bluuebunny/crossref_metadata_2025 huggingface dataset:
Long-term survival and reversal of iron overload after marrow transplantation in dogs with congenital hemolytic anemia
Authors: Weiden PL , Hackman RC , Deeg HJ , Graham TC , Thomas ED , Storb R
Published: October 2019

Severe hemolytic in Basenji dogs secondary to pyruvate kinase deficiency was corrected by marrow transplantation from hematologically normal littermates. These dogs have now been followed for more than 5.5 yr. Essentially normal hematopoiesis has persisted, and the dogs remain in good health without cirrhosis or osteosclerosis. Furthermore, hepatic iron overload present before transplantation has gradually decreased. These results in dogs suggest that marrow transplantation could prevent the morbidity and mortality of severe hemolytic anemia and associated iron overload in man.
Quantitation of three types of gamma chain of HbF by high pressure liquid chromatography; application of this method to the HbF of patients with sickle cell anemia or the S-HPFH condition
Authors: Huisman TH , Altay C , Webber B , Reese AL , Gravely ME , Okonjo K , Wilson JB
Published: October 2019

A modification of a high pressure liquid chromatographic (HPLC) procedure is described that enables the complete separation and quantitation of the A gamma T, A gamma I, and G gamma chains in human fetal hemoglobin. The method, which is fast and accurate, requires 5 to 2000 micrograms Hb F. The purity of the Hb F is not essential and admixture of up to 70% adult Hb does not interfere with the determination. The method has been applied to the Hb F of 64 Black SS patients and 7 persons with the Hb S-HPFH (G gamma A gamma type) conditions. (A) Both “adult” G gamma to A gamma (2:3) and “newborn” G gamma to A gamma (3:2) ratios were observed in adult SS patients, 8 yr and older. Only 12% of the SS patients had the “newborn” ratio. This high G gamma to A gamma ratio may be due to a modification of the genetic switch mechanism that regulates the change of this ratio after birth. (B) Intermediate G gamma to A gamma ratios were only found in young SS patients, 5 yr of age or less. The results suggest a delayed switch of the newborn leads to adult ratio in sickle cell anemia. (C) The A gamma T chain was present in only 6% of all SS patients. One patient is homozygous for this variant chain. (D) Three of the 7 subjects with Hb S-HPFH were positive for the A gamma T chain. Its percentage was low, which suggests that the A gamma T chain gene is in trans of the HPFH determinant. (E) Quantitation of the three gamma chain types is also possible in the Hb F from Hb S heterozygotes with (nearly) normal Hb F levels. Such an analysis is useful for an evaluation of genetic conditions involving variations in the production of (different types of) Hb F.
Prevention of graft rejection following bone marrow transplantation
Authors: Gale RP , Ho W , Feig S , Champlin R , Tesler A , Arenson E , Ladish S , Young L , Winston D , Sparkes R , Fitchen J , Territo M , Sarna G , Wong L , Paik Y , Bryson Y , Golde D , Fahey J , Cline M
Published: October 2019

Bone marrow transplantation from an HLA-identical sibling is increasingly used in the treatment of severe aplastic anemia. One major problem with this approach is graft rejection that occurs in 25%-60% of patients conditioned for transplantation with cyclophosphamide. At most transplant centers it has been difficult to accurately identify patients at high risk for graft rejection. We studied a conditioning regimen of cyclophosphamide (200 mg per kg) and low-dose total body irradiation (3 Gy; equivalent to 300 rad) in 23 consecutive unselected patients with aplastic anemia followed for a minimum of 6 mo. There was only one episode of graft rejection. Graft-versus-host disease and interstitial pneumonitis were not increased by the more intensive conditioning regimen. Actuarial survival was 61% at 1 yr and 49% at 2.5 yr. Cyclophosphamide and low-dose total body irradiation is an effective conditioning regimen in patients with aplastic anemia. It may be particularly useful when accurate predictive tests of graft rejection are not available as is the case in most transplant centers.
Role of kidney in the catabolic clearance of human platelet antiheparin proteins from rat circulation
Authors: Bastl CP , Musial J , Kloczewiak M , Guzzo J , Berman I , Niewiarowski S
Published: October 2019

Stimulated platelets release at least two antiheparin proteins: platelet factor 4 (PF4) and low affinity platelet factor 4 (LA-PF4) from which beta-thromboglobulin (beta TG) is derived. We have found previously marked elevation of LA-PF4/beta TG antigen in platelet poor plasma of patients with chronic renal failure, whereas levels of PF4 remained normal. Therefore, we examined the role of the kidneys in the metabolic clearance of LA-PF4/beta TG and PF4. The supernates of aggregates of thrombin-stimulated human platelets were injected into sham operated control rats, nephrectomized rats, and into rats with acute ureteral ligation. The disappearance of human LA-PF4/beta TG antigen and PF4 in rat plasma determined by specific radioimmunoassays followed biphasic exponential curves. The half-lives (t1/2) for the fast and slow components of LA-PF4 in control rats were 6.4 and 68.4 min. Nephrectomy significantly increased these times to 9.7 and 144 min, while ureteral ligation resulted in no significant change. Comparison of the level of LA-PF4/beta TG antigen and of creatinine in aorta and in renal vein showed 25%-30% extraction of these compounds by the kidney. Less than 0.1% of the total LA-PF4 antigen injected was recovered in the urine of control rats. In contrast to these results, the clearance of PF4 was not affected by nephrectomy. In conclusion: (1) functional renal tissue is necessary for normal clearance of LA- PF4/beta TG, but renal excretion does not play a major role in its elimination suggesting that the protein is catabolized by the kidney; and (2) catabolic clearance of PF4 does not depend on functioning kidney tissue.
Electrophoretic mobility distributions distinguish hairy cells from other mononuclear blood cells and provide evidence for the heterogeneity of normal monocytes
Authors: Petty HR , Ware BR , Liebes LF , Pelle E , Silber R
Published: October 2019

The electrophoretic mobility distributions of hairy cells, normal monocytes. CLL, and normal lymphocytes isolated from blood were determined by electrophoretic light scattering. Values obtained for hairy cells, 1.52 X 10(-4) cm2/V . sec, were indistinguishable from that of normal monocytes. The mobility of CLL lymphocytes was similar to that of normal B cells. After exposure to neuraminidase, hairy cells revealed a homogeneous distribution with a reduced mobility of 0.55 X 10(-4) cm2/V . sec, while normal monocytes showed a heterogeneous distribution of electrophoretic mobilities suggestive of subpopulations. The electrokinetic behavior of hairy cells thus differs from that or normal and CLL lymphocytes before, and from that of monocytes after, treatment with neuraminidase. The hairy cell therefore possesses a distinct pattern of surface charge properties that clearly distinguish it from the circulating B cells, T cells, or monocytes.
Erythropoiesis in vitro: enhancement by neuraminidase
Authors: Rowley PT , Ohlsson-Wilhelm BM , Farley BA
Published: October 2019

Neuraminidase treatment of human fetal liver or adult marrow cells prior to culture results in an increased number of erythroid colonies and bursts. No increase occurs in the number of nonerythroid colonies. The number of bursts having more than eight subunits is increased preferentially. Individual burst subunits are also enlarged. Neuraminidase-treated cells yield erythroid bursts when cultured in concentrations of erythropoietin insufficient to produce bursts from untreated cells. It is proposed that (1) neuraminidase treatment of adult and fetal cell mixtures specifically stimulates differentiation of erythroid precursors, (2) the preferential stimulation of erythroid bursts having many subunits suggests a preferential susceptibility of more primitive BFU-Es, and (3) neuraminidase treatment enhances the response of erythroid precursors to erythropoietin.
Inhibition of platelet phospholipid methylation during platelet secretion
Authors: Shattil SJ , McDonough M , Burch JW
Published: October 2019

A pathway for the synthesis of membrane phosphatidylcholine involving the N-methylation of phosphatidylethanolamine has been detected in several types of mammalian cells. Furthermore, it has been implicated in the coupling of agonist binding to cell response. We examined whether human platelets exhibit this synthetic pathway and whether platelet agonists influence its activity. When washed platelets were incubated with 0.15 microM L-[methyl-3H]methionine at 37 degrees C, they incorporated methyl-3H into their phospholipids linearly at the rate of 1 pmole/10(9) platelets/hr. When incubated with 20 microM radiolabeled methionine, they incorporated about 15 pmole/10(9) platelets/hr. The radioactivity was found predominantly in phosphatidyl- N-monomethylethanolamine, phosphatidyl-N,N-dimethylethanolamine, and phosphatidylcholine. Thrombin caused an immediate (within 15 sec) and sustained (up to 30 min) decrease in the rate and extent of N- methylation of platelet phospholipids. This was accounted for by a decrease in synthesis of methylated phospholipids rather than an increase in their degradation. This thrombin effect correlated with serotonin release and could be dissociated from platelet aggregation and prostaglandin synthesis. Thrombin also decreased the synthesis of phosphatidylcholine when choline was used as the radiolabeled substrate. Other agonists such as epinephrine, adenosine diphosphate (ADP), or A23187 also decreased phospholipid methylation under conditions in which they stimulated serotonin release. These data demonstrate that platelets are capable of synthesizing phosphatidylcholine from phosphatidylethanolamine by N-methylation and that agonists perturb this pathway as they induce platelet secretion. The precise role of phospholipid methylation in either resting or stimulated platelets remains to be established.
Impaired platelet response to thromboxane-A2 and defective calcium mobilization in a patient with a bleeding disorder
Authors: Lages B , Malmsten C , Weiss HJ , Samuelsson B
Published: October 2019

Platelet aggregation, secretion, and thromboxane formation induced by various agonists, including arachidonate, prostaglandin-G2 (PGG2), and thromboxane-A2 (TxA2), were examined in a patient with a bleeding disorder who was previously reported to have a TxA2-related defect. Aggregation and 14C-5HT secretion were decreased, and no TxB2 formation occurred in response to adenosine diphosphate (ADP), epinephrine, or collagen. Arachidonate-induced aggregation and TxB2 formation, and PGG2- induced aggregation (but not TxB2 formation) were impaired at low agonist concentrations. The patient's platelets did not aggregate in response to TxA2 generated from arachidonate in normal platelets, but were capable of synthesizing TxA2 from both arachidonate and PGG2. In addition, aggregation and secretion induced by low concentrations of the ionophore A23187 were impaired in platelet-rich plasma (PRP) and in gel-filtered platelets in the absence of extracellular calcium; these responses became normal at higher A23187 concentrations or, in GFP, at low A23187 concentrations in the presence of exogenous calcium. These findings indicate that the TxA2 defect in this patient does not result from a thromboxane synthetase deficiency, but may be due to impaired mobilization of platelet calcium, and thus are consistent with the possibility that TxA2 may act as a calcium ionophore.
A radioreceptor assay for quantitating plasma factor VIII/von Willebrand's protein
Authors: Kao KJ , Pizzo SV , McKee PA
Published: October 2019

A sensitive and precise radioreceptor assay for determining plasma levels of human factor VIII/von Willebrand's factor (FVIII/vWF) has been developed by taking advantage of the FVIII/vWF receptor sites on human platelets. Paraformaldehyde-fixed platelets, which were processed and then stored, retained FVIII/vWF binding activity and therefore could be used as a convenient source of receptors. The human plasma samples to be tested were initially filtered on 4% agarose columns to concentrate the FVIII/vWF protein in the void volume and to remove the factor(s) that interferes with the assay. The percent recovery of FVIII/vWF in the pooled eluent was measured by the recovery of added trace 125I-FVIII/vWF. The coefficients of intra- and interassay variation were 6% and 10%, respectively. The plasma FVIII/vWF concentrations determined by the assay for pooled normal plasma, hemophilia A plasma, and plasmas from two patients with von Willebrand's disease were 16.3 +/- 0.5, 52.6 +/- 1.5, 6.8 +/- 0.8, and 3.2 +/- 0.2 microgram/ml, respectively. The range of plasma FVIII/vWF concentrations varied between 8.3 microgram/ml and 24.9 microgram/ml for 10 normal adults. The plasma FVIII/vWF concentrations determined by the radioreceptor assay correlated well with levels measured by the ristocetin-induced platelet aggregation method, thus demonstrating the functional relevancy of the radioreceptor assay for plasma FVIII/vWF.
Fetal liver cells produce both fetal and adult erythrocytes in semiallogeneic radiation chimeras: possible influence of adult environment
Authors: Mouchiroud G , Blanchet JP
Published: October 2019

Two kinds of erythrocytes are released in the blood of irradiated adult hybrid mice grafted with parental fetal liver cells: fetal antigen- bearing erythrocytes (Ft+ cells) and adult-type Ft- erythrocytes. Both are of parental origin, as determined by immune lysis using histocompatibility alloantigens. The latter cells make up all the recipient's red blood cells 2 mo after receipt of the graft, Ft+ cells then being no longer detected. The transient duality of erythropoiesis in irradiated adults grafted with fetal liver cells has been confirmed by studying the kinetics of CFU-E populations, as characterized by their ability to give rise to Ft+ or Ft- erythrocytes. The results are discussed in terms of environmental factors that influenc erythroid differentiation.
Embryonic hemoglobins in man and other mammals
Authors: Fantoni A , Farace MG , Gambari R
Published: October 2019

This report attempts to review the present state of research on embryonic hemoglobins in humans and other mammals from a cytologic, molecular, and clinical point of view. In all mammals, the yolk sac is the site responsible for the “primitive” erythropoiesis, which produces an erythroid population characterized by peculiar cytologic features and globin gene expression. Morphological and molecular events that underlie prenatal erythropoiesis are described herein giving rise to questions regarding biology at large (i.e., the differential activity of genes capable of similar functions); molecular biology of eukaryotic genes (i.e., globin gene organization and structural subtleties); and clinical hematology (i.e., syndromes associated with the appearance of embryonic hemoglobins.
Procoagulant activity of sarcoma sublines with different metastatic potential
Authors: Colucci M , Giavazzi R , Alessandri G , Semeraro N , Mantovani A , Donati MB
Published: October 2019

It has been suggested that cancer cell procoagulant activity influences metastasis formation by promoting fibrin deposition around tumors. We have investigated the procoagulant activity of various tumor cell sublines with different metastatic capacity derived from metastatic nodules of a murine fibrosarcoma. All the cells tested possessed a marked thromboplastin-like activity; they were, however, heterogeneous as regards the degree of procoagulant activity; the two cell lines with virtually no metastatic capacity showed 6--8 times higher procoagulant activity than the cells from the parent line; in contrast, the procoagulant activity of the two sublines with higher metastatic capacity did not differ significantly from that of the parent line. These findings support the hypothesis that fibrin is part of a defence reaction against cancer cell invasiveness.
Stromal colony-stimulating activity production and myeloid colony- forming cells in human hemopoietic and nonhemopoietic bone marrow
Authors: Schwartz RS , Greenberg PL
Published: October 2019

In order to evaluate the role of the stromal bone marrow microenvironment in regulating granulopoiesis, we have examined the capacity of adult human proximal hemopoietic (PH) and distal nonhemopoietic (DNH) long bone to produce colony-stimulating activity (CSA), characterized the cellular sources of CSA, and quantitated the colony-forming cells (CFU-GM) of marrow from these sites. Stromal elements were obtained from slices of cancellous bone. PH bone marrow stroma contained CFU-GM concentrations similar to aspirated PH marrow and significantly more CFU-GM than DNH bone marrow: 20.7 +/- 4.8/10(5) cells and 25.8 +/- 12.0/mg bone versus 0.81 +/- 0.34/10(5) cells and 0.02 +/- 0.01/mg bone (p less than 0.001). Conditioned media prepared from PH and DNH bone were quantitated for CSA by their ability to promote in vitro granulocyte colony formation of nonadherent human marrow cells. Stromal CSA production was destroyed by freeze--thawing and was radioresistant (4400 rad). Of DNH stromal cells, 15%--30% were monocyte-macrophage, but the slow absolute numbers of these cells suggested alternative CSA cellular sources in distal bones. PH stroma produced significantly more CSA than DNH bone stroma: 0.72 +/- 0.10 versus 0.30 +/- 0.06 U/mg bone (p less than 0.01). The CSA concentration gradient between PH and DNH bones may contribute to the regulation of granulopoiesis in marrow and to the absence of hemopoiesis distally.
Effect of ascorbate on abnormal neutrophil, platelet and lymphocytic function in a patient with the Chediak-Higashi syndrome
Authors: Weening RS , Schoorel EP , Roos D , van Schaik ML , Voetman AA , Bot AA , Batenburg- Plenter AM , Willems C , Zeijlemaker WP , Astaldi A
Published: October 2019

A diminished chemotactic response was observed with the neutrophils of a patient with the Chediak-Higashi syndrome, who was not in the accelerated phase of the disease. An abnormally low release of myeloperoxidase from these cells during phagocytosis was also noted; this resulted in a decreased iodination capacity and probably also caused the defect in the intracellular killing of bacteria by the neutrophils. The level of cyclic AMP in these cells was elevated, but decreased after treatment with ascorbate either in vitro or in vivo. During ascorbate therapy, the bactericidal activity of the neutrophils normalized, whereas the chemotactic response remained low. Nevertheless, the patient had significantly less infections during ascorbate therapy. The bleeding tendency, due to a storage-pool disorder of the Chediak-Higashi platelets, was unaffected by treatment with ascorbate. The patient's lymphocytes did not display any activity in antibody-dependent lymphocytotoxicity. This defect was not affected by treatment with ascorbate either.
Relationship of a leukemia-associated antigen to the presence of lymphoblasts in the peripheral blood in children with acute lymphocytic leukemia
Authors: Morgan E , Hsu CC
Published: October 2019

Peripheral blood samples from 57 children with newly diagnosed E- rosette-negative, surface-immunoglobulin negative acute lymphocytic leukemia (ALL) were studied for the presence of a leukemia-associated antigen (ALLA). Ficoll-Hypaque separated cells were tested using a rabbit antiserum to human null lymphoblasts and an indirect immunofluorescent assay. The percentage of ALLA-positive cells were compared to the percentage of lymphoblasts determined by differential counts of a Wright-Giemsa-stained smear of a concurrently obtained peripheral blood sample. The mean ratio of percentage of lymphoblasts to percentage of ALLA-positive cells was 0.90. However, in 13 patients, the ratio of percent of ALLA-positive cells to percent of lymphoblasts was equal to or greater than 2:1. In the blood of 6 additional children (5 newly diagnosed, 1 relapsed patient) in whom no morphologically identifiable lymphoblasts were detected. ALLA-positive cells were present (7%-49%). These results indicate that testing for ALLA-positive cells in a sensitive technique for detection of leukemic cells in children with ALLA-positive ALL.
Regeneration of hemopoietic precursor cells in spleen organ cultures from irradiated mice: influence of genotype of cells injected and of the spleen microenvironment
Authors: von Melchner H , Lieschke GJ
Published: October 2019

The regeneration of hemopoietic precursor cells (colony-forming cells, CFC) was monitored in spleen organ cultures from lethally irradiated mice injected with 10(7) normal syngeneic or allogeneic bone marrow cells. The important role of the microenvironment in supporting hemopoiesis was confirmed by the failure of mutant S1/S1d spleens to support CFC regeneration in organ cultures. However, the extent and quality of the CFC regeneration was clearly dependent on the genetic properties of the injected cells. Evidence for this was obtained from the regeneration patterns of various CFC types in organ cultured spleens derived from different mouse donor-recipient strain combinations (CBA/CBA, CBA/C57BL, CBA/BALB/c, C57BL/C57BL, C57BL/CBA, C57BL/BALB/c) that maintained the differences in the bone marrow frequency of various CFC types characteristic of the donor strain.
Purification of erythroblastic nests
Authors: Macario AJ , Dugan C , Perez-Lloret IL , Conway de Macario E
Published: October 2019

We describe a method for preparing purified erythroblastic nests in large numbers (approximately 10(6)/run) in three steps: (1) induction of splenic erythropoiesis in mice, (2) preparative differential centrifugation for the removal of erythrocytes and single cells from spleen cell suspensions, and (3) sedimentation in an isokinetic gradient of Ficoll 400 in Joklik's modification of minimum essential medium. Viability of isolated EN is very high, as demonstrated by the trypan blue exclusion and in vitro erythrocyte formation methods.
Rapid destruction of newly synthesized excess beta-globin chains in HbH disease
Authors: Sancar GB , Cedeno MM , Rieder RF
Published: October 2019

A subject with HbG Philadelphia-HbH disease exhibited an unusually high alpha/beta synthesis ratio; when peripheral blood was tested in vitro on several occasions, ratios of 0.63 - 0.89 were obtained after incubations of 30–120 min. HbH amounted to 5%-8% of the circulating hemoglobin. Rapid destruction of excess newly synthesized beta-globin was demonstrated in kinetic and pulse-chase experiments. After 2 min of incubation, the alpha/beta synthesis ratio was 0.48; this figure rose to 0.89 by 30 min. The zero time alpha/beta ratio was estimated to be 0.35. The degradation of beta-chains was calculated to proceed at approximately one-half the rate of beta-globin synthesis; this result was confirmed by the loss of 50% of the specific activity in beta- chains during 9 min of a chase experiment following a 10-min radioactive pulse. The results suggest that efficient proteolysis may be responsible, in some blacks, for the low levels of excess beta- globin chains in HbH disease as well as for the mildness of the clinical disorder.
Terminal differentiation of human promyelocytic leukemic cells in primary culture in response to retinoic acid
Authors: Breitman TR , Collins SJ , Keene BR
Published: October 2019

The recent finding that retinoic acid induces terminal granulocytic differentiation of the human promyelocytic leukemia cell line, HL-60, prompted an investigation of the sensitivity to this inducer of human myelocytic leukemia cells in primary suspension culture. Of the 21 leukemic specimens, only cells from the two patients with acute promyelocytic leukemia differentiated in response to retinoic acid. After an incubation period of 5--7 days in 1 microM retinoic acid, the cells from these two patients showed extensive morphological and functional maturation. Thus, because it appears that retinoic acid specifically induces granulocytic differentiation of leukemic promyelocytes, this compound may have therapeutic utility in the treatment of acute promyelocytic leukemia.
On the preservation of contractile proteins during storage of human platelets
Authors: Lucas RC , Lawrence J , Stracher A
Published: October 2019

A study has been undertaken to determine the rate at which stored platelets lose their ability to respond to stimuli and to establish whether this decrease in function could be ascribed to the storage- induced proteolysis of prominent platelet proteins observed by others. Platelet concentrates were stored at 4 degrees C and 25 degrees C for up to 14 days, and their ability to secrete and aggregate in response to appropriate stimuli was determined at 6, 96, and 192 hr after venipuncture. At each time point the protein complement of the platelets was also monitored by SDS-polyacrylamide gel electrophoresis to assess the extent of intracellular protein degradation. Platelets from concentrates stored at either temperature exhibited a decreased ability to respond to stimuli as storage time increased. After 8 days of storage at 4 degrees C and up to 9 days at 25 degrees C, no proteolysis of major platelet proteins was observed; however, complete loss of platelet function was observed. This strongly indicates that a decrease in platelet function should not be causally linked to degraded contractile-structural proteins and that extending the functional life of platelets during storage is still an attainable goal since proteolysis is not the inevitable result of short-term storage.
The effect of alpha-thalassemia on the expression of the beta- thalassemia/HPFH heterozygote in a black family
Authors: Beutler E , Turner E , Kuhl W
Published: October 2019

A 2-yr-old black girl presented with a thalassemic clinical picture and was found to have nearly 100% fetal hemoglobin in her red cells. Pedigree analysis indicated that she was a heterozygote for the hereditary persistence of fetal hemoglobin gene and for a beta O- thalassemia gene. A brother, who also had nearly 100% fetal hemoglobin in his red cells, manifested, in contrast to his sister, no anemia and only minimal splenomegaly. Examination of the family's alpha-globin loci using the restriction endonuclease Eco Rl demonstrated that the brother had a single alpha-locus deletion that he had inherited from his mother. The mild clinical manifestations of this boy are consistent with the often expressed view that excess alpha chains may contribute significantly to the hematologic manifestation of beta-thalassemia.
Recategorizing childhood acute lymphoblastic leukemia with monoclonal antibodies to human T cells
Authors: Sondel PM , Borcherding W , Shahidi NT , Ganick DJ , Schultz JC , Hong R
Published: October 2019

The lymphoblasts of three patients with childhood acute lymphoblastic leukemia (ALL) were analyzed for their immunologic surface markers. Blasts from two of these patients did not form rosettes with sheep erythrocytes and the third did so marginally, suggesting these patients had non-T-cell leukemia. These blasts were also tested with monoclonal antibodies that detect thymocyte differentiation markers, and all three patients were highly reactive with at least two of these reagents. We anticipate the availability of multiple standardized monoclonal reagents will necessitate a recategorization of ALL phenotypes. Some of these leukemic phenotypes may not correspond to normal stages of lymphoid differentiation. Therefore, we suggest that it may be inappropriate to attempt to identify and categorize leukemic cells by the pathways of normal differentiation.
The factor VIII complex: structure and function
Authors: Hoyer LW
Published: October 2019

Normal human plasma contains a complex of two proteins that are important in hemostasis and coagulation. The factor VIII procoagulant protein (antihemophilic factor) and the factor VIII-related protein (von Willebrand factor) are under separate genetic control, have distinct biochemical and immunologic properties, and have unique and essential physiologic functions. While the nature of their interaction and the details of the biochemical structures remain to be determined, the information now available permits a preliminary understanding of the molecular defects in hemophilia and von Willebrand's diseases.
The heritable nature of clonal characteristics in acute myeloblastic leukemia
Authors: McCulloch EA , Buick RN , Curtis JE , Messner HA , Senn JS
Published: October 2019

Marked patient-to-patient variation is observed when blood or marrow from AML patients is examined using colony methods in culture. Concentrations of the progenitors of colonies change with time during the course of the disease. We asked whether blast progenitor properties were more stable. We measured blast cell self-renewal and drug sensitivity (adriamycin and cytosine arabinoside) repeatedly in the courses of seven AML patients. These properties were found to be stable or slowly evolving. We conclude that capacity for renewal and sensitivities to certain chemotherapeutic drugs are heritable characteristics in leukemic clones.
Distribution of complement receptor subtypes in non-Hodgkin's lymphomas of B-cell origin
Authors: Cossman J , Jaffe ES
Published: October 2019

Surface receptors specific for either the C4b (CR1) or C3d (CR2) component of complement were examined on the neoplastic cells from 30 cases of non-Hodgkin's lymphoma of B-cell origin and on cells derived from 9 normal lymphoid tissues. Lymphocyte suspensions from non- neoplastic peripheral blood, tonsils, and lymph node contained three categories of complement receptor lymphocytes (CRL): cells with receptors for both C4b and C3d (CR1+, CR2+); cells with receptors for C4b but not C3d (CR1+, CR2-), and cells with receptors for C3d but not C4b (CR1-, CR2+). The mean of the proportion of total CRL expressing receptors only of C3d (CR1-, CR2+) was 0.35 for non-neoplastic tissues and 0.28 for malignant lymphomas of follicular center cell (FCC) origin. However, the proportion of cells with this phenotype was significantly higher in well differentiated lymphocytic lymphomas (WDL) and chronic lymphocytic leukemia (CLL) (0.65) and in intermediately differentiated lymphocytic lymphomas (IDL) (0.59). Histologic compartmentalization of the CRL subtypes was observed in frozen sections of normal lymphoid tissue. CR1+ cells were present in lymphoid follicles interfollicular areas, and in splenic red pulp. CR2+ cells were confined to lymphoid follicles. These findings strongly suggest that complement receptor phenotypes may be useful markers of B-cell differentiation.
Immunoblastic sarcoma of T-cell versus B-cell origin: I. Clinical features
Authors: Levine AM , Taylor CR , Schneider DR , Koehler SC , Forman SJ , Lichtenstein A , Lukes RJ , Feinstein DI
Published: October 2019

Within the Lukes-Collins classification system of malignant lymphoma, a tumor of large transformed lymphocytes, termed immunoblastic sarcoma (IBS), is described. This morphological type would have been included within the “histiocytic” category of Rappaport. Immunoblastic sarcoma may be of B-lymphocytic or T-lymphocytic origin. Since differences or similarities of these two variants have not yet been described, we reviewed the case histories of 35 such patients, all of whom had immunologic marker studies performed. Nineteen patients had T-cell IBS (T-IBS), whereas 16 had B-cell IBS (B-IBS). Median age for both groups was approximately 50 yr. A history of prior immune disorder was found in 31% of B-IBS and 16% of T-IBS cases. Prior lymphoproliferative malignancy was noted in 21% of T-IBS and 13% of B-IBS patients. All T- IBS patients first presented because of lymphadenopathy, whereas 56% of B-IBS cases initially presented because of extranodal disease. Systemic “B” symptoms were common in both. Similarly, most patients had widespread disease (stage III or IV) at diagnosis. Clinically suspected hepatic (p = 0.05) and retroperitoneal node (p = 0.01) involvement were more often found in T-IBS. Forty-one percent of T-IBS patients demonstrated polyclonal hypergammaglobulinemia, a finding seen in no B- IBS patient (p = 0.02). Although not statistically significant because of small numbers of patients, data on therapy and survival suggest that IBS of B-cell type may be successfully treated with aggressive, multiagent chemotherapy, while alternative approaches appear warranted in T-cell disease.
Clinical pharmacology of deoxycoformycin
Authors: Major PP , Agarwal RP , Kufe DW
Published: October 2019

Deoxycoformycin (DCF) is an inhibitor of adenosine deaminase (ADA). Twenty-one courses of DCF were administered to 13 patients ranging in age from 15 to 78 yr. Eight patients had T-cell disorders, and five patients had non-T-cell malignancies. The i.v. bolus dose was escalated from 5 to 30 mg/sq m/day, and the duration of the courses ranged from 1 to 5 days. The DCF plasma half-life ranged from 4.9 to 6.2 hr and was independent of dose. The dose-limiting toxicities involved the central nervous system (CNS) and the kidneys. Other toxicities included bronchitis, decreases in hematocrit, arthralgias, and myalgias. Mortality was encountered in three patients. These toxic effects may have been secondary to the accumulation of the metabolites adenosine and deoxyadenosine. Deoxyadenosine and adenosine were both detectable in plasma (10(-6) M) and in urine (10(-3) M). Two partial remissions were observed: one in a patient with T-cell ALL and another in a patient with mycosis fungoides. Minimal responses characterized by either declines in peripheral blast counts or partial resolution of adenopathy were observed in five other patients. No responses were observed in six patients. These observations suggest that DCF is effective in the treatment of T-cell lymphoid malignancies.
Biochemical and biophysical characterization of glucocorticoid receptors in normal lymphoid tissue
Authors: McCaffrey R , Lillquist A , Bell R
Published: October 2019

3H-triamcinolone acetonide labeled glucocorticoid receptors in normal lymphoid tissues can be resolved into two component by DEAE chromatography: peak I elutes at 0.04 M salt and peak II is 0.22 M salt. By glycerol gradient centrifugation, peak I is 3.5S and peak II 8.5S. Peak I binds to DNA and chromatin, while peak II binds to neither. After heat activation, peak II alters its coefficient of sedimentation to 3.5S and on DEAE rechromatography changes its elution position to 0.04 M salt (peak I area) and acquires affinity for DNA. Glucocorticoid receptors in lymphoblastic leukemia cells can now be characterized using these techniques and compared to receptors in normal lymphoid cells.
Chromosome studies in adult T-cell leukemia in Japan: significance of trisomy 7
Authors: Ueshima Y , Fukuhara S , Hattori T , Uchiyama T , Takatsuki K , Uchino H
Published: October 2019

Chromosomes were studied in cells from 15 japanese patients with adult T-cell leukemia (ATL). Mitoses were obtained from unstimulated peripheral blood in 12 patients and a lymph node in one patient. In two other patients, mitotic cells were obtained solely from peripheral blood stimulated with phytohemagglutinin (PHA). Chromsomally abnormal cells were seen in 14 of the 15 patients. The abnormal cells had a modal number of chromosomes in near diploid range in 12 patients, and in near triploid and tetraploid range in the remaining 2 patients, respectively. Eight of the nine patients analyzed by Q-banding had clonal chromosome abnormalities. The most common abnormality was trisomy no.7 or 7q, which was seen in 5 cases and has been primarily observed in lymphoid neoplasms. A 14q+ marker chromosome was found in two patients and a Dq+ in one patient; loss of a sex chromosome was found in three patients. Most chromosomes were involved in gains, losses, or structural rearrangements, but abnormalities of no. 11, which have been frequently found in lymphoid malignancies, was not observed in our series. The significance of these chromosome abnormalities is discussed.
Cytotoxic chemotherapy for cancer in Felty's syndrome: role of lithium carbonate
Authors: Pazdur R , Rossof AH
Published: October 2019

A 61-yr-old white man with Felty's syndrome, who had previously undergone splenectomy, presented for cytotoxic chemotherapy. Random granulocyte counts remained low, prohibiting the initiation of such treatment. A trial of lithium carbonate was instituted, resulting in prompt elevation of granulocyte counts into the normal range. Cytotoxic chemotherapy was then administered, and fluctuations of neutrophil counts similar to those of hematologically normal individuals were observed.
Granulopoietic effects of human bone marrow fibroblastic cells and abnormalities in the "granulopoietic microenvironment"
Authors: Greenberg BR , Wilson FZD , Woo L
Published: October 2019

The in vitro granulopoietic effects of adherent bone marrow fibroblastic cells (FC) were studied in normal humans and in patients with acute myelogenous leukemia (AML) and myeloproliferative disorders (MPD). To determine their influence on granulopoiesis, we established FC in liquid-phase cultures, overlaid the adherent FC with normal bone marrow cells in agar, and subsequently measured the growth of CFU-C. When using target marrows containing few spontaneous colonies, increased numbers of CFU-C were found above the FC obtained from normals. No growth greater than controls was observed in those areas lacking FC. If target marrows contained large numbers of spontaneous CFU-C, actual inhibition of colony formation was produced by FC co- incubation. In contrast to normals, FC obtained from untreated AML and MPD patients typically failed to enhance granulopoiesis. Regardless of source, FC were not synergistic with the effects of placenta- conditioned media (typically being inhibitory) for colony number, but were synergistic for colony size. Conditioned media obtained from FC cultures did not enhance colony formation and actually inhibited spontaneous colony formation. Thus, microenvironmental abnormalities in interactions between “stromal cells” and hematopoietic progenitors may be important in the pathogenesis and clinical expression of hematopoietic malignancies in humans.
Surface phenotype of Japanese adult T-cell leukemia cells characterized by monoclonal antibodies
Authors: Hattori T , Uchiyama T , Toibana T , Takatsuki K , Uchino H
Published: October 2019

We studies the surface phenotype and the functional activities of leukemic cells from three patients with Japanese adult T-cell leukemic (ATL) using the panel of OK and anti-Tac monoclonal antibodies, which react with differentiation antigens and define functionally distinct T- cell subsets or activated and terminally differentiated T cells. The phenotype of ATL cells were determined to be OKT1+T3+T4+T10+T5-T8-Okla1- , although cells from two patients suppressed pokeweed mitogen (PWM) induced normal B-cell differentiation, and cells from all patients lacked helper activity in this system. In addition, after cultivation with PWM, ATL cells from all patients were reactive with anti-Tac monoclonal antibody, and cells from one patient were reactive with OKlal. These findings suggest that ATL cells arise from peripheral mature T-cell subsets and also suggest that the transition of surface phenotype of ATL cells to functionally mature and activated T cells occurs in culture.
Plasma and urine beta-thromboglobulin concentration in patients with deep vein thrombosis
Authors: de Boer AC , Han P , Turpie AG , Butt R , Zielinsky A , Genton E
Published: October 2019

Plasma and urine beta-thromboglobulin (BTG) were measured in 52 patients with established deep vein thrombosis (DVT) and in 100 patients with clinically suspected DVT but with a negative venogram. Both plasma BTG (geometric mean 54: 95% range 12--239 ng/ml) and urine BTG (0.25; 0.03--3.1 ng/ml) were significantly elevated (p less than 0.005) in patients with DVT compared to symptomatic patients with a negative venogram (plasma BTG 32, 9--112 ng/ml; urine BTG 0.12, 0.02-- 0.58 ng/ml). Sensitivity (35%) and specificity (80%) of the plasma BTG assay for the diagnosis of DVT were low. The urine BTG assay had a sensitivity of 37% but a specificity of 100%. There was a significant correlation between plasma and urine BTG (r = 0.68, p less than 0.005). Serial BTG measurements were made in urine (40 patients) and plasma (20 patients) from high-risk neurosurgical cases who were screened with 125I-fibrinogen leg scanning and impedance plethysmography. BTG was elevated postoperatively and returned to normal within 2 or 3 days, but rose again in 10 patients in association with the development of DVT. The rise of BTG preceded the uptake of 125I-fibrinogen and lasted for only a few days. The return to normal of BTG was not related to treatment with anticoagulants. While measurement of BTG in plasma and urine is of limited value in the clinical diagnosis of venous thrombosis, the data indicate platelet activation occurs in venous thrombosis, but is maximal or perhaps limited to the initial phase of thrombus development.
Molecular and cell biologic aspects of erythropoiesis in long-term bone marrow cultures
Authors: Dexter TM , Testa NG , Allen TD , Rutherford T , Scolnick E
Published: October 2019

In long-term marrow cultures, proliferation and differentiation of hemopoietic stem cells occurs for several months. Normally, only the most primitive erythroid progenitor cells are produced (the BFU-E). Following treatment with anemic mouse serum (AMS) or normal mouse serum plus erythropoietin, the BFU-E mature into CFU-E, which then go to produce mature nonnucleated red cells. This development is associated with the production of adult type hemoglobin. Furthermore, erythropoiesis and granulopoiesis occur in association with discrete cellular elements of the adherent cell layer in the long-term culture. Following treatment with AMS, erythropoiesis is enhanced while granulopoiesis is depressed, with no apparent competition at the stem cell or progenitor cell level.
Membrane fluidity changes accompanying phagocytosis in normal and in chronic granulomatous disease polymorphonuclear leukocytes
Authors: Ingraham LM , Boxer LA , Haak RA , Baehner RL
Published: October 2019

We have studied membrane fluidity changes in polymorphonuclear leukocytes (PMN) during phagocytosis. Membrane fluidity was assessed by electron spin resonance (ESR) using a nitroxide-substituted stearic acid analog (5DS) as a spin probe. PMN from normal subjects and from 3 CGD patients (2 males, 1 female) were incubated in Kreb's Ringers phosphate with or without opsonized zymosan. ESR spectra were obtained and the order parameter (S), which is inversely related to membrane fluidity, was calculated. Without zymosan addition, S for normal (0.638) and for CGD (0.635) were not significantly different (p less than 0.35). The S values indicate that under resting conditions the molecular environment of the CGD membrane is similar to that of normal PMN membranes. However, with addition of opsonized zymosan, the normal, but not the CGD, PMN showed a significant increase (CGD, S = 0.638; normal, S = 0.647; p less than 0.001). This change in S for the normals is consistent with a more restricted movement of 5DS. Treatment of normal PMN with a mixture of scavengers specific for H2O2 (catalase, 1600 U/ml), O2-.(superoxide dismutase, 100 micrograms/ml), and for HO., (sodium benzoate, 1mM) during zymosan stimulation gave S values similar to those of resting cells. Catalase alone also lowered S value, suggesting that H2O2 was instrumental in causing the initial S value increase. This idea was supported by studies in which CGD cells were incubated with zymosan in the presence of glucose oxidase, an enzyme that catalyzes glucose oxidation resulting in the direct reduction of molecular oxygen to H2O2. Our results indicate that reduced O2 by- products, particularly H2O2, can cause altered biophysical properties of PMN membrane during phagocytosis.
Immature Megakaryocytes in the Mouse: Morphology and quantitation by acetylcholinesterase staining
Authors: Long MW , Williams N
Published: October 2019

Three types of immature megakaryocytes, detected by their morphological properties, have been characterized in bone marrow of normal C57BL/6 mice. Morphological classification of these cells was carried out by determining (1) presence and relative amount of acetylcholinesterase, (2) cell size, (3) nuclear/cytoplasm ratio, and (4) nuclear shape. The immature megakaryocytes were classified as: (A) cells distinguished by a round nucleus (10.6 +/- 1.1 mu diameter; mean +/- SEM), which had the highest nucleus / cytoplasm ratio and lowest content of acetylcholinesterase; (B) cells with an indented nucleus (13.0 +/- 1.9 mu diameter), which had increased acetylcholinesterase content and reduced nucleus/cytoplasm ratio compared to the round-nucleus cell type; and (C) lobed-nucleus cells (14.5 +/- 2.9 mu diameter), which showed further increase in acetylcholinesterase content and reduction in nucleus/cytoplasm ratio. Increased numbers of immature megakaryocytes were detected, indicating that a proportion of these cells are undetected using conventional staining techniques. Based on the observed alterations in size, acetylcholinesterase content, and nuclear complexity, it was concluded that these cells constitute part of a progressive maturation sequence intermediate between the progenitor cell (CFU-Mk) and mature easily recognizable megakaryocytes.
Phenotypic characterization of human bone marrow granulocyte-macrophage forming progenitor cells
Authors: Levine MN , Fay JW , Jones NH , Metzgar RS , Haynes BF
Published: October 2019

Cell surface antigens of the human bone marrow CFU-C have been studied. Human marrow cells were incubated with a variety of monoclonal antisera and complement prior to culture in semisolid media. By using indirect immunofluorescent studies, the percentage of bone marrow cells binding the antibodies was determined. The CFU-C phenotype is HLA+, la+, 4F2+, 3A1-, and DUALL-1-. This study provides information that is useful in the study of myeloid cell ontogeny and necessary for the use of some of these reagents in the treatment of bone marrow cells prior to human bone marrow transplantation in various clinical settings.
Failure of desmopressin to lower serum sodium or prevent crisis in patients with sickle cell anemia
Authors: Charache S , Walker WG
Published: October 2019

An analogue of arginine vasopressin (desmopressin, DDAVP) was evaluated for production chronic hyponatremia and prevention of sickle cell crisis. With sodium restriction (100 meq Na + / day) and water loading ( greater than 3 liters/day), persistent hyponatremia could not be achieved, nor could crises be prevented or aborted. Patients would not comply with a regimen of lower salt and higher fluid intake. More rigorous treatment might be practical during acute sickle cell crises, and a regimen similar to that used here might be more effective in children, whose renal concentrating mechanisms are still intact.