaly and spleen dysfunction in pediatric patients. Scant data exist on possible correlations between spleen morphology and function in HS and SCD. This study aimed to assess the histologic and morphometric features of HS and SCD spleens, to get possible correlations with disease pathophysiology. In a large series of spleens from SCD, HS, and control patients, the following parameters were considered: (i) macroscopic features, (ii) lymphoid follicle (LF) density, (iii) presence of perifollicular marginal zones, (iv) presence of Gamna-Gandy bodies, (v) density of CD8-positive sinusoids, (vi) density of CD34-positive microvessels, (vii) presence/distribution of fibrosis and smooth muscle actin (SMA)-positive myoid cells, and (viii) density of CD68-positive macrophages. SCD and HS spleens had similar macroscopic features. SCD spleens had lower LF density and fewer marginal zones than did HS spleens and controls. SCD also showed lower CD8-positive sinusoid density, increased CD34-positive microvessel density and SMA-positive myoid cells, and higher prevalence of fibrosis and Gamna-Gandy bodies. HS had lower LF and CD8-positive sinusoid density than did controls. No significant differences were noted in red pulp macrophages. By multivariate analysis, most HS spleens clustered with controls, whereas SCD grouped separately. A multiparametric score could predict the degree of spleen changes irrespective of the underlying disease. In conclusion, SCD spleens display greater histologic effacement than HS, and SCD-related changes suggest impaired function due to vascular damage. These observations may contribute to guide the clinical management of patients.


Subject(s)
Anemia, Sickle Cell/pathology , Spherocytosis, Hereditary/pathology , Spleen/pathology , Anemia, Sickle Cell/complications , Anemia, Sickle Cell/physiopathology , Anemia, Sickle Cell/surgery , Biomarkers/analysis , Biopsy , Child , Child, Preschool , Female , Hemolysis , Humans , Immunohistochemistry , Italy , Male , Microvessels/pathology , Retrospective Studies , Severity of Illness Index , Spherocytosis, Hereditary/complications , Spherocytosis, Hereditary/physiopathology , Spherocytosis, Hereditary/surgery , Spleen/blood supply , Spleen/physiopathology , Spleen/surgery , Splenectomy
9.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 24(1): 229-32, 2016 Feb.
Article in Chinese | MEDLINE | ID: mdl-26913426

ABSTRACT

OBJECTIVE: To establish a new method for detection of red blood cell osmotic fragility by using flow cytometry. METHODS: The hypotension salt solution of different concentrations (0.70 ml normal saline+0.3 ml deionized water, 0.60 ml normal saline+0.40 ml deionized water and 0.55 ml normal saline+0.45 ml deionized water) were prepared with normal saline and deionized water, in which the red blood cells were suspended, and the residual red blood cells were detected by flow cytometer. RESULTS: There was no significant difference in percentage of residual red blood cells between different time points detected by flow cytometer in 3 different hypotonic salt solutions. The percentage of residual red blood cells in B+C+D+E+F+G detected time region was different among 3 NaCl dilution groups. The percentage of residual red blood cells in normal control was lower than that in hemoglobinopathy group. The percentage of residual red blood cells in hereditary spherocytosis (HS) group was obviously lower than that in hemoglobinopathy and normal control groups. The comparison of 3 different dilution concentrations found that the second concentration (0.60 ml normal saline+0.40 ml deionized water) is more suitable to screen HS by FC500 flow cytometer. CONCLUSION: The detection of red cell osmotic fragility by using flow cytometry is a simple, rapid, objective and economic way that can be an effective screening method for diagnose the HS.


Subject(s)
Erythrocytes/cytology , Flow Cytometry , Osmotic Fragility , Humans , Spherocytosis, Hereditary/physiopathology
10.
Am J Hematol ; 90(4): 339-45, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25641515

ABSTRACT

Red blood cells (RBCs) are deformable and flow through vessels narrower than their own size. Their deformability is most stringently challenged when they cross micrometer-wide slits in the spleen. In several inherited or acquired RBC disorders, blockade of small vessels by stiff RBCs can trigger organ damage, but a functional spleen is expected to clear these abnormal RBCs from the circulation before they induce such complications. We analyzed flow behavior of RBCs in a microfluidic chip that replicates the mechanical constraints imposed on RBCs as they cross the human spleen. Polymer microchannels obtained by soft lithography with a hydraulic diameter of 25 µm drove flow into mechanical filtering units where RBCs flew either slowly through 5- to 2-µm-wide slits or rapidly along 10-µm-wide channels, these parallel paths mimicking the splenic microcirculation. Stiff heated RBCs accumulated in narrow slits seven times more frequently than normal RBCs infused simultaneously. Stage-dependent retention of Plasmodium falciparum-infected RBCs was also observed in these slits. We also analyzed RBCs from patients with hereditary spherocytosis and observed retention for those having the most altered mechanical properties as determined by ektacytometry. Thus, in keeping with previous observations in vivo and ex vivo, the chip successfully discriminated poorly deformable RBCs based on their distinct mechanical properties and on the intensity of the cell alteration. Applications to the exploration of the pathogenesis of malaria, hereditary spherocytosis, sickle cell disease and other RBC disorders are envisioned.


Subject(s)
Biomimetics/methods , Erythrocytes/cytology , Hemorheology , Microfluidic Analytical Techniques/methods , Spherocytosis, Hereditary/pathology , Spleen/blood supply , Biomimetics/instrumentation , Equipment Design , Erythrocytes/parasitology , Erythrocytes/ultrastructure , Humans , Microcirculation , Microfluidic Analytical Techniques/instrumentation , Plasmodium falciparum/isolation & purification , Spherocytosis, Hereditary/physiopathology , Spleen/ultrastructure
12.
Curr Opin Hematol ; 21(3): 186-92, 2014 May.
Article in English | MEDLINE | ID: mdl-24572988

ABSTRACT

PURPOSE OF REVIEW: This review discusses the unexpected role of red blood cell (RBC) adhesiveness in the pathophysiology of two red cell diseases, hereditary spherocytosis and polycythemia vera, and two 'nonerythroid' disorders, central retinal vein occlusion and Gaucher disease. These pathologies share common clinical manifestations, that is vaso-occlusion and/or thrombotic events. RECENT FINDINGS: Recently, the direct involvement of RBC adhesion to the vascular endothelium has been demonstrated in the occurrence of vaso-occlusive events, in particular in sickle cell disease (SCD). Several erythroid adhesion molecules and their ligands have been identified that belong to different molecular classes (integrins, Ig-like molecules, lipids...) and are activated by a variety of signaling pathways. Among these, the laminin receptor, Lutheran/basal cell adhesion molecule, which is activated by phosphorylation, appears to play a central role in several pathologies. SUMMARY: RBC adhesiveness might be involved in complications such as the vaso-occlusive crisis in SCD, thrombosis in polycythemia vera, splenic sequestration in hereditary spherocytosis, occlusions in central retinal vein occlusion and bone infarcts in Gaucher disease. Characterization of this pathological process at the cellular and molecular levels should prove useful to develop new therapeutic approaches based on the blockade of RBC abnormal interactions with vascular endothelium and/or circulating blood cells.


Subject(s)
Cell Adhesion/physiology , Erythrocytes/physiology , Polycythemia Vera/physiopathology , Spherocytosis, Hereditary/physiopathology , Endothelium, Vascular/physiopathology , Humans
13.
Rev. cuba. hematol. inmunol. hemoter ; 28(4): 310-326, oct.-dic. 2012.
Article in Spanish | CUMED | ID: cum-54062

ABSTRACT

La esferocitosis hereditaria es la anemia hemolítica congénita más frecuente en la población caucásica. Tiene una amplia variabilidad clínica y desde el punto de vista hematológico se caracteriza por anemia y presencia de esferocitos en la lámina periférica. Su base fisiopatológica está determinada por el defecto de algunas de las proteínas que conforman la membrana eritrocitaria, por el efecto del bazo sobre los hematíes anómalos y otros factores. A la luz de los conocimientos actuales, la interpretación dinámica del proceso requiere adentrarse en los estadios iniciales de la hematopoyesis, pues desde etapas tan tempranas como la enucleación del eritroblasto en la formación del reticulocito, hasta posibles procesos inflamatorios tardíos, pudieran modular la expresión de la enfermedad. Se hace una revisión de las características estructurales y funcionales de la membrana eritrocitaria, así como algunos aspectos generales de las propiedades del hematíe para facilitar la comprensión de los eventos que tienen lugar a partir del compromiso molecular de las proteínas que conforman la membrana(AU)


Hereditary spherocytosis is the most common congenital hemolytic anemia among Caucasian population. It has wide clinical variety and from the haematological point of view, it is characterized by the presence of spherocytes anemia in peripheral lamina. Its pathophysiological defect is determined by some of the proteins that make up the red cell membrane due to the effect on erythrocytes of abnormal spleen, and other factors. In view of current knowledge, the dynamic interpretation of this process requires delving into the early stages of hematopoiesis, since the expression of this disease could modulate from early stages of erythroblast enucleation in reticulocyte formation until late potential inflammatory processes. A review was made on the structural and functional characteristics of the erythrocyte membrane, as well as some general aspects of the properties of the red cell to facilitate understanding of events which take place through proteins molecular involvement forming the membrane(AU)


Subject(s)
Humans , Male , Female , Spherocytosis, Hereditary/epidemiology , Spherocytosis, Hereditary/genetics , Spherocytosis, Hereditary/physiopathology , Membrane Proteins/therapeutic use
14.
Rev. cuba. hematol. inmunol. hemoter ; 28(4): 310-326, oct.-dic. 2012.
Article in Spanish | LILACS | ID: lil-663857

ABSTRACT

La esferocitosis hereditaria es la anemia hemolítica congénita más frecuente en la población caucásica. Tiene una amplia variabilidad clínica y desde el punto de vista hematológico se caracteriza por anemia y presencia de esferocitos en la lámina periférica. Su base fisiopatológica está determinada por el defecto de algunas de las proteínas que conforman la membrana eritrocitaria, por el efecto del bazo sobre los hematíes anómalos y otros factores. A la luz de los conocimientos actuales, la interpretación dinámica del proceso requiere adentrarse en los estadios iniciales de la hematopoyesis, pues desde etapas tan tempranas como la enucleación del eritroblasto en la formación del reticulocito, hasta posibles procesos inflamatorios tardíos, pudieran modular la expresión de la enfermedad. Se hace una revisión de las características estructurales y funcionales de la membrana eritrocitaria, así como algunos aspectos generales de las propiedades del hematíe para facilitar la comprensión de los eventos que tienen lugar a partir del compromiso molecular de las proteínas que conforman la membrana


Hereditary spherocytosis is the most common congenital hemolytic anemia among Caucasian population. It has wide clinical variety and from the haematological point of view, it is characterized by the presence of spherocytes anemia in peripheral lamina. Its pathophysiological defect is determined by some of the proteins that make up the red cell membrane due to the effect on erythrocytes of abnormal spleen, and other factors. In view of current knowledge, the dynamic interpretation of this process requires delving into the early stages of hematopoiesis, since the expression of this disease could modulate from early stages of erythroblast enucleation in reticulocyte formation until late potential inflammatory processes. A review was made on the structural and functional characteristics of the erythrocyte membrane, as well as some general aspects of the properties of the red cell to facilitate understanding of events which take place through proteins molecular involvement forming the membrane


Subject(s)
Humans , Male , Female , Spherocytosis, Hereditary/epidemiology , Spherocytosis, Hereditary/physiopathology , Spherocytosis, Hereditary/genetics , Membrane Proteins/therapeutic use
15.
Mo Med ; 109(3): 195-8, 2012.
Article in English | MEDLINE | ID: mdl-22860286

ABSTRACT

Transgenic mice represent a unique opportunity in biomedical research to discover the genes underlying disease and understand how manipulating the function of single genes and proteins alters physiology in a whole animal system. These advances in biomedical research may accelerate the time between when basic discoveries are made and when the research can be 'translated', that is, when the research will positively impact the lives of patients. The purpose of this article is to present some examples of promising mouse models of human diseases.


Subject(s)
Biomedical Research/methods , Disease Models, Animal , Translational Research, Biomedical , Anemia/genetics , Anemia/physiopathology , Animals , GATA1 Transcription Factor , Humans , Mice , Mice, Transgenic , Muscular Dystrophy, Duchenne/genetics , Muscular Dystrophy, Duchenne/physiopathology , Spherocytosis, Hereditary/genetics , Spherocytosis, Hereditary/physiopathology
16.
Am J Hematol ; 86(12): E73-6, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21953840

ABSTRACT

Vascular complications, including pulmonary hypertension (PH), have been reported to occur following splenectomy for various disorders,including hereditary spherocytosis (HS). We performed a prospective cross-sectional study of 36 adults with HS (78% with prior splenectomy)utilizing echocardiography to estimate tricuspid regurgitant jet velocity (TRV) as well as measurement of N-terminal pro-B-type natriuretic peptide (NT-proBNP) to screen for PH. No participant with HS hada significantly elevated TRV or NT-proBNP level, despite a median 25-year interval since splenectomy (95% confidence interval for point prevalence 0, 0.097). Although our study was limited by a small sample size, it appears that persons with HS, following splenectomy, appear unlikely to be at significantly increased risk of developing PH to the degree reported for thalassemia and sickle cell disease


Subject(s)
Hypertension, Pulmonary/epidemiology , Postoperative Complications/epidemiology , Spherocytosis, Hereditary/physiopathology , Splenectomy/adverse effects , Adult , Aged , Aged, 80 and over , Cross-Sectional Studies , Early Diagnosis , Female , Humans , Hypertension, Pulmonary/diagnostic imaging , Hypertension, Pulmonary/etiology , Male , Middle Aged , Natriuretic Peptide, Brain/blood , Postoperative Complications/diagnostic imaging , Postoperative Complications/etiology , Prevalence , Prospective Studies , Protein Precursors/blood , Risk Factors , Spherocytosis, Hereditary/blood , Spherocytosis, Hereditary/surgery , Texas/epidemiology , Tricuspid Valve/diagnostic imaging , Ultrasonography , Young Adult
17.
Blood Cells Mol Dis ; 46(2): 166-70, 2011 Feb 15.
Article in English | MEDLINE | ID: mdl-21138793

ABSTRACT

Hereditary spherocytosis (HS) is usually classified as mild, moderate or severe using conventional features, namely, hemoglobin (Hb) concentration, reticulocyte count and bilirubin levels, which do not always contribute to an adequate clinical classification. The aim of our study was to establish the importance of some laboratory routine parameters, as markers of HS clinical outcome, by studying a control group (n=26) and unsplenectomized HS patients (n=82) presenting mild, moderate or severe HS. We performed a basic hematologic study and evaluated the reticulocyte count, bilirubin, erythropoietin (EPO) and soluble transferrin receptor (sTfR) levels; the osmotic fragility (OFT) and criohemolysis tests (CHT); the ratios Hb/MCHC (mean cell hemoglobin concentration), Hb/RDW (red cell distribution width) and MCHC/RDW, were calculated. Hb changed significantly in accordance with HS severity, but not reticulocytes or bilirubin. We found that MCHC, RDW, EPO, sTfR, OFT, CHT and the calculated ratios were significantly changed in patients, and, therefore, were valuable as complementary diagnostic tools for HS. Moreover, RDW, Hb/MCHC, Hb/RDW and MCHC/RDW changed significantly with worsening of HS; thus, they are also good markers for the clinical outcome of HS. In conclusion, we propose the use of these routine parameters as useful to complement the analysis of HS severity.


Subject(s)
Biomarkers/blood , Severity of Illness Index , Spherocytosis, Hereditary/physiopathology , Bilirubin/blood , Case-Control Studies , Erythrocyte Volume , Erythrocytes , Erythropoietin/blood , Hemoglobins/analysis , Humans , Osmotic Fragility , Receptors, Transferrin/blood , Reticulocyte Count , Reticulocytes , Spherocytosis, Hereditary/blood , Spherocytosis, Hereditary/classification , Spherocytosis, Hereditary/diagnosis
18.
Ann Hematol ; 90(7): 759-68, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21181161

ABSTRACT

Hereditary spherocytosis (HS) is characterised by weakened vertical linkages between the membrane skeleton and the red blood cell's lipid bilayer, leading to the release of microparticles. All the reference tests suffer from specific limitations. The aim of this study was to develop easy to use diagnostic tool for screening of hereditary spherocytosis based on routinely acquired haematological parameters like percentage of microcytes, percentage of hypochromic cells, reticulocyte counts, and percentage of immature reticulocytes. The levels of haemoglobin, mean cell volume, mean corpuscular haemoglobin concentration, reticulocytes (Ret), immature reticulocytes fraction (IRF), hypochromic erythrocytes (Hypo-He) and microcytic erythrocytes (MicroR) were determined on EDTA samples on Sysmex instruments from a cohort of 45 confirmed SH. The HS group was then compared with haemolytical disorders, microcytic anaemia, healthy individuals and routine samples (n = 1,488). HS is characterised by a high Ret count without an equally elevated IRF. All 45 HS have Ret >80,000/µl and Ret(10(9)/L)/IRF (%) greater than 7.7 (rule 1). Trait and mild HS had a Ret/IRF ratio greater than 19. Moderate and severe HS had increased MicroR and MicroR/Hypo-He (rule 2). Combination of both rules gave predictive positive value and negative predictive value of respectively 75% and 100% (n=1,488), which is much greater than single parameters or existing rules. This simple and fast diagnostic method could be used as an excellent screening tool for HS. It is also valid for mild HS, neonates and ABO incompatibilities and overcomes the lack of sensitivity of electrophoresis in ankyrin deficiencies.


Subject(s)
Diagnostic Tests, Routine/methods , Erythrocytes, Abnormal/chemistry , Reticulocytes/chemistry , Spherocytosis, Hereditary/blood , Spherocytosis, Hereditary/diagnosis , Adolescent , Adult , Aged , Child , Child, Preschool , Erythrocyte Indices , Erythrocyte Volume , Female , Hemoglobins/analysis , Humans , Infant , Infant, Newborn , Male , Middle Aged , Spherocytosis, Hereditary/physiopathology , Young Adult
19.
Ann Hematol ; 90(5): 509-13, 2011 May.
Article in English | MEDLINE | ID: mdl-21063708

ABSTRACT

In the present study, we questioned the role of oxidative stress in hereditary spherocytosis (HS), where red blood cells (RBC) have a shortened survival due to primary deficiency in membrane proteins. Using flow cytometry techniques, we showed that RBC derived from 17 HS patients of seven families generate more reactive oxygen species, membrane lipid peroxides, and less reduced glutathione than normal RBC. Following in vitro incubation of HS-RBC from seven patients with a fermentation bioproduct of Carica papaya (fermented papaya preparation (FPP)) with known antioxidative properties, oxidative stress markers were significantly reduced. Similar results were obtained following treatment with FPP for 3 months of 10 adult HS patients, as well as decreased tendency to undergo hemolysis. The hemoglobin levels increased by >1 g/dl, mean corpuscular hemoglobin concentration decreased by >1 g/dl, and the reticulocyte count decreased by 0.93%. Concomitantly, lactic dehydrogenase decreased by 17% and indirect bilirubin by 50%. A significant decrease in malonyldialdehyde was also detected. These data indicate that oxidative stress plays an important role in the pathophysiology of HS which can be ameliorated by an antioxidant such as FPP. Additional clinical trials with FPP and other antioxidants are warranted.


Subject(s)
Antioxidants/therapeutic use , Carica/chemistry , Hemolysis/drug effects , Oxidative Stress/drug effects , Phytotherapy , Plant Preparations/therapeutic use , Spherocytosis, Hereditary/drug therapy , Adolescent , Adult , Erythrocyte Indices/drug effects , Erythrocytes/drug effects , Female , Fermentation , Glutathione/blood , Humans , Lipid Peroxides/blood , Male , Oxidation-Reduction , Reactive Oxygen Species/blood , Reticulocyte Count , Spherocytosis, Hereditary/blood , Spherocytosis, Hereditary/physiopathology , Young Adult
20.
Hematology ; 14(3): 164-7, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19490762

ABSTRACT

Hereditary spherocytosis (HS) is an inherited membranopathy characterized by phenotypic and genotypic heterogeneity. This study describes the clinico-hematological profile of 70 HS patients diagnosed at a tertiary care center in North India over a period of five years. Patients commonly presented with intermittent jaundice (82.9%), pallor (80%) and dark colored urine (11.4%). The common signs were splenomegaly (92.9%), hepatomegaly (50%), cholelithiasis or choledocholithiasis (36.8%) and hemolytic facies (10%). Family history was contributory in 28.6% patients. Blood transfusion (BT) requirement was present in 35.7% patients. Unconjugated and conjugated hyperbilirubinemia was seen in 89.1 and 10.9% patients respectively. At presentation, the hemoglobin ranged from 3-14 g/dl with a mean of 9.37 g/dl (SD2.43). Spherocytes were seen in 88.6% and incubated Osmotic fragility test (OFT) was positive in 88.2% patients. The Eosin-5-maleimide (EMA) flow cytometric test was done in 28 patients. Mean fluorescence intensity (MFI) for normal subjects was 11861.5 (SD-883.51) and for confirmed HS patients was 7949.3 (SD1304.1). Taking the MFI range of 5341.1-10 557.5 for HS, eight cases of suspected HS/undiagnosed hemolytic anemia with a negative (n=5) or equivocal (n=3) incubated OFT were diagnosed as HS. An increase in HbF level was seen in 10 cases ranging from 2.1 to 17.7% with a mean of 5.66%, three of these had associated beta thalassaemia trait. Twelve patients (17%) underwent splenectomy and 91% of them did not require any BT post-splenectomy. Among the patients treated conservatively 49% had persisting pallor and 16.3% had transfusion requirement.


Subject(s)
Spherocytosis, Hereditary/blood , Spherocytosis, Hereditary/diagnosis , Adolescent , Adult , Child , Child, Preschool , Eosine Yellowish-(YS)/analogs & derivatives , Female , Flow Cytometry , Follow-Up Studies , Hematologic Tests , Humans , India , Infant , Infant, Newborn , Male , Middle Aged , Spherocytes/pathology , Spherocytosis, Hereditary/physiopathology , Spherocytosis, Hereditary/therapy , Young Adult
SELECTION OF CITATIONS
SEARCH DETAIL
...