bluuebunny/crossref_metadata_2025 huggingface dataset:
Polymerase chain reaction amplification applied to the determination of β‐like globin gene cluster haplotypes
Authors: Sutton Millicent , Bouhassira Eric E. , Nagel Ronald L.
Published: February 2007

We report here on the application of the polymerase chain reaction (PCR) technique for the determination of the β‐like gene cluster haplotypes. Seven fragments containing each one of the following polymorphic sites—Xmnl 5′ to the Gγ gene, Hindlll in the IVSII of Gγ and A γ gene, Hincll 3′ and inside the γ gene, Hinfl 5′ of the β gene—and Hpal 3′ of the β gene—are amplified using the PCR technique. Each amplified fragment is subsequently digested with the appropriate enzyme, analyzed by electrophoresis on agarose gel containing ethidium bromide, and visualized under ultraviolet light. This technique has the advantages of rapidity, safety, and cost‐effectiveness.
Antinuclear antibodies in patients with chronic idiopathic autoimmune thrombocytopenia followed 2–30 years
Authors: Panzer Simon , Penner Edward , Graninger Wolfgang , Schulz Elisabeth , Smolen Josef S.
Published: February 2007

We determined whether serological investigations can assist to distinguish between chronic idiopathic autoimmune thrombocytopenia (cAITP) and immune‐mediated throm‐bocytopenia in patients at risk to develop systemic lupus erythematosus (SLE); 82 patients were seen in this institution for the evaluation of immune thrombocytopenia. In nine of these patients a diagnosis of SLE was made at presentation or shortly thereafter. In the remaining 73 patients with an established diagnosis of cAITP we obtained serum samples at a median disease duration of 7 years; 45 patients were investigated again 2 years later. cAITP patients suffered from their disease for up to 30 years (median 7 years, range 2–30 years) without developing clinical features of SLE. Elevated antinuclear antibodies were found in all patients with a diagnosis of SLE but also in seven patients with established cAITP. All patients who had thrombocytopenia in association with SLE had additional antibodies to anti‐nDNA and/or antibodies to soluble cellular antigens. Neither in the initial, nor in the follow‐up investigation, such antibodies were discernible in patients with cAITP. We conclude that patients with cAITP do not develop SLE and do not form antibodies to defined cellular antigens, even if suffering from the disease for many years. The ANA titer does not help to differentiate cAITP from thrombocytopenia of SLE. A differentiation, however, between the two diseases is possible by the combined screening for antinuclear antibodies, anti‐nDNA, and antibodies to defined soluble cellular antigens.
Resolution of pure red cell aplasia and lymphoma: Response to intravenous gammaglobulin and combination chemotherapy
Authors: Vukelja Svetislava J. , Krishnan Jayashree , Link Christina M. , Salvado August J. , Knight Robert D.
Published: February 2007

Pure red cell aplasia (PRCA) rarely occurs in nonthymic lymphoproliferative disorders. This report describes the clinical course and therapy of a patient with a history of Sjogren syndrome who developed well‐differentiated lymphocytic lymphoma associated with PRCA and severe hemolytic anemia. Life‐threatening hemolytic anemia combined with the presence of multiple antibodies and lack of erythroid precursors was treated successfully with a single dose of intravenous gammaglobulin. A sustained, complete remission and normalization of the bone marrow was achieved following six courses of an aggressive chemotherapy regimen. Thus, occasionally low‐grade lymphomas can produce life‐threatening complications, requiring a more aggressive therapeutic intervention than those routinely applied.
Chronic myelogenous leukemia and sweet syndrome
Authors: Cohen Philip R. , Kurzrock Razelle
Published: February 2007

We report a 64‐year‐old woman with chronic myelogenous leukemia of 3 years duration who developed Sweet syndrome. Improvement in her blood counts after hydroxyurea was not associated with a decrease in size of the skin lesions. However, the cutaneous lesions of Sweet syndrome quickly resolved with systemic prednisone. Sweet syndrome has only been documented in the literature for five other chronic myelogenous leukemia patients. The characteristics, treatment, and differential diagnosis of this disorder in chronic myelogenous leukemia patients are reviewed.
Simultaneous lupus anticoagulant and anticardiolipin assays and clinical detection of antiphospholipids
Authors: Rosove Michael H. , Brewer Petronella M. C. , Runge Alice , Hirji Karim
Published: February 2007

Lupus anticoagulants and/or anticardiolipin antibodies were detected in 100 patients with autoimmune disorders, thrombosis, or pregnancy loss. Significant agreement between tests for these two antipnospholipid activities was lacking. Performing both assays is thus important in maximizing the likelihood of detecting antiphospnolipids that may have clinical relevance.
Selected indices of micronutrient status in adult patients with sickle cell anemia (SCA)
Authors: Tangney C. C. , Phillips G. , Bell R. A. , Fernandes P. , Hopkins R. , Wu S.‐M.
Published: February 2007

In 24 adults with hemoglobin SS followed at the Duke University Comprehensive Sickle Cell Center, we have studied the following nutritional parameters: reduced ascorbic acid; dehydroascorbic acid; alpha and beta carotenes; cryptoxanthin; and alpha and gamma tocopherols in whole blood, washed red blood cells, plasma, or serum. In the same population we also examined reduced glutathione (GSH) and oxidized gluthathione (GSSG). Fifteen of these 24 patients also were interviewed for usual dietary intakes using a 28‐day dietary history. Data obtained from patients with hemoglobin SS, sickle cell anemia (SCA) were compared to those found for seven healthy normal black adults of similar age. Plasma alpha tocopherol levels were significantly lower in SCA individuals than those of the controls (P< 0.004). Alpha and gamma tocopherol levels in sickle RBCs were significantly higher than those from RBC suspensions of control subjects (P< 0.007, andP< 0.001, respectively). All serum values for carotenoids examined, specifically, beta carotene, alpha carotene, and cryptoxanthin were also markedly depressed when compared to those of healthy controls (P< 0.001,P< 0.002, andP< 0.001, respectively). No other statistically significant differences were found between the two groups for any of the remaining variables, including dietary estimates. Dietary analyses suggest that dietary intakes of SCA individuals exceeded the recommended daily allowances (RDA) of all macro‐ and micronutrients measured, and intakes of most nutrients exceeded those of black controls interviewed. These results suggest that in individuals with SCA, several micronutrients vital to maintaining reducing capacity are present in diminished quantities in plasma/serum. These anomalies exist in SCA patients even though their intake of these micronutrients are similar to those of healthy black men and women.
Splenectomy in patients with AIDS
Authors: Mathew Anna , Raviglione Mario C. , Niranjan Usha , Sabatini Maria T. , Distenfeld Ariel
Published: February 2007

To establish the indications for splenectomy in patients with human immunodeficiency virus (HIV) infection we retrospectively analyzed 12 patients who underwent splenectomy. Patients with HIV infection who had immune thrombocytopenic purpura (ITP) were excluded as they had no splenomegaly and a definite indication for splenectomy exists in some of these patients. All 12 patients were anemic; 6 were thrombocytopenic and 6 leukopenic. All patients had splenomegaly and all were febrile. At surgery 3 patients were found to have Mycobacterium avium intracellulare (MAI) infection; 2 had splenic abscess due to Salmonella group D; 1 each had cytomegalovirus (CMV) splenitis and localized Kaposi's sarcoma (KS) of the spleen. No definite histopathologic diagnosis could be made in five patients, all of whom had evidence of extramedullary hematopoiesis. The degree of splenic enlargement did not correlate with the outcome. Both clinical and hematologic improvements were achieved in patients with splenic abscess and in patients who had splenomegaly, anemia, and thrombocytopenia. The presence of either of these findings constitutes an indication for splenectomy. Anemia and/or leukopenia without thrombocytopenia failed to improve; the presence of MAI and active CMV infection also resulted in failure. The presence of either of these conditions may be considered a contraindication to splenectomy.
Development of anti‐idiotypic antibodies in a patient with a factor VIII autoantibody
Authors: Tiarks Cheryl , Pechet Liberto , Humphreys Robert E.
Published: February 2007

Antibodies against coagulation factor VIII were detected in a 33‐year‐old who developed severe bleeding 2 months postpartum. After treatment with plasmapheresis and immunosuppressive agents, the inhibitor was no longer detectable, but anti‐idiotypic antibodies were detected, as demonstrated by binding in immunoradiometric assays, and by in vitro neutralization of the initial factor VIII autoantibodies. The anti‐idiotypic titer subsequently declined but was still detectable 1 year later. Three commercial immunoglobulin preparations and pooled multiparous IgG also bound to (but did not neutralize) the patient's factor VIII antibody. These studies suggest that anti‐idiotypic antibodies, arising in the face of immunosuppressive therapy, might suppress autoimmunity to factor VIII and confirm the presence of anti‐idiotypic antibodies in pooled normal immunoglobulins.
Case report: Allergic reaction with immune hemolytic anemia resulting from chlorambucil
Authors: Thompson‐Moya Luis , Martin Thomas , Heuft Hans‐Gert , Neubauer Andreas , Herrmann Richard
Published: February 2007

In a 73‐year‐old female with chronic lymphocytic leukemia we observed an acute illness with shivering, high fever, and hemolytic anemia following chlorambucil administration. Reexposure in a controlled clinical situation suggested an allergic drug reaction. In an in vitro assay, we were able to demonstrate chlorambucil as the causative agent of immune hemolysis.
Use of recombinant activated factor VII for treatment of a retropharyngeal hemorrhage in a hemophilic patient with a high titer inhibitor
Authors: Macik B. Gail , Hohneker John , Roberts H. R. , Griffin Anne M.
Published: February 2007

Treatment options during an acute hemorrhage for a hemophilic patient who has an inhibitor (antibody) to factor VIII (FVIII) are limited. If the inhibitor titer is high, even massive doses of FVIII are not sufficient to neutralize the antibody. Likewise, immunoadsorption techniques or plasmapheresis cannot remove enough antibody to permit treatment with FVIII. Allergic reactions and cross‐reacting inhibitors complicate therapy with porcine FVIII. Prothrombin complex concentrates (PCC) may be effective but the mechanism is unclear. The theory that activated factor VII (FVIIa) is the active principle in PCC prompted our treatment of a patient with a recombinant FVIIa (rFVIIa) product (NOVO Industries). The patient presented with a large retropharyngeal hemorrhage and an initial inhibitor titer of 129 Bethesda units (BU). Despite Autoplex therapy (100U/kg), tracheal compression by the hematoma increased and asphyxiation was imminent. RFVIIa therapy (60 μg/kg) was substituted for Autoplex and nine doses were given without complication. The hemorrhage was controlled. By 18 hr breathing was normal and swallowing and speech were greatly improved. Clinically, the patient dramatically responded to rFVIIa. In addition, the purity, the lack of known infectious agents, and the ease of administration make rFVIIa a potentially attractive new therapy. Use of this product also promises to further our understanding of in vivo hemostasis.
Post‐transfusion purpura and isoimmune neonatal thrombocytopenia in the same family
Authors: Cobos Everardo , Gandara David R. , Geier Larry J. , Kirmani Saeeda
Published: February 2007

Post‐transfusion purpura and isoimmune neonatal thrombocytopenia are rare and unusual syndromes leading to severe thrombocytopenia. In both disorders the PLA1 platelet antigen is involved in the pathogenesis. A 41‐year‐old woman with an obstetrical history of isoimmune neonatal thrombocytopenia in two of her children developed post‐transfusion purpura following a transfusion of packed red blood cells. Despite the apparent link in these two disorders associated with the PLA1 platelet antigen, this is the first reported case of post‐transfusion purpura and isoimmune neonatal thrombocytopenia occurring in the same family.
Carrier detection for hemophilia B: Evaluation of multiple polymorphic sites
Authors: Mariani G. , Chistolini A. , Hassan H. J. , Gallo E. , Xigen Gu , Papacchini M. , Di Paolantonio T. , Fantoni A.
Published: February 2007

DNA analysis was performed in families with hemophilia B. Restriction fragment length polymorphisms (RFLPs) produced by endonucleases Taql, Xmnl, and Ddel were studied by two factor IX genomic probes, F9(Vlll) and F9(Xlll). Fifty‐seven subjects from ten families were investigated; of them, 31 were carriers (11 obligate and 20 potential). Of the potential carriers, ten displayed laboratory features allowing for a phenotypic diagnosis of heterozygosity. Segregation analysis of the markers was informative in 19/20 potential carriers, which belong to nine of the ten studied families. Among the potential carriers, Taql allowed the carriership assessment in 15 (78.9%), Xmnl in 15 (94.7%), and Ddel in two (10.4%). Diagnosis was not possible in one family since a homozygosity in the key individuals with all the employed enzymes (Taql, Xmnl, Ddel, + BamHI) was found. Hemophilia B syndrome in two families likely results from a new mutation. In one family, a first‐trimester prenatal diagnosis was performed. The use of RFLP analysis allowed us to improve genetic counseling as compared with the phenotypic evaluation by clotting factor assays. Indeed, evaluation of RFLP increased by 26% the carriership assessment of the potential carriers of the hemophilia B trait.
VAD‐based regimens as primary treatment for multiple myeloma
Authors: Alexanian Raymond , Barlogie Bart , Tucker Susan
Published: February 2007

An alternating VCAD‐VAD regimen, combining vincristine‐doxorubicin by continuous infusion with cyclophosphamide and pulse dexamethasone, or VAD alone, was given to 175 previously untreated patients with multiple myeloma. The response rate with primary VAD‐based regimens of 55% was virtually identical to the 54% in comparable patients treated previously with similar programs by using bolus vincristine‐doxorubicin. Despite responses to VAD that were more rapid in onset than any previous treatment, remission and survival times were similar. This may be due to major differences in drug sensitivity between progenitor and differentiated plasma cells. A VAD‐based regimen seems better for newly diagnosed patients when rapid control of multiple myeloma is necessary.
Orthotopic liver transplantation in a patient with combined hemophilia A and B
Authors: Delorme Michael A. , Adams Paul C. , Grant David , Ghent Cameron N. , Walker Irwin R. , Wall William J.
Published: February 2007

A 38‐year‐old man with severe factor IX and mild factor VIII deficiencies complicated by cirrhosis secondary to chronic non‐A non‐B hepatitis underwent orthotopic liver transplantation as treatment for both the cirrhosis and his congenital coagulopathy. Intraoperative hemostasis was obtained with factor VII‐depleted prothrombin complex concentrate and fresh frozen plasma. Factor VIII and factor IX levels were assayed frequently in the perioperative period, and both returned to normal within 24 hr and remained normal postoperatively. Liver transplantation can be considered as definitive therapy for hemophilia A and/or B with transfusion‐related liver disease.
Multiple myeloma: Relationship between survival and cellular morphology
Authors: Pasqualetti Paolo , Casale Raffaele , Collacciani Antonio , Abruzzo Benjamin P. , Colantonio Domenico
Published: February 2007

Myeloma cells in bone marrow aspirates from 121 patients with multiple myeloma at time of diagnosis were evaluated, by assigning them into morphological categories. Cox's model was used to calculate the coefficients for total myeloma cells and “plasmablasts” in respect to survival. Applying these coefficients, the Multiple Myeloma Cellular Score (MMCS) was calculated. MMCS significantly divides the patients into three stages. In fact, there are significant differences both between mean survivals (P< 0.001) and between survival curves (P< 0.00001). There is also a significant correlation (P< 0.00001) between the calculated MMCS and survival in the whole patient group.
Induced expression of A monocytoid B lymphocyte antigen phenotype on the reh cell line
Authors: Patel Bimal , Mohammad Ramzi M. , Blaustein John , Al‐Katib Ayad
Published: February 2007

A human common acute lymphoblastic leukemia (ALL) cell line, REH, was treated in vitro with γ‐interferon (γ‐IFN) or 12‐O‐tetradecanoylphorbol 13‐acetate (TPA). Untreated (control) and treated cells were analyzed for changes in growth patterns, morphology, cytochemistry, surface phenotype, and terminal transferase (TdT) activity. TPA but not γ‐IFN induced further maturation of REH cells along the B‐cell lineage. There was a dramatic decrease in CALLA expression, loss of TdT activity, induction of Leu M5, and increase in Leu 14 expression. TPA also induced monocytoid morphological features on REH cells. Enzymatically, the induced cells strongly expressed acid phosphatase (tartrate sensitive), α‐naphthol acetate esterase (NAE), and periodic acid Schiff (PAS). We conclude that TPA induced monocytoid B‐lymphocyte features on REH cells within the B‐cell lineage, which should not be confused with monocytes/macrophage. The phenotype of cells in this stage is Leu 14 +, Leu M5 +, BL1 +, Leu 12 +, AcP +, PAS +, NAE +, CALLA‐, TdT‐, MO1‐, and MO2‐.
Precise quantitation of palgg: A new radiometric microtechnique
Authors: Schwartz Kenneth A. , Gauger John A. , Davis John M.
Published: February 2007

We report the development of a radiometric assay for platelet‐bound IgG that is both sensitive and quantitative. The assay utilized 96‐well millititer plates incorporating a 0.2 μm filter membrane in the bottom. A 125l‐labeled monoclonal antihuman IgG, as a secondary antibody, detected the platelet‐bound human IgG. Since 5 ± 106 platelets were used for each assay, tests for platelet‐bound IgG can be performed on persons with severe thrombocytopenia. For the detection of circulating antiplatelet alloantibodies, as little as 10 μl of platelet‐free plasma per assay is required.

Antiplatelet IgG was quantitated by using anti‐PlA1 antibody that was purified with affinity and elution and DEAE chromatography. This purified antiplatelet antibody was labeled with 125l and was used to determine the binding ratio of secondary antibody to primary antibody. Under our standard conditions, this ratio was found to be stable at approximately 0.35 over the sensitivity range of the assay. The assay can detect approximately 200 molecules of human IgG per platelet (0.1 ng of secondary antibody bound per 5 ± 106 platelets). It has a linear range from ) to 7,000 molecules per platelet.

Quantitation of anti‐PlA1 binding for platelets stored for up to 6 months under refrigeration showed no change in number of PlA1 binding sites. Clinical studies showed that 18 of 19 ITP patients had an increased number of IgG molecules per platelet as did patients with malignancy and drug‐induced immune thrombocytopenia. Patients who had received multiple platelet transfusions had antiplatelet antibody in their plasma. Normal amounts of PAlgG were observed in platelets and plasma of patients with nonimmune thrombocytopenia. The advantages of this method are that it is: 1) a more precise quantitation of PAlgG via direct measurement of binding ratio with PlA1 antibody; 2) performed with small amounts of platelets and plasma; 3) both sensitive and specific; and 4) reliably reproducible with both fresh and stored platelets.
Alternation of two doses of intravenous gammaglobulin in the maintenance treatment of patients with immune thrombocytopenic purpura: More is not always better
Authors: Bussel James B. , Fitzgerald‐Pedersen Julie , Feldman Charles
Published: February 2007

Intravenous infusion of gammaglobulin (IVGG) has been shown to be effective in the maintenance treatment of children and adults with immune thrombocytopenic purpura (ITP). The objective of this treatment is not to increase the platelet count into the normal range but rather to keep the platelet count above a safe level, usually 20,000—30,000/μI. This use requires periodic infusion whenever the platelet count falls. The dose of gammaglobulin in previous studies was 1 gm/kg/Infusion in the majority of patients. This study compared doses of 0.5 and 1.0 gm/kg/Infusion by alternating the two doses in the maintenance treatment of 11 patients with ITP. There was no significant difference in the duration of response following single IVGG infusion between these two doses. This finding suggests that there is no advantage to the greater dose of IVGG and will substantially lower the cost of as well as facilitating IVGG maintenance treatment. It was not possible to determine whether a lower dose would impact adversely on the curative potential of IVGG. This effect of IVGG has never been proved and, if it exists, may be mediated by a different mechanism from the acute platelet response.
Durable complete remissions after 2′‐deoxycoformycin treatment in patients with hairy cell leukemia resistant to interferon alpha
Authors: Blick Mark , Lepe‐Zuniga Jose L. , Doig Robert , Quesada Jorge R.
Published: February 2007

The efficacy of interferon alpha in treatment of hairy cell leukemia is well established. Our experience with low doses of 2′‐deoxycoformycin, a competitive inhibitor of adenosine deaminase, showed that it was also effective against hairy cell leukemia in 10 of 11 patients studied whose disease was resistant to interferon alpha. Ten patients entered complete remission after five to 12 doses of 2′‐deoxycoformycin (4 mg/m2 every other week), and one patient did not respond. No relapses were observed after median followup of 18.5 months; unmaintained complete remissions lasted from more than 10 to more than 30 months. The study demonstrated that 2′‐deoxycoformycin induces a high rate of durable, unmaintained complete remissions in patients with hairy cell leukemia resistant to interferon alpha.
Treatment of platelet‐alloimmunization with cyclosporin a in a patient with aplastic anemia
Authors: Yamamoto Masahiro , Ideguchi Hiroshi , Nishimura Junji , Nawata Hajime , Inaba Shoichi , Tokunaga Kazuo , Kiyokawa Hiroyuki , Maeda Yoshiaki
Published: February 2007

The development of alloantibodies to platelets is a major problem in the supportive management of thrombocytopenia in patients with severe aplastic anemia. We report here a case of aplastic anemia refractory to platelet transfusion. An immunosuppressant, cyclosporin A, which was used for the therapy of aplastic anemia, modulated alloimmunization to platelets in this patient, followed by repeated platelet transfusion. The treatment reduced platelet alloantibodies detected by anti‐human immunoglobulin lymphocytotoxicity test, with change of the CD4/CD8 ratio in T lymphocytes in peripheral blood. These results suggest the usefulness of cyclosporin A for the prevention of platelet alloimmunization.
Effect of histocompatibility factors on pulmonary retention of indium‐111‐labeled granulocytes
Authors: Dutcher Janice P. , Riggs Charles , Fox Jeremiah J. , Johnston Gerald S. , Norris Dottie , Wiernik Peter H. , Schiffer Charles A.
Published: February 2007

Granulocyte transfusions are associated with a number of side effects including febrile transfusion reactions and occasionally pulmonary infiltrates. There is evidence that the presence of preformed antibodies may be a cause of these complications. In this study, allogeneic 111lndium‐labeled granulocytes were used to evaluate the pulmonary retention of radioactivity in alloimmunized and non‐alloimmunized patients in an attempt to assess antibody effect on granulocyte migration. After injection of labeled allogeneic granulocytes into neutropenic patients, the ratios of lung to heart activity were calculated for the first 30 min of scanning. There was significantly greater retention of radioactivity from cells in the lungs of patients who were alloimmunized, having both lymphocytotoxic (anti‐HLA) and leukoagglutinating antibodies, compared to the activity in the lungs of non‐alloimmunized patients (P < 0.001) or of patients receiving autologous granulocytes (P < 0.001). This study demonstrates that labeled, mismatched granulocytes may be retained in the lungs for a significantly longer time in patients with preformed antibodies. This implies that transfusion of large numbers of such mismatched granulocytes, i. e., granulocyte transfusions, may also be retained in the lungs of alloimmunized patients, which could lead to pulmonary compromise. Therefore, granulocyte transfusions from random donors should not be given to alloimmunized patients.
No beneficial effect of splenectomy in hereditary high red cell membrane phosphatidylcholine hemolytic anemia: Clinical and membrane studies of 20 patients
Authors: Otsuka Akiyo , Sugihara Takashi , Yawata Yoshihito
Published: February 2007

The clinical features and red cell membrane characteristics of 20 patients with hereditary high red cell membrane phosphatidylcholine hemolytic anemia (HPCHA) were studied in relation to the effect of splenectomy. After splenectomy, anemia worsened and the extent of increased hemolysis in these patients was unchanged, indicating a contraindication for splenectomy. Concomitant with these results in clinical hematology, marked stomatocytic changes, increased red cell phosphatidylcholine content, and enhanced sodium transport, which were observed before splenectomy, were not improved by splenectomy.
Surface light chain phenotype in indolent lymphomas: Lack of prognostic significance
Authors: Vandermolen Louis A. , Duffey Patricia L. , Cossman Jeffrey , Jaffe Elaine S. , Longo Dan L.
Published: February 2007

The indolent follicular and diffuse lymphomas are neoplasms of B‐cell origin. In several other B‐cell neoplastic disorders, including multiple myeloma, hairy cell leukemia, and chronic lymphocytic leukemia, the light chain isotype of the surface immunoglobulin has been reported to have prognostic significance. Patients with tumors expressing λ light chains usually fare more poorly than those with k light chain‐bearing tumors. We analyzed the clinical data and immunologic phenotype of 101 patients with indolent lymphoma. Eighty‐nine of the 101 patients demonstrated surface immunoglobulin of only one light chain type (k‐47, λ‐42). Patients in both groups were matched for known prognostic factors. There were no significant differences in disease‐free survival or overall survival between the two groups or within histologic subtypes. These results indicate that the immunologic light chain phenotype in indolent lymphoma, unlike other B‐cell neoplasms, is not a prognostic indicator of survival.
Neutrophilic myelofibrosis presenting as philadelphia chromosome negative BCR non‐rearranged chronic myeloid leukemia
Authors: Stewart Keith , Carstairs Kelvin C. , Dubé Ian D. , Keating Armand
Published: February 2007

Chronic myeloid leukemia consists of Philadelphia chromosome positive disease in 90% of cases, and a further 5%, although Philadelphia chromosome negative, exhibit bcr gene rearrangements consistent with the disease. The remaining 5% of cases have a heterogeneous clinical picture with a course unlike that of classical chronic myeloid leukemia, and may belong to different pathologic entities.

We report five cases belonging to the latter group, initially identified as Philadelphia chromosome negative, bcr non‐rearranged chronic myeloid leukemia, that developed progressive leucocytosis, absolute monocytosis, myelodysplasia, extramedullary hematopoiesis, and had evidence of myelofibrosis. These cases may represent a distinct clinical entity characterized by neutrophilic myelofibrosis, which can be identified prospectively by clinical and pathologic criteria. Standard therapy for treating chronic myeloid leukemia or idiopathic myelofibrosis may not be appropriate for this group.
Patient with pyruvate kinase deficiency developed acute myelogenous leukemia
Authors: Leki Ryuji , Miwa Shiro , Fujii Hisaichi , Kudoh Shouji , Kimura Hitoshi , Takaku Fumimaro
Published: February 2007

A 44‐year‐old previously healthy male was diagnosed as anemic and treated at a nearby hospital. A year later, he was admitted to our hospital for precise examination of the progression of refractory anemia. Blood examination showed hemolytic anemia and more detailed examination of this hemolysis revealed pyruvate kinase deficiency. The activity was 15.1% of a normal control. Pyruvate kinase activities of his family members were normal or slightly decreased. Ten months after admission to our hospital, 2% of blast cells with Auer's body in the peripheral blood were noted which increased progressively. The bone marrow contained 17.3% of blast cells with a subsequent diagnosis of acute myelogenous leukemia being made.

Although intensive chemotherapy was performed, the patient achieved a brief remission and died 2 years after admission to our hospital.
Diagnosis of cobalamin deficiency I: Usefulness of serum methylmalonic acid and total homocysteine concentrations
Authors: Allen Robert H. , Stabler Sally P. , Savage David G. , Lindenbaum John
Published: February 2007

The serum cobalamin assay is the primary diagnostic test for cobalamin deficiency. It appears to be an excellent screening test since most patients with clinically confirmed cobalamin deficiency have low levels. Recent studies indicate that the clinical picture of cobalamin deficiency is much more diverse than previously believed. It is also apparent that many patients with low serum cobalamin concentrations are not cobalamin deficient. Thus, there is a need for additional diagnostic tests to further distinguish patients with low serum cobalamin levels who are actually cobalamin deficient and will benefit from lifetime treatment from those who are not deficient and will not benefit. Serum levels of methylmalonic acid and total homocysteine have been shown to be markedly elevated in most patients with cobalamin deficiency, and total homocysteine concentrations are markedly elevated in most patients with folate deficiency. The levels of these metabolites fall to normal if these patients are treated with the appropriate vitamin but remain essentially unchanged if the wrong vitamin is administered. These observations demonstrate that serum methylmalonic acid and total homocysteine levels are useful in diagnosing patients with cobalamin and folate deficiency and in distinguishing between these two vitamin deficiencies.
Subtle and atypical cobalamin deficiency states
Authors: Carmel Ralph
Published: February 2007

Evidence for cobalamin (vitamin B12) deficiency usually involves some combination of low serum cobalamin levels, clinical abnormalities (classically, megaloblastic anemia and neurologic defects), metabolic abnormalities, and response to therapy. However, cobalamin deficiency may often display few of the expected clinical findings. Identification of the underlying cause is also important in the diagnosis of deficiency, and its value may be particularly great when the expression of deficiency is subtle. The cause of cobalamin deficiency is usually malabsorptive, but may sometimes be limited to malabsorption of food cobalamin while free cobalamin is absorbed normally. Nongastroenterologic entities may sometimes also be found. All of these considerations allow the proposal of four patterns of cobalamin deficiency. The first type is classical deficiency; typical megaloblastic anemia with or without neurologic dysfunction occurs because of classical cobalamin malabsorption such as lack of intrinsic factor (pernicious anemia). The second type consists of classical cobalamin malabsorption in which the cobalamin deficiency is expressed subtly rather than in classical fashion. There is no megaloblastic anemia and sometimes the only evidence of deficiency may be metabolic. In the third type, cobalamin deficiency is expressed classically but is attributable to a subtle or atypical cause, such as food‐cobalamin malabsorption. In the fourth type, deficiency is both expressed subtly and arises from subtle or atypical causes. Such presentations require further investigation but are a challenging expansion of our understanding and recognition of cobalamin deficiency.
Function of vitamin B12 in the central nervous system as revealed by congenital defects
Authors: Hall Charles A.
Published: February 2007

The 13 cases of methylcobalamin (MeCbl) deficiency presenting in early infancy have all been developmentally delayed, and the majority have had seizures, hypotonia, lethargy, and microcephaly. The CNS injury appears to occur during the first 6 months of postnatal life. The same symptoms are seen in acquired cobalamin (Cbl) deficiency in the same age group. MRI performed at age 18–19 months and after 13–14 months of large amounts of Cbl, in two cases showed delayed myelination, most pronounced in the cerebrum. Isolated MeCbl deficiency is the consequence of cblE and G mutations where the lesion is of a single Cbl‐dependent enzyme, the methyltransferase. One effect of a deficiency of MeCbl, and of the associated failure of the methionine synthase reaction, is, therefore, an impairment of myelination of the brain of the newborn. The slow, but usually incomplete, improvement in psychomotor status after years of treatment with Cbl may be related to the eventual myelination. However, the hypotonia, lethargy, and impaired responsiveness react to treatment with Cbl within 24–48 hours, which suggests an expression of MeCbl deficiency on the CNS distinct from the delayed myelination. Although there is much to be learned, it is now clear that a normally functioning Cbl‐dependent methyl transferase is required for development and function of the human brain.
Low holotranscobalamin II is the earliest serum marker for subnormal vitamin B12 (cobalamin) absorption in patients with AIDS
Authors: Herbert Victor , Fong Warren , Gulle Veronica , Stopler Tracy
Published: February 2007

In AIDS, as previously found in pernicious anemia (PA), the earliest serum marker of subnormal vitamin B12 (cobalamin) absorption, and therefore of negative B12 balance, is low serum holotranscobalamin II (holo‐TC II; B12‐TC II) despite normal total serum B12 level, normal serum homocysteine, and normal classic (oral free radio‐B12) Schilling test. This may be accompanied by subtle and insidious damage to hematopoietic, immunologic, neuropsychiatric, nutritional and alimentary systems, confirmed by correction on therapeutic trial with B12 therapy. Our studies suggest such selective B12 deficiency occurs in about half of the HIV‐1 infected, in part due to frequent depression of B12 absorption by HIV‐1 attack on the gastric mucosa and/or opportunistic infection attack on the small bowel, and in part due to a telescoping of the continuum of the stages of negative B12 balance in relation to damage to B12 delivery by the infective and/or systemic disease process. In AIDS, when total serum B12 is normal despite tissue depletion of B12, if the classic Schilling test does not reveal subnormal food B12 absorption, the food Schilling test does. We hypothesize that DNA‐synthesizing cells of the hematopoietic, immunologic, neurologic and other systems which have surface receptors solely for holo‐TC II, and which have low B12 stores, rapidly become dysfunctional due to B12 deficiency when holo‐TC II is low, while cells (such as liver cells) which also have surface receptors for holohaptocorrin (B12‐haptocorrin) remain B12‐replete. We believe this to be another example of the concept of selective nutrient deficiency in one cell line but not another.
Application of the avidin‐biotin‐peroxidase complex technique for ultraimmunocytochemical characterization of leukemic cells
Authors: Yano Takahiro , Ohtake Ken‐Ichi , Kameda Kazuo , Kizaki Masahiro , Ogawa Tetsuhei
Published: February 2007

For simultaneous demonstration of cellular ultrastructure, myeloperoxidase activity, and presence of a membrane‐bound antigen in a given blood cell, we examined three different fixatives: periodate‐lysine‐paraformaldehyde (PLP) and paraformaldehyde and glutaraldehyde for their applicability to preembedding electron microscopic immunocytochemistry using monoclonal antibodies and the avidin‐biotin‐peroxidase complex (ABC) technique. This procedure was examined in samples from 3 normal volunteers and 29 patients with acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), lymphosarcoma cell leukemia (LSCL), blastic phase of chronic myelogenous leukemia (CML‐BC), or other unclassified leukemias. PLP fixation preserved the immunoreactivity of surface glycoproteins as well as immunoglobulins to the most satisfactory extent. Leukemic cells fixed with PLP maintained their fine structural details, so that we could identify their cytoplasmic organelles, although glutaraldehyde produced the best preservation of cellular ultrastructure. In three patients with ALL, our method revealed that a significant portion of blasts possessed both lymphoid surface antigens and peroxidase‐positive cytoplasmic granules. Our method was also useful in identifying the lineage of peroxidase‐negative leukemic cells, including monoblastic leukemia and megakaryoblastic leukemia cells. Ultraimmunocytochemistry using PLP fixation and the ABC technique may be a promising strategy for determining the nature of blastic cells that remain unclear after a conventional work‐up, for characterizing leukemic cells in patients with a relatively low blast cell count in the bone marrow or peripheral blood, and for estimating the presence and frequency of leukemia with multilineage expression.
Properties of the exchangeable splenic platelets released into the circulation during exercise‐induced thrombocytosis
Authors: Chamberlain Kevin G. , Tong Michael , Penington David G.
Published: February 2007

The human spleen normally retains about one‐third of the body's platelets in an exchangeable pool which can be released into the circulation by α‐adrenergic stimulation. Some previous investigators concluded that the splenic platelet population was enriched in a subpopulation of large, young, dense platelets (megathrombocytes) but more recent research suggests that platelet size, age, and density are largely independent variables. In this investigation the properties of the splenic platelets were studied after their release into the circulation by acute strenuous exercise in 11 normal subjects. The exercise caused a rise in mean platelet count from 245 ± 49 to 328 ± 71 × 109/L—a net increase of 24 ± 6% after correction for haemoconcentration. The mean platelet volume (MPV) of citrated platelets increased from 6.38 ± 0.78 to 6.59 ± 0.68 fL after exercise (P< 0.01)—a rise of 3.7 ± 4.1% suggesting that the MPV of the splenic platelet population was about 20% greater than that of the normal circulating population. The age distribution of the platelets was studied by measuring the platelet monoamine oxidase (MAO) activity several days after irreversible inhibition by tranylcypromine, when the young platelets had normal MAO activity but the older platelets had only 20% of normal activity. The mean platelet MAO activity did not change after exercise, indicating that the age distributions of the circulating and splenic populations were very similar. The platelet contents of several putative markers of platelet age (sialic acid, serotonin, β‐thromboglobulin, β‐N‐acetylglucosaminidase) were also unchanged after exercise. Modal platelet density decreased slightly but not significantly after exercise. The splenic platelet population has a larger MPV but appears to have similar age and density distributions to the basal circulating population.
Effectiveness of the cryosupernatant fraction of plasma in the treatment of refractory thrombotic thrombocytopenic purpura
Authors: Byrnes John J. , Moake Joel L. , Klug Panpit , Periman Phillip
Published: February 2007

Many patients with thrombotic thrombocytopenic purpura (TTP) satisfactorily respond to plasma therapy (plasmapheresis and/or plasma infusion). Some, however, respond either not at all or only transiently and incompletely. Evidence indicates that platelets and endothelial cell‐derived unusually large von Willebrand factor (ULvWF) multimers, as well as the largest vWF multimers in plasma, form thrombi which are deposited in the microvascular circulation. Accordingly, platelet transfusions are avoided unless there is intra‐cranial or other life‐threatening hemorrhage. The largest plasma multimers of vWF are, however, replenished by the infusion of large volumes of whole plasma. We postulated that under conditions of massive plasma replacement, plasma depleted of the largest multimers of vWF might be preferable for the treatment of TTP episodes. The largest vWF multimers sediment in the cryoprecipitate, and the cryoprecipitate‐poor fraction of plasma (cryosupernatant) is depleted of these forms. Seven patients responding inadequately to intensive plasma therapy were switched to receive cryosupernatant in place of whole plasma. All patients improved quickly following this change in therapy, and the TTP syndrome resolved in all seven.
Bone marrow in vitro growth and cytogenetic studies in patients with fab‐classified primary myelodysplastic syndromes
Authors: Swolin Birgitta , Rödjer Stig , Westin Jan
Published: February 2007

Thirty‐eight consecutive patients with a FAB‐classified primary myelodysplastic syndrome (MDS) were investigated for in vitro growth of colony‐forming units for granulocyte‐macrophage precursors (CFU‐GM) and cytogenetic analysis of bone marrow cells. Abnormal CFU‐GM growth was found in 30 patients (79%), and clonal chromosome abnormalities were found in 13 patients (34%). The eight patients who showed normal CFU‐GM growth were either cytogenetically normal (n = 5), or had a 5q‐ deletion (n = 3) as single or dominating karyotypic abnormality. Among the 30 patients with reduced or no colony growth, ten patients had a clonal chromosome abnormality.

Leukemia developed in eight patients. None of them grew any CFU‐GM colonies, and three of them were cytogenetically abnormal at the time of diagnosis of MDS.

Analysis of the bone marrow in vitro growth for CFU‐GM and the karyotype in patients with MDS emphasizes the close relationship between these disorders and manifest acute leukemia. Subgroups of MDS may be defined by a cytogenetic classification (e.g., the 5q‐ syndrome), and the CFU‐GM growth pattern can be of value for predicting leukemic transformation.
Development of 5q‐ myelodysplasia in a patient with sarcoidosis
Authors: Tunkel Allan R. , Sebastianelli Kenneth J. , Pandit Niraj , Brody Jerome I.
Published: February 2007

Sarcoidosis has been observed in association with numerous blood dyscrasias including lymphoma, leukemia, and multiple myeloma. This report describes a patient with sarcoidosis and a refractory anemia whose bone marrow karyotype showed deletion of the long arm of chromosome 5, consistent with a myelodysplastic syndrome. Concurrent sarcoidosis and myelodysplasia may relate to the continued availability of cytokines as a consequence of repeated macrophage, T‐cell, and B‐cell interactions, with evolution to the 5q‐ abnormality. This association may merit specific attention in the future approach to the diagnostic evaluation in certain patients with sarcoidosis.
Desferrioxamine improves neutrophil phagocytosis in thalassemia major
Authors: Cantinieaux Brigitte , Hariga Chantal , Fondu Pierre , Ferster Alina , Toppet Michèle
Published: February 2007

The aims of the present study are: first, to assess the toxic role of serum from thalassemic patients in phagocytosis of PMN from healthy controls, and second, to seek to determine whether serum and cellular disturbances of polymorphonuclear neutrophils (PMN) phagocytosis, observed in thalassemic patients, can be prevented and/or corrected by use of desferrioxamine (DFX).

Two kinds of in vitro incubations–‐without or with DFX–‐were performed. PMN or serum from thalassemic patients or from healthy controls was used. First, a phagocytosis defect of 3 different bacteria species was induced in PMN from healthy controls by incubation in thalassemic serum. Second, DFX could prevent, already at 1 μM, the phagocytic defect induced in normal PMN by the incubation with thalassemic serum, with disappearance of the toxic role of thalassemic serum at higher concentrations. Third, improvement of the phagocytosis defect of PMN from thalassemic patients was also observed at 1 μM of DFX for the 3 bacteria species. Normalization was obtained at higher concentrations for gram‐negative bacteria.

In vivo studies revealed, after a 3 hr subcutaneous infusion of DFX into 3 thalassemic patients, an improvement of the phagocytosis results and a decrease of the Prussian Blue reactivity of the PMN.

It is concluded first that an iron‐mediated defect in phagocytosis can be induced in normal neutrophils by incubation in serum from thalassemic patients, and second that a precautious and intensive chelation therapy seems to be advantageous for increasing PMN defense against infectious agents. Special care must nevertheless be taken in order to detect rapidly opportunistic (such as Yersinia) infections.
Sequential intermediate‐dose cytosine arabinoside and mitoxantrone for patients with relapsed and refractory acute myelocytic leukemia
Authors: Paciucci Paolo Alberto , Cuttner Janet , Holland James F.
Published: February 2007

Based on in vitro evidence of time‐dependent synergistic kill of HL‐60 leukemia cells exposed to Ara‐C and mitoxantrone, 44 patients with relapsed or refractory AML and 3 with blastic CML were treated with a timed sequence of both drugs. There were 25 females and 22 males, with a median age of 53 (range 21–75). Of 31 patients with relapsed AML, 24 had one prior remission, 6 had two and 1 had three. Of these, 15 had failed a second reinduction attempt. Thirteen patients were primarily refractory to induction with Ara‐C plus daunorubicin. Each dose of Ara‐C, 500 mg/m2, was followed after 6 hr by mitoxantrone, 5 mg/m2, and the sequence was repeated four to six times (44–68 hr) in different cohorts of patients. All but two patients (one with blastic CML and one in relapse and refractory) are evaluable for response and toxicity. Of 16 patients in relapse without prior reinduction 7 achieved CR and 3 PR (62% response rate); there were 3 CR in the 14 patients who were in relapse and refractory (21% response rate) and 4 CR and 1 PR (35% response rate) in the 14 patients with primary anthracycline resistance. Five of seven patients previously exposed to mitoxantrone achieved CR. Response lasted from 2 to 42 months, with two patients alive and in continuing remission at 34 and 42 months. Average marrow recovery was seen after 25 days and time to remission was 30 days. Six patients died in induction (four from sepsis and two from the tumor lysis syndrome) and 21 had progessive disease. Chemotherapy was well tolerated with minor nausea and vomiting in 13 patients, moderate in 20, and severe in 2. Most patients did not have evidence of drug‐induced mucositis: it was minor in 9 and moderate in 2. Renal dysfunction was attributable to the use of nephrotoxic antibiotics. Hepatic dysfunction was reversible and was minor in 10 patients, moderate in 13, and severe in 3. Sequential, timed administration of intermediate‐dose Ara‐C and mitoxantrone is an active and well‐tolerated antileukemic regimen.
Alloimmune neonatal thrombocytopenia associated with incidental maternal thrombocytopenia
Authors: Burrows Robert F. , Kelton John G.
Published: February 2007

In this report, we describe a mother with mild incidental thrombocytopenia who delivered a severely thrombocytopenic infant having alloimmune thrombocytopenia. This coincidental association should be considered because the recognition and proper diagnosis of alloimmune neonatal thrombocytopenia is important, not only in the current pregnancy, but also in future pregnancies.