bluuebunny/crossref_metadata_2025 huggingface dataset:
Phagocytosis-induced chemotaxis receptor cycling in neutrophils is mediated by thiol oxidation
Authors: Lane TA , Lamkin GE
Published: October 2019

We previously demonstrated that neutrophil (PMN) phagocytosis of opsonized zymosan (OPZ) caused oxygen-dependent inhibition of chemotactic peptide receptor (CPR) binding using the ligand 3H-formyl- methionyl-leucyl-phenylalanine (3H-FMLP). In the current studies we sought to determine the mechanism of CPR inhibition by OPZ. We found that 3 mM cysteine and 5 mM dithiothreitol (DTT) did not decrease PMN phagocytosis, but abolished CPR inhibition by OPZ. Furthermore, incubation of PMN with DTT after OPZ partially restored the suppressed CPR. PMN CPR saturation studies with 3H-FMLP on PMN incubated with or without DTT after phagocytosis indicated that DTT restored receptor numbers to 92% +/- 6% of prephagocytosis values and also improved CPR affinity for 3H-FMLP. In additional studies we found that the cell penetrating thiol reagents mercuric chloride (HgCl2), N-ethylmaleimide (NEM), and diamide, but not the nonpenetrating agents p- chloromercuriphenylsulfonic acid and p-chloromercuribenzoate, caused rapid, dose-dependent, DTT-inhibitable suppression of up to 85% CPR binding. CPR inhibition by HgCl2 and NEM was irreversible. PMN CPR saturation studies showed that NEM decreased total receptor numbers, not CPR affinity. The effect of NEM was not inhibited by receptor occupancy at 4 degree C or at 37 degree C. These studies indicate that: (1) intact thiol groups are a requirement for CPR ligand-binding; (2) the thiol groups are presumptively located on the interior side of, or within hydrophobic portions of, the plasma membrane and are not part of the 3H-FMLP binding site; (3) thiol oxidation-reduction induces reversible alteration in CPR binding; (4) phagocytosis-induced modulation of the CPR is due, in part, to thiol oxidation. These studies suggest that postphagocytic, thiol-mediated reduction in CPR binding may play a physiologic role in the inhibition of PMN chemotaxis at inflammatory sites. The state of CPR-associated thiols may be an important determinant of CPR binding under physiologic conditions other than phagocytosis.
Nucleic Acids of Rabbit Reticulocytes
Authors: BURT N. S. , MURRAY R. G. E. , ROSSITER R. J.
Published: October 2019

1. The concentration of acid soluble phosphorus, lipid phosphorus and nucleic acid phosphorus was determined in the red cells and polymorphonuclear leukocytes of the rabbit. The concentration of acid soluble phosphorus was of the same order in both types of cells, but there was more lipid phosphorus and nucleic acid phosphorus in the polymorphonuclear leukocytes than in the red cell. Over 80 per cent of the nucleic acid in white cells was of the desoxypentose type.

2. Immature cells were produced in the peripheral blood stream of rabbits either by the administration of phenylhydrazine or by successive bleedings. Either treatment caused a fall in the concentration of hemoglobin, the red cell count and the hematocrit with an increase in the mean corpuscular hemoglobin. There was also an increase in the concentration of acid soluble phosphorus and lipid phosphorus and a great increase in the concentration of nucleic acid phosphorus which was coincident with the increase in number of immature cells as judged by the reticulocyte count. Almost all of the excess nucleic acid was of the pentose type.

3. The basophilia of the immature cells disappeared after treatment with ribonuclease or after hydrolysis with normal hydrochloric acid. Desoxyribonuclease was without effect.

4. These results suggest that the basophilic substance of the immature rabbit red cell, and presumably the reticulum of the reticulocyte, is of cytoplasmic rather than nuclear origin.
Activity of Megakaryocytes in the Postoperative State
Authors: KERHULAS A. A. , OHLER ROBERT L. , WARREN RICHARD , BELKO JOHN S.
Published: October 2019

Simultaneous bone marrow smears and platelet counts on the peripheral blood were made preoperatively and on the third and the tenth days after operation in 14 patients. Observations of the activity of the megakaryocytes of the bone marrow on the third day were inconclusive and require further observation. There was, however, a significant increase in activity on the tenth day.

It is postulated that the thrombocytosis which occurs on the tenth postoperative day is the result of increased activity of the megakaryocytes of the bone marrow induced by the attempt of the body to replenish depleted platelet reserves.
Platelet effects on tissue factor and fibrinolytic inhibition of cultured human fibroblasts and vascular cells
Authors: Smariga PE , Maynard JR
Published: October 2019

Platelets stimulate tissue factor, the initiator of the extrinsic coagulation pathway, and increase fibrinolytic inhibition in fibroblasts grown in vitro. Cellular tissue factor increases an average of 2.8-fold over the control levels after a 6-hr incubation with platelets, and no activity is present in the media. Fibrinolytic inhibition is stimulated in both the fibroblasts and their media in the presence of platelets and accumulates throughout a 24-hr incubation. Neither leukocytes nor erythrocytes stimulate these changes. Both tissue factor and fibrinolytic inhibition increases are dependent on platelet concentration and are blocked by inhibitors of RNA or protein synthesis. Control smooth muscle cells have higher tissue factor and fibrinolytic inhibition than fibroblasts, but their response to the presence of platelets is similar. Confluent monolayers of endothelial cells have very low levels of tissue factor that are not altered by the presence of platelets. However, the ability of endothelial cells to inhibit fibrinolysis is enhanced by the presence of platelets. The fraction that stimulates tissue factor and fibrinolytic inhibition is distinct from platelet-derived growth factor and from the fraction that enhances leukocyte tissue factor. It is associated with an insoluble, nonmitogenic fraction that is not inactivated by phospholipase C, or diisopropylfluorophosphate, nor is it chloroform:methanol extractable. Platelets are a physiologic modulator for both cellular tissue factor and the fibrinolytic system in vitro.
Decreased number of circulating BFU-Es in paroxysmal nocturnal hemoglobinuria
Authors: Rotoli B , Robledo R , Luzzatto L
Published: October 2019

In order to quantitate early erythroid progenitor cells in paroxysmal nocturnal hemoglobinuria (PNH), we have cultured peripheral blood mononuclear cells from 7 PNH patients in a 0.8% methylcellulose medium containing erythropoietin, 2 U/ml. In our experimental conditions, the number of erythroid colonies obtained per 5 X 10(5) mononuclear cells plated was 20.1 +/- 1.9 (SEM) in normal subjects and 2.8 +/- 0.56 (SEM) in PNH patients. In plates from PNH subjects, 38 of 117 showed no growth of erythroid colonies, whereas plates from normal subjects always had colonies. Our findings suggest that PNH patients, despite their hemolytic condition, have a depleted erythroid precursor compartment, and this may play a major role in the pathogenesis of their anemia.
Use of asplenic rabbits to demonstrate that platelet age and density are related
Authors: Corash L , Shafer B
Published: October 2019

An experimental model using asplenic rabbits injected with radioactive amino acids has been developed to examine the density distribution of newly released platelets. Platelets from asplenic rabbits had a larger mean volume and greater protein content than those from asplenic animals. Radiolabel, indicative of new platelets, is preferentially incorporated into the most dense platelets during the early postinjection period. Platelets of intermediate density also demonstrated augmented early label incorporation compared to the lightest cells, In contrast, during the late postinjection phase, there is preferential labeling of the least dense platelets. The specific radioactivity of heavy and light platelets rises to approximately equal levels, and falls abruptly, but the peak activities occur at distinctly separate times consistent with the concept of platelet density modification during platelet aging. Although platelet biologic properties are broadly distributed, these studies support the concept that on the average young platelets have an increased density and become less dense as they circulate.
Cell competence for industion of differentiation by insulin and other compounds in myeloid leukemic clones continuously cultured in serum- free medium
Authors: Symonds G , Sachs L
Published: October 2019

Clones of myeloid leukemic cells varying in their competence for induction of differentiation have been continuously grown in serum-free medium. In the medium used, which contained transferrin, the growth rates of these cells were nearly similar to those found in serum- containing medium. The clones also maintained in this medium their competence for induction of differentiation by the normal macrophage and granulocyte differentiation-induction protein MGI-2, the steroid dexamethasone, and lipopolysaccharide. In contrast to the results with these inducters, some clones continuously cultured in a serum-free medium showed a gain of inducibility by insulin and another clone a gain of inducibility by the tumor promoter 12-O-tetradecanoylphorbol-13- acetate in low serum and serum-free medium. Induction of differentiation by these two compounds was therefore inhibited in these clones by the presence of serum. It is suggested that serum-free medium may also show the existence of other inducers of differentiation not detected in serum-containing medium and that these results are relevant to the possible therapeutic use of compounds such as insulin for the induction of normal differentiation in leukemic cells in vivo.
Potentiation of fibroblast growth by nodular sclerosing Hodgkin's disease cell cultures
Authors: Newcom SR , O'Rourke L
Published: October 2019

Cell cultures were established from 8 lymph nodes replaced by nodular sclerosing Hodgkin's disease. Serum-containing and serum-free conditioned media from these cultures potentiated fibroblast growth and were found to be consistently more potent than fibroblast growth factor, 100 ng/ml, every other day. Both a proliferative response and transformation-like growth were observed using BALB/c 3T3 cells, human diploid fibroblasts, and human embryonic fibroblasts as target cells. The Hodgkin's disease growth factor(s) was not produced by fibroblasts or lymphocytes in the Hodgkin's cultures and was most potent when the Hodgkin's cultures had been enriched with Hodgkin's giant cells. Removal of normal macrophages decreased the proliferative activity but did not eliminate it or nonadherent growth of 3T3 cells in agar. Control cultures of 6 nonmalignant lymph nodes, a Lennert's lymphoma, a mixed cellularity Hodgkin's disease lymph node, and a malignant histiocytosis cell line suggested that among lymph node disorders, this feature may be relatively specific for nodular sclerosing Hodgkin's disease.
Purification of colony-stimulating factor by affinity chromatography
Authors: Waheed A , Shadduck RK
Published: October 2019

Studies were undertaken to determine whether L-cell-derived colony-stimulating factor (CSF) could be purified by a single step affinity chromatographic technique. A quantity of 100 X 10(6) units of purified anti-CSF was coupled to cyanogen bromide activated Sepharose 4B; colony assays revealed complete binding of the antibodies to the gel. Three 10- liter pools of serum-free L-cell CSF were concentrated by ultrafiltration, applied to the gel, and eluted with a low pH, high molarity buffer. Recovery of CSF ranged from 68%-100% with greater than 1000-fold decreases in protein content. Specific activity of the purified CSF ranged from 2.8 to 5.9 X 10(7) U of CSF/mg protein. Following iodination, each purified pool of CSF revealed a major 63,000- dalton peak of radioactivity that comigrated with CSF activity in SDS- acrylamide gels. In addition, several smaller peaks of 50,000 and 40,000 molecular weight were also detected. Approximately two-thirds of the purified CSF was adherent to concanavalin-A with elution by a competing sugar. Electrophoretic mobility was retarded by incubation with neuraminidase. These chromatographic studies confirm the observation that CSF is a glycoprotein but also suggest variable degrees of glycosylation of the molecule. This chromatographic technique should prove useful in the rapid purification of large quantities of CSF for physiologic and biochemical characterization.
Enzymologic classification of acute leukemias: nonspecific esterase markers distinguish myeloid and lymphoid varieties
Authors: Yourno J , Burkart P , Lizzi F , Tartaglia A
Published: October 2019

Nonspecific esterase zymograms of purified leukemic cells from a case of acute myelomonocytic leukemia (AMML) and from a case of acute non-B, non-T lymphocytic leukemia (ALL) were produced by isoelectric focusing and staining with alpha-naphthyl acetate (alpha NA) or alpha-naphthyl butyrate (alpha NB) substrate. A “myeloid” zymogram was found with AMML cells, which closely matched control monocyte and granulocyte zymograms. On the other hand, nonspecific esterase of ALL showed a striking departure from the zymogram pattern of control B lymphocytes and T lymphocytes. An intense reactivity with a very low isoelectric point accounted for most of the ALL nonspecific esterase activity. No corresponding reactivity or relatively small amounts thereof were seen in other zymograms. Conversely, few of the isoenzymes that were prominent in zymograms of control lymphocytes were apparent above trace levels in ALL zymograms. Thus, zymogram analysis of nonspecific esterases clearly differentiated the myeloid leukemia from the lymphoid leukemia and provided a potential marker for each. The AMML cells appeared well enough differentiated with respect to nonspecific esterases as to be similar to mature cells of like lineage. It is plausible that the ALL cells, however, were arrested at an earlier stage of esterase expression as reflected by the associated atypical species.
Correlation of occupation and karyotype in adults with acute nonlymphocytic leukemia
Authors: Golomb HM , Alimena G , Rowley JD , Vardiman JW , Testa JR , Sovik C
Published: October 2019

Seventy-four adult patients with acute nonlymphocytic leukemia (ANLL) were classified retrospectively as to whether or not they had had occupational exposure to insecticides, chemicals and solvents, or petroleum products. Fifty-eight patients were considered nonexposed and 16 were considered exposed. The chromosome banding pattern was abnormal in 37 of the 74 patients (50%). Twenty-five of the 58 (43%) nonexposed patients had a clonal chromosome abnormality, compared with 12 of the 16 (75%) exposed patients (p = 0.02). Only 2 of 23 (8.7%) females with an abnormal karyotype were exposed, whereas 10 of 14 (71%) males with an abnormal karyotype were exposed. Either a -5/5q- or a -7/7q- was present in 67% of the exposed patients with a chromosome abnormality, compared to 28% of the aneuploid nonexposed patients. The -7/7q- abnormality was present in 7 of the 12 (58.3%) exposed patients, versus 5 of the 25 (20%) nonexposed patients with abnormal karyotypes (p less than 0.05). The -5/5q- anomaly was observed in 4 of the 12 (33%) exposed patients and in 4 of the 25 (16%) nonexposed abnormal patients. Our study supports the observation that a subset of patients with ANLL de novo have a history of occupational exposure and a unique pattern of chromosome abnormalities.
Effects of neuraminidase on O2 consumption and release of O2 and H2O2 from phagocytosing human polymorphonuclear leukocytes
Authors: Suzuki H , Kurita T , Kakinuma K
Published: October 2019

Neuraminidase type X (NMD-type-X, Sigma Chemical Co., St. Louis, Mo.), which is obtained from a further purification of neuraminidase type V (NMD-type-V, Sigma), markedly enhanced the release of O2- and H2O2 from phagocytosing human polymorphonuclear leukocytes (PMN). In contrast, O2 consumption by NMD-type-X-treated PMN was identical to that of untreated PMN. Morphological observations suggested that the enhancement of O2- and H2O2 release was caused by excessive release of the oxygen metabolites into the extracellular medium from incompletely formed phagocytic vacuoles as was observed with cytochalasin-B-treated cells. Our observations are in contrast to the previous reports of Tsan et al. that showed complete inhibition of both O2- and H2O2 release from phagocytosing PMN by the treatment with NMD-type-V.
Evaluation of AMSA in previously treated patients with acute leukemia: results of therapy in 109 adults
Authors: Legha SS , Keating MJ , McCredie KB , Bodey GP , Freireich EJ
Published: October 2019

AMSA was evaluated in the treatment of 109 adults with previously treated acute leukemia. Of the 102 evaluable patients, 82 had AML, 17 ALL, and 3 CML in blastic phase. A number of different dose schedules of AMSA were explored, and we conclude that the optimum dose of AMSA for remission induction in acute leukemia is 120 mg/sq m/day for 5 days. Complete remissions were observed in 23 (28%) patients with AML and in 1 patient with ALL. Patients who achieved complete remission were maintained on AMSA using a dose of 30–40 mg/sq m/day for 5 days repeated at 4-wk intervals. The median duration of complete remission was 12 wk (3–59 wk), and the responders survived significantly longer than the failures (27 wk versus 8 wk, p = 0.002). The side effects associated with AMSA therapy included mild nausea and vomiting, stomatitis, diarrhea, phlebitis, alopecia, and myelosuppression-related infections. Our results indicate that AMSA is a useful new antileukemic agent for the treatment of relapsed acute leukemia and appears to have activity comparable to that of the currently available drugs, such as cytarabine and the anthracycline antibiotics.
Presence of mixed colony-forming cells in long-term hamster bone marrow suspension cultures: response to pokeweed spleen conditioned medium
Authors: Eastment CE , Ruscetti FW , Denholm E , Katznelson I , Arnold E , Ts'o PO
Published: October 2019

In contrast to the murine system, long-term hamster bone marrow suspension cultures maintain proliferation of both pluripotent and committed stem cells in the absence of an adherent layer and without addition of exogenous factors, such as hydrocortisone. Addition of pokeweed-mitogen-stimulated hamster spleen conditioned medium (SCM) to these long-term suspension cultures produces an increase in the number of mixed colonies assayed in soft-agar, These mixed colonies, which contained four cell lineages--granulocytic, erythroid, megakaryocytic, and macrophage--could be generated from cells grown in suspension for over 6 mo. Addition of SCM also induces an initial rapid expansion of the myeloid compartment, and this expansion results in 70% of the cells being terminally differentiated granulocytes. In contrast, addition of SCM to hamster bone marrow cultures containing both adherent cells and hematopoietic stem cells produced no change in the number of mixed colonies generated in the culture. This system allows the in vitro study of the process of stem cell proliferation and differentiation and also provides a means to examine the relationship of adherent and supernatant bone marrow populations.
Cyclosporin-A abrogates transfusion-induced sensitization and prevents marrow graft rejection in DLA-identical canine littermates
Authors: Storb R , Deeg HJ , Atkinson K , Weiden PL , Sale G , Colby R , Thomas ED
Published: October 2019

Previous studies had shown that marrow graft rejection generally did not occur in untransfused dogs given 900-rad total body irradiation and hemopoietic grafts from DLA identical littermates (only 1 of 59 rejected), but was seen in all instances after three preceding transfusions of whole blood from the marrow donor on days -24, -17, and -10 before transplantation (19 of 19 rejected). The present study was undertaken to investigate whether immunization by 3 preceding transfusions of whole blood from the DLA-identical littermate marrow donor could be abrogated by administration of the immunosuppressive agent cyclo-sporin-A, 20 mg/kg/day intramuscularly on days -5 to 0. Seven of 10 dogs showed sustained marrow engraftment. 2 failed to engraft, and 1 dog died to early to be evaluated. It was concluded that immunization to non-DLA antigens by preceding whole blood transfusions could be abrogated in most cases by a short-course of cyclosporin-A before total body irradiation and marrow transplantation, resulting in successful and sustained marrow engraftment.
Surface membrane vesicles from mononuclear cells stimulate erythroid stem cells to proliferate in culture
Authors: Dainiak N , Cohen CM
Published: October 2019

In order to examine the contribution of cell surface materials to erythroid burst-promoting activity (BPA), we separated media conditioned by a variety of human cell types into pellets and supernatants by centrifugation. When added to serum-restricted cultures of nonadherent human marrow cells, pellets contained about half of the total stimulatory activity. Freeze-fracture electron microscopy of the pellets revealed the presence of unilamellar membrane vesicles ranging from 0.10 to 0.40 microM in diameter. The amount of BPA in culture increased with added vesicle concentration in a saturable fashion. Preparation of leukocyte conditioned medium (LCM) from 125I-wheat germ agglutinin labeled cells and studies comparing the glycoprotein composition of vesicles with that of leukocyte plasma membranes suggest that LCM-derived vesicles are of plasma membrane origin. Moreover, partially purified leukocyte plasma membrane preparations also contained BPA. While disruption of vesicles by freezing/thawing and hypotonic lysis did not alter BPA, heat, trypsin, or pronase treatment removed greater than 65% of BPA, implying that vesicle surface rather than intravesicular molecules express BPA. Results of BPA assays performed in two-layer clots indicated that proximity to target cells is required for vesicle BPA expression. We conclude that membrane vesicles spontaneously shed from cell surfaces may be important regulators of erythroid burst proliferation in vitro.
The kinetics of iron uptake by isolated intestinal cells from normal mice and mice with sex-linked anemia
Authors: Peppriell JE , Edwards JA , Bannerman RM
Published: October 2019

No significant differences of Km or Vmax could be demonstrated for the initial rate (0–3 min) of iron uptake by preparations of isolated duodenal mucosal epithelial cells from normal or sex-linked anemic (sla) mice. Isolated cells from sla mice do, however, take up significantly more iron than normal cells after 20-min incubation. Upon reincubation with various types of mobilizing agents, the amount of iron released from, and the amount of iron remaining in, cells from sla mice was significantly elevated compared to control values. The percentage of iron released was the same for both types of cells, for all mobilization media employed. The current data, in conjunction with previous studies that showed deficient mucosal to serosal transport of iron in everted duodenal sacs from sla mice, suggest that the lesion in these mice may be located between the mucosal cell and the vascular compartment rather than in the mucosal cell.
T-cell lymphoma and the Chediak-Higashi syndrome
Authors: Argyle JC , Kjeldsberg CR , Marty J , Shigeoka AO , Hill HR
Published: October 2019

The majority of patients with Chediak-Higashi syndrome (CHS) develop a lymphoproliferative disorder during the accelerated phase of the disease. Controversy exists regarding the benign versus malignant nature of this cellular proliferation. For the first time, we have characterized the immunologic cell markers on the cellular infiltrate in a lymph node from a patient with CHS. The infiltrate was composed almost entirely of T cells, with histopathologic features consistent with a non-Hodgkin's T-cell lymphoma.
Association between HLA-D region antigens and disease-free survival in childhood non-T, non-B acute lymphocytic leukemia
Authors: Casper JT , Marrari M , Piaskowski V , Lauer SJ , Duquesnoy RJ
Published: October 2019

The frequency of three serologically defined HLA-D region antigens--DR, MB, and MT--was determined in a group of 74 children with non-T, non-B acute lymphocytic leukemia (ALL). Statistically, there were no significant differences in the frequency of any antigen in these ALL patients as compared with a panel of 85 normal controls. However, significant differences in HLA-DR frequencies were observed between patients who relapsed or who remained disease-free during a 30-mo period of chemotherapy. An increased incidence of relapse was associated with DR5, while disease-free remission during chemotherapy was associated with DR7. Life table analysis also demonstrated that DR5 was significantly associated with a decrease in disease-free survival in these patients. These data suggest that HLA-associated genetic factors may influence the responses of ALL patients to chemotherapy.
Peripheral blood myeloid progenitor cell cultures in patients with hypereosinophilic syndrome (CFU-eos in hypereosinophilic syndrome)
Authors: Bjornson BH , Harley JB , Andre-Schwartz J , Fauci AS , Desforges JF
Published: October 2019

Myeloid progenitor cell cultures (CFU-C) were established in a double- layer agar system with peripheral blood mononuclear cells from 13 patients with the hypereosinophilic syndrome (HES). Normal controls produced 49% +/- 3.5% eosinophil colonies; results in 7 of the 13 HES patients were within the normal range, while in 5, the proportion of eosinophil colonies was greater than 3 standard deviations above the normal mean, and in 1 patient there was a low proportion of eosinophil colonies. The production of an increased proportion of eosinophil colonies correlated with more aggressive disease. Experiments in which normal progenitor cells were cultured over feeder layers of mononuclear cells demonstrated that cells of 3 of the 5 patients had an excess production of eosinophil colony-stimulating activity. When HES patients progenitor cells were cultured over normal feeder layers, 2 of the 5 patient samples continued to produce an increased proportion of eosinophil colonies, suggesting that these patients have an excess proportion of progenitor cells committed to eosinophil differentiation. Thus, the results demonstrated heterogeneity of growth characteristics for the HES patients. None, however, had the colony growth characteristic of acute or chronic myelogenous leukemia.
Activation of factor VIII by factor IXa
Authors: Rick ME
Published: October 2019

Thrombin causes an increase in factor VIII coagulant (VIII:C) activity, which is followed by a decay of VIII:C activity to below baseline levels. It has been suggested that a similar interaction of trace amounts of thrombin and factor VIII is a necessary prerequisite before factor VIII can participate in the coagulation cascade. In the current study, factor IXa, a serine protease with structural similarities to thrombin, is shown to cause an increase and subsequent fall in VIII:C in a manner qualitatively similar to the reaction with thrombin. The reaction is inhibited by a human inhibitor to factor IX and the interaction appears to involve only VIII:C, since factor-VIII-related protein (VIII:RP) is not changed on polyacrylamide gel electrophoresis (PAGE) or radioimmunoassay during the reaction. Phospholipid increases the activation of factor VIII by factor IXa, and high concentrations of diisopropylfluorophosphate and hirudin inhibit the reaction. The physiologic significance of the interaction of factor IXa with factor VIII is not entirely clear since the concentration of factor IXa needed for activation is much greater than the concentration of thrombin required for similar activation of factor VIII. Factor IXa is most likely to play a role in the intrinsic cascade acting as an initial activator of factor VIII, since factor IXa precedes thrombin in this clotting sequence. In addition, factor IXa may be important wherever relatively high local concentrations of factor IXa and factor VIII occur, particularly in the presence of phospholipid, which may serve to localize the coagulation factors.
Platelet-type von Willebrand's disease: characterization of a new bleeding disorder
Authors: Miller JL , Castella A
Published: October 2019

An autosomally transmitted bleeding diathesis sharing some, but not all, features previously described in von Willebrand's disease (vWd) was studied in five patients representing three generations of a single family. Bleeding times in the upper normal range in conjunction with low-normal platelet counts, normal factor VIII coagulant activity and VIII-related antigen, decreased VIII-ristocetin cofactor activity, selective decrease of the higher molecule weight factor VIII/von Willebrand factor (VIII/vWF) multimers, and increased ristocetin- induced platelet agglutination at low ristocetin concentrations were characteristic. Binding of patient VIII/vWF to washed normal platelets was within normal limits, whereas binding of normal VIII/vWF to patient platelets was significantly increased (p less than 0.001 at 0.6 mg/ml ristocetin). This disorder accordingly appears to involve an intrinsic platelet abnormality affecting platelet-VIII/vWF interactions. It is proposed that the concept of vWD be broadened to include patients with this abnormality, which may appropriately be called “Platelet-type von Willebrand's disease.”
Allogeneic bone marrow transplantation for chronic myelogenous leukemia in chronic or accelerated phase
Authors: Champlin R , Ho W , Arenson E , Gale RP
Published: October 2019

Eight patients with Ph1-positive chronic myelogenous leukemia (CML) in chronic or accelerated phase received high-dose cyclophosphamide, total body irradiation, and bone marrow transplantation from an HLA-identical sibling donor. All patients had prompt engraftment and achieved complete hematologic remission. Six patients remain alive and in continuous remission with a normal bone marrow karyotype 3–20+ mo posttransplant. One patient died from cytomegalovirus interstitial pneumonitis. Only one patient who was transplanted in accelerated phase relapsed 6.5 mo posttransplant and died in blast crisis. High-dose combined modality therapy is capable of producing sustained complete remissions in patients with CML treated during chronic or accelerated phase.
Cumulative experience with a simplified solid-phase radioimmunoassay for the detection of bound antiplatelet IgG, serum auto-, allo-, and drug-dependent antibodies
Authors: Faig D , Karpatkin S
Published: October 2019

A simplified, sensitive, solid-phase radioimmunoassay employing 125I- staphylococcal protein A has been developed that is capable of detecting bound antiplatelet IgG as well as serum auto-, allo-, and drug-dependent antiplatelet antibodies. The simplified assay employs a ratio of test over control platelet counts per minute (cpm) for detection of positive results. All reagents are commercially available. The assay can be performed with as little as 10(6) washed platelets (10 microliters of whole blood) that have been stored for as long as 8 wk at 4 degrees C in microtiter plates. The assay time, employing stored platelets, is 4 hr. Bound platelet IgG is positive in 93% of 46 thrombocytopenic patients with autoimmune disease and correlates inversely with their platelet count, r = -0.65, p less than 0.001. The ability of this assay to detect serum antibody was studied with a rabbit anti-human platelet antibody capable of giving optimal immunoprecipitation with solubilized platelet membranes at a tier of 1:10. The present assay increases the sensitivity of antibody detection 256-fold to a titer of 1:2560. Human serum antiplatelet membrane antibody was positive in 2 of 2 patients with anti-PLA-1 antibody (titers of 1:256 and greater than 1:64); 7 of 12 multiply transfused patients who were refractory to platelet transfusion (2 had titers of greater than 1:256 and greater than 1:32); 5 of 19 patients with autoimmune thrombocytopenic purpura (2 had titers of 1:64 and 1:32); and 10 of 14 patients with clinical histories of drug-dependent antiplatelet antibody (2 had titers of 1:1280 for quinidine and 1:384 for phenazopyridine).
Myeloma protein kinetics following chemotherapy
Authors: McLaughlin P , Alexanian R
Published: October 2019

The effects of chemotherapy were evaluated in 43 multiple myeloma patients with high monoclonal globulin levels in both serum and urine. In responding patients, Bence Jones protein excretion declined more rapidly and markedly than the serum myeloma protein. Bence Jones protein excretion was reduced by 50% within 2 mo in responders, remained unchanged in nonresponders, and declined with a slow halving time of 2–7 mo in patients with partial disease control. Three patients with clinical resistance to treatment showed atypical protein changes and progressively more Bence Jones protein excretion relative to their serum monoclonal component. Serial measurements of Bence Jones protein excretion provided an early and reliable index of tumor mass change in patients with multiple myeloma.
Nonrandom chromosome abnormalities in angioimmunoblastic lymphadenopathy
Authors: Kaneko Y , Larson RA , Variakojis D , Haren JM , Rowley JD
Published: October 2019

Cytogenetic and pathologic studies were performed on six patients with angioimmunoblastic lymphadenopathy (AILD). All six had diffuse lymphadenopathy; five had fever, four had weight loss, and four had a diffuse erythematous rash. All patients except one had a polyclonal elevation of immunoglobulin. All patients had diagnostic findings in lymph node (LN) and bone marrow (BM) biopsies. Two patients died of progressive AILD; one patient died after transformation of AILC to immunoblastic sarcoma (IBS); one patient died of gastrointestinal bleeding of unknown cause. The remaining two patients, who have achieved complete remission with intensive chemotherapy, are alive 20 and 8 mo after the diagnosis; one of these had AILD and the other, both AILD and IBS. Despite diagnostic BM biopsy findings, none of the patients had chromosome abnormalities in their BM cells. In studying LN cels of 5 patients, however, we found chromosome abnormalities in each; clonal abnormalities were detected in two, both clonal and nonclonal abnormalities in two, and only nonclonal single-cell abnormalities in one. An extra chromosome 3, seen in four patients, was clonal in two and nonclonal in the two others. Cells with +5, +15, +19, +21, +22 were seen in two patients. All patients had 50% or more normal dividing cells in their LN. The mosaicism of unrelated abnormal cells in their LN. The mosaicism of unrelated abnormal karyotypes that was seen in four patients suggests that this malignant tumor is not necessarily monoclonal in its early stages, but that one clone may be selected and predominate in the late stage. Because nonrandom acquired clonal chromosome abnormalities are a consistent feature of malignancies, our data suggest that AILD may be a malignant disease despite its original description as a benign proliferative process. Therefore, it may require aggressive chemotherapy.
Characteristic abnormality of deoxyribonucleoside triphosphate metabolism in megaloblastic anemia
Authors: Iwata N , Omine M , Yamauchi H , Maekawa T
Published: October 2019

To elucidate the biochemical basis of megaloblastic hematopoiesis, the cellular content and metabolism of deoxyribonucleoside triphosphates (dNTPs) were investigated using the bone marrow cells from nine patients with untreated vitamin B12 deficiency and one with folic acid deficiency. The marked imbalance among four dNTPs was noted in all patients. dTTP was invariably elevated rather than depressed. The most striking abnormality, however, was the excessive accumulation of dCTP, which represented the consistent feature exclusive for megaloblastic anemia. Purine nucleotides were also involved to a lesser extent. The apparent turnover pattern of the dTTP pool of megaloblastic anemia marrow cells, in the presence or absence of hydroxyurea, did not differ significantly from that of normoblastic hematopoiesis. The megaloblastic cells assimilated exogenous thymidine into dTTP pool in vitro with enhanced efficiency. It was suggested that the excessive accumulation of dCTP may be related more closely to the pathogenesis of megaloblastic hematopoiesis than to the presumed but not proved deficiency of dTTP.
Neutrophil adhesiveness during prostacyclin and heparin hemodialysis
Authors: Spagnuolo PJ , Bass SH , Smith MC , Danviriyasup K , Dunn MJ
Published: October 2019

We evaluated neutrophil adhesive function in patients undergoing chronic hemodialysis using either prostacyclin or heparin as antithrombotic agents. Patients underwent successive hemodialyses with prostacyclin (4 ng/kg/min) and heparin. There were no significant differences noted in neutrophil adhesive function during either dialysis: transient neutropenia developed in each case; impaired neutrophil adhesiveness to plastic developed during both dialyses; neutrophil aggregation was diminished when compared to predialysis responses during both dialyses. Furthermore, the number of circulating Fc-receptor-bearing neutrophils fell significantly during both prostacyclin and heparin hemodialysis. Our study demonstrates that substitution of prostacyclin for heparin in doses that do not cause hypotension, does not prevent neutropenia or alter the diminished neutrophil adhesiveness that occurs during heparin hemodialysis.
Specific factor VIII-related antigen fragmentation: an in vivo and in vitro phenomenon
Authors: Montgomery RR , Johnson J
Published: October 2019

Factor VIII concentrates have been shown to have reduced ability to correct the bleeding time defect in von Willebrand's disease and to have abnormal mobility of their VIIIR:Ag on crossed immunoelectrophoresis. This report concerns the partial characterization of a fragment of VIIIR:Ag that lacks some of the normal antigenic determinants present on VIIIR:Ag. It is present in commercial factor VIII concentrates, but not in cryoprecipitate, and is recovered from the plasma of hemophilic patients who have been infused with these concentrates. The fragment is also produced during disseminated intravascular coagulation. Although it has a similar mobility on SDS agarose electrophoresis to the smallest multimer of VIIIR:Ag, they are not immunologically identical.
Inactivation of factor VIII coagulant activity by two different types of human antibodies
Authors: Gawryl MS , Hoyer LW
Published: October 2019

Human antibodies that inactivate factor VIII procoagulant activity (VIII:C) are heterogeneous in their kinetic properties. We report there here the properties of four type I and four type II antibodies classified according to Biggs et al. Type I antibodies have second- order inactivation kinetics and completely destroy VIII:C when present in high concentration; type II antibodies have more complex kinetics and do not completely inactivate VIII:C even when tested undiluted. The latter properties correspond to the in vivo finding in some patients that there is detectable VIII:C, even though there is also a significant inhibitor titer. It has been suggested that the antibody- antigen complex in these patients retains some VIII:C activity. This is unlikely, however, since protein-A-Sepharose (PAS) did not adsorb any VIII:C activity from mixtures of type II antibodies with normal human plasma. An alternate possibility, reduced VIII:C inactivation due to a steric effect the factor-VIII-related protein (VIIIR, von Willebrand factor), appears to be a more important factor, since three of four type II antibodies had inactivating properties like type I antibodies when they were tested with separated VIII:C instead of plasma. Although the fourth type II antibody did not completely inactivate separated VIII:C the residual coagulant activity was adsorbed from this mixture by PAS. These data indicate that type II anti-VIII:C react with different antigenic determinants than type I antibodies and that these determinants are partially blocked in the factor VIII complex by VIIIR.
Modified LSA2-L2 treatment in 53 children with E-rosette-positive T- cell leukemia: results and prognostic factors (a Pediatric Oncology Group Study)
Authors: Pullen DJ , Sullivan MP , Falletta JM , Boyett JM , Humphrey GB , Starling KA , Land VJ , Dyment PG , Vats T , Duncan MH
Published: October 2019

In an attempt to improve the poor outlook for children with T-cell leukemia (T-ALL), the Southwest Oncology Group, Pediatric Division, used a modified LSA2-L2 multidrug regimen to treat 53 patients with E- rosette-positive T-ALL. This regimen was chosen because of its demonstrated efficacy in T-cell (mediastinal) non-Hodgkin's lymphoma. Complete remission (CR) rate was 88%. Range of follow-up for those patients remaining in CR is 24–49 mo (median 39 mo). Life table analysis estimates that 40% (SE 8.3%) of all patients who started induction therapy will remain failure-free at 3 yr. For patients achieving CR, 46% (SE 9%) are projected to remain in both marrow and extramedullary CR at 3 yr. Median failure-free duration was 13 mo, but only 1 patient has relapsed beyond 16 mo. Twenty-nine percent of initial relapses were isolated CNS relapses. The following presenting factors did not relate significantly to outcome: hemoglobin, platelet count, uric acid, race, and mediastinal mass. Age greater than 10 yr was a poor prognosis indicator only in the less than 50,000/microliter WBC group. Sex was not a significant factor after adjusting for WBC. WBC was the most important prognostic factor: 19% (SE 8%) of patients with WBC greater than 50,000/microliter are projected to remain failure- free at 3 yr as compared to 67% (SE 11%) of patients with WBC less than 50,000/microliter. Although the overall results are better than those previously reported for pediatric patients with T-ALL, the long-term failure-free rate remains low for patients presenting with greater than 50,000/microliter WBC.
Con-A-stimulated superoxide production by granulocytes: reversible activation of NADPH oxidase
Authors: Cohen HJ , Chovaniec ME , Wilson MK , Newburger PE
Published: October 2019

Stimulation of granulocyte (PMN) superoxide (O2-) production by concanavalin-A (Con-A) can be monitored continuously in the spectrophotometer. Both the rate of activation and final activity of the O2--generating system is dependent on the concentration of Con-A. Alpha methylmannoside (alpha MM) can prevent Con-A, but not phorbol myristate acetate (PMA) or zymosan, induced O2- production. Alpha MM inhibits both the rate of activation and the final rate of O2- production. When alpha MM is added after the attainment of a maximal rate of O2- production with Con-A, O2- production continues for another minute before it ceases. When PMA is added to such treated cells, it restores O2- production. Although the inhibition of O2- production by alpha MM on previously activated cells requires time, most of the bound concanavalin-A is removed immediately after the addition of alpha MM. Treatment of cells with L-1-tosylamido-2-phenylethyl-chloromethyl ketone (TPCK) prevents activation of PMN by Con-A to a greater extent than it does for either PMA or zymosan. TPCK has no effect on the binding of Con-A. TPCK, when added after Con-A, will inactivate O2- production by the cells. The addition of PMA after TPCK treatment restores O2--generating activity. Membrane-enriched particles from PMN activated with Con-A, alpha MM, and PMA demonstrate that the change in O2- production seen by whole cells is due to an alteration of the activity of the NADPH oxidase. Thus, Con-A stimulation of human PMN O2- production can be prevented and reversed by the addition of either alpha MM or TPCK and that PMA can reactivate Con-A and either alpha MM- or TPCK-treated cells. The activation, inactivation, and reactivation occur as a result of changes in the plasma membrane NADPH-dependent O2-- generating enzyme.
Subcellular localization of hydrogen peroxide production in human polymorphonuclear leukocytes stimulated with lectins, phorbol myristate acetate, and digitonin: an electron microscopic study using CeCl3
Authors: Ohno YI , Hirai KI , Kanoh T , Uchino H , Ogawa K
Published: October 2019

The ultrastructural H2O2-producing site in human polymorphonuclear leukocytes (PMN) stimulated with soluble stimuli was studied using a CeCl3-technique. CeLlular aggregation and formation of small vacuoles were observed when PMN were stimulated with 100 microgram/ml concanavalin-A, 1 mg/ml phytohemagglutinin, or 100 microgram/ml wheat germ agglutinin for 10 min at 37 degrees C. Electron-dense deposits formed from the reaction of H2O2 and CeCl3 were observed on the contact surface of the plasma membrane of aggregated PMN stimulated with lectins. Treatment with 5 microgram/ml cytochalasin-B before lectin- stimulation induced an enhanced formation of vacuoles, degranuLation, rounding of the contour, cellular aggregation, and enhancement of the deposits. Phorbol myristate acetate (PMA; 100 ng/ml) induced strong leukocyte aggregation, the formation of multiple huge vacuoles, degranulation, and H2O2 production at almost all of the contact surface between adjoining PMN and between PMN and erythrocytes, mononuclear cells, or thrombocytes. In PMN stimulated with digitonin (B microgram/ml), vacuolar formation, degranulation, multiple projections on the surface, and H2O2 production on the whole surface membrane were demonstrated. It is shown that cellular aggregation and cell-to-cell contact have an important role in the induction of O2- production induced by lectins or PMA and that O2- production induced by the detergent is not dependent on leukocyte aggregation.
Hemolytic anemia in hereditary pyrimidine 5'-nucleotidase deficiency: nucleotide inhibition of G6PD and the pentose phosphate shunt
Authors: Tomoda A , Noble NA , Lachant NA , Tanaka KR
Published: October 2019

We evaluated the erythrocytes of two patients with hereditary pyrimidine 5′-nucleotidase deficiency. Significant findings included an increased reduced glutathione content, increased incubated Heinz body formation, a positive ascorbate cyanide test, and decreased intraerythrocytic pH. The pentose phosphate shunt activity of the patients' red cells as measured by the release of 14CO2 from 14C-1- glucose was decreased compared to high reticulocyte controls. Glucose-6- phosphate dehydrogenase (G6PD) activity in hemolysates from control erythrocytes was inhibited 43% by 5.5 mM cytidine 5′-triphosphate (CTP) and 50% by 5.5 mM in uridine 5′-triphosphate (UTP) at pH 7.1. CTP was a competitive inhibitor for G6P (Ki = 1.7 mM) and a noncompetitive inhibitor for NADP+ (Ki = 7.8 mM). Glutathione peroxidase, glutathione reductase, and 6-phosphogluconate dehydrogenase were not affected by these compounds. Pentose phosphate shunt activity in control red cell hemolysate at pH 7.1 was inhibited to a similar degree by 5.5 mM CTP or UTP. Since the intracellular concentrations of G6P and NADP+ are below their KmS for G6PD, these data suggest that high concentrations of pyrimidine 5′-nucleotides depress pentose phosphate shunt activity in pyrimidin 5′-nucleotidase deficiency. Thus, this impairment of the pentose phosphate pathway appears to contribute to the pathogenesis of hemolysis in pyrimidine 5′-nucleotidase deficiency hemolytic anemia.