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International Journal of Hematology Mar 2011Although the therapeutic efficacy of β(654)-thalassaemia treatment using a combination of RNAi and antisense RNA to balance the synthesis of α- and β-globin chains...
Although the therapeutic efficacy of β(654)-thalassaemia treatment using a combination of RNAi and antisense RNA to balance the synthesis of α- and β-globin chains has been demonstrated previously, and the safety of lentiviral delivery remains unclear. Herein, we used the same β(654)-thalassaemia mouse model to develop a therapy involving direct delivery of siRNA and antisense RNA plasmids via intravenous injection to simultaneously knock down α-globin transcript levels and restore correct β-globin splicing. The amount of α-globin mRNAs in siRNA-treated MEL cells decreased significantly, and the properly spliced β-globin mRNA was restored in HeLaβ(654) cells transfected with pcDNA-antisense plasmid. Furthermore, treatment of β(654)-thalassaemic mice with siRNA and antisense RNA plasmids resulted in significant reduction of poikilocytosis and reticulocyte counts in blood samples, decreased nucleated cell populations in bone marrow, and reduced intrasinusoidal extramedullary haematopoiesis loci and iron accumulation in liver. RT-PCR analysis revealed that treatment resulted in down-regulation of α-globin mRNA synthesis by ~50% along with an increase in the presence of normally spliced β-globin transcripts, indicating that the phenotypic changes observed in β(654)-thalassaemic mice following treatment resulted from restoration of the balance of α/β-globin biosynthesis.
Topics: Animals; Bone Marrow Cells; Down-Regulation; Genetic Vectors; HeLa Cells; Hematopoiesis, Extramedullary; Humans; Mice; Mice, Mutant Strains; Plasmids; RNA Splicing; RNA, Antisense; RNA, Small Interfering; alpha-Globins; beta-Globins; beta-Thalassemia
PubMed: 21369857
DOI: 10.1007/s12185-010-0727-1 -
Blood Cells, Molecules & Diseases Mar 2011We report on a truncated α-spectrin chain, spectrin(Exeter), associated with ellipto-poikilocytosis. Analysis of erythrocyte membranes of affected individuals revealed...
We report on a truncated α-spectrin chain, spectrin(Exeter), associated with ellipto-poikilocytosis. Analysis of erythrocyte membranes of affected individuals revealed a truncated α-spectrin chain with normal amounts of spectrin dimer. In the proband and her father, one haploid set of α-spectrin cDNA lacked exons 11 and 12, leading to partial deletion of repeats α4 and α5 (83 amino acids) of the α-spectrin chain. In one allele of genomic DNA, a 3567bp deletion starting in intron 10 and ending in intron 12 of the SPTA1 gene was found. The common polymorphic SPTA1 α(LELY) allele was found in trans to the SPTA1αExeter allele in the proband. The proband had inherited the SPTA1Exeter allele from her father and the αLELY allele from her healthy, asymptomatic mother. This is the first report of an interstitial deletion in the SPTA1 gene associated with ellipto-poikilocytosis.
Topics: Alleles; Amino Acid Sequence; Base Sequence; Child; Elliptocytosis, Hereditary; Exons; Female; Humans; Infant; Introns; Male; Molecular Sequence Data; Sequence Deletion; Spectrin
PubMed: 21212007
DOI: 10.1016/j.bcmd.2010.12.006 -
International Journal of Laboratory... Apr 2011Hereditary spherocytosis (HS) and hereditary pyropoikilocytosis (HPP, severe form of hereditary elliptocytosis) are unrelated red cell disorders caused by defects in...
INTRODUCTION
Hereditary spherocytosis (HS) and hereditary pyropoikilocytosis (HPP, severe form of hereditary elliptocytosis) are unrelated red cell disorders caused by defects in distinct regions of the red cell cytoskeleton. The high predictive value of the eosin-5-maleimide (EMA)-binding test for the diagnosis of HS is because of its interaction with transmembrane proteins band 3, Rh protein, Rh glycoprotein and CD47, which are reduced on HS red cells. Our study was undertaken to determine why EMA-labelled HPP red cells were previously found to give much lower fluorescence readings than HS.
METHODS
Flow cytometry was used to determine the relative amounts of monoclonal antibodies bound to red cells from normal adults, HS and HPP groups. Confocal microscopy was used to visualise the overall staining pattern of the red cells with selected antibodies.
RESULTS
In flow cytometry, HPP red cells gave lower antibody binding to the four EMA-reactive membrane proteins than HS red cells and bound less antibody to glycophorins A and C, and CD59. Confocal images of Rh protein and band 3 immunostaining revealed a greater number of HPP red cells having partial or no fluorescence than in HS and normal controls.
CONCLUSION
Lesser amounts of EMA-reactive membrane proteins were detected in HPP than HS red cells, thus confirming their lower fluorescence readings in the EMA-binding test. The concomitant reduction in glycophorins A and C, and CD59 in HPP could have caused cellular contraction, resulting in poikilocytosis.
Topics: Adult; Elliptocytosis, Hereditary; Eosine Yellowish-(YS); Erythrocyte Membrane; Erythrocytes; Humans; Membrane Proteins; Protein Binding; Spherocytosis, Hereditary
PubMed: 21054813
DOI: 10.1111/j.1751-553X.2010.01270.x -
Indian Journal of Physiology and... 2010Anaemia, a frequently encountered clinical entity, is commonly overlooked and considered secondary to underlying illness but it alters the length and quality of life. A...
Anaemia, a frequently encountered clinical entity, is commonly overlooked and considered secondary to underlying illness but it alters the length and quality of life. A number of epidemiologic studies have shown a correlation between white blood cell (WBC) counts and ischaemic events. Elevated leucocytes is considered a risk factor for cardiovascular and ischaemic diseases. Since anaemia is associated with hypoxia and ischaemia, it may be possible that it may affect white blood cells (WBCs). The present study is planned to estimate total and differential (TLC and DLC) leucocyte counts, neutrophil to lymphocyte ratio (N/L ratio) and platelet counts in anaemia. Reticulocyte counts (P < 0.001) and erythrocyte sedimentation rate (P < 0.001) were higher in anaemic vs controls. TLC was insignificantly higher. On differential count neutrophils (%) (P < 0.01) and basophils (%) (P < 0.001) were higher, eosinophils and monocytes were less (P < 0.001), without any alterations in lymphocyte counts in anaemic subjects. Poikilocytosis and anisocytosis of RBC is also present in patients of anaemia. Increased N/L ratio (P < 0.001) and decreased platelet counts (P < 0.05) was observed in cases of anaemia compared to controls. Leucocytes are altered in cases of anaemia.
Topics: Adult; Anemia; Female; Humans; Leukocyte Count; Male; Young Adult
PubMed: 21046926
DOI: No ID Found -
Acta Chirurgica Iugoslavica 2010In healthy population, uric acid comprises the major component of 10-20% of renal stones. Extreme hiperuricaemia is seen in cancer patients with tumour lysis syndrome...
In healthy population, uric acid comprises the major component of 10-20% of renal stones. Extreme hiperuricaemia is seen in cancer patients with tumour lysis syndrome (TLS) which is classically associated with haematological malignancies with rapid tumour growth rates such as acute lymphoid leukaemia and high grade lymphomas. Primary melofibrosis (Agnogenic myeloid metaplasia-AMM) is a chronic myeloproliferative disease characterized by splenomegaly, a leukoerythroblastic blood picture, teardrop poikilocytosis and varying degrees of marrow fibrosis. Due to the increased extramedullary haematopoiesis, hiperuricemia may occur. However, TLS in patients with AMM is, according to the available literature, described just in one patient. In this paper we present a case of a 47-year-old male patient who was admitted to the hospital with symptoms of fatigue and small amount of urine, and clinical signs of plethora and enlarged spleen. The laboratory findings showed leuko-and erythrocytosis, increased levels of urea-BUN (32 mmol/l) and creatinine (766 mmol/l) as well as uric acid (920 mmol/l). The immediate abdominal ultrasound confirmed extreme splenomegaly, but also showed bilateral hydronephrosis of grade II-III with two stones in proximal part of right ureter and one in proximal part of left ureter as well as empty bladder. Stones were not seen on plain film. Since the patient was in complete anuria, with further rapid elevation of BUN and creatinine levels, bilateral ureteral stents were applicated together with extensive hydration, urine alkalization and administration of allopurinol which resulted in the complete recovery of kidney function. The bone marrow biopsy was also performed and histopathological diagnosis was: Hypercellulary phase of AMM.
Topics: Humans; Hyperuricemia; Male; Middle Aged; Primary Myelofibrosis; Ureteral Obstruction
PubMed: 20949707
DOI: 10.2298/aci1002079c -
Tuberkulez I Bolezni Legkikh 2009One hundred and twenty-three female patients aged 16 to 49 years with pulmonary tuberculosis were examined and followed up. Of them, 83 (67.5%) patients in whom... (Comparative Study)
Comparative Study
One hundred and twenty-three female patients aged 16 to 49 years with pulmonary tuberculosis were examined and followed up. Of them, 83 (67.5%) patients in whom tuberculosis had occurred and run in the presence of iron-deficiency anemia (IDA) made up a study group and 40 (32.5%) patients in whom tuberculosis had occurred without IDA formed a control group. In the women from the study group, the clinical manifestations of the disease had its own peculiarities: a less pronounced temperature reaction, meager expectoration, severe signs of hypoxia (dyspnea), and chest pain and 97% were found to have abnormally shaped red blood cells (poikilocytosis). Inclusion of Tardiferon into complex treatment aids in enhancing the efficiency of treatment for tuberculosis in the presence of IDA, promoting to a greater extent the regression of the clinical manifestations of the disease and the achievement of abacillation than cicatrization.
Topics: Adolescent; Adult; Anemia, Iron-Deficiency; Antitubercular Agents; Delayed-Action Preparations; Drug Combinations; Drug Therapy, Combination; Female; Ferrous Compounds; Fertility; Follow-Up Studies; Humans; Iron; Middle Aged; Mucins; Treatment Outcome; Tuberculosis, Pulmonary; Young Adult
PubMed: 20099387
DOI: No ID Found -
International Journal of Dermatology Aug 2009A 24-year-old man was referred to our clinic in August 2003 with complaints of weakness, dizziness, and bilateral knee pain of 3 years' duration. Bilateral digital...
A 24-year-old man was referred to our clinic in August 2003 with complaints of weakness, dizziness, and bilateral knee pain of 3 years' duration. Bilateral digital clubbing had been found on routine physical examination during his military service 4 years earlier. There were no cardiorespiratory or abdominal symptoms. There was no compromise in the activities of everyday life. The patient was not a chronic smoker. In the family history of the patient, his brother had been diagnosed with pachydermoperiostosis in another center 2 years earlier, but did not return to the hospital for a follow-up investigation of myelofibrosis. On physical examination, the patient showed marked drumstick clubbing of the hands (Fig. 1), and a pale general appearance. The causes of digital clubbing are shown in Table 1 (Fawcett RS, Linford S, Stulberg DL. Nail abnormalities: clues to systemic disease. Am Fam Physician 2004; 69: 1417-1424). Deep nasolabial folds were seen on the face. Skin hypertrophy, cutis verticis gyrata, and seborrhea on the face were also observed. The patient also complained of hyperhidrosis. Examination of the cardiovascular system was normal. There was bilateral swelling of the ankle and knee (Fig. 2). Hepatosplenomegaly was found on abdominal examination. Investigations showed hypochromic microcytic anemia [hemoglobin, 8.58 g/dL (normal, 12.2-18.1 g/dL); hematocrit, 28.1% (normal, 37.7-53.7%); white blood cell count, 3430/mm(3) (normal, 4600-10,200/mm(3)); neutrophils, 2470/mm(3) (normal, 2000-6900/mm(3)); lymphocytes, 820/mm(3) (normal, 600-3400/mm(3)); platelets, 162,000/mm(3) (normal, 142,000-424,000 mm(3)); mean corpuscular volume, 73.7 fL (normal, 80-97 fL)]. Anisocytosis, poikilocytosis, microcytosis, and hypochromia were observed on peripheral blood examination, and the erythrocyte sedimentation rate was 37 mm/h. The serum C-reactive protein level was 50.1 mg/L (normal, 0-5 mg/L). Biochemical parameters, including serum calcium, phosphate, alkaline phosphates and liver function tests, were found to be within the normal range. The causes of secondary hypertrophic osteoarthropathy associated with pulmonary, rheumatologic, endocrine, cardiac, and gastroenterologic disorders were excluded. Growth hormone level and thyroid function tests were normal. Antinuclear antibody, TORCH [Toxoplasma immunoglobulin M (IgM), rubella IgM, cytomegalovirus IgM, herpes simplex IgM] panel, and markers of hepatitis were negative. Serum Igs and rheumatoid factor were found to be within the normal range. There was subperiosteal new bone formation on bilateral knee X-ray (Fig. 3). Radiography of the chest, pulmonary function tests, arterial blood gas, and echocardiography were normal. Abdominal ultrasonography revealed hepatosplenomegaly. Amyloid deposition was not determined in rectal biopsy. Reticulin-type myelofibrosis was found on bone marrow biopsy (Figs 4 and 5). In the cytogenetic study, monosomy 22 was detected in four of 20 metaphase plates.
Topics: Biopsy; Chromosomes, Human, Pair 22; Humans; Knee Joint; Male; Monosomy; Osteoarthropathy, Primary Hypertrophic; Osteoarthropathy, Secondary Hypertrophic; Primary Myelofibrosis; Young Adult
PubMed: 19659869
DOI: 10.1111/j.1365-4632.2008.03774.x -
Blood Cells, Molecules & Diseases 2009One of the most commonly inherited anemias in man is Hereditary Spherocytosis (HS) with an incidence of 1 in 2000 for persons of Northern European descent. Mouse models...
One of the most commonly inherited anemias in man is Hereditary Spherocytosis (HS) with an incidence of 1 in 2000 for persons of Northern European descent. Mouse models of HS include spontaneous inherited hemolytic anemias and those generated by gene targeting. The Neonatal anemia (Nan) mouse is a novel model of HS generated by N-ethyl-N-nitrosurea mutagenesis and suffers from a severe neonatal anemia. Adult Nan mice have a lifelong hemolytic anemia with decreased red blood cell numbers, hematocrit, and hemoglobin, but elevated zinc protoporphyrin levels. Blood smears taken from Nan mice show a hypochromic anemia characterized by poikilocytosis, anisocytosis and polychromasia. The Nan phenotype can be transferred by bone marrow transplantation indicating that the defect is intrinsic to bone marrow. The hemolytic anemia in adult Nan mice can be identified by osmotic fragility testing. Examination of the erythrocyte membrane skeleton proteins (EMS) reveals a global deficiency of these proteins with protein 4.1a being completely absent. The Nan locus maps to mouse Chromosome 8 and does not co-localize with any known EMS genes. The identification of the Nan gene will likely uncover a novel protein that contributes to the stability of the EMS and may identify a new mutation for HS.
Topics: Anemia, Hemolytic; Animals; Bone Marrow Transplantation; Chromosome Mapping; Chromosomes, Mammalian; Disease Models, Animal; Erythrocyte Membrane; Erythrocytes; Male; Membrane Proteins; Mice; Mice, Inbred C57BL; Microscopy, Electron, Scanning
PubMed: 19409822
DOI: 10.1016/j.bcmd.2009.03.009 -
Bulletin of Environmental Contamination... Jul 2009Acute (4 day) and short-term (7 day) toxicity studies (at 1/5th and 1/10th of LC(50)) of textile dye wastewaters and their selected ingredients (azo dye methyl red and...
Acute (4 day) and short-term (7 day) toxicity studies (at 1/5th and 1/10th of LC(50)) of textile dye wastewaters and their selected ingredients (azo dye methyl red and heavy metals Cd, Cu, Ni and Zn) were made on a freshwater fish Gambusia affinis under laboratory conditions. LC(50) value was found to be the lowest in four cases, and the EC(50) value for reduction in erythrocyte counts in the remaining four tests. Thus, the reduction in erythrocyte counts to the 50% level was similar in sensitivity to fish mortality. The short-term toxicity studies revealed significant disorders in erythrocyte morphology (poikilocytosis) and its counts to be the better indices for toxicity monitoring in the absence of fish mortality.
Topics: Animals; Biological Assay; Coloring Agents; Cyprinodontiformes; Environmental Monitoring; Erythrocyte Count; Erythrocytes; Fresh Water; Industrial Waste; Metals, Heavy; Textile Industry; Toxicity Tests; Water Pollutants, Chemical
PubMed: 19322506
DOI: 10.1007/s00128-009-9711-y -
Journal of Environmental Biology Nov 2008The influence of increased zinc concentrations (0.1, 0.5, 1.0, 1.5 and 2.0 mg(-1) ZnSO4 x 7H2O) on the total number and the morphology of the erythrocytes, as well as...
The influence of increased zinc concentrations (0.1, 0.5, 1.0, 1.5 and 2.0 mg(-1) ZnSO4 x 7H2O) on the total number and the morphology of the erythrocytes, as well as the processes related to their formation and destruction in the spleen of Carassius gibelio were investigated ex situ. It was found that zinc concentrations caused pathological alterations in the erythrocytes that were not identical in the different concentrations-poikilocytosis; ruptures in cell membranes in the concentrations of 0.5mg(-1) and 1.5 mg(-1); cells with double nuclei (symplasts); in the concentration of 1.0 mg(-1); in the highest concentrations (1.5 mg(-1) and 2.0 mg(-1)) presence of erythrocytes at initial stage of atypical mitotic division. Against the background of those various alterations, the total number of the erythrocytes in the peripheral blood increased simultaneously with the increase of zinc concentrations (p < 0.001). Morphological alterations in the spleen were also observed, indicating a compensational tendency against the toxic influence of zinc upon the fish erythrocytes-hyperplasia of the red pulp and lack of hemosiderin. These results show that the alterations in the total number and the morphology of the erythrocytes are connected with the relevant compensatory histopathological alterations in the spleen. The use of the ascertained alteration could be valuable in monitoring zinc-polluted waters.
Topics: Animals; Carps; Erythrocytes; Spleen; Zinc
PubMed: 19297988
DOI: No ID Found