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Haematologica Mar 2008beta-thalassemia is one of the most common genetic diseases in the world and requires extensive therapy. Lentiviral-mediated gene therapy has been successfully exploited...
BACKGROUND
beta-thalassemia is one of the most common genetic diseases in the world and requires extensive therapy. Lentiviral-mediated gene therapy has been successfully exploited in the treatment of beta-thalassemia and showed promise in clinical application. Using a human beta-globin transgenic mouse line in a beta-thalassemia diseased model generated with a lentiviral-mediated approach, we investigate the stable therapeutic effect on a common thalassemia syndrome.
DESIGN AND METHODS
Human beta-globin gene lentiviral vector was constr ucted, followed by subzonal microinjection into single-cell embryos of beta(IVS-2-654)-thalassemia mice to generate a transgenic line. Human beta-globin gene expression was examined with RT-PCR, Western-blotting and ELISA. The hematologic parameters and tissue pathology were investigated over time in founder mice and their off-spring.
RESULTS
Transgenic mice with stable expression of the lentivirus carrying human beta-globin gene were obtained. A marked improvement in red blood cell indices and a dramatic reduction in red blood cell anisocytosis, poikilocytosis and target cells were observed. Nucleated cell proportion was greatly decreased in bone marrow, and splenomegaly with extramedullary hematopoiesis was ameliorated. Iron deposition in liver was also reduced. There was a two-fold increase in the survival rate of the beta(IVS-2-654) mice carrying human beta-globin transgene. Significantly, the germline integration of the lentiviral construct was obtained and stable hematologic phenotype correction was observed over the next two generations of the transgenic mice.
CONCLUSIONS
The generation of human beta-globin transgenic mice in a beta(IVS-2-654)-thalassemia mouse mediated with lentiviral vectors provides a useful model and offers an attractive means to investigate the transgenic stable therapeutic effect in beta-thalassemia.
Topics: Animals; Bone Marrow; Disease Models, Animal; Female; Genetic Therapy; Genetic Vectors; Germ-Line Mutation; Globins; Humans; Lentivirus; Liver; Male; Mice; Mice, Mutant Strains; Mice, Transgenic; Microinjections; Mutagenesis, Insertional; Recombinant Fusion Proteins; Spleen; Virus Integration; beta-Thalassemia
PubMed: 18268280
DOI: 10.3324/haematol.12010 -
American Journal of Hematology Jan 2003Myelofibrosis is characterized by reticulin fibrosis of the bone marrow with resulting features of myelophthisis. Besides hematopoietic malignancies and other neoplasms...
Myelofibrosis is characterized by reticulin fibrosis of the bone marrow with resulting features of myelophthisis. Besides hematopoietic malignancies and other neoplasms involving the bone marrow, myelofibrosis has been described in association with autoimmune disorders, especially systemic lupus erythematosus. We describe the clinicopathologic features of a primary form of autoimmune myelofibrosis (AIMF) in patients who do not have systemic lupus erythematosus or another well-defined autoimmune syndrome. Absence of marked splenomegaly, peripheral blood cytopenias with mild teardrop poikilocytosis and leukoerythroblastosis, bone marrow lymphoid aggregates, and presence of autoantibodies are some of the salient features of primary AIMF. AIMF should especially be differentiated from chronic idiopathic myelofibrosis, a neoplastic myeloproliferative disease. Primary AIMF appears to have an excellent prognosis, with all patients reported in this series responding to a short course of corticosteroid therapy.
Topics: Adrenal Cortex Hormones; Adult; Aged; Autoantibodies; Autoimmune Diseases; Bone Marrow; Comorbidity; Diagnosis, Differential; Female; Fever; Fibrosis; Humans; Immunosuppressive Agents; Male; Megakaryocytes; Middle Aged; Primary Myelofibrosis; Spleen; Sweating
PubMed: 12508261
DOI: 10.1002/ajh.10258 -
The Journal of Clinical Investigation Aug 1981Erythrocytes from three patients with severe hemolytic anemia, marked erythrocyte fragmentation, and elliptocytic poikilocytosis, were studied in terms of both their...
Erythrocytes from three patients with severe hemolytic anemia, marked erythrocyte fragmentation, and elliptocytic poikilocytosis, were studied in terms of both their membrane protein composition and their mechanical characteristics. Erythrocytes from the patients' parents and one minimally affected and one normal sibling were also studied. Morphologic observations implied that the severely affected patients suffered from homozygous hereditary elliptocytosis because erythrocytes of both parents and the one minimally affected sibling showed moderate elliptocytosis on smear, whereas those of an unaffected sibling had normal morphology. The parallel findings of markedly reduced levels of band 4.1 in the erythrocyte membrane proteins of the patients and an intermediate reduction in the cells of the parents and the putative heterozygous sibling, suggest that the elliptocytic shape of the cells was related to the reduced levels of band 4.1. Additional studies showed marked abnormalities in cellular deformability and membrane fragility in the erythrocytes from the homozygous patients. Importantly, these changes were also closely proportional to the reduced levels of band 4.1, suggesting a central role for this protein in the maintenance of normal membrane stability and normal cell shape. It seems likely that this role for band 4.1 is intimately related to its known biochemical connection to the "membrane skeleton" through its linkage with spectrin and actin.
Topics: Actins; Blood Proteins; Cytoskeletal Proteins; Cytoskeleton; Elliptocytosis, Hereditary; Erythrocyte Membrane; Erythrocytes; Humans; Membrane Proteins; Neuropeptides; Pedigree; Protein Binding; Spectrin
PubMed: 6894932
DOI: 10.1172/jci110275 -
Korean Journal of Pediatrics Nov 2016Herein, we report a rare case of hemolytic anemia with reticulocytopenia following intravenous immunoglobulin therapy in a young infant treated for Kawasaki disease. A...
Herein, we report a rare case of hemolytic anemia with reticulocytopenia following intravenous immunoglobulin therapy in a young infant treated for Kawasaki disease. A 2-month-old boy presented with fever lasting 3 days, conjunctival injection, strawberry tongue, erythematous edema of the hands, and macular rash, symptoms and signs suggestive of incomplete Kawasaki disease. His fever resolved 8 days after treatment with aspirin and high dose infusion of intravenous immunoglobulin. The hemoglobin and hematocrit decreased from 9.7 g/dL and 27.1% to 7.4 g/dL and 21.3%, respectively. The patient had normocytic hypochromic anemia with anisocytosis, poikilocytosis, immature neutrophils, and nucleated red blood cells. The direct antiglobulin test result was positive, and the reticulocyte count was 1.39%. The patient had an uneventful recovery. However, reticulocytopenia persisted 1 month after discharge.
PubMed: 28018448
DOI: 10.3345/kjp.2016.59.11.S60 -
Journal of Veterinary Internal Medicine 1992The hematologic toxicity of doxorubicin, 30 mg/m2 body surface area (BSA) every 21 days to a cumulative dose of 300 mg/m2, was evaluated in six cats. Complete blood and...
The hematologic toxicity of doxorubicin, 30 mg/m2 body surface area (BSA) every 21 days to a cumulative dose of 300 mg/m2, was evaluated in six cats. Complete blood and platelet counts were performed daily during the first treatment cycle. They were monitored before treatment for all remaining cycles, and at the average neutrophil nadir (day 8) starting with cycle 4. Significant poikilocytosis developed after the first treatment and remained throughout the study, although anemia did not occur. No other red blood cell abnormalities were seen. Platelet counts remained within the reference range throughout the first treatment cycle, but mild thrombocytopenia (88,000-288,000/uL) was found in 11.3% of subsequent complete blood counts (CBCs). Thrombocytosis was seen in 30.9% of CBCs. Neutropenia did not occur during the first treatment cycle although neutrophil counts did decrease, with the nadir occurring between days 8 and 11. All neutrophil counts returned to pretreatment values by day 14. Neutropenia was documented after 14 of 46 (30.4%) doxorubicin treatments, and was associated with fever in 5 cats (10.9%). All fevers responded to oral antibiotic therapy. Neutropenia that lasted more than 14 days developed in two cats, necessitating dosage reduction to 25 mg/m2 BSA. At the dose used in this study, doxorubicin administration was associated with acceptable hematologic toxicosis in most cats.
Topics: Animals; Blood Platelets; Cat Diseases; Cats; Doxorubicin; Erythrocytes; Female; Leukocyte Count; Male; Neutropenia; Neutrophils; Platelet Count; Thrombocytopenia
PubMed: 1432901
DOI: 10.1111/j.1939-1676.1992.tb00352.x -
Blood Feb 1976A child with congenital hemolytic anemia, extreme microcytosis and bizarre red cell morphology has been studied. Splenectomy at the age of 21 mo greatly improved the...
A child with congenital hemolytic anemia, extreme microcytosis and bizarre red cell morphology has been studied. Splenectomy at the age of 21 mo greatly improved the hemolytic anemia, although red cell morphology was unchanged. Aniso- and poikilocytosis were marked on a stained smear, and there were many small hyperchromatic cells of irregular shape. The MCV of 25 cu mu was very low and the MCHC was normal. Osmotic fragility of fresh blood was increased, and postsplenectomy blood showed a fraction of extremely fragile cells. Concentration and fluxes of Na+ and K+ were normal, except K+ efflux, which was stimulated by external Ca2+. Inward Ca2+ movement into the patient's red cells was elevated three- to fourfold above red cells of the same mean age. Red cell Ca2+ concentration was raised 2.5 times normal and most of the Ca2+ was localized in the stroma. Red cell lipid, sialic acid, and ouabain-binding sites, all per milliliter of cells, were increased by 16%-23%, and, since these substances estimate the amount of membrane, it was likely that Ca2+ content per unit of membrane area was at least twice normal. Deformability of the cells, as judged by their filterability was markedly impaired. It was concluded that the red cell membrane was defective, and an increased membrane Ca2+ content was associated with reduced deformability, hemolysis, and distorted red cell morphology in this syndrome.
Topics: Anemia, Hemolytic, Congenital; Calcium; Cations, Monovalent; Cell Membrane; Cell Membrane Permeability; Child, Preschool; Erythrocytes, Abnormal; Filtration; Humans; Lipids; Male; Osmotic Fragility; Potassium; Sialic Acids; Sodium; Time Factors
PubMed: 1244919
DOI: No ID Found -
Veterinary Research Forum : An... 2014An 8-day-old newborn female twin Holstein calves with a history of weakness, anorexia, emaciation and convulsion were presented to Tabriz University Veterinary Teaching...
An 8-day-old newborn female twin Holstein calves with a history of weakness, anorexia, emaciation and convulsion were presented to Tabriz University Veterinary Teaching Hospital. On admission, the calves were febrile and recumbent. Physical examination revealed many ticks from the external body surface of the animals, right and left prescapular lymphadenopathy, severe opisthotonos, nystagmus, pedaling, blindness, hyperpnea and hypersthenia. Buccal and vaginal mucous were pale and no other physical abnormalities were diagnosed. Fecal flotation, complete blood count, bone marrow aspiration, cerebrospinal fluid (CSF) analysis, necropsy and histopathological examination were performed. Fecal flotation showed no helminth eggs or coccidial oocysts. On blood smears obtained from the earlap, >70 percent of erythrocytes were infected with piroplasms organisms and schizonts were obvious in smears of lymphocytes lymph node. Blood count revealed a lymphopenia, poikilocytosis, anisocytosis and non-regenerative anemia (packed cell volume; mean, 16%). Histopathological examination revealed Arthus reaction through the walls of cerebral blood vessels, which resulted in local necrosis of the brain. Analysis of CSF showed no abnormality in appearance or biochemical and cell counts. Although the calves were treated with a single intramuscular injection of buparvaquone and oxytetracycline once daily they did not respond to the treatment and died. In conclusion, the present cases showed a rare cerebral form of theileriosis by vertical transmission that confirmed by the presence of piroplasms on blood films and multinuclear schizonts on lymph node aspiration smears, gross and histopathological examinations and unsuccessful treatment in a newborn twin Holstein calves.
PubMed: 25568725
DOI: No ID Found -
Blood Jun 1986Hemolytic anemia with red cell fragmentation, poikilocytosis, and elliptocytosis was discovered in a 6-week-old black infant. Both parents and a brother of the...
Hemolytic anemia with red cell fragmentation, poikilocytosis, and elliptocytosis was discovered in a 6-week-old black infant. Both parents and a brother of the propositus had compensated mild Hereditary Elliptocytosis (HE). Elliptocytosis was prominent in the proband's father with the presence of numerous rod-shaped cells whereas, in the proband's mother, elliptocytosis was less marked and cells were less elongated than in the father. The proband's red cells fragmented at 45 degrees C instead of 49 degrees C for control cells. Both the parents' and brother's red cells fragmented at 47 degrees C. The deformability of the proband's red cells was markedly reduced when measured with the ektacytometer; the red cells of both the proband's parent and brother exhibited an intermediate decrease in red cell deformability. Spectrin self-association was defective in the propositus as well as in his parents and brother. Limited tryptic digestion of the proband's spectrin, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), revealed a complete absence of the normal 80,000 dalton alpha I domain and the presence of an abnormal 65,000 dalton peptide. Two-dimensional isoelectric focusing/SDS-PAGE of limited tryptic digests of spectrin from both the proband's parents and brother revealed a decrease in the normal 80,000 alpha I domain and the presence of the 65,000 peptide variant. On the basis of biochemic studies performed on the patients' spectrin, we concluded that the proband had homozygous HE, having inherited the structural defect of spectrin present in a heterozygous state in each of his parents. On a clinical and morphologic level, homozygous HE imitates two other forms of congenital hemolytic anemia associated with a spectrin self-association defect: HE with pycnocytosis in infancy and Hereditary Pyropoikilocytosis. This report emphasizes the importance of confronting clinical and rheological as well as biochemical investigations in studying and discussing different entities.
Topics: Electrophoresis, Polyacrylamide Gel; Elliptocytosis, Hereditary; Erythrocyte Deformability; Female; Homozygote; Hot Temperature; Humans; Infant; Isoelectric Focusing; Male
PubMed: 3708157
DOI: No ID Found -
Clinico-haematological alterations and therapeutic management of tick borne fever in cross bred cow.Journal of Parasitic Diseases :... Dec 2018A 5 years old crossbred cow was brought to the Veterinary Clinical Complex of Lala Lajpat Rai University of Veterinary and Animal Sciences Hisar with history of...
A 5 years old crossbred cow was brought to the Veterinary Clinical Complex of Lala Lajpat Rai University of Veterinary and Animal Sciences Hisar with history of progressive weakness, pale mucous membrane, anorexia, high fever (105 °F), tachycardia, laboured breathing and coffee coloured urine. Analysis of haematological parameters revealed severe anaemia, leucocytopenia, marked poikilocytosis of erythrocytes. Blood smear examination showed presence of signet ring shaped organisms, piroplasms and condensed dot forms of in the stained erythrocytes. Further animal was treated with buparvaquone @ 2.5 mg/kg b.wt deep I/M in neck region and long acting oxytetracycline at 25 mg/kg b. W. slow I/V daily in normal saline solution for 5 days. Berenil (Diminazene aceturate 5%) injection was also administered @ 1 ml/20 kg b.wt. I/M along with supportive therapy. Clinical signs started to subside 3 days post treatment. Complete recovery was achieved by 4 weeks post treatment however animal succumbed to death due to immunosuppression.
PubMed: 30538366
DOI: 10.1007/s12639-018-1034-7 -
Journal of the National Medical... Nov 1985Sarcoidosis is a recently identified cause of functional asplenia that can be diagnosed by radionuclide imaging. A 31-year-old woman with a five-year history of...
Sarcoidosis is a recently identified cause of functional asplenia that can be diagnosed by radionuclide imaging. A 31-year-old woman with a five-year history of histologically compatible sarcoidosis was found to have nonvisualization of the spleen on technetium 99m sulfur colloid (radiopharmaceutical) liver-spleen scan. This scintigraphic finding was accompanied by poikilocytosis and Howell-Jolly bodies in the peripheral blood smear. A subsequent gallium 67 citrate scan reflected an abnormal increase in concentration of activity in the spleen, suggesting an active inflammatory process.Based upon this constellation of findings, it was concluded that acquired functional asplenia is the result of reticuloendothelial cell replacement via infiltration of the spleen by epithelioid cell granulomas of active sarcoidosis. This case also illustrates the reversibility of functional asplenia of sarcoidosis following adrenocorticosteroid therapy. Functional asplenia in sarcoidosis is now found to have a recognizable radionuclide imaging pattern.
Topics: Adult; Female; Humans; Sarcoidosis; Spleen; Splenic Diseases; Ultrasonography
PubMed: 3908697
DOI: No ID Found