bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of short-term hypoxia on platelet counts of mice
Authors: McDonald TP , Cottrell M , Clift R
Published: October 2019

Recent studies have shown that long-term hypoxia causes decreased platelet counts in mice and short-term hypoxia increased platelet counts. In an attempt to explain the mechanism that increases platelet counts of mice after exposure to short-term hypoxia, we measured platelet counts, total circulating platelet counts (TCPC), total circulating platelet masses (TCPM), percentages of 35S incorporation, and platelet sizes. Platelet counts, as well as TCPC and TCPM of mice, increased after 1–3 days of hypoxia, but these values were decreased after 6–7 days of hypoxia. Although platelet counts were increased in hypoxic mice, the percentage 35S incorporation into platelets and platelet sizes did not show a concurrent increase. After 6 days of hypoxia, average platelet diameters began to increase as platelet counts decreased. Splenic release did not account for the increase in platelet counts of mice after short-term hypoxia. It seems possible, therefore, that megakaryocytes “shed” platelets into the circulation in response to hypoxia. The platelets that enter the circulation in response to short-term hypoxia are smaller and incorporate less 35S than platelets that are produced in response to acute thrombocytopenia.
Hairy-cell leukemia with T-cell features
Authors: Cawley JC , Burns GF , Nash TA , Higgy KE , Child JA , Roberts BE
Published: October 2019

A case of clinically and hematologically typical hairy-cell leukemia has been presented in which, at the various times of testing, 52%–95% of peripheral blood and 73% of splenic mononuclear cells formed spontaneous sheep erythrocyte (E) rosettes. Many of the rosetting cells were shown to be typical morphologic hairy cells by light and electron microscopy. It was found that 70%–75% of peripheral blood mononuclear cells stained with an anti-T antiserum, and this antiserum also abolished E-rosette formation. A variable percentage of peripheral blood mononuclear cells was also shown to bear surface (IgDK) and internal (IgMK and IgGK) immunoglobulins. Additional B-cell features demonstrated included possession of the P29/34 la-like antigen and formation of mouse rosettes. It was demonstrated by a variety of blocking and inhibition studies that the E-rosette formation was not attributable to chance antigen specificity of the surface membrane immunoglobulin. These marker studies suggest that this is a case of true hybrid cell HCL. Despite these unusual marker characteristics, the patient showed no distinctive clinical or hematologic features.
Regulation of megakaryopoiesis in long-term murine bone marrow cultures
Authors: Williams N , Jackson H , Sheridan AP , Murphy MJ Jr , Elste A , Moore MA
Published: October 2019

Megakaryocytes and their precursor cells were sustained in mouse bone marrow suspension cultures for over 4–6 wk. Megakaryocyte precursor cells were detected by their capacity to form colonies of megakaryocytes in semisolid agar cultures. Colony formation was dependent on the presence of medium conditioned by a myelomonocytic leukemic cell line (WEHI-3CM). Megakaryocytes from the liquid and semisolid cultures were identified by cytoplasmic acetylcholine esterase and by ultrastructural analysis. The suspension medium from the bone marrow liquid cultures which sustained megakaryopoiesis was not directly acitive in stimulating megakaryocyte colony formation in the semisolid agar cultures, but potentiated the number of colonies detected when WEHI-3CM was present. Bone marrow-conditioned medium increased the sensitivity of megakaryocyte progenitor cells to the stimulus in WEHI-3CM. Addition of the activities present in the two sources produced a quantitative assay for the detection of mouse megakaryocyte progenitor cells. These studies showed: (1) that no inductive regulator of in vitro clones of megakaryocytes was present in the supernatants from the long-term marrow cultures and, (2) that at least two factors were necessary for the induction of megakaryocyte progenitors to proliferate and differentiate in semisolid cultures in vitro.
Hb Potomac (101 Glu replaced by Asp): speculations on placental oxygen transport in carriers of high-affinity hemoglobins
Authors: Charache S , Jacobson R , Brimhall B , Murphy EA , Hathaway P , Winslow R , Jones R , Rath C , Simkovich J
Published: October 2019

Blood from a woman with unexplained erythrocytosis had increased oxygen affinity, but no abnormality could be detected by electrophoresis or chromatography of her hemolysate. Separation of the tryptic peptides of her beta chains disclosed two half-sized peaks in the regions of beta T- 11. The faster of these was abnormal, with the structure beta 101 Glu replaced by Asp. The new hemoglobin was called “Potomac.” Three of the proband's four surviving siblings and both of her children were carriers. Differences in the ratio of carrier: normal children born to male of female carriers of 23 other high-affinity hemoglobins were not significant. The high proportion of carriers in this kindred was probably due to chance alone, and not because high maternal oxygen affinity interfered with oxygen transport to fetuses with normal hemoglobin.
Determinants of fecal and urinary iron excretion in desferrioxamine- treated rats
Authors: Hershko C
Published: October 2019

Radioiron excretion in the urine and feces after continuous subcutaneous infusion of desferrioxamine (DF) was studied in normal and hypertransfused rats in whom reticuloendothelial and parenchymal iron stores had been labeled by selective 59Fe probes. The existence of two alternative pathways for the chelation of iron in vivo by DF was indicated by the results. The first pathway was intracellular chelation in hepatocytes with chelated iron excreted through the bile only. The second pathway, which was extracellular, could only be activated after saturation of the transferrin iron binding capacity, with the chelated iron excreted by the kidneys. The last mechanism was probably identical with the mode of action of diethylenetriaminepentaacetic acid (DTPA). Further studies will be required in order to establish whether the enhancement in iron chelation obtained by the continuous infusion of DF in patients with complete saturation of circulating transferrin may or may not be related to the extracellular mechanism of iron chelation in hypertransfused rats.
Lymphocyte receptors for concanavalin A in Hodgkin disease
Authors: Aisenberg AC , Weitzman S , Wilkes B
Published: October 2019

The number of lymphocytes with mobile receptors for concanavalin A (Con A) on their surface membrane (forming visible caps after the addition of fluorescein-conjugated Con A) was determined in the peripheral blood of 53 patients with Hodgkin disease. Of 29 individuals studied prior to treatment, the level of capped cells was found to be below the normal range in 9 of 13 in stages I and IIA, 6 of 8 in stage IIIA, and all 8 in stages IIIB and IV. Even among patients in remission 2 yr after successful treatment the level was below the lower normal limit in 9 of 16. The number was also reduced in 7 of 8 individuals with recurrent lymphoma. The level of lymphocytes that cap with Con A may prove to be a more sensitive measure of active Hodgkin disease than the total peripheral lymphocyte count or the level of T cells. This lymphocyte parameter merits further study as a correlate in vitro of cellular immunity.
Three stages of erythropoietic progenitor cell differentiation distinguished by a number of physical and biologic properties
Authors: Gregory CJ , Eaves AC
Published: October 2019

Previous studies have shown that erythroid precursors at sequential stages of differentiation along the red cell pathway can be distinguished by differences in the size and maturation kinetics of the colonies to which they give rise in vitro. Using criteria based on these two parameters, it is thus possible to identify three distinct erythroid progenitor cell populations in the mouse, known as day 8 BFU- E, day 3 BFU-E, and CFU-E. These cell types have now been shown to differ in a number of other respects, including progenitor cell size, sensitivity to cycle-active agents, response to plethora, and effects of the W/Wv genotype. In addition, a comparison of the differences found between day 8 BFU-E and day 3 BFU-E on one hand and those distinguishing day 3 BFU-E and CFU-E on the other provides support for the view that early erythropoietic cell differentiation involves a series of changes that take place long before competence to synthesize hemoglobin becomes manifest.
Osmotic fragility of peripheral blood lymphocytes in chronic lymphatic leukemia and malignant lymphoma
Authors: Resnitzky P , Reichman N
Published: October 2019

The osmotic fragility (OF) of peripheral blood lymphocytes from patients with chronic lymphatic leukemia (CLL) and non-Hodgkin malignant lymphoma (ML) was investigated employing an automatic recording method and compared with that of lymphocytes from healthy subjects and from patients suffering from various non-neoplastic diseases. The curves from CLL and ML showed a pattern of increased lymphocyte OF compared with those of the two control groups, and the difference was statistically significant ( less than 0.001). In CLL the increase in OF was more pronounced than in ML, and the shape of the curve was different from that in the other groups. The employment of peripheral blood lymphocyte OF as an additional diagnostic parameter in the diagnosis of CLL and ML is suggested.
Neutrophil transfusion: effect of storage and of collection method of neutrophil blood kinetics
Authors: Price TH , Dale DC
Published: October 2019

The kinetics in blood of autologous neutrophils collected by phlebotomy, filtration leukapheresis (FL), or intermittent-flow centrifugation (IFC), labeled with 32P-diisopropylfluorophosphate, and stored at 4 degrees C for up to 2 days were measured in 41 normal subjects. Mean initial recovery for unstored IFC cells was 34.0%, compared to 7.9% for unstored FL cells. Blood half-lives were 4.1 and 2.7 hr for unstored IFC and FL cells, respectively. With neutrophils collected by phlebotomy and stored in whole blood for 1–2 days, posttransfusion recoveries and blood half-times were significantly decreased. Storage of both IFC and FL preparations resulted in only moderate kinetic abnormalities in comparison to the unstored cells. These studies indicate that the ability of unstored IFC cells to circulate is basically normal, whereas that of unstored FL cells is significantly impaired. The data further suggest that these neutrophil concentrates might be stored for 1–2 days prior to transfusion.
B lymphocyte antigens in the differential diagnosis of human neoplasia
Authors: Kadin ME , Billing RJ
Published: October 2019

Human B lymphocyte antigens (HBLA) were detected with fluorescent- labeled antibodies on malignant cells of 102 patients with Hodgkin disease and other lymphomas, plasma cell myeloma, and nonlymphoreticular neoplasms including carcinomas of the breast, lung, and ovary, soft tissue sarcomas, and neuroblastoma. HBLA were present in Hodgkin disease and other lymphomas of B cell or histiocyte derivation. They were absent in plasma cell myeloma and nonlymphoreticular neoplasms. Absorption studies revealed that malignant T cells had smaller amounts of HBLA, usually not detected by immunofluorescence. Expression of HBLA was dependent on both cell differentiation and origin. Detection of HBLA enabled immunologic distinction of Reed-Sternberg and other lymphoma cells from morphologically similar cells of nonlymphoreticular origin. The rapidity, reproducibility, and economy of the immunofluorescence test make this a useful clinical tool for the differential diagnosis of lymphoma from other malignant disorders in man.
Isoelectric focusing of human hemoglobin: its application to screening, to the characterization of 70 variants, and to the study of modified fractions of normal hemoglobins
Authors: Basset P , Beuzard Y , Garel MC , Rosa J
Published: October 2019

Isoelectric focusing on slabs of acrylamide gel was adapted for the screening of abnormal hemoglobins, the characterization of 70 human variants, and the study of minor fractions of normal hemoglobin. The screening method was as fast and inexpensive as conventional techniques, allowed the simultaneous analysis of some 50 samples of whole blood, and yielded resolution superior to that obtained by other methods with hemolysates. Among the 70 variants, 31 mutants could not be separated from HbS by cellulose acetate electrophoresis. The characterization technique of electrofocusing allowed us to distinguish between most variants. Only one mutant, Hb Galveston, could be confused with HbS. Hb Koln, the most frequent unstable mutant, exhibited a special pattern. HbA1C was separated from HbA. Preliminary results indicate that quantitation of HbA1C by gel scanning is feasible.
Study of a kindred with partial deficiency of red cell 2,3- diphosphoglycerate mutase (2,3-DPGM) and compensated hemolysis
Authors: Travis SF , Martinez J , Garvin J Jr , Atwater J , Gillmer P
Published: October 2019

A kindred with partial deficiency of red cell 2,3-diphosphoglycerate mutase (2,3-DPGM) was studied. The propositus presented with indirect hyperbilirubinemia, normal hemoglobin (15.8 g/dl), and elevated reticulocyte count (4.6%). The red cell 51Cr survival was decreased (tau1/2 16 days). Incubated osmotic fragility was normal; autohemolysis was increased and corrected with glucose and ATP. The P50 was 18.5 mm Hg (normal 25.5 +/- 3), but the stability, electrophoresis, and fingerprinting of hemoglobin were normal. The concentration of 2,3- diphosphoglycerate (2,3-DPG) was reduced to 43% of normal. Red cell 2,3- DPGM was decreased to 59% of normal; 2,3-DPG phosphatase was similarly decreased. All red cell glycolytic and hexose monophosphate shunt enzymes, glycolytic intermediates other than 2,3-DPG, and glucose consumption and lactate production were normal. Five family members showed similar hematologic findings. The deficiency appears to be secondary to decreased enzyme synthesis and to be inherited as an autosomal dominant trait in this family. Partial deficiency of 2,3-DPGM should now be considered in the differential diagnosis of compensated hemolysis associated with increased oxygen affinity.
Production of eosinophil-rich guinea pig peritoneal exudates
Authors: Pincus SH
Published: October 2019

A variety of methods for production of eosinophil-rich guinea pig peritoneal exudates were tested in order to procure large numbers of purified eosinophils. Repeated peritoneal lavage with saline did not produce peritoneal eosinophilia, nor did intraperitoneal injection of dextran beads. Mast cell degranulating agents polymyxin B and, to a lesser degree, compound 48/80 produced significant peritoneal eosinophilia after 6–8 wk of weekly intraperitoneal injections (p less than or equal to 0.05). After more than 8 wk of treatment a mean of 52.1 x 10(6) eosinophils comprising an average of 62% of the peritoneal exudate cells was recovered from polymyxin B-treated guinea pigs. However, polymyxin B-induced peritoneal eosinophilia was abolished by pretreatment with the antihistamine diphenhydramine. Metiamide, an H2 receptor antagonist, did not block the production of peritoneal eosinophils. This is a highly satisfactory method for preparing peritoneal eosinophils; however, the mechanism of action remains to be clearly elucidated.
Physiologic features of hemolysis axxociated with altered cation and 2,3-diphosphoglycerate content
Authors: Albala MM , Fortier NL , Glader BE
Published: October 2019

A hemolytic disorder characterized by altered RBC cation composition (increases Na, decreases K), reduced monovalent cation content (decreased Na + K/liter RBC), and decreased levels of 2,3- diphosphoglycerate (2,3-DPG) is described. The etiology of these RBC abnormalities was not elucidated following extensive laboratory evaluation, although two important physiologic principles were manifested by this case: (1) Hemolysis was relatively well compensated (41% hematocrit) despite a significantly decreased RBC survival (51 Cr t 1/2 = 10.5 days). This effect presumably was due to reduced 2,3-DPG content (1.9 mumol/ml RBC) and the associated increase in whole blood oxygen affinity (P50 = 19.6 mm hg). (2) RBC size and water content were normal in spite of marked cation depletion. This anomaly was thought to reflect the osmotic effects of reduced polyvalent anion (2,3-DPG) content.
Inhibitor of hematopoietic cell proliferation derived from a human leukemic cell line
Authors: Olofsson T , Cline MJ
Published: October 2019

A continuously growing human myeloid leukemia cell line (K562) produced a potent high-molecular-weight inhibitor of hematopoietic cell proliferation. It was most active against myeloid stem cells (CFU-C) and proliferating T lymphocytes; it was less active against erythroid precursors (CFU-E) and did not inhibit fibroblasts or established lines of epithelioid cells or B lymphocytes. Inhibition of CFU-C was by direct interaction rather than by modulation of production of colony- stimulating activity and probably occurred at restricted points in the cell cycle. Inhibition could, within limits, be reversed by washing the target cells. Production of inhibitors of hematopoiesis is not a general property of established cell lines, and only two have thus far been identified in screening of 30 such lines.
Measurement of Donath-Landsteiner antibody-producing cells in idiopathic nonsyphilitic paroxysmal cold hemoglobinuria (PCH) in children
Authors: Miyagawa Y , Yamada S , Komiyama A , Akabane T
Published: October 2019

2-yr-old girl with a nonsyphilitic type of paroxysmal cold hemoglobinuria (PCH) is reported. We applied the plaque-forming cell (PFC) assay to the measurement of Donath-Landsteiner (DL) antibody- producing cells in the patient. Some of the circulating B lymphocytes formed plaques in our assay system. Initially the number of PFC was 1442 +/- 225/10(6) lymphocytes and DL titer was 1:16. Thereafter the PFC decreased in number as the DL titer decreased. Eight weeks later circulating PFC were not found but the DL titer was still 1:1 positive. The PFC assay appears to be useful as a new method for obtaining rapid imformation as to the disease status of PCH.
Effects of temperature on granulocyte preservation
Authors: McCullough J , Wieblen BJ , Peterson PK , Quie PG
Published: October 2019

With the increasing use of granulocyte transfusion it is becoming important to determine if granulocytes can be preserved for a few days. If so, the optimum storage conditions must be identified. We studied the function in vitro of granulocytes collected as they would be for transfusion by continuous-flow centrifuge leukapheresis (CFCL) and filtration leukapheresis (FL). Granulocytes collected by CFCL maintained normal ability to phagocytose and kill bacteria after 48 hr and normal chemotaxis after 24 hr of storage at 20 degrees--24 degrees C. Neither 1 degrees--6 degrees C nor 37 degrees C were as effective in preserving chemotactic response. Agitation of the granulocyte suspension during storage caused reduced bacterial killing and chemotaxis. Granulocytes collected by FL functioned very poorly after 24 hr storage at all temperatures studied. These studies suggest that it may be possible to store CFCL granulocytes at 20 degrees--24 degrees C for 24 hr. FL granulocytes should not be stored at all.
Comparison of intracellular bactericidal activities of human neutrophils and eosinophils
Authors: DeChatelet LR , Migler RA , Shirley PS , Muss HB , Szejda P , Bass DA
Published: October 2019

Eosinophils from a patient with hypereosinophilia were observed to phagocytize radiolabeled E. coli or S. aureus as effectively as neutrophils from a normal control. This was observed at a number of bacteria/cell ratios and a various time intervals following initial challenge and was confirmed by direct microscopic examination. In spite of comparables rates of phagocytosis, the eosinophils were consistently less capable of killing the bacteria. This correlates with an inability of eosinophil peroxidase to catalyze the peroxidase-H2O2-CI--mediated decarboxylation of amino acids; in contrast, both eosinophil and neutrophil peroxidases showed similar capabilities to iodinate protein in vitro. These data are compatible with the importance of the chloride- mediated mechanism in the bactericidal activity of intact phagocytes.
Chromosome analysis of spleen and/or lymph nodes of patients with chronic myeloid leukemia (CML)
Authors: Stoll C , Oberling F , Flori E
Published: October 2019

Origin and spread of the chromosomally abnormal cells that appear in chronic myeloid leukemia (CML) after transformation are unknown. Spleen and lymph node may be involved. In 16 patients with CML splenectomy and/or adenectomy were performed before or during the blastic crisis of the disease, followed by a chromosomal analysis of the cells from the removed organ. At the same time, the chromosomes of the blood cell and of the bone marrow were also analyzed. Analyses were done with R banding. The results show that an extramedullary clonal development with duplication of the Ph1 chromosome and other features occurred. From a cytogenetic standpoint, acute blastic phase of CML is frequently characterized by an increased number of chromosomes owing to preferential gain of additional chromosomes. This, then, would clearly point to extamedullary acute transition in CML.
Measurement of sickling by controlled temperature increase
Authors: Chang H , Nagel RL
Published: October 2019

Previous experiments to study the rate of red cell sickling have employed rapid mixing apparatus for SS cells with dithionite and have shown that the half-time of sickling is quite rapid, on the order of seconds. An alternative approach is to slow down the rate by taking advantage of the negative temperature coefficient of the process. We developed a method in which deoxygenation of a cell suspension is carried out at 0 degrees C. A linear temperature gradient to 37 degrees C is applied, and a gradual increase in the percentage of sickled cells is observed. At a heating rate of 1.5 degrees C/min the temperature at which half of the cells became sickled was 19 degrees C for SS cells treated with dithionite, 22 degrees C for SC cells, 28 degrees C for AS cells, 22 degrees C for cyanate-treated SS cells, and 23 degrees C for SS cells in the presence of 0.1 M butylurea. Thus this method promises to be useful for the study of sickling rates and the screening of potential anti-sickling agents.
Erythroid regulatory factors: correlation of detectable titers in vivo and in vitro
Authors: Dunn CD , Lange RD
Published: October 2019

Titers of erythroid regulatory factors (ERF), as measured in vivo with a standard erythrocythemic mouse technique, showed a highly significant correlation with those obtained in vitro with a fetal mouse liver cell (FMLC) assay. The results from the assay in vitro were somewhat higher than would be predicted from the assay in vivo. Nevertheless, the FMLC technique has been found to be a valid and useful technique particularly in the study of serum ERF titers.
Identification of dihydropteridine reductase in human platelets
Authors: Abelson HT , Gorka C , Beardsley GP
Published: October 2019

Normal human platelets were shown to contain the enzyme dihydropteridine reductase. The enzyme was not found in a variety of other cells of hematogenous origin. Partial purification and kinetic and physical data indicated that the platelet enzyme is similar to that previously characterized from liver. Dihydropteridine reductase is important for the regeneration of tetrahydrobiopterin, a required cofactor in hydroxylation reactions involved in biogenic amine formation. The presence of the enzyme may indicate that some synthesis de novo of serotonin and/or catecholamines occurs in platelets, as opposed to a purely storage and transport function. In addition, screening for hyperphenylalaninemia due to dihydropteridine reductase deficiency may become feasible by assaying platelets for enzyme activity.
Effects of local anesthetics on human platelets: filopodial suppression and endogenous proteolysis
Authors: Nachmias VT , Sullender JS , Fallon JR
Published: October 2019

Agents that affect platelet shape may be useful in understanding the mechanism of shape change; for this reason the effects of local anesthetics are worthy of further study. Local anesthetics cause platelets to retract filopodia. At short time intervals (up to about 30 min) and low concentrations of the drugs, the filopodia are reextended when the platelets are gel filtered with eluant free of anesthetic. At longer time intervals (1–2 hr) or higher drug concentrations, the retraction becomes irreversible. When the polypeptide composition of the total platelet lysate was examined on SDS gels, proteolysis of two high molecular weight bands was seen when the suppression became irreversible. These polypeptides, estimated as 250,000 and 230,000 daltons, were major components of a precipitate that formed when platelets were lysed at low ionic strength and were also enriched in a “cytoskeletal” preparation made by lysing platelets attached to glass beads and analyzing the adherent residue. Electron micrographs of platelets lysed on surfaces showed an intermeshed network of filaments to be a major component of the residue. The results suggest that the proteins comprised of these bands may be part of the cytoskeletal system and that their integrity may be necessary for the platelet to reextend filopodia following suppression.
Inhibition of granulocyte erythrophagocytosis by HLA antisera
Authors: Nusbacher J , MacPherson JL , Gore I Jr , Grinberg R
Published: October 2019

The ability of HLA antisera to inhibit granulocyte erythrophagocytosis (EP) of opsonized red blood cells (RBC) was evaluated. Human granulocytes (PMN) were separated from heparinized whole blood by the Ficoll-Isopaque technique and suspended in McCoy's medium. EP occurred when the PMN were incubated with opsonized RBC. For six HLA antibody specificities evaluated, prior incubation of PMN with HLA antisera resulted in significant inhibition of EP without cytolysis when the PMN donor was positive for the specific HLA antigen, but not when the donor was negative for the antigen. The inhibition was time- and dose- dependent. Prior absorption of HLA antisera with HLA-specific platelets reduced or abolished the inhibition. An example of anti-NB1 also inhibited EP in 4 individuals. These data suggest that HLA antibody may adversely affect granulocyte phagocytic function. Inhibition of EP might be useful in evaluating compatibility prior to granulocyte transfusion.
Action of thrombin on surface glycoproteins of human platelets
Authors: Mosher DF , Vaheri A , Choate JJ , Gahmberg CG
Published: October 2019

We labeled surface glycoproteins of human platelets by the neuraminidase-galactose oxidase/borotritiide and the periodate/borotritiide methods. When labeled platelets were treated with 1-nM thrombin, a minor glycoprotein weighing 68,000–85,000-d was lost from the surface, and a soluble glycoprotein weighing 57,000- 68,000-d was found in the supernatant. Treatment of platelets with ADP, collagen, or the calcium ionophore A23187 did not cause loss of the 68,000–85,000-d glycoprotein from platelet surfaces or appearance of the 57,000–68,000-d glycoprotein in the supernatant. However, trace amounts of the intact 68,000–85,000-d glycoprotein were found in the supernatants of platelets that were not treated with thrombin. The numerous effects of thrombin on platelets could be initiated by cleavage and release the thrombin-sensitive glycoprotein.
Induction of myeloperoxidase deficiency in granulocytes in lead- intoxicated dogs
Authors: Caldwell KC , Taddeini L , Woodburn RL , Anderson GL , Lobell M
Published: October 2019

Lead interferes with heme synthesis in erythrocytes and has a deleterious effect on red cell membranes. We measured myeloperoxidase (MPO) enzyme activity in the granulocytes of dogs fed increasing quantities of lead. Concurrently, iodination capability and in vitro bactericidal activity were measured. Blood lead levels were monitored. Three of 4 dogs poisoned with lead developed significant decreases in MPO enzyme activity in their granulocytes. The decline in MPO activity correlated with cumulative lead toxicity as judged by blood lead levels and clinical signs of lead poisoning. Iodination ability in all 4 dogs decreased with cumulative lead toxicity. After discontinuation of lead administration, recovery of granulocyte MPO activity preceded recovery of iodination ability. This observation suggests the possibility of separate effects of lead on iodination ability and MPO activity. Moderate impairment of bactericidal capacity developed in 3 of 4 dogs with severe lead poisoning. Clinical infections were not observed during the course of the study.
Chromosome aberrations in B lymphocytes of atomic bomb survivors
Authors: Kamada N , Kuramoto A , Katsuki T , Hinuma Y
Published: October 2019

Although chromosome aberrations in T lymphocytes and bone marrow cells have been reported in atomic bomb survivors, the presence of chromosome abnormalities has not been demonstrated in B lymphocytes because of the technical difficulties involved in B-lymphocyte separation. A method for detecting chromosome aberrations in B lymphocytes was established by “stimulation” of B lymphocytes with Epstein-Barr virus (EBV) instead of “separation” of B lymphocytes by rosette formation. The EBV- stimulated lymphocytes were isolated as single colonies in soft agar and transferred to liquid culture for further cell growth. The EBV- stimulated B lymphocytes of two heavily exposed survivors showed 50% and 12.5% chromosome abnormalities 30 yr after exposure to the effects of the atomic bomb. The former patient seemed to have a karyotypically abnormal clone of B lymphocytes in vivo. The method used in this study and the evidence of chromosome aberrations in B lymphocytes for long periods after radiation exposure will be useful and important in elucidating the malignant processes of acute lymphocytic leukemia, B- cell lymphoma, and multiple myeloma among high-risk groups having a history of accidental or therapeutic exposure to radiation or radiomimetic drugs.
Fetal characteristics of erythrocytes in sickle cell anemia: an immunofluorescence study of individual cells
Authors: Maniatis A , Papayannopoulou T , Bertles JF
Published: October 2019

In a group of disease states that includes sickle cell anemia (SS disease), two fetal erythrocyte markers, Hb F and i antigen, persist into adulthood. Using the technique of single-cell immunofluorescence, we determined the expression of l-i antigens and the presence of Hb F within populations of erythrocytes. Subjects tested included normal adults, normal newborns, patients with SS disease, and individuals with sickle cell trait. We classified erythrocytes reacting to anti-i as i cells and those reacting to anti-l as l cells, a terminology analogous to that used to identify erythrocytes containing increased amounts of Hb F as F cells. The expression of l and i antigens within populations of both normal and SS erythrocytes was found to be heterogeneous. The proporations of both i cells and l cells in all SS patients studied exceeded those found in normal adults, and an overall stronger-than- normal reactivity of individual SS cells to the two antibodies was observed. Proportions of F cells showed no correlation with proportions of i cells; and with double fluroescence staining for both Hb F and i, a significant proportion of each total SS red cell population was found to carry only one or the other marker. These findings confirm and clarify on a cellular level our previous demonstration, by means of quantitative hemagglutination, that there is increased expression of both l and i by whole populations of SS erythrocytes. In addition, we provide here new information on the expression of l and i within populations of normal human erythrocytes.
Hemoglobin Crete (beta 129 ala leads to pro): a new high-affinity variant interacting with beta o -and delta beta o -thalassemia
Authors: Maniatis A , Bousios T , Nagel RL , Balazs T , Ueda Y , Bookchin RM , Maniatis GM
Published: October 2019

Hemoglobin Crete, beta129 (h7)ala leads to pro, is a new mutant hemoglobin (Hb) with high oxygen affinity that was discovered in a Greek family in various combinations with beta- and deltabeta- thalassemia. The propositus, who presented an unusual clinical picture of an “overcompensated” hemolytic state, with erythrocytosis, splenomegaly, abnormal red cell morphology, and marked erythroid hyperplasia, appeared doubly heterozygous for Hb Crete and deltabeta- thalassemia. His red cells contained 67% Hb Crete and 30% Hb F, and the combination of these two hemoglobins resulted in a blood P50O2 of 11.2 mm Hg. A brother with Hb Crete trait (38% Hb Crete, 56% Hb A, blood P50O2 23.0 mm Hg) did not have significant erythrocytosis. Purified Hb Crete was heat-unstable and exhibited a high oxygen affinity, and a normal Bohr effect. We postulate that the beta 129 proline substitution disrupts the H helix, perturbing nearby residues involved in alpha 1 beta 1 contact sites of the Hb tetramer.
Neutrophil-mediated antibody-dependent killing of herpes-simplex-virus- infected cells
Authors: Siebens H , Tevethia SS , Babior BM
Published: October 2019

The ability of neutrophils to serve as effectors of antibody-dependent cell-mediated cytotoxicity (ADCC) was studied using herpes-simplex- virus-infected (HSV-infected) human embryonic lung cells as targets. Killing of infected cells by neutrophils required HSV-immune serum and did not take place in the presence of nonimmune serum or in the absence of serum. Uninfected cells were not killed. ADCC was evident by 30 min and reached a value of 66% by 4 hr at an effector:target ratio of 100:1. Cells from patients with chronic granulomatous disease were as effective as normal cells, and this indicates that ADCC could be mediated by processes that are independent of the respiratory burst.
Prednisone-responsive aplastic anemia: a mechanism of glucocorticoid action
Authors: Bagby GC Jr , Goodnight SH , Mooney WM , Richert-Boe K
Published: October 2019

We performed serial agar cultures (CFU-C) using marrow cells from a patient with prednisone-responsive aplastic anemia and from five patients with prednisone-resistant aplasia. Colony growth was decreased in all patients. Cortisol (10(-7)-10(-4)M) significantly enhanced colony growth in the prednisone-responsive patient but fauled to enhance colony growth in the remaining five patients. Further studies in the responsive patient indicated that (1) colony growth was enhanced by depleting T lymphocytes from the marrow cells, (2) colon growth T- depleted marrow cells was inhibited by autologous peripheral blood lymphocytes (PBL), (3) cortisol failed to enhance colony growth of T- depleted marrow cells, (4) PBL and PBL-conditioned medium inhibited colony growth of both autologous and allogeneic marrow cells, but neither cortisol-treated PBL nor T-depleted PBL were inhibitory. Serial cultures in the responsive patient showed that colony growth normalized during remission when “suppressor” cells were absent and that colony growth was subnormal during a later relapse when cortisol-resistant “suppressor” cells were present. Therefore, in this prednisone- responsive patient, cortisol-sensitive T lymphocytes suppressed granulopoiesis in vitro. Our observations suggest that aplastic anemia in this patient is immunologically mediated and that prednisone therapy enhanced hemopoiesis in vivo by inhibiting the “suppressor” T lymphocytes.
A chemotactic inhibitor produced by blast cells and present in the serum of a patient with acute lymphoblastic leukemia
Authors: Blumenfeld W , Territo M
Published: October 2019

A chemotactic factor inhibitor (CFI) was found in the serum of a patient with acute lymphoblastic leukemia. The factor was characterized as heat-labile, with activity against complement-dependent chemotactic factors and against the chemotactic factor produced by Escherichia coli. Normal sera and sera from other patients with acute leukemia also demonstrate heat-labile inhibitory activity against complement- dependent chemotactic factors but not against the E. coli-derived chemotactic factor. Supernatants from cultured lymphoblasts of the patient also possessed CFI activity similar in character to that found in his serum. It is suggested that in vivo the lymphoblasts were responsible for the chemotactic defect observed in his serum, presumably by manufacturing and releasing a chemotactic factor inactivator.
Monoclonal immunoglobulin rescue from a patient with chronic lymphocytic leukemia and autoimmune hemolytic anemia
Authors: Sikora K , Krikorian J , Levy R
Published: October 2019

In order to test whether autoimmune hemolytic anemia in a patient with chronic lymphocytic leukemia was due to autoantibody production by the neoplastic clone, somatic cell hybrids were constructed between the patient's leukemic cells and a mouse myeloma line. Light-chain- restricted human immunoglobulin was secreted by 4 of 11 of the established hybrid lines. No binding of this immunoglobulin to red cell surface antigens could be detected. Thus, we conclude that the autoantibody in this patient represented a concomitant reactive response and was not the product of the malignant clone of B cells.
A case of generalized amyloidosis associated with cyclic neutropenia
Authors: Shiomura T , Ishida Y , Matsumoto N , Sasaki K , Ishihara T , Miwa S
Published: October 2019

A case of generalized amyloidosis associated with cyclic neutropenia is presented. A 24-yr-old female with cyclic neutropenia died from intestinal obstruction caused by necrosis and perforation of the small intestine. Post-mortem examination revealed generalized amyloidosis involving almost all organs. Amyloid deposits were prominent, especially in the alimentary tract, kidneys, spleen, and small blood vessels. As has been suggested in gray collie dogs with congenital cyclic neutropenia known to develop secondary amyloidosis in adulthood, an increase of antigenic stimulation during the intermittent bouts of acute infections may be one of the factors responsible for the development of secondary amyloidosis in this case. Although the association of amyloidosis and cyclic neutropenia in man has rarely been described, it is probable that amyloidosis is not a rare complication of human cyclic neutropenia, considering that patients with this hematologic disorder are chronically exposed to excessive antigenic stimulation.
Hand-mirror cell leukemia associated with mental retardation: immunologic, chromosome, and morphological studies
Authors: Stern R , Widirstky ST , Wurster-Hill DH , Allen RD , Smith KA , Cornwell GG , Cornell CJ
Published: October 2019

Cycochemical, morphological, immunologic, and cytogenetic studies were carried out on hand-mirror cells (HMC) from a mentally retarded patient with a constitutional chromosome abnormality, 46,XX,r(21), and acute lymphoblastic leukemia. Scanning electron and differential interference contrast microscopy showed microspikes on the uropodia, but little evidence of cellular motility, despite formation and disappearance of individual uropodia in cell suspensions. The cells rosetted with sheep erythrocytes, suggesting T-cell origin. Cells derived from the bone marrow (80% HMC) showed a high degree of polyploidy (60%) and a bimodal chromosome number of 49 (49,XX,+10,-21, +3 rings) and 94 (6 no. 10, 3 no. 18, 2 no. 21 chromosomes, 3 ring chromosomes, plus 4 copies of each other chromosome).
Immune suppression of erythropoiesis in transient erythroblastopenia of childhood
Authors: Koenig HM , Lightsey AL , Nelson DP , Diamond LK
Published: October 2019

Serum and IgG from four children with transient erythroblastopenia of childhood (TEC) was tested to see what effect it would have on development of erythroid colonies from bone marrow mononuclear cells. Serum and IgG specimens obtained at the time of diagnosis uniformly suppressed erythroid colony development from CFU-E. Washed bone marrow mononuclear cells from a child with TEC failed to grow in the presence of his own serum, but grew normally in the presence of isologous serum. Serum specimens obtained from patients after recovery from TEC had no effect on erythroid colony development. The anemia of TEC appears to be due to transient immune suppression of erythroid colony development.
Pseudo-Chediak-Higashi anomaly in a case of acute myeloid leukemia: electron microscopic studies
Authors: Tulliez M , Vernant JP , Breton-Gorius J , Imbert M , Sultan C
Published: October 2019

The formation and fine structure of giant granules in neutrophil promyelocytes of a patient with a variant of acute myelogenous leukemia were investigated by electron microscopy. The patient presented with large lymph nodes and disseminated intravascular coagulation (DIC). By light microscopy, numerous giant granules, resembling those of Chediak- Higashi syndrome (CHS), were present, but Auer bodies could not be found. By electron microscopy, these giant granules were seen to be formed by fusion of azurophilic granules, as in CHS; however, they were different from the large granules of CHS, since they contained numerous microcrystalline structures like those of Auer bodies. However, the crystalline cores of these granules exhibited a periodicity different from that of Auer bodies of acute promyelocytic leukemia. This clinical and hematologic syndrome (giant granules, enlarged lymph nodes, and DIC may represent a variant of acute promyelocytic leukemia.
Serum lactic dehydrogenase as a prognostic tool for non-Hodgkin lymphomas
Authors: Ferraris AM , Giuntini P , Gaetani GF
Published: October 2019

Serum LDH levels have been found to be significantly increased in non- Hodgkin lymphoma (NHL) patients, both histiocytic and lymphocytic. The duration of survival of NHL negatively correlates with the level of serum lactic dehydrogenase (LDH), and statistical analysis reveals that patients with lower levels of LDH have a longer survival rate than the patients with higher LDH activity, irrespective of their histologic classification. The analysis of the results by the Test for Trend in Prognosis allows us to establish that the correlation of the rate of survival and LDH levels is independent from other clinical parameters.
AET-treated platelets: their usefulness for platelet antibody detection and an examination of their altered sensitivity to immune lysis
Authors: Cimo PL , Gerber SA
Published: October 2019

2-Aminoethylisothiouronium bromide (AET) increases the sensitivity of blood cells to complement-mediated immune lysis. We compared the sensitivities of untreated or AET-treated platelets to immune lysis induced by different types of platelet antibody in the 51Cr platelet lysis test. AET platelets were 8–16 times more sensitive to autoantibody and alloantibody, but 8–16 times less sensitive to drug- dependent antibody. AET-platelets bound similar amounts of alloantibody but less drug-dependent antibody, and they lysed at higher complement dilutions than did untreated platelets. AET-platelets detected 10 of 25 autoantibodies, 9 of 9 alloantibodies, and 5 of 8 drug-dependent antibodies. Untreated platelets detected 1 of 25, 6 of 9, and 7 of 8 of these respective platelet antibodies. The use of AET-platelets in the 51Cr platelet lysis test increases its sensitivity for detecting non- drug-dependent platelet antibodies. AET-platelets resemble paroxysmal nocturnal hemoglobinuria (PNH) platelets in their enhanced sensitivity to complement-mediated lysis. They differ from PNH platelets in their insensitivity to immune lysis induced by drug-dependent antibodies and, in this respect, are similar to Bernard-Soulier syndrome platelets.
Plasma beta-thromboglobulin: differentiation between intravascular and extravascular platelet destruction
Authors: Han P , Turpie AG , Genton E
Published: October 2019

To elucidate the usefulness of beta-thromboglobulin (beta TG) in the differentiation of the mechanism of thrombocytopenia, plasma beta TG concentration was measured in one patient with amegakaryocytic thrombocytopenia, four patients with autoimmune thrombocytopenia (ATP), two patients with thrombotic thrombocytopenia (TTP), and one patient with thrombocytopenia secondary to disseminated intravascular coagulation (DIC). Plasma beta TG was not measurable in amegakaryocytic thrombocytopenia, was normal in ATP, and was increased in TTP and DIC. These data indicate that in thrombocytopenic patients, increased plasma beta TG concentration may result from intravascular platelet consumption with release of platelet constituents in contrast to extravascular platelet destruction by the macrophage-monocyte system.
Willebrand factor in hemostasis in the in vitro bleeding time
Authors: Kimura A , Bowie EJ , Campbell RJ , Fass DN
Published: October 2019

Heparinized porcine blood and plasma, at constant hydrostatic pressure, was allowed to flow through a 5-mm incision in a small piece of porcine skin. Changes in the exuded blood volume were measured, and the incision site was examined microscopically. When normal blood flowed through either normal or von Willebrand skin, the exuded blood volume decreased gradually and eventually stopped. Microscopic examination revealed a platelet plug in the incision site. This plug was positive for Willebrand factor when examined by immunofluorescence. In contrast, the blood from von Willebrand pigs continued to flow constantly, and a platelet plug was not seen. The delayed in vitro hemostasis in von Willebrand blood was corrected to the normal range by the addition of either normal plasma or partially purified Willebrand factor. Normal blood, in which the Willebrand factor was immunologically inhibited, showed delayed hemostasis. For this in vitro system, it appeared that plasmatic Willebrand factor played an essential role in hemostasis.
Unicellular or multicellular origin of human granulocyte-macrophage colonies in vitro
Authors: Singer JW , Fialkow PJ , Dow LW , Ernst C , Steinmann L
Published: October 2019

The assumption that human granulocyte-macrophage colonies have a unicellular origin and thus are true clones has been directly tested. Cells from seven females heterozygous for the common glucose-6- phosphate dehydrogenase (G-6-PD) gene (GdB) and the variant GdA were cultured in semisolid medium for granulocyte-macrophage colony growth and the enzyme type of individual colonies was determined. When the colony density was less than 20/dish, more than 95% of colonies had either type A or type B G-6-PD, but not both. At colony densities greater than 30/dish, between 15% and 75% of colonies had both enzyme types and therefore arose from more than one cell. These results are consistent with a unicellular origin for the colonies only when they are cultured at low densities. With increasing colony density, there was a greater frequency of colonies with both type A and type B activity, suggesting that accurate enumeration of committed stem cells can only be performed at low colony concentrations.
Incidence and patterns of bone marrow and blood involvement by lymphoma in relationship to the Lukes-Collins classification
Authors: Foucar K , McKenna RW , Frizzera G , Brunning RD
Published: October 2019

Pretreatment lymph nodes, bone marrow, and blood were examined in 176 cases of non-Hodgkin's lymphoma. By the criteria of the Lukes and Collins functional--morphological classification, 158 (90%) were B-cell lymphomas and 17 (10%) were T-cell lymphomas. Bone marrow involvement was present in 53% of cases: 51% of B-cell types and 65% of T-cell types. Marrow involvement was most frequent in small lymphocyte (B) (89%), convoluted lymphocyte (60%), and small cleaved follicular center cell (FCC) lymphomas (55%). The pattern of bone marrow involvement was most frequently focal paratrabecular in B-cell lymphomas and diffuse in T-cell lymphomas. Blood involvement was present in 50% of cases with bone marrow lymphoma and generally reflected extensive bone marrow disease. There was a higher incidence of both bone marrow and blood involvement in pediatric patients than in adults.