bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the role of in vivo proteolysis (fibrinogenolysis) in prolonging the thrombin time of human umbilical cord fibrinogen
Authors: Galanakis DK , Mosesson MW
Published: October 2019

These studies have been directed at evaluating the role played by proteolysis (fibrinogenolysis) in vivo in prolonging the thrombin time of human umbilical cord (“fetal”) fibrinogen. The aggregation rate of cord fibrin compared with that from adult plasma is always delayed when the reaction is carried out under conditions of relatively high ionic strength (e.g., 0.29); this difference is not apparent at relatively low ionic strength (e.g., 0.09). In addition, as assessed by turbidimetric techniques, the maximum absorbance attained by cord fibrin is considerably less than that attained by adult fibrin. Coagulable fibrinogen catabolites (i.e., fraction I-5) are present in cord plasma and, like their counterparts from adult plasma, lack various portions of the COOH-terminal region of the A alpha chain. However, their presence in plasma does not explain the behavioral differences between cord and adult fibrin. Moreover, differences revealed by turbidimetric comparison of cord and adult fibrin from plasma fraction I-2 persist in fibrin from fraction I-5; it therefore appears that the COOH-terminal region of the A alpha chain does not contain the structure(s) accounting for the unique behavior of “fetal” fibrinogen.
Effect of heterologous antibody on human platelets
Authors: Colman RW , Schreiber AD
Published: October 2019

The effect of heterologous anti-human platelet antibody on human platelet function was examined in the presence and absence of whole plasma as an in vitro model for antibody-induced immune damage to cells. Heterologous IgG anti-human platelet antibody mediated platelet aggregation and released serotonin from both platelets in plasma and from platelets isolated by gel filtration and increased the availability of platelet acid phosphatase in a dose-response fashion. Anti-platelet antibody failed to release beta-glucuronidase (lysosomal enzyme marker) or cause lactic dehydrogenase loss (cytolysis). The effect of the antiplatelet antibody on platelets proceeded in the absence of complement. The active molecule in the anti-platelet antiserum was isolated in the IgG fraction and all three indi cators of platelet injury were mediated by purified monomeric IgG. Thrombin was not required for the antibody-mediated effects, as three thrombin inhibitors failed to block the reaction. EDTA was an effective inhibitor, suggesting a cation requirement; however, as little as 38 muM calcium was sufficient for effective platelet aggregation and release. The inability of acetylsalicylic acid to inhibit the effect of the antiplatelet antibody suggests that heterologous antibody (IgG) induced platelet alteration proceeds by a different mechanism than that mediated by ADP and epinephrine and does not involve endogenous platelet prostaglandin synthesis.
Kappa and lambda light chain disease: survival rates and clinical manifestations
Authors: Shustik C , Bergsagel DE , Pruzanski W
Published: October 2019

Ninety-seven patients with light chain disease (LCD) were studied. The median survival from diagnosis was 30 mo for 52 patients with kappa-LCD and 10 mo for 45 patients with lambda-LCD (p less than 0.0007). A lower proportion of kappa-LCD patients (15.7%) than lambda-LCD patients (42.2%) died within the first 6 mo after diagnosis. The survival of the remaining patients with kappa-LCD was still much longer than of those with lambda-LCD (p = 0.022). The shorter survival of lambda-LCD patients could not be ascribed to an increased incidence of recognized manifestations indicating a poor prognosis (e.g., anemia, hypercalcemia, azotemia, low albumin, the extent of osteolytic lesions, or proteinuria), the incidence of amyloidosis, the clinical stage of the disease at diagnosis, or the response to treatment, and remains unexplained. A comparison of the clinical manifestations of LCD with those of other myelomas revealed some differences. LCD patients were slightly younger than IgA and IgG patients but older than IgD patients. A 1:1 ratio of males to females was similar to the ratios in IgA and IgG myeloma, but differed from the 3:1 ratio reported for IgD myeloma. Plasma-cell leukemia developed in 7/97 LCD patients, an incidence that was higher than has been reported in other myelomas. The initial BUN was more than or equal to 30 mg/100 ml in 54 of 95 LCD patients, an incidence that was higher than has been reported for IgA and IgG myeloma, but lower than the incidence in IgD myeloma. The incidence of amyloidosis in LCD (23 of 97 patients) was similar to that reported for IgA and IgG myeloma, but less than the incidence in IgD myeloma.
Evidence that functional subunits of antihemophilic factor (Factor VIII) are linked by noncovalent bonds
Authors: Poon MC , Ratnoff OD
Published: October 2019

Partially purified human antihemophilic factor (AHF, factor VIII), when treated with high concentrations of salt, has been shown to dissociate into two components: one, of relatively low molecular weight, possesses procoagulant activity, and the other, of higher molecular weight, forms precipitates with heterologous antiserum against AHF and supports ristocetin-induced platelet aggregation. The ease of separation suggests that the two components in the native state might be held together by noncovalent bonds. Earlier observations do not exclude the possibility that the subunits may be covalently bonded in nature but might be severed by plasma proteolytic enzymes during laboratory manipulation. The issue was examined by preparing partially purified AHF from fresh human plasma in the presence of protease inhibitors, including benzamidine, soybean trypsin inhibitor, epsilon-aminocaproic acid, heparin, and hirudin. Under these conditons, gel filtration in the presence of 0.25 M calcium chloride and 0.001 M benzamidine resulted in its separation into two components, having properties identical to those separated in the absence of these protease inhibitors. The inhibitor mixture blocked generation and action of streptokinase- and kaolin-activated plasmin from plasma, and protected both plasma AHF and partially purified AHF from the action of thrombin. Surface-induced activation of PTA (factor XI) was partially inhibited, and that of Christmas factor (factor IX) was completely inhibited. This observation provides further evidence that in the native state the high- and low-molecular-weight components of preparations of antihemophilic factor are held together by noncovalent bonds.
Immunosuppressive therapy of Factor VIII inhibitors
Authors: Hultin MB , Shapiro SS , Bowman HS , Gill FM , Andrews AT , Martinez J , Eyster EM , Sherwood WC
Published: October 2019

Immunosuppressive therapy was used in seven hemophiliac and three nonhemophiliac patients with factor VIII inhibiors. Permanent disappearance of the inhibitor occurred in three hemophiliac and two nonhemophiliac patients following treatment with cyclophosphamide and factor VIII. Critical factors influencing the response to therapy may include both the titer and duration of the inhibitor and the degree of intervening factor VIII exposure prior to immunosuppressive therapy. Two severe hemophiliacs with low titer inhibitors that disappeared without specific therapy are also reported.
Further characterization of the circulating cell in chronic lymphocytic leukemia
Authors: Schultz EF , Davis S , Rubin AD
Published: October 2019

Peripheral lymphocytes from normal individuals and from patients with chronic lymphocytic leukemia (CLL) were cultured in vitro for 1–7 days. The growth response to phytohemagglutinin (PHA) was quantitated by the incorporation of tritiated uridine into RNA nucleotide during a 2-hr pulse with the radioisotope. While the maximum response in PHA- stimulated normal cultures appeared at 2–3 days, CLL cultures required 5–7 days to develop their maximal response, which was 50%-60% of the normal magnitude. Dilution of the number of normally reactive lymphocytes by culturing them with totally unreactive, mitomycin- treated cells produced a normal 72-hr maximal response, no matter what proportion of unreactive cells was included in the PHA-stimulated cultures. In addition, the response of peripheral lymphocytes from patients with myeloblastic leukemia, where large numbers of unreactive myeloblasts diluted the normal small lymphocytes, a depressed reaction occurred at the anticipated 2–3 days. Nylon fiber-adherent lymphocytes consisting of 85% immunoglobulin (Ig)-bearing cells responded minimally to PHA, but showed no evidence of a delay. When isolated from CLL patients, both fiber-adherent cells (Ig-bearing) as well as non-fiber- adherent (sheep erythrocyterosetting) cells responded to PHA in a delayed fashion. Similarly, a case of CLL, in which 93.5% of the circulating lymphocytes bore sheep red blood cell receptors, showed its peak response to PHA at 7 days. Therefore, using surface marker criteria considered characteristic of normal T cells and B cells, the delayed response to PHA on the part of CLL lymphocytes was independent of thymic or nonthymic origin.
A diffusible stimulator of eosinophilopoiesis produced by lymphoid cells as demonstrated with diffusion chambers
Authors: Miller AM , McGarry MP
Published: October 2019

Previous experiments have indicated that eosinophilopoiesis is stimulated in lymphoid cell-dependent eosinophil responses to certain antigens. In order to study if the potential for this stimulation of eosinophilopoiesis is a function of lymphoid cells and can be expressed on challenge with the eosinophilia-inducing antigen, the diffusion chamber technique for the culture in vivo of murine hemopoietic cells has been modified. A quadrachamber diffusion assembly allows for the simultaneous maintenance in the same host of four cell populations, pairs of which are separated by a cell-impermeable Millipore diffusion membrane of defined porosity. Spleen cells for chambers were from normal mice and mice primed with tetanus toxoid; secondary challenge induces eosinophilia. These spleen cells were placed transfilter from isogeneic bone marrow cells and cultured in vivo for 6 days in normal mice that received tetanus toxoidintraperitoneally following chamber- assembly implant. The marrow cell transfilter from spleen cells of primed-donor origin exhibited significantly greater eosinophilopoiesis than contiguous-chamber marrow transfilter from normal spleen cells. Such stimulated eosinophilopoiesis was independent of total chamber marrow cellularity. The data indicated that antigen-stimulated lymphoid cells may be the source of an eosinophilopoietic factor.
Evidence for early recruitment of granulocyte precursors during high- dose methotrexate infusions in mice
Authors: Pinedo HM , Chabner BA , Zaharko DS , Bull JM
Published: October 2019

The effects of constant exposure to high concentrations of methotrexate in vivo on the committed stem cell (CFU-C) were studied by in vitro culture of mouse bone marrow. Bone marrow samples were obstained from animals receiving a continuous infusion, and were cultured in a methotrexate-free semisolid gel system. The effects of methotrexate infusion on the pluripotent stem cell population (CFU-S) were studied as well. Constant exposure to 10(-5) M methotrexate produced a rapid decrease in total nucleated cells per femur, reaching 35% of control at 12 hr and remaining at approximately this level throughout 48 hr of drug infusion. A decrease in the number of both CFU-C and CFU-S per femur was observed, which paralleled the drop in nucleated cells during the first 24 hr. However, in contrast to an additional drop in the number of CFU-S, an increase of CFU-C number per femur was observed from 24 to 48 hr. These data indicated a self-limited cell kill of nucleated bone marrow cells, and suggested recruitment of CFU-C from the CFU-S pool between 24 and 48 hr of infusion despite continued methotrexate infusion.
Granulopoiesis in chronic myeloid leukemia. II. serial cloning of blood and bone marrow cells in agar culture
Authors: Olofsson T , Olsson I
Published: October 2019

Colony-forming cells (CFC) and CFC in S-phase were assayed in chronic myeloid leukemia (CML). A correlation was found between leukocyte counts and CFC of blood, suggesting that the leukocytosis of CML depended on expansion of the committed granulopoietic stem cell compartment. Serial studies performed in four cases demonstrated a decrease of the CFC in S-phase during early stages of developing leukocytosis, which was consistent with the operation of growth control mechanisms. During later stages, serial studies revealed that sudden increments of CFC S-phase coincided with rapidly growing leukocytosis, which was consistent with leukemic cell populations escaping growth control. H-thymidine labeling indices for differentiated precursor cells showed slight variations not coinciding with variations of the CFC S-phase fraction. Cyclic oscillations of the white cell count were observed in one case. The white cell count and the fraction of CFC in S- phase displayed a direct relationship, indicating that the occurence of cycles was not likely to be due to a negative feedback mechanism elaborated by mature granulocytes. In another case, marked cycling of the CFC S-phase fraction was found without distinct oscillations of the white cell count. The present work has emphasized the necessity for serial assays of parameters of cell kinetics in vitro in CML since changing relationships were found between stem cell and differentiated cell kinetics during different phases of the disease.
Factors which affect erythropoiesis in partially nephrectomized and sham-operated rats
Authors: Anagnostou A , Vercellotti G , Barone J , Fried W
Published: October 2019

Current concepts of the pathogenesis of anemia in uremic animals are derived mainly from the results of studies performed either in vitro or in bilaterally nephrectomized animals. These data may not be applicable to the situation which exists in more chronically uremic animals. In 1932, Chanutin and Ferris showed that removal of five-sixths of the renal mass caused rats to become uremic and to remain so for a prolonged period of time. Rats made uremic in this manner were utilized as models for studying the pathogenesis of the anemia of uremia. Removeal of five-sixths of the renal mass of rats caused their BUNs to rise to over 100 mg/100 ml and to remain at this level for over 3 wk. The hematocrits of these uremic rats fell from 42% to approximately 30% in 3 wk. Erythropoietin (Ep) production immediately fell to a barely detectable level postoperatively and did not increase significantly in 3 wk, although the renal remnant hypertrophied. Extrarenal Ep production also remained at a low level and did not increase during the 3-wk observation period. The response of plethoric uremic rats to 2 units of Ep was as great (in some experiments greater) as that of sham- operated ones. A surprising finding was that plethoric uremic rats, injected with saline rather than with Ep, incorporated more 59Fe into their red blood cells than did sham-operated ones. This finding suggested that in uremic rats erythropoiesis was less markedly suppressed by plethora than it was in non-uremic rats.
Distribution of spermidine and spermine in blood from cystic fibrosis patients and control subjects
Authors: Cohen LF , Lundgren DW , Farrell PM
Published: October 2019

Previous studies have shown an abnormality of the spermidine-to- spermine (Spd/Spm) ratio in whole blood of cystic fibrosis homo-and heterozygotes. To investigate Spd and Spm distribution amoung blood components as a possible cause of the abnormality, blood was fractionated using Rabinowitz's glass bead technique and Boyum's Ficoll- Hypaque method. Free (unconjugated) polyamines were extracted with perchloric acid and quantitated on an amino acid analyzer. In controls, mean +/- SEM concentrations in nmoles/10(9) cells of Spd and Spm, respectively, were 1.02 +/- 0.08 and 0.894 +/- 0.28 for erythrocytes; 126 +/- 31 and 357 +/- 105 for lymphocytes; 36 +/- 16 and 240 +/- 33 for granulocytes; and less than 0.5 and less than 0.5 nmoles/ml for plasma. When converted to the concentration in whole blood, it was found that greater than 90% of Spd and over 70% of Spm was associated with erythrocytes. While the higher cellular concentration in leukocytes was not unexpected, the fact that Spd and Spm in whole blood were primarily associated with erythrocytes was a new finding. Comparison with controls revealed that the Spd/Spm ratio in both whole blood and erythrocytes was significantly higher in the group of cystic fibrosis patients.
Immunologic characterization of canine factor VIII
Authors: Benson RE , Dodds WJ
Published: October 2019

Canine factor VIII (FVIII) preparations isolated from cryoprecipitates by gel chromatography were pooled to provide one batch of antigen for simultaneous immunization of two rabbits and a goat. The goat and rabbit antisera had similar FVIII-neutralizing titers, but the latter had seven to ten times more precipitating titer for FVIII-related antigen (FVIII-RA). Absorption with material low in FVIII had little effect on the precipitating titer of the rabbit antibody, but it abolished the precipitating capacity of the goat antibody and caused a 20% reduction in the neutralizing titer of both antisera. Results obtained in the Laurell assay with the two different antisera were similar. This finding was true whether the FVIII-RA levels were reduced, normal, or elevated, as well as for heat-treated and frozen- thawed plasmas. Both antisera were neutralized by the same canine plasma to a similar extent. Analysis of FVIII concentrates by crossed immunoelectrophoresis suggested that canine FVIII-RA was heterogeneous, with slow-and fast-migrating components. The presence of more than one antigenic site on the FVIII complex was also supported by the disparity between the FVIII-neutralizing and -precipitating titers of goat antiserum and by the demonstration that FVIII-RA, FVIII-neutralizing antigen, and procoagulant activity varied independently.
Duplication of part of the long arm of chromosome 1 in marrow cells of a treated case of myelomatosis
Authors: Spriggs AI , Holt JM , Bedford J
Published: October 2019

In a case of classical myelomatosis treated with melphalan, a clone of cells with a chromosomal abnormality was found in the bone marrow during remission. There was good reason to think that the hemopoietic cells, rather than plasmacytoma cells, were implicated. Although the clone persisted, no evidence of leukemia developed over a period of observation of 2 yr. The anomaly was interpreted as a duplication of part of the long arm of chromosome 1, which appeared to involve the segment q21 to q31.
Marrow regeneration after mechanical depletion
Authors: Brecher G , Tjio JH , Smith WW , Haley JE
Published: October 2019

The origin of marrow regeneration after mechanical depletion was reinvestigated in mouse chimeras. The results were compatible with the local origin of stem cells from remnants of incompletely removed marrow, but not with their origin from a common precursor of both bone and hemopoietic cell lines. In transplanted femurs depleted by a modified technique of in vivo evacuation of marrow, hemopoietic regeneration failed to occur. The presence of hemopoietic stem cells in the Haversian canals was thus excluded. The demonstration of ample hemopoiesis with minimal bone formation in nondepleted controls in which bone marrow initially became necrotic provided new evidence that osteogenesis was not a prerequisite of hemopoietic regeneration.
Size distribution, electronic recognition, and counting of human blood monocytes
Authors: Loos H , Blok-Schut B , Kipp B , van Doorn R , Meerhof L
Published: October 2019

During a study on the separation of human blood monocytes from lymphocytes, a method was developed to recognize and count monocytes by electronic means. Lightscattering (Cytograf, Bio/Physics), and changes in electrical resistance (Channelyzer, Coulter) were used to size mononuclear leukocytes directly in cell suspensions. Both methods revealed a size distribution profile in which two populations of mononuclear leukocytes could be distinguished. The largest cells were virtually eliminated after phagocytosis of iron particles. We confirmed that these cells were monocytes by three different criteria: the intracellular lysozyme activity, the number of phagocytes, and the percentage of cells with kidney-shaped nuclei. The highly significant correlations we found showed that monocytes could be recognized and counted by electronic sizing. For this method, purified mononuclear leukocyte preparations had to be used, since the presence of erythrocytes, platelets, and polymorphonuclear cells interfered. Statistical analysis revealed that electronic sizing permitted discrimination of differences in monocyte content of 4.5%, with a probability of 95%. It was calculated that this sensitivity of electronic monocyte counting was about three times higher than the sensitivity of microscopic methods. Since 100,000 cells can be sized within a few seconds, not only the efficiency of the preparation but also minor changes in the size of monocytes and lymphocytes introduced during the isolation can be followed.
A simplified in vitro classification for prognosis in adult acute leukemia: the application of in vitro results in remission-predictive models
Authors: Spitzer G , Dicke KA , Gehan EA , Smith T , McCredie KB , Barlogie B , Freireich EJ
Published: October 2019

Previous classification in vitro of adult acute leukemia incorporating morphology has been complex and difficult to understand. We have devised a simplified classification based solely on leuekemic proliferation in vitro. Seventy-six patients with adult acute leukemia previously untreated were included in this study and received identical chemotherapy. Three groups were recognized. The complete remission rate was 76% in the 21 patients with no leukemic growth in vitro (Group 1), 75% in 36 patients with leukemic cell growth but aggregates of 20 cells or less (Group 2), and only 21% in the 15 patients with aggregates of greater than 20 (Group 3). There was a highly significant difference in complete remission rates between Group 3 and the other two groups (p less than 0.001). Linear logistic regression analysis demonstrated the independence of the growth in vitro from other prognostic variables. A predictive model utilizing the in vitro result more accurately predicted for remission, both retrospectively and prospectively, than a model constructed with presently known prognostic parameters. The cause of death in failures suggested that this system detects resistance to the chemotherapy.
Fetal erythropoiesis following bone marrow transplantation
Authors: Alter BP , Rappeport JM , Huisman TH , Schroeder WA , Nathan DG
Published: October 2019

“Fetal” erythrocytes are present in older children and certain adults with hematologic disorders. To determine if regenerating bone marrow produces such cells, we examined the blood of seven allogeneic bone marrow transplant recipients. Six patients were engrafted with donor cells, while on e patients recovered autologous bone marrow after rejection of a marrow transplant. All seven patients had fetal hemoglobin levels of up to 10% by 100 days after transplant. In three patients, the Ggamma to Agamma ratio in the fetal hemoglobin was “newborn”, while in one it was “adult”. Gamma chain synthesis in blood and bone marrow never exceeded 20% of total non-alpha globin synthesis. The fetal hemoglobin was heterogeneously distributed in the cells. High titer i antigen also appeared. All fetal characteristics declined by 200 days. Erythropoiesis during bone marrow recovery appears to be associated with an accelerated, albeit partial, recapitulation of ontogeny.
Kinetics of mobilization of neutrophils and their marrow pool in protein-calorie deficiency
Authors: Suda AK , Mathur M , Deo K , Deo MG
Published: October 2019

Migration of marrow neutrophils under basal conditions and their mobilization, following subcutaneous implantation of cover slips, were investigated in groups of protein-deficient rats, using 3HTdR with sequential autoradiography of the peripheral blood smears. Animals fed a protein-rich diet served as controls. The pattern of appearance of labeled neutrophils in the blood was identical in the two groups under basal conditions. However, a higher percentag of labeled neutrophils appeared earlier in the blood following cover slip implantation in the deficient rats as compared to controls. The inflammatory exudate on the cover slips was low throughout the period of observation in deficient animals, with a delay in the appearance of monocytes. A pool of mature neutrophils resides in the bone marrow. It is proposed that in PCM there is atrophy of all neutrophil compartments, including that of the marrow pool, associated with a proportionate reduction in the efflux of cells from one compartment to another. This proportionate reduction in efflux would explain the normal kinetics of migration of neutrophils under basal conditions in the deficient rats in spite of a reduction in the marrow pool. On the other hand, in cover slip-implanted deficient rats, appearance of larger numbers of labeled neutrophils in the blood is attributed to a reduction in size of the marrow pool. This hypothesis is substantiated by the experiments in protein-deficient monkeys in which estimation of the marrow pool of neutrophils revealed a marked reduction in the deficient animals. It is further proposed that a diminuation of the marrow pool of neutrophils and retarded mobilization of cells at the site of inflammation are important mechanism responsible for the increased susceptibility of the malnourished host to infections.
Phosphorylation in erythrocyte membranes from abnormally shaped cells
Authors: Greenquist AC , Shohet SB
Published: October 2019

Erythrocyte protein phosphorylation was examined in membrane preparations of 25 patients with hereditary spherocytosis (HS). Reduced phosphorylation in substrate polypeptides was observed in 22 HS erythrocyte membranes for both splenectomized and nonsplenectomized patients. A reduction in labeling was also observed in a polypeptide which was labeled only in the presence of cAMP. No reduction was observed in membranes from immunologically acquired spherocytosis cells or from cells from patients with hereditary elliptocytosis. Heating membranes to 45 degrees C caused negligible inhibition of membrane protein phosphorylation, while heating to 50 degrees C extensively inhibited phosphorylation. Dephosphorylation of membrane protein that occurred in isolated membranes was not dependent upon cAMP.
Inhibition and potentiation of platelet function by lysolecithin
Authors: Joist JH , Dolezel G , Cucuianu MP , Nishizawa EE , Mustard JF
Published: October 2019

The effects of lysolecithin (LPC) on aggregation, serotonin release, shape, and lysis of rabbit, pig, or human platelets in platelet-rich plasma (PRP) or Tyrode albumin solution were examined during prolonged incubation. LPC added to citrated or heparinized PRP from humans or rabbits at a final concentration above 100 muM caused instantaneous inhibition of platelet aggregation induced by adenosine diphosphate (ADP), epinephrine (human PRP only), collagen, or thrombin. The inhibitory effect of LPC was found to be partially reversible over a period of 60–90 min. LPC at final concentrations above 30 muM also caused inhibition of ADP-, collagen-, and thrombin-induced aggregation and collagen- and thrombin-induced release of serotonin in suspensions of rabbit, pig, or human platelets. With washed platelets, the inhibitory effect not only rapidly disappeared but was followed by transient potentiation of aggregation and serotonin release. This potentiating effect of LPC was most pronounced when thrombin was used as stimulus. Both inhibition and potentiation were observed at concentrations of LPC that did not cause a significant change in platelet shape or loss from platelets of lactic dehydrogenase. Inhibition and potentiation were also observed when platelets were added to suspending medium containing LPC, although considerably higher concentrations of LPC were required under these conditions. Potentiation was not observed when LPC was added to citrated or heparinized rabbit or human PRP or to washed rabbit platelets suspended in a medium containing 4% bovine serum albumin. It seemed likely that some or all of the observed effects of LPC on platelet function were due to structural modification of the platelet membrane insufficient to result in gross membrane damage or platelet lysis. In addition, the results of experiments using 14C-LPC seemed to indicate that the observed potentiating effect of LPC on platelet function may be related to its rapid uptake and metabolism by the platelets.
Immunologic differences in human isoferritins: implications for immunologic quantitation of serum ferritin
Authors: Hazard JT , Yokota M , Arosio P , Drysdale JW
Published: October 2019

Immunologic differences in several tissue isoferritin populations have been investigated using antibodies developed in rabbits against human liver ferritin and in guinea pigs against HeLa cell ferritin. Our results indicate that the quantity of ferritin measured by radioimmunoassays is markedly affected by the isoferritin and subunit composition of the ferritin sample as well as by the nature of the antiferritin antibodies. Possible applications for the selective quantitation of different serum ferritin populations are discussed.
Thrombocytotic suppression of megakaryocyte production from stem cells
Authors: Goldberg J , Phalen E , Howard D , Ebbe S , Stohlman F Jr
Published: October 2019

Megakaryocytopoiesis in the spleens of lethally irradiated mice transplanted with marrow cells was suppressed by platelet transfusions. In one group of experiments, animals were irradiated and transfused with bone marrow cells on day O. They were then given either no treatment, platelets, platelet-poor plasma, or saline on days 0, 2, 4, 6, and 8, and then were sacrificed on day 10. Megakaryocytes per section in the spleens of mice receiving platelets were 24%-48% of the values in the groups given plasma, saline, or bone marrow only. The number of pure megakaryocyte colonies was also diminished by platelet hypertransfusion. Another experiment examined the effect of platelets or plasma administered on days 1 and 2 or days 6 and 7 after irradiation and bone marrow transfusion. Hypertransfusion on days 6 and 7 was as effective in suppressing megakaryocytopoiesis as hypertransfusion every other day for 10 days. Animals given platelets or plasma only on days 1 and 2 did not have any significant change in their megakaryocyte number. These results implied that committed megakaryocyte precursors were more sensitive to inhibition by increased platelet levels than pluripotential stem cells. Further experiments with plethoric animals indicated that different levels of erythropoietin did not account for the effects of platelet hypertransfusion. The findings could be explained by inhibition of cell proliferation or of differentiation of megakaryocyte precursors by increased platelet levels.
Hematopoietic stem cells: effect of preirradiation, bleeding, and erythropoietin on thrombopoietic differentiation
Authors: Goodman R , Grate H , Hannon E , Hellman S
Published: October 2019

A method of measuring differentiation of stem cells towards platelets is described using syngeneic bone marrow injected into lethally irradiated mice. Fourteen days after such injection, the platelet counts are found to be proportional to the number of bone marrow cells injected and can be used as a measure of platelet progenitors. Perturbation of the milieu in which the transplanted marrow is placed by host preirradiation, bleeding, or erythropoietin administration leads to enhanced thrombopoiesis. It has been shown previously that similar perturbation favors erythropoiesis at the expense of granulopoiesis. The data from these and other experiments appear to be consistent, with a model of the stem cell compartment as a continuum with proliferative activity increasing as commitment is restricted. These functions vary inversely with the capacity for self-renewal. The various stem cell assays measure different ranges of stem cells, but overlap within this continuum.
Formation of lipid inclusions in normal human leukocytes
Authors: Lutas EM , Zucker-Franklin D
Published: October 2019

Normal peripheral blood leukocytes left standing at room temperature develop large inclusions which increase in number and size with time of incubation. The formation and nature of these inclusions were investigated. At 0 hour the structures were present in only 1% of the cells, whereas at 48 hr, they were in virtually all neutrophils and monocytes. Ultrastructurally, the globules measured 0.5–1.5 mu in diameter and they were usually not membrane bound. Histochemical analysis indicated that they were lipid in nature. The inclusions were separated on a sucrose density gradient and their isolation was confirmed by electron microscopy. Thin-layer chromatography of the isolated globules suggested that they consisted predominantly of triglycerides. Since it is known that the cells synthesize triglycerides at rest, it was postulated that the structures may represent the storage form for free fatty acids which may be utilized for membrane synthesis during phagocytosis.
Immunodeficiency in patients with non-Hodgkin lymphomas
Authors: Jones SE , Griffith K , Dombrowski P , Gaines JA
Published: October 2019

Seventy-one previously untreated patients with non-Hodgkin lymphomas were studied with several readilyvailable tests of immune function: number of peripheral blood lymphocytes, serum immunoglobulins, and delayed hypersensitivity to six recall antigens. The results were correlated to histology (Rappaport classification), stage (Ann Arbor classification), the presence of symptoms, and survival. As a group, 38 patients with diffuse lymphomas exhibited marked impairment in reactivity to five of six antigens (p less than 0.03 to p less than 0.001). In addition, lymphopenia and reduced levels of serum IgA were found in association with diffuse histiocytic lymphoma. Among patients with diffuse lymphoma, lymphocyte number and skin test reactivity tended to be greater in those with localized disease or without constitutional symptoms, and survival was superior for patients free of symptoms (p less than 0.01). As a group, 33 patients with nodular lymphoma had normal numbers of lymphocytes, lower levels of serum IgG and IgA, and significant impairment of reactivity to two antigens (streptokinase-streptodornase and mumps; p less than 0.01); reactivity to three other antigens (Candida albicans, coccidiodin, and tuberculin) was normal. Survival for patients with nodular lymphoma was superior (p less than 0.01) compared to those with diffuse lymphomas. In summary, severe immunodeficiency was found in patients with diffuse lymphoma (particularly diffuse histiocytic lymphoma), and definite but much less severe immunodeficiency was characteristic of patients with nodular lymphoma.
The Philadelphia chromosome in human macrophages
Authors: Golde DW , Burgaleta C , Sparkes RS , Cline MJ
Published: October 2019

Three patients with chronic myelocytic leukemia in different phases of the natural history of the disease were studied. Their bone marrow cells were cultured under conditions favoring macrophage proliferation, and parallel cytogenetic and cytochemical studies were performed. All cell metaphases examined contained the Ph1 chromosome at a time when more than 80% of these metaphases were in identifiable macrophages. We conclude that the mononuclear phagocyte cell line contains the abnormal chromosome in Ph1-positive chronic myelocytic leukemia.
Polycythemia vera: hormonal modulation of erythropoiesis in vitro
Authors: Golde DW , Bersch N , Cline MJ
Published: October 2019

We performed in vitro culture studies examining the interaction of erythropoietin with red cell progenitors in polycythemia vera. Bone marrow was obtained from five patients with typical disease and from five healthy volunteers, and assayed for erythroid colony formation (CFU-E) by the methylcellulose technique. In cultures without added erythropoietin, a mean eightfold greater cloning efficiency was noted with the polycythemia vera marrows, as compared to normal. There was prominent stimulation of colony formation by erythropoietin, and the shape of the erythropoietin dose-response cruves appeared to be similar in both patients and controls. Anti-erythropoietin antibody reduced the number of CFU-E in cultures not containing added erythropoietin, but did not eliminate them. Dexamethasone (10(-9) M) caused a consistent increase in CFU-E in the patients' cultures. These studies provide evidence for functional erythropoietin and glucocorticosteroid receptor mechanisms on erythroid precursors in polycythemia vera. The observations are consistent with a concept of this disease as a disorder of hematopoietic stem cells in which peripheral erythrocytosis is caused by an expanded erythroid progenitor compartment which maintains responsiveness to hormonal modulation.
Treatment of juvenile chronic myeloid leukemia with sequential subcutaneous cytarabine and oral mercaptopurine
Authors: Lilleyman JS , Harrison JF , Black JA
Published: October 2019

Three cases of the juvenile type of chronic myeloid leukemia are described, all of which have shown a clinical and hematologic response to repeated cycles of sequential subcutaneous cytarabine and oral mercaptopurine. It appears that this regime, as described, may afford some control of a disease process previously supposed to be unresponsive to chemotherapy. This control, however, is likely to be reflected in an improvement in well-being, rather than overall survival time. Patient 1 survived 26 mo from diagnosis, and patients 2 and 3 are alive and well 12 and 9 mo from diagnosis, respectively.
Cell size and plamitate-1-14c turnover of rabbit marrow fat
Authors: Trubowitz S , Bathija A
Published: October 2019

Adipose tissue is a major component of normal rabbit marrow. Morphological considerations suggest an active role for this tissue in hematopoiesis. This hypothesis was tested by injecting 50 micronCi of palmitate-1–14C intravenously into fed, hematologically normal New Zealand rabbits. The animals were sacrificed 24 hr later and the femoral marrow removed. Samples of subcutaneous and perinephric fat were taken for comparison. The fat cells were isolated by the Rodbell method and the diameters measured. Incorporation of the 14C-palmitate in the triglyceride fraction was determined and the composition of the fatty acids was measured by gas chromatography. The mean diameter of the marrow fat cell was 46 micronm (mean cell volume 55 pl); the mean diameter of the perinephric fat cell 70 micronm (mean cell volume 200 pl). 14C-Palmitate turnover per gram triglyceride was some fivefold greater in the marrow fat; however, when expressed on a cell basis, the turnover for the marrow and perinephric fat cell was similar. The marrow fat contained a higher concentration of unsaturated fatty acids. These findings suggest that there is greater lipolysis and lesser storage in the marrow fat than in the perinephric.
Circulation of donor lymphocytes after blood transfusion in man
Authors: Schechter GP , Whang-Peng J , McFarland W
Published: October 2019

Atypical lymphocytes (ATL) appear in the circulation of a large proportion of patients during the first week following a blood transfusion. In order to determine the source of these ATL, cytogenetic studies were performed on the peripheral blood leukocytes of ten adult patients who received fresh blood from donors of the opposite sex. Nine of the ten patients had spontaneously dividing mononuclear cells of the recipient or host karyotype circulating during the latter part of the first week after transfusion. In two patients, the spontaneously dividing cells were of donor as well as of host origin. Six patients had circulating phyothemagglutinin-responsive lymphocytes of the donor karyotype noted from 1–7 days after transfusion. These findings lend support to our hypothesis that the increase in circulating atypical lymphocytes seen 1 wk after transfusion represents the counterpart in vivo of the in vitro mixed leukocyte reaction. The dividing donor cells may represent a subclinical graft-versus-host reaction.
Pretreatment cytokinetic studies in 94 children with acute leukemia. Relationship to other variables at diagnosis and to outcome of standard treatment
Authors: Murphy SB , Aur RJ , Simone JV , George S , Mauer AM
Published: October 2019

The relationship of the pretreatment bone marrow mitotic index (MI) and in vitro 3H-thymidine labeling index (LI) to other variables present at diagnosis and to the outcome of standard therapy was examined in a series of 94 children with acute leukemia (71 ALL, 23 AML). The range of observed values at diagnosis was extremely broad, from less than 1% labeled cells to more than 25%. The median LI% for all patients studied was 5.2%; the median MI% was 0.3%. The values for AML and ALL did not differ significantly despite older age, higher initial white blood cell (WBC) counts, and poorer response to therapy of AML patients. The initial MI and LI were positively correlated with each other, but unrelated to age or initial WBC count in either ALL or AML. However, the LI and MI were significantly higher (p less than 0.01) in the group of 10 children of 54 studied with ALL whose blasts formed spontaneous rosettes with sheep red blood cells at 37 degrees C. The initial LI and MI were not shown to be related to the likelihood of attaining a complete response or to the length of remission in either ALL or AML. There was thus no evidence that either the initial MI or LI% of marrow blasts was of any prognostic significance in children with acute leukemia.
Abnormalities in granule formation in acute myelogenous leukemia
Authors: Bainton DF , Friedlander LM , Shohet SB
Published: October 2019

Granule formation was investigated in differentiating neutrophils of a patient with acute myelogenous leukemia (AML) by means of the combined techniques of electron microscopy and peroxidase cytochemistry. Two important pathologic features were observed: first, an abnormal concentration and packaging of peroxidase into Auer rods in leukemic promyelocytes, and second, the presence of Auer rods surrounded by single-unit membranes in some mature polymorphonuclear leukocytes (PMN). An additional unexpected finding was the discovery of two distinct populations of PMN circulating concurrently; a minor (less than 5%) normal one that contained both peroxidase-positive azurophilic and peroxidase-negative specific granules and a major abnormal one characterized by the absence of specific granules. None of these abnormalities was observed during the two remissions of this patient's disease. During relapse a “hiatus leukemicus” occurred, which also revealed two populations of cells, a majority population of leukemic blasts, and a minority population of a few normal PMN. These findings documented several developmental abnormalities in the differentiating cells of myelogenous leukemia and also suggested that concurrent normal and abnormal populations of PMN may be a helpful diagnostic feature of a leukemic process.
Characteristics of blast crisis in chronic granulocytic leukemia
Authors: Rosenthal S , Canellos GP , DeVita VT Jr , Gralnick HR
Published: October 2019

The terminal phase of most patients with Ph1-positive chronic granulocytic leukemia (i.e., blast crisis) resembles acute leukemia. The clinical and hematologic features of blast crisis in 73 patients with chronic granulocytic leukemia have been reviewed. Two major morphological subgroups, lymphoblastic and myeloblastic, were identified. The lymphoblastic group in general had more profound thrombocytopenia and a greater number of blasts, while the myeloblastic group had more severa anemia. Extramedullary leukemia was documented in 27 patients. In 12 patients extramedullary leukemia preceded or occurred simultaneously with blast crisis in the bone marrow and peripheral blood. On the basis of this study we present hematologic criteria for the diagnosis of blast crisis and emphasize the importance of extramedullary leukemia in heralding the onset of blast crisis.
Depressed in vitro lymphocyte responses to PHA in patients with Hodgkin disease in continuous long remissions
Authors: Case DC Jr , Hansen JA , Corrales E , Young CW , Dupont B , Pinsky CM , Good RA
Published: October 2019

Twenty consecutive patients with Hodgkin disease in continuous complete remission and off treatment for at least 5 yr (range 5–25 yr, median 9 yr) were studied with a battery of immunologic parameters. Skin test reactivity to four common antigens, sensitization to 2,4- dinitrochlorobenzene, absolute lymphocyte count, relative percentage of T cells (as measured by spontaneous rosette formation with sheep erythrocytes) and B cells (as measured by immunofluorescence with polyvalent antiserum), and absolute number of T and B cells were normal when compared with controls. However, the mean value of lymphocyte response in vitro to the mitogen phytohemagglutinin for the study population was significantly decreased (p less than 0.001) when compared with the controls. This abnormality in response to mitogen could not be correlated with age, sex, stage, symptoms, histologic subclassification, or previous treatment. The data suggest the existence of a persisting cell-mediated immune defect in the circulating lymphocytes in patients with long-standing Hodgkin disease that might otherwise be considered “cured.”
Antibody response to intravenous immunization following splenic tissue autotransplantation in Sprague-Dawley rats
Authors: Schwartz AD , Dadash-Zadeh M , Goldstein R , Luck S , Conway JJ
Published: October 2019

The response to intravenous challenge with sheep erythrocytes was determined in Sprague-Dawley rats following autotransplantation of splenic tissue into the subcutaneous tissue, peritoneal cavity, or a surgically created omental pouch. There was a marked rise in heterophil antibody titer following intravenous challenge in ten control animals, and no rise in titer in nine of ten asplenic animals. Heterophil antibody titers increased in all of the animals with transplants, but the response in some was less than that in control animals. Thus autotransplanted splenic tissue demonstrated immunologic responses similar to those of normal, intact spleen.
Eosinophil colony formation in semisolid cultures of human bone marrow cells
Authors: Dresch C , Johnson GR , Metcalf D
Published: October 2019

A method for in situ staining and scoring of eosinophil colonies and clusters in the whole culture dish has been developed for agar cultures of human marrow cells. Cultures stimulated by human placental conditioned medium were found to develop the same proportion of eosinophil, neutrophil, and monocyte colonies as cultures stimulated by conventional peripheral white cell underlayers. Eosinophil colonies and clusters always contained pure populations of eosinophils, and no mixed colonies of eosinophils with other cells were observed. Eosinophil colony formation was delayed in onset relative to neutrophil or monocyte colony formation, but by day 14 eosinophil colonies comprised 20% of all colonies. Eosinophil colonies were absent in cultures of marrow cells from patients with acute myeloid leukemia in relapse, but eosinophil colonies and clusters were found with higher than normal frequency in cultures from acute leukemic patients in remission. A high frequency of eosinophil colonies and clusters was also observed in cultures from three patients with idiopathic thrombocytopenic purpura. The new staining procedure should permit a wide range of studies on eosinophil precursor populations in the marrows of patients with various diseases.