bluuebunny/crossref_metadata_2025 huggingface dataset:
Measurement of human serum erythropoietin by erythroid colony‐forming technique using fetal mouse liver cells
Authors: Okamoto Takahiro , Kanamaru Akihisa , Hara Hiroshi , Nagai Kiyoyasu
Published: February 2007

A quantitative bioassay for serum erythropoietin in anemic patients was established with erythroid colony‐forming technique using methyl cellulose. Fetal mouse liver contains many erythropoietin‐dependent erythroid colony‐forming cells (CFU‐E) with the concentration as well as sensitivity to erythropoietin being the highest at 13–14 days of gestation. Linearity in the erythropoietin response curve for the number of colonies from CFU‐E in fetal livers of 13–14 days gestation was obtained for erythropoietin concentrations in the culture medium of 3.9 to 125 mU/ml. When newborn liver cells were used instead, the linearity occurred at 31.2 to 250 mU/ml. Measurement of erythropoietin by the erythroid colony‐forming technique for sera from patients with various anemias was compared with that of the assay method using radioiron incorporation into heme in the suspension culture of fetal mouse liver cells. There was a good correlation between the values of both assay methods (r = 0.856, p < 0.001). Also, levels of erythropoietin measured by this assay method showed the expected inverse relationship to the hemoglobin concentration in the sera from the patients.
A case of chronic aneosinocytosis
Authors: Telerman Adam , Amson Robert B. , Delforge Alain , Collard Elizabeth , Stryckmans Pierre A.
Published: February 2007

A 59‐year‐old woman, after complete recovery from an episode of drug‐induced agranulocytosis, was found to sustain a chronic absence of recognizable mature and immature eosinophils in blood and bone marrow during a follow‐up period of 8 years. Her bone marrow and peripheral blood cells cultured in vitro were able to produce normal numbers of eosinophil colonies. The present disorder was thus not caused by a lack of EO‐CFU‐C, nor by a defect in EO‐CSA, nor by an immunologically mediated mechanism acting on mature or immature eosinophils or on EO‐CSA producing cells. It is suggested that the marrow was not able to differentiate normally along the eosinophil pathway, possibly owing to a microenvironmental defect or, less likely, an intrinsic stem cell defect.
Phosphofructokinase (PFK) deficiency due to a catalytically inactive mutant M‐type subunit
Authors: Zanella A. , Mariani M. , Meola G. , Fagnani G. , Sirchia G.
Published: February 2007

A case of M‐type PFK deficiency due to the synthesis of a structurally abnormal and catalytically inactive M‐subunit was reported. PFK activity was reduced (39% of normal) in red cells, normal in leukocytes and platelets, and absent in muscle. The red cell enzyme was not inhibited by antiserum to human muscle PFK and displayed normal biochemical properties (Km for ATP and fructose‐6‐phosphate, storage stability at + 4°C and −80°C, optimum pH, electrophoretic pattern and molecular weight).

The complete lack of PFK activity in muscle was confirmed on both histological preparations and muscle extracts. Double immunodiffusion analysis using an antinormal M‐PFK serum revealed that the enzyme molecule was present and immunologically identical with normal, although it was catalytically inactive. The muscle abnormality was also confirmed by electromyography, ischemic exercise testing, histochemistry and electron microscopy. Moreover, PFK activity was investigated in myoblast cultures maintained up to 25 days, and it was found to be absent.
Phenotypic heterogeneity of human T‐cell malignancies: Demonstration by monoclonal antibodies and cytochemical markers
Authors: Knowles Daniel M. , Halper James P. , Machin Genevieve A. , Byeff Peter , Mertelsman Roland , Chess Leonard
Published: February 2007

The present study sought to delineate the phenotypic heterogeneity of the human T‐cell malignancies. Twenty T‐cell neoplasms were investigated for reactivity with the OKT hybridoma monoclonal antibodies and expression of acid α‐naphthyl acetate esterase (ANAE), β‐glucuronidase (BG), and acid phosphatase (AP) activity. Twelve cases (Mycosis fungoides, Sezary syndrome, cutaneous T‐cell lymphoma, chronic lymphocytic leukemia) were OKT3+T4+, ie, expressed the phenotype commonly associated with mature T‐helper cells. These cases were further divisible into ANAE+BG+ (6 cases), ANAE‐BG+ (5 cases), and ANAE‐BG‐ (1 case) phenotypes. In contrast to the 12 OKT3+T4+ cases, the remaining 8 cases showed considerable inter‐ and intratumor heterogeneity with respect to reactivity with the OKT antibodies. Six of these cases (acute lymphoblastic leukemia, lymphoblastic lymphoma) expressed phenotypes consistent with various intrathymic stages of T‐cell differentiation. Five of the latter 6 cases were AP+BG+ANAE‐, analogous to the majority of normal cortical thymocytes; an OKT3+T4‐T8+T10+ neoplasm was ANAE+, analogous to normal medullary thymocytes. Two cases expressed the previously undescribed OKT3+T4‐T8‐T10+ phenotype. These studies demonstrate that the T‐cell malignancies are divisible into phenotypes which correspond to normal maturational stages of T‐cell differentiation and functionally distinct T‐cell subsets. Phenotypic analysis of the human T‐cell malignancies may provide a basis for understanding their biological heterogeneity and may aid in the identification of transitional stages of T‐cell differentiation and minor T‐cell subsets.
Familial T‐cell lymphoblastic lymphoma: Association with von recklinghausen neurofibromatosis and gardner syndrome
Authors: Kaplan Joseph , Cushing Barbara , Chang Chung‐Ho , Poland Ronald , Roscamp Jacqueline , Perrin Eugene , Bhaya Nirmala
Published: February 2007

A family is described in which three of seven siblings developed a T‐cell lymphoblastic lymphoma. Tumor cells formed rosettes with sheep erythrocytes, lacked surface Ig, and expressed human T‐ but not B‐lymphocyte antigens. They lacked the enzyme terminal transferase suggesting a lymph node rather than thymic T‐cell origin. Autopsy findings supported this conclusion. All three sibs, like their father, had numerous cafe'‐au'lait spots indicative of Von Recklinghausen neurofibromatosis. One child had a subcutaneous fibroma, and another had multiple colonic polyps and exostoses characteristic of Gardner syndrome. Both are autosomal dominant conditions known to predispose to malignancies. The genetic factors responsible for these conditions may also predispose to the development of “postthymic” T‐cell lymphoblastic lymphoma.
Acute myeloid leukemia as the first hematologic manifestation of fanconi anemia
Authors: Auerbach Arleen D. , Weiner Michael A. , Warburton Dorothy , Yeboa Kwame , Lu Li , Broxmeyer Hal E.
Published: February 2007

A six‐year‐old girl with Fanconi anemia (FA) developed acute myeloid leukemia (AML) as the first hematologic manifestation of the syndrome. She remains in remission 18 mo after diagnosis although her management is complicated by unusual sensitivity to chemotherapeutic agents. Marrow cells studied prior to initiation of leukemia therapy showed increased chromosome breakage and an abnormal clone in which a number 7 and a number 8 chromosome were replaced by two marker chromosomes. During the present remission her cultured lymphocytes, bone marrow cells, and fibroblasts showed increased “spontaneous” chromosome breakage as well as enhanced sensitivity to the clastogenic effect of the difunctional alkylating agent diepoxybutane (DEB), features characteristic of FA. Eight months into remission 50% of her marrow cells comprised an abnormal clone, which was monosomic for the number 7 chromosome but had both copies of number 8; in addition a variable number of unique marker chromosomes were present in clonal as well as nonclonal cells. This same marrow sample, upon culture, showed an abnormal growth pattern of CFU‐GM, absence of detectable CFU‐GEMM and BFUe, non‐responsiveness of CFU‐GM to inhibition by acidic isoferritins, increased bone marrow acidic isoferritin inhibitory activity, and absence of detectable bone marrow cell‐derived GM‐CSF. The implications of these findings to leukemogenesis in FA are discussed.
Rosette formation between chronic T‐lymphocytic leukemia cells and macrophages: A facsimile of normal T‐lymphocyte—macrophage cooperation
Authors: Jennette J. Charles , Benson John D. , Wilkman Alice S.
Published: February 2007

Neoplastic T lymphocytes from a patient with chronic T‐lymphocytic leukemia formed rosettes with macrophages. It is proposed that this phenomenon represents retention by these abnormal cells of the normal ability of T lymphocytes to associate physically with macrophages. Recognition of rosettes between benign macrophages and leukemia or lymphoma‐derived lymphocytes may indicate T‐cell lineage of the neoplastic lymphoproliferation.
Aryl sulfatase a in acute nonlymphocytic leukemia
Authors: Pillai M. V. , James G. W. , King M. E. , Fox F. W. , Choi S. C.
Published: February 2007

To ascertain whether estimation of urinary aryl sulfatase A (ASA) is a reliable noninvasive technique for monitoring disease activity in adult acute nonlymphocytic leukemia, we studied the excretion pattern of this enzyme during active disease, partial remission, and complete remission in ten patients and compared the results with those of ten healthy volunteers. There was no significant difference in enzyme excretion between complete remission cases and the control group. Patients with partial remission and active disease had significantly different urinary enzyme levels from each other and from the control and complete remission groups.

Estimation of urinary ASA is a useful noninvasive method of monitoring disease activity in adult acute nonlymphocytic leukemia.
Neutrophil marrow cellularity in neutropenia
Authors: Dancey John T. , Brubaker Leonard H.
Published: February 2007

In order to identify individuals in whom marrow abnormalities might be contributing to or responsible for neutropenia, we quantitatively examined the number and distribution of cells comprising neutrophil marrow in patients with blood neutrophils less than 2,000/μl. Neutrophil marrow cellularity was determined from ferrokinetic estimation of normoblast numbers and neutrophil‐normoblast ratios obtained from marrow biopsy sections. Only two of 30 patients exhibited the change in cellularity expected of a normal marrow responding to removal of circulating neutrophils: reduced numbers of segmented cells, an expanded mitotic pool, and a normal ratio of metamyelocytes and band forms to promyelocytes and myelocytes. Twenty‐three patients had basal mitotic pool size or increased numbers of segmented marrow cells despite neutropenia, a hypoplastic mitotic pool, or a reduction in the number of metas and bands relative to promyelocytes and myelocytes. The results in individual patients were consistent with hypoplasia, subnormal proliferative or release responses, loss of cells during ontogeny, or combinations thereof. In five cases the results could not be so classified. Clinical observations seldom predicted marrow cellularity. Diverse disorders of marrow function appear to be common among neutropenic patients. Neutropenia constitutes a rich field for study of neutrophil marrow physiology.
The role of autologous helper and suppressor T cells in the regulation of human granulopoiesis
Authors: Barr Ronald D. , Stevens Catherine A.
Published: February 2007

Helper and suppressor T cells play a physiological role in immunoregulation and possibly in other biological systems. Previous studies have suggested that unfractionated T cells influence human blood cell formation in vitro. Subpopulations of such cells (TM and TG cells) were prepared by immune ox erythrocyte resetting techniques. Co‐cultures were established in semi‐solid agar with autologous null lymphocytes as a source of granulocyte—macrophage progenitor cells (CFU‐GM). TM (helper) cells produced an increase and TG (suppressor) cells a decrease in the expression of CFU‐GM as reflected in the number of GM colonies generated by comparison with cultures of null lymphocytes alone. These data point to a contribution from T lymphocyte subpopulations to the physiological regulation of human granulopoiesis.
Long survival in sickle cell anemia
Authors: Shurafa Muhammad S. , Prasad Ananda S. , Rucknagel Donald L. , Kan Yuet Wai
Published: February 2007

Ten patients with sickle cell anemia surviving beyond the age of 40 were compared to 11 subjects with sickle cell anemia who died before that age. Hematologic and biochemical data as well as clinical and physical parameters of the two groups were compared. There was no statistically significant difference between the two groups with respect to the degree of anemia, severity of hemolysis, or hemoglobin A2 levels. A statistically significant difference was found between the two groups with respect to fetal hemoglobin, red cell zinc, and secondary sexual characteristics. Although the long survivors had fewer crises per year than the short survivors, the long‐term complications such as leg ulcer, congestive heart failure, and aseptic necrosis of the hip were more common in the older patients. Cerebrovascular accidents were the cause of death in 9/11 short survivors and are absent in the long survivors. The α/β chain synthesis ratio was normal in the long‐survivor group and α gene mapping in five subjects in that group revealed the genotype αα/αα in four and — α/αα in one. The older patients as a group had higher red cell zinc values. The secondary sexual characteristics were also better developed in the older subjects. The overall significance of zinc status and of a higher HbF on longevity of sickle cell anemia patients remains unknown.
Gamma‐chain heterogeneity of fetal hemoglobin in nonblack β‐ and δβ‐thalassemia and hpfh heterozygotes and homozygotes
Authors: Efremov G. D. , Ibarra B. , Gurgey A. , Sukumaran P. K. , Altay C. , Huisman T. H. J.
Published: February 2007

The fetal hemoblobin (Hb F) of a few hundred nonblack patients with a heterozygosity or homozygosity for β‐thalassemia (β‐thal), δβ‐thalassemia (δβ‐thal), and some forms of the hereditary persistence of Hb F (HPFH) was isolated by DEAE‐cellulose chromatography and further characterized by high‐pressure liquid chromatography. Quantitative data for the three types of γ chain (AγT, AγI, and G‐γ) were compared with those obtained for the Hb F from black patients with similar conditions.

The Gγ chain levels in nonblack β‐thal heterozygotes varied greatly and did not fall into two distinct groups with high or low levels, as has been observed in blacks. The level of the AγT chain in AγT heterozygotes did not differ significantly when this anomaly was in cis or in trans to the β‐thal determinant. Beta‐thalassemia homozygotes from Turkey and Yugoslavia, had Gγ values varying between 40% and 80%. Only 13 of 34 patients carried the AγT gene. Nine were AγT heterozygotes with an AγT/total Aγ level averaging 39% and four were AγT homozygotes.

The Gγ chain levels in nonblack δβ‐thal heterozygotes varied between 28% and 46%. An additional AγT chain heterozygosity in cis to the δβ‐thal determinant demonstrated that over 90% of the γ chains is produced by genes in cis to this anomaly. Analyses of members of two relatively large families with β‐thal, δβ‐thal, and the AγT chain heterozygosities and homozygosities occurring in different combinations allowed a more or less quantitative evaluation of the production of γ‐chain genes in cis or in trans to either of the two types of thalassemia determinants. Such calculations were possible both in simple heterozygotes and in persons with the β‐δβ‐thalassemia condition.
Activated partial thromboplastin time after heparin removal (aPTT/HR) in a new scheme of anticoagulant monitoring
Authors: Branson Herman E. , Gupta Naveen , Slater Lewis M. , Fagin Awilda , Armentrout Steven A.
Published: February 2007

Activated partial thromboplastin time after heparin removal (aPTT/HR), a test employing anion exchange chromatography, was devised as an alternative to the prothrombin time after heparin removal (PT/HR) to monitor simultaneous anticoagulation with heparin and coumarins. The potential utility of the aPTT/HR was assessed by performing parallel PTs and aPTTs on 62 consecutive plasmas from coumarin‐treated outpatients. All samples had 0.2 units/ml of heparin added and then removed to see if the maneuver influenced therapeutic group assignment. In no instance did reassignment occur. A conditional Irwin‐Fisher test (P = 0.000604) and a special multinomial trial analysis (P = 0.002) indicated that the aPTT would be at least comparable to the PT for following coumarin antithrombotic prophylaxis. Since the heparin removal procedure had no influence on therapeutic categorization, the same statistical proof could be applied to the relationship between aPTT/HR and PT/HR. This study indicates that the aPTT can be used to monitor all stages of heparin and/or coumarin anticoagulation.
Red cell enzyme activity during blood storage and reactivation of phosphofructokinase
Authors: Noble Nancy A. , Tanaka Kouichi R. , Myhre Byron A. , Johnson Delores E.
Published: February 2007

Activities of 21 red cell enzymes were measured in 15 units of CPD‐A1 anticoagulated blood before and after 35 days of storage. Paired t‐tests revealed that five of the enzyme activities decreased significantly after 35 days of storage (P < 0.05) while four increased significantly after storage (P < 0.05). The most striking change was in phosphofructokinase (PFK), which decreased an average of 33% (P < 0.005). Mean lactate dehydrogenase activity increased to 128% of its zero‐day activity (P < 0.005). Percent red cell survival, determined by reinfusion of a 51Cr‐tagged aliquot of stored blood into the autologous donor, was highly correlated with 35‐day ATP levels (r = 0.97; P < 0.001). No significant relationships between two measures of red cell survival and any of the nine significantly altered enzymes were found.

Studies of reactivation of PFK in hemolysates using 4 mM concentrations of numerous compounds indicate that the compounds or combinations of compounds used which reactivate PFK significantly all contain at least one high‐energy phosphate group. These data suggest that loss of PFK activity during storage may be due to loss of organic phosphates. Whether the changes described have an important role in blood transfusion remains to be determined.
Acquired cyclic neutropenia: Successful treatment with prednisone
Authors: Rodgers George M. , Shuman Marc A.
Published: February 2007

A previously healthy woman developed severe, periodic neutropenia after ingestion of phenylbutazone. Oscillations in the monocyte count and hemoglobin concentrations also occurred. The neutropenic episodes were associated with severe bacterial infections requiring hospitalization. Lithium induced a transient interruption in the neutropenia, but continued use was ineffective. Prednisone in a dosage of 100 mg daily successfully interrupted the neutrophil cycling and prevented infection. The patient has remained in remission on 10 mg of prednisone on alternate days.
Iron overload in hereditary spherocytosis: Association with hla‐linked hemochromatosis
Authors: Edwards Corwin Q. , Skolnick Mark H. , Dadone Mary M. , Kushner James P.
Published: February 2007

Heavy iron loading is a rare clinical finding in patients with hereditary spherocytosis. A pedigree was studied in which the proband, a 38‐year‐old man, had both hereditary spherocytosis and overt hemochromatosis. He had never received blood transfusions. The 8‐year‐old son of the proband also had hereditary spherocytosis and mildly increased iron stores. The 39‐year‐old brother of the proband did not have spherocytosis but did have overt hemochromatosis. Since hemochromatosis is transmitted as an HLA‐linked autosomal recessive disorder, HLA haplotypes serve as markers of hemochromatosis alleles. In this pedigree only individuals with two hemochromatosis alleles (homozygosity) had heavy iron loads, whether hereditary spherocytosis was present or not. The presence of hereditary spherocytosis may have contributed to the magnitude of the iron loading but was not a major factor. Our findings suggest that nontransfusional hemochromatosis found in association with hereditary spherocytosis is due primarily to homozygosity for hemochromatosis.
Chronic lymphocytic leukemia: A monoclonal disease
Authors: Solanki Dilip L. , McCurdy Paul R. , Macdermott Richard P.
Published: February 2007

A black woman with chronic lymphocytic leukemia (CLL) was found to have monoclonal B lymphocytes with one type of surface immunoglobulin and one variant of G6PD (glucose‐6‐phosphate dehydrogenase) (G6PD A). Erythrocytes and T cells contained both G6PD A and G6PD B and hence were of polyclonal origin. The CLL cells in this patient likely arose from a developmental stage later than the step of differentiation into T and B lymphocytes. Furthermore, her erythrocytes did not arise from a stem cell affected by the CLL process.
Transfer of pelger‐huet anomaly by bone marrow transplantation
Authors: Barton Ann M. , Macleod Beth C. , Thomas E. D.
Published: February 2007

A marrow transplant was performed on a patient with refractory acute myelomonocytic leukemia using marrow from a sibling donor with congenital Pelger‐Huet anomaly (P‐H). The patient, free of P‐H prior to transplant, subsequently acquired the anomaly, thus providing a useful means of demonstrating engraftment of donor cells.
Prediction of platelet production during chemotherapy of acute leukemia
Authors: Bessman David
Published: February 2007

Mean megakaryocyte ploidy, mean platelet volume, and platelet count were measured during 17 courses of chemotherapy for acute nonlymphocytic leukemia. During the myelosuppression from chemotherapy, all three variables fell; during recovery, megakaryocyte ploidy rose 1–2 days before platelet volume, which in turn rose 1–2 days before platelet count. Serial platelet volumes and counts of these patients were compared to the nomogram of the inverse nonlinear relation between platelet count and platelet size in reference subjects. Platelet volume became inappropriately small before platelet count fell substantially and remained small through most of the thrombocytopenic nadir. The end of the nadir was predictable 1–2 days after platelet volume increased to lie congruent with the reference nomogram. Changes in thrombopoiesis appear to occur sequentially in megakaryocyte ploidy, platelet volume, and platelet count. Changes in platelet count, and therefore the appearance of duration of the thrombocytopenic nadir, can be predicted by 1–2 days with platelet volume and 3–4 days with megakaryocyte ploidy. As platelet count rose, despite the continuing predominance of “young” platelets, MPV fell, suggesting that megakaryocyte stimulation as well as platelet age affects platelet size.
Further studies on the relationship between platelet buoyant density and platelet age
Authors: Boneu B. , Vigoni F. , Boneu A. , Caranobe C. , Sie P.
Published: February 2007

The relationship between platelet buoyant density and platelet age was investigated in eight human subjects submitted to an autologous chromium labeled platelet survival study. Platelets were isolated after isopycnic centrifugation using either discontinuous isoosmotic stractan gradients (five subjects), or various continuous and linear isoosmolar gradients (three subjects). A paradoxical radioactivity enrichment of the dense platelets and a premature loss of radioactivity in the light platelets were observed. These results are explained by a shift of the radioactivity distribution curve toward higher densities during the 3–4 days after platelet injection, while the standard deviation of the distribution was conserved throughout the platelet life span. These results suggest that young platelets are heterogeneous and slightly less dense than the total platelet population.
Sideroblastic anemia terminating in myelofibrosis
Authors: Lukowicz Daniel F. , Myers Thomas J. , Grasso Joseph A. , Albala Maurice M.
Published: February 2007

Two patients with primary acquired sideroblastic anemia who eventually developed myelofibrosis with myeloid metaplasia are reported. Splenectomy was performed in one patient because of increasing blood transfusion requirements, and splenic histology revealed both myeloid metaplasia and ringed sideroblasts. The second patient showed a partial response of the sideroblastic anemia to pyridoxine therapy during the early stages of development of the myelofibrosis. Myelofibrosis with myeloid metaplasia may represent a nonspecific response of the marrow tissue to sideroblastic anemia as to other primary hematologic disorders.
Hemoglobin osler: Report of a new family with exercise studies before and after phlebotomy
Authors: Butler William M. , Spratling Larry , Kark John A. , Schoomaker Eric B.
Published: February 2007

Five members of a new family with hemoglobin Osler are described. Exercise studies were performed before and after phlebotomy in one patient. These studies demonstrated that anaerobic threshold was directly related to hematocrit. Our data suggest that the elevation of red cell mass seen with this high oxygen affinity hemoglobin is an appropriate physiologic response which improves tissue oxygen delivery. Despite the increase in whole blood viscosity associated with erythrocytosis, our data do not support the use of therapeutic phlebotomy to improve exercise performance in patients with hemoglobin Osler.
Absence of prolonged benefit of initial leukapheresis therapy for hairy cell leukemia
Authors: Golomb Harvey M. , Kraut Eric H. , Oviatt David L. , Prendergast Edward J. , Stein Richard S. , Sweet Donald L.
Published: February 2007

Four patients with the leukemic phase of hairy cell leukemia were treated with leukapheresis. Two patients failed to respond, and the other two had only transient responses; hematologic improvement lasted one month in one case and four months in the second. The patient with a four‐month response underwent a second series of leukapheresis resulting in a response lasting at least 8 months. Two of the four patients subsequently had an adequate trial of prolonged chlorambucil therapy and continued to have a clinical response. We conclude that leukapheresis has little to offer to the majority of hairy cell patients for the long‐term management of post‐splenectomy patients who develop the leukemic phase. However, an occasional patient can have a transient, and even, rarely, a prolonged response.
Automated analysis of reticulocytes using fluorescent staining with both acridine orange and an immunofluorescence technique
Authors: Seligman Paul A. , Allen Robert H. , Kirchanski Stefan J. , Natale Peter J.
Published: February 2007

We have used an automated system to measure serial reticulocyte samples from phlebotomized rats and rats treated with the chemotherapeutic agent actinomycin‐D. In performing the automated samples, we utilized two different methods. The first method employed a fluorescent dye, acridine orange, which stains RNA in a manner similar to supravital stains presently used in performing reticulocyte counts. The second method is unique in that we utilized an immunofluorescent assay with antitransferrin recptor antibody which identifies the transferrin receptor found on the cell surface of reticulocytes but not on mature red cells. Both methods gave reproducible values in estimating the increase in reticulocytes seen after phlebotomy and the initial reticulocytopenia followed by recovery of the peripheral blood reticulocyte count seen after treatment with actinomycin‐D. Also, since the antitransferrin receptor antibody appears to recognize the more immature reticulocytes, both methods taken together may provide an estimate of the maturity of a population of reticulocytes.
Quantitation of hemoglobin biosynthesis with agarose isoelectric focusing
Authors: Leary Anne G. , Porter Pamela N. , Ogawa Makio
Published: February 2007

Isoelectric focusing in polyacrylamide gels has been widely used to separate fetal (HbF) and adult (HbA) hemoglobins, and the relative synthesis of HbA and HbF has been estimated by fluorography or autography of dried gels. In order to measure the absolute synthesis of hemoglobin, we developed a system that utilizes isoelectric focusing in agarose and quantitates the total radioactivity of separated hemoglobins. After isoelectric focusing, the protein bands are individually eluted from the agarose and the radioactivity measured by liquid scintillation counting. We used this technique to study the synthetic capabilities of erythroid precursors at sequential times in culture. As previously reported, the relative ratio of HbF decreased over time in culture. However, our results clearly revealed that the absolute synthesis of HbF did not decrease until there was a parallel decrease in hemoglobin A, and that changes in the relative ratio occur because of disproportionate increases in HbA. This methodology, allowing independent evaluation of the radioactivity in synthesized hemogrlobin, enabled us to gain new insight into the biosynthetic capabilities of erythroid precursors in clonal cell culture.
The in vitro effects of vincristine on peripheral blood leukocyte progenitor cells (CFU‐C) in patients in blast crisis of chronic granulocytic leukemia: Correlation with clinical response
Authors: Goldberg Jack , Zamkoff Kenneth W. , Nelson Douglas A. , Davey Frederick R. , Gottlieb Arlan J.
Published: February 2007

The in vitro sensitivity of circulating progenitor cells (CFU‐C) of 20 patients in blast crisis of chronic granulocytic leukemia (CGL) to vincristine was correlated with the clinical response to vincristine in vivo. Eleven patients who achieved either a good or partial clinical response displayed a reduction in the number of colonies or clusters formed by their peripheral blood leukocytes in a double layer agar culture assay following incubation with vincristine. The CFU‐C of five of six patients who failed to respond clinically to vincristine and prednisone were not suppressed following incubation with up to 12 μM vincristine. Three additional patients were not evaluable due to early post‐treatment deaths. In vitro assay of the effects of vincristine on CFU‐C appears to have predictive value for in vivo response in blast crisis of CGL.
Two cases of phosphofructokinase deficiency associated with congenital hemolytic anemia found in Japan
Authors: Tani Kenzaburo , Fujii Hisaichi , Takegawa Susumu , Miwa Shiro , Koyama Wataru , Kanayama Masaaki , Imanaka Akihiro , Imanaka Fumio , Kuramoto Atsushi
Published: February 2007

Two kindreds of phosphofructokinase (PFK) deficiency associated with congenital nonspherocytic hemolytic anemia and mild myopathy were found in Japan. Both probands had jaundice, gallstones, and slight to moderate degree of exercise intolerance. They showed decreased level of red cell PFK activity and no increase of blood lactate in forearm ischemic exercise test. We studied these probands' red cell PFKs by partial purification and condensation. Muscle type isozyme of PFK in both cases was not demonstrable in starch gel electrophoresis and DEAE‐Sephadex chromatography. The clinical symptoms are considered to be due to a defect of muscle type isozyme.
An unusual autoimmune hemolytic anemia in a patient with immunoblastic sarcoma
Authors: Freedman John , Cheng Lim Fang , Murray David , Myers Robert
Published: February 2007

This report describes a patient with immunoblastic sarcoma and autoimmune hemolytic anemia with reticulocytopenia and red cell hypoplasia. Both IgG and IgM red cell autoantibodies were present in the patient's serum, and both antibodies had the uncommon specificity of anti‐IT. This report appears to be the first to describe IgG auto anti‐IT and autoimmune hemolytic anemia in a patient who did not have Hodgkin's disease; the laboratory findings and clinical course are described. The reticulocytopenia and red cell hypoplasia may have been immunologically mediated. The patient responded to plasma exchange, corticosteroids, and chemotherapy.
An unusual neurologic reaction following polyvalent pneumococcal vaccine in a patient with hairy cell leukemia
Authors: Friedland Michael L. , Wittels Edward G.
Published: February 2007

A patient with hairy cell leukemia experienced two episodes of an unusual neurologic disturbance associated with vaccination. The association with polyvalent pneumococcal vaccine has not previously been noted.
Membrane proteins on human megakaryocytes and platelets identified by monoclonal antibodies
Authors: Thiagarajan Perumal , Perussia Bice , De Marco Luigi , Wells Katherine , Trinchieri Giorgio
Published: February 2007

We describe five monoclonal antibodies that react with four discrete antigens present on human platelets. Antibodies B2.12 and B59.2 precipitate the glycoprotein IIb‐IIIa complex from radiolabeled platelet membrane extracts and inhibit platelet aggregation induced by adenosine diphosphate (ADP), collagen, or epinephrine. The antigen recognized by the two antibodies is present on megakaryocytes but either absent entirely or expressed in small amounts on platelets from Glanzmann's thrombasthenic patients. The antigen recognized by antibody B37.3 is absent from thrombasthenic platelets. Antibody B1.12 reacts with an antigen shared by platelets and 20% of peripheral blood lymphocytes and is a potent inducer of platelet aggregation. Antibody B2.10 reacts specifically with platelets and megakaryocytes but does not affect platelet functions. Thus, these reagents are useful tools in diagnostic and functional studies of both normal and abnormal platelets.
Autoimmune thrombocytopenic purpura. Comparison of three different methods for the detection of platelet antibodies
Authors: Vizcaíno Gilberto J. , Diez‐Ewald María
Published: February 2007

An evaluation of three techniques for detection of antiplatelet antibodies in ATP is made. The platelet factor 3 availability after immunoinjury and the immunohystochemical technique using immunoperoxidase were more sensitive than 14C‐5HT release from platelets for this purpose. The percentage of positive cases was similar in acute and chronic ATP patients while it significantly fell in cases in remission. A high proportion of patients in clinical remission had normal platelet aggregation, despite the persistence of antiplatelet antibodies.
Lymphocytotoxic antibody is a predictor of response to random donor platelet transfusion
Authors: Hogge Donna E. , Dutcher Janice P. , Aisner Joseph , Schiffer Charles A.
Published: February 2007

The transfusion records of 189 patients with acute leukemia were analyzed to correlate lymphocytotoxic antibody (LCTAb) levels with response to a series of random‐donor platelet transfusions (Tx). Twenty‐one patients were studied twice at times of different LCTAb levels. All transfusions were given when patients were clinically stable without disseminated intravascular coagulation, bleeding, temperature > 101°F, or splenomegaly. The mean 1‐hr and 24‐hr corrected count increments (CCI) for all patients with negative LCTAb were 16,100 and 12,000, and values for patients with positive LCTAb were 5,600 and 2,600 (P < 0.0005). Thirteen patients had intermediate LCTAb (10–20%) and a variable response to Tx. Of the 137 patients with negative LCTAb levels 106 (77%) had good mean CCI of > 10,000 at 1 hr and > 7,500 at 24 hr following transfusion. In contrast of 60 patients with positive LCTAb (> 20% cytotoxicity), 53 (88%) had poor CCI of < 10,000 at 1 hr and < 7,500 at 24 hr after transfusion. Only 4 patients with positive LCTAb had a good response to random donor platelets at both 1 and 24 hrs. Eighteen patients had negative LCTAb with a high 1‐hr and low 24‐hr CCI. Thirteen of these had a history of positive LCTAb and in 9 there was an anamnestic rise following transfusion. Nine of 137 patients had negative LCTAb with low 1‐hr and 24‐hr CCI. LCTAb is highly predictive of response to random donor platelets. Cytotoxicity to > 20% of tested lymphocytes virtually precludes a good CCI at 1 and 24 hr.
Erythroblastic transformation of chronic granulocytic leukemia: A clinical and cytogenetic case study
Authors: O'Malley F. M. , Garson O. M. , McGrath K. M. , Elliott P. J. , Whiteside M. G.
Published: February 2007

A 33‐year‐old male with Philadelphia chromosome‐positive chronic granulocytic leukemia developed an erythroblastic transformation of the disease that was characterized cytogenetically by marked hyperdiploidy with multiple numerical and structural abnormalities. It is suggested that such bizarre chromosome abnormalities may be helpful in the diagnosis of this variety of transformation.
Decreased osmotic fragility in heritable canine myopathy
Authors: Abhold Raymond H. , Hegreberg Gerald A. , Magnuson James A.
Published: February 2007

Osmotic fragility was examined in red blood cells from dogs with a heritable muscle disorder that clinically resembles a muscular dystrophy. Several erythrocyte abnormalities have been reported in partients with certain forms of muscular dystrophy and it is thought that these changes reflect genetically induced alterations in the plasma membrane. It is believed that the examination of erythrocytes may eventually lead to the understanding of membrane involvement in muscle disorders. In this study, the mean osmotic fragility was found to be significantly lower in affected cells than in normal cells. These differences were maintained regardless of changes in incubation temperature (5°, 20°, or 35°C) and pH (6.5. 7.0, 7.5. or 8.0). Quantitative analysis of glycolytic metabolites and adenine nucleotide concentrations revealed little variance between erythrocytes from normal and affected animals. Similarly, the pattern of membrane protein phosphorylation in intact crythrocytes from affected animals did not differ from that observed when erythrocytes from normal animals were examined. Of the red cell indices measured, the erythrocyte count in affected animals was moderately increased, but both the mean corpuscular volume and mean corpuscular hemoglobin content were significantly reduced. From these data it is concluded that the decrease in osmotic fragility cannot be explained by differences in cell metabolism or energy production. However, the decrease in affected cell mean corpuscular volume and mean corpuscular hemoglobin content may be correlated with the decrease in osmotic fragility in a manner similar to that observed in the hemolytic disorder of beta‐thalassemia.
Sustained post‐transfusion granulocyte count increments following transfusion of leukocytes obtained from donors with chronic myelogenous leukemia
Authors: Schiffer Charles A. , Aisner Joseph , Dutcher Janice P. , Wiernik Peter H.
Published: February 2007

Leukocyte transfusions from patients with chronic myelogenous leukemia (CML) and elevated WBC counts were given to 14 patients with acute leukemia for the treatment of 16 infectious episodes. The WBCs were not irradiated to determine if engraftment with production of granulocytes would occur following infusion of immature myeloid elements. No recipient was alloimmunized by clinical and serologic criteria. High leukocyte yields were obtained using a variety of differential centrifugation techniques with a mean WBC yield/transfusion of 95 × 109 (range 19–275). A mean of 2.5 transfusions (range 1–11) were administered/recipient with a mean total dose of 235 × 109 WBC/transfusion episode (range 50–590). Seven patients had granulocyte counts greater than 500/μl for four or more days (range 4–11 days) following the last transfusion. Ph1 chromosomes were documented in 2 of 4 patients tested 2–8 days following transfusion. Leukocyte alkaline phosphatase scores were increased (>150) in 5/5 recipients tested post‐transfusion demonstrating that production of this enzyme can be induced in CML granulocytes. Except for one severe transfusion reaction, there were no significant side effects and no recipient developed signs of graft versus host disease. All patients with sustained increments demonstrated rapid clinical improvement including 3 severely infected, poor risk patients undergoing initial induction therapy. These 3 patients all achieved complete remission with no evidence of the Ph1 chromosome. Because of the high dose of WBC which can be collected and the salutary effect of continued leukocyte production, CML WBC may be the preparation of choice for selected, non‐alloimmunized, severely infected patients.
Thrombocytopenic purpura with giant platelets and ultrastructural platelet defects
Authors: Vizcaíno Gilberto J. , Diez‐Ewald Maria
Published: February 2007

The case of a patient with thrombocytopenia and giant platelets is presented. Platelet function tests including bleeding time, aggregation, availability of PF3, and serotonin uptake and release were abnormal. A bone marrow aspirate showed megakaryocytes that while not increased in number, were young in appearance. Electron microscopy of platelets revealed a dilated surface‐connecting canalicular system and a prominent membrane complex. No similar findings were encountered among the patient's relatives. The various diagnostic possibilities in this patient are discussed.
Protein kinases in normal human blood cells
Authors: Tuy Françoise Phan Dinh , Henry Joëlle , Rosenfeld Claude , Kahn Axel
Published: February 2007

Protein kinases active on basic and acidic artificial substrates were investigated in normal human erythrocytes, platelets, polymorphonuclear and mononuclear cells. These two types of protein kinases were partially purified by affinity chromatography, then assayed for their enzymatic activity using [γ‐32P] ATP or GTP as phosphoryl donor. Partially purified kinases active on acidic substrates were subjected to high‐performance liquid chromatography (HPLC). Protein kinases active on basic sustrates were analyzed by cellulose acetate electrophoresis of crude cellular extracts and the influence of 3′5′ cyclic AMP was studied. Three forms of casein‐phosvitin kinases could be distinguished according to their molecular weight (165 K, 38 K, and 31 K). The 165 K species, in contrast to the light species, can use GTP instead of ATP as phosphoryl donor and corresponds to the “so‐called” casein kinase 2. This form is very sensitive to proteolysis and, when partial purification is performed without the addition of various antiproteolytic agents, it is degraded into 120–135 K and 105–115 K active species; this artefactual degradative process is especially active in platelet extracts. As many as eight different active bands of histone and protamine kinases can be separated by cellulose acetate electrophoresis, several of them being stimulated by cyclic AMP. Isozymic patterns of protein kinases, levels of activity on the different substrates, and utilization of ATP and GTP were found to be specific for each cell type. These results suggest the possibility of using protein kinases as markers for cell differentiation.
Studies on the sequestration of chemically and enzymatically modified erythrocytes
Authors: Smedsrød Bård , Aminoff David
Published: February 2007

We have developed an experimental animal model to establish the patterns of sequestration of untreated, as well as chemically and enzymatically modified red blood cells (RBC). The intraperitoneal route of transfusion provides a useful way of transferring large numbers of untreated RBC into small animals and assuring their introduction into the circulation within 24 hr. Moreover, this route “filters” some types of modified erythrocytes, eg, glutaraldehyde treated RBC. From the pattern of sequestration, the RBC in the peritoneal cavity then pass through the liver where other types of modified RBC are sequestered, eg, after trypsin, pronase, protease, or sialidase treatment. Some modified RBC show a preference for the spleen as the site of sequestration, eg, galactose oxidase or N‐ethylmaleimide treated RBC. These appear in the spleen despite intraperitoneal transfusion. Relevant to this study is the observation that in the rat old RBC are sequestered both by liver and spleen, while asialo‐erythrocytes are sequestered by liver only. A possible reason for this difference is discussed in the text.
Alterations of nuclear size profiles in AKR/J mice with spontaneous or transplanted leukemia
Authors: Schwartz G. N. , Patrene K. D. , Boggs S. S.
Published: February 2007

Electronic sizing with a Coulter Counter was used to measure the frequency and number of cells with a large nuclear volume in tissues from AKR/J mice with spontaneous, long passaged, or first passage leukemia. The mean percentage of cells with a large nuclear volume was 1% in the blood, 4% in the spleen, 10% in the bone marrow, and 5% in the thymus of nonleukemic AKR/J mice. The percentage of cells with a large nucleus increased to 14–83% in all four tissues from AKR/J mice with spontaneous leukemia. In mice with a long passaged leukemia, a progressive increase in the percentage of cells with a large nucleus in the spleen and blood was associated with an increase in the number of cells/tissue with a large nucleus. However, in the bone marrow, an increase in percentage of cells with a large nuclear volume appeared to be the result of a decrease in the number of cells/humerus with a small nuclear volume. In the thymus, there was only a slight increase in the percentage of cells with a large nucleus. In mice with a first passage leukemia, there was a progressive increase, until death of the animals (21 ± 1.9 days), in thymus weight and in the percentage of cells with a large nucleus. Also, the number of cells with a large nucleus increased in the spleen and bone marrow until day 12. Thereafter, there was a decrease in the total number of cells and number of cells with a large nucleus.
Myeloid differentiation antigen defined by a monoclonal antibody IF10
Authors: Yamada K. , Okabe N. , Saito H. , Suzuki R. , Kumagai K.
Published: February 2007

A monoclonal antibody of IgM class that defines the antigen present on human peripheral blood granulocytes was produced and characterized. This monoclonal antibody (IF10) was made from a single fusion between P3‐X63‐Ag8‐U1 (P3U1) myeloma cells and splenoctyes from a BALB/c mouse immunized against human cultured monocytoid cell line THP‐1 cells. IF10‐defined antigen(s) was expressed on the cells of granulocyte lineage such as peripheral blood granulocytes and metamyelocytes, myelocytes, promyelocytes, and a part of myeloblasts in the normal bone marrow, whereas it was not detected on resting and activated T lymphocytes, B lymphocytes, adherent monocytes, and thymocytes. The IF10‐defined antigen was also expressed on cultured monocytoid cell lines as well as myeloid and myeloid/erythroid (K‐562) cell lines. T lymphoblastoid cell lines, particularly those representing the T cells at an early thymocyte level, and a part of null cell lines were also reactive to IF10. Therefore, IF10 may be a unique monoclonal antibody that defines an antigen(s) which is expressed on almost whole stages of granulocytes and early stages of macrophages and T‐cell lineages, and possibly during very early stages of erythroid lineage.
Requirements for growth of human erythroid progenitors in nutrient agar
Authors: Januszewicz E. , Cooper I. A. , Bradley T. R.
Published: February 2007

Human erythroid bursts and neutrophil colonies from fetal liver, term cord blood, and adult bone marrow were grown in nutrient agar, and the colonies were scored using a fixed whole plate method. Human placental conditioned medium was shown to contain burst promoting activity, and low oxygen tension markedly enhanced growth of erythroid bursts. Morphological analysis of colonies showed that eosinophil colonies were more numerous in bone marrow than fetal liver and conversely macrophage and neutrophil‐macrophage colonies were more numerous in fetal liver. Erythroid bursts were more numerous and larger in fetal liver. Human fetal liver cells do not produce endogenous colony‐stimulating factor.