bluuebunny/crossref_metadata_2025 huggingface dataset:
Partial immunologic reconstitution of a patient with acquired agammaglobulinemia: a transient phenomenon accompanying therapeutic plasmapheresis
Authors: Wenz B , Rubinstein A
Published: October 2019

A patient with acquired agammaglobulinemia was treated with plasmapheresis. The rationale for this procedure was based on the presence of a cytotoxic autoantibody with specificity for helper (TH2-) T lymphocytes. Plasmapheresis reduced the autoantibody concentration to undetectable levels, which resulted in an increase number of helper T cells. These T cells provided normal in vitro helper activity. Plasmapheresis did not correct a concomitant suppressor T-cell defect, and the clinical remission ended during the fifth month of exchange therapy.
Monoclonal CLL B-cells may be induced to grow in an in vitro B-cell colony assay system
Authors: Perri RT , Kay NE
Published: October 2019

A simple reproducible in vitro B-cell colony assay system was used to evaluate B-cell growth in controls and patients with chronic lymphocytic leukemia (CLL). All six CLL patients studied formed B-cell colonies. The number of colonies was significantly less in patients than controls (66 +/- 18 versus 127 +/- 8). CLL colonies were shown to be monoclonal and appeared to reflect the circulating malignant B-cell clone in our patient group, while the six controls studied formed polyclonal B-cell colonies. Wright-Giemsa staining showed typical plasma cells to have developed in the controls but not in the patients. Cells from CLL patients retained a more lymphoid appearance. It is believed that investigations with this B-cell assay will provide the means for further in vitro evaluation of malignant B-cell proliferation in other lymphoproliferative disorders.
Large granular lymphocytes in human peripheral blood: ultrastructural and cytochemical characterization of the granules
Authors: Grossi CE , Cadoni A , Zicca A , Leprini A , Ferrarini M
Published: October 2019

Large granular lymphocytes (LGL) are defined as nonadherent mononuclear cells with cytoplasmic azurophilic granules, avid receptors for the Fc portion of IgG, and cytotoxic functions (NK or ADCC activities). In the present study, the granules of LGL isolated from human peripheral blood have been analyzed by enzyme cytochemistry and electron microscopy. It had been found that: (1) in the single cells, granules at different stages of maturation could be detected: in addition, packaging of the granules took place in the proximity of the Golgi apparatus, which is similar to that seen in secretory cell types. (2) Acid phosphatase (AP) was observed within the granules and the vesicles located in the Golgi area: the Golgi apparatus identified through its thiamine pyrophosphatase-positivity was consistently negative for AP. (3) Alpha naphthyl-acetate esterase (ANAE) activity was localized in the granules as well as on the membrane of LGL and monocytes. (4) The ANAE activity of LGL was of the monocytic and not of the lymphocytic type, as shown by NaF inhibition. (5) The LGL granules, although identifiable as primary lysosomes, were not involved in the process of phagocytosis, since LGL failed consistently to ingest latex particles or opsonized red cells.
A case of weak blood group B expression (Bm) associated with abnormal blood group galactosyltransferase
Authors: Yoshida A , Yamato K , Dave V , Yamaguchi H , Okubo Y
Published: October 2019

The mechanisms of unusually weak A and B blood group expressions have not been well understood. Since the human blood group A and B substances are produced by the action of blood group GalNAc transferase and Gal transferase, respectively, the mechanism may be elucidated by examining the properties of the blood group transferases and membranes of the subjects with the abnormality. We examined a case associated with very weak B activity in red blood cells, an absence of the B agglutinin in serum, and an existence of the H and B substances in saliva, i.e., a case commonly classified as Bm. More than 85% of H sites remained unglycosylated in the subject's red cell membranes. The blood group Gal transferase activity in the subject's plasma and red cell membranes was about 50% of that of normal. The pH-activity profile and the Michaelis constants for UDP-Gal and 2'-fucosyllactose of the subject's enzyme were distinctively different from that of normal enzyme. These findings led us to conclude that the weak B activity in the present Bm case was due to a direct mutation in B gene resulting in formation of variant B enzyme with low affinity to UDP-Gal and insufficient galactosylation of H sites in the subject.
Severe-glucose-6-phosphate dehydrogenase (G6PD) deficiency associated with chronic hemolytic anemia, granulocyte dysfunction, and increased susceptibility to infections: description of a new molecular variant (G6PD Barcelona)
Authors: Vives Corrons JL , Feliu E , Pujades MA , Cardellach F , Rozman C , Carreras A , Jou JM , Vallespi MT , Zuazu FJ
Published: October 2019

Molecular, kinetic, and functional studies were carried out on erythrocytes and leukocytes in a Spanish male with G6PD deficiency, congenital nonspherocytic hemolytic anemia (CNSHA), and increased susceptibility to infections. G6PD activity was absent in patient's red cells and was about 2% of normal in leukocytes. Molecular studies using standard methods (WHO, 1967) showed G6PD in the patient to have a slightly fast electrophoretic mobility at pH 8.0 with otherwise normal properties (heat stability at 46 degrees C, apparent affinity for substrates, optimum pH, and utilization of substrate analogues). Other tests showed the patient's granulocytes to engulf latex particles normally, but to have impaired reduction of nitroblue tetrazolium and ferricytochrome-c as well as reduced iodination. Chemotaxis and random migration of the patient's granulocytes were normal as were myeloperoxidase, leukocyte alkaline phosphatase (LAP), and ultrastructural features. The molecular characteristics of G6PD in the patient differed from those of all previously reported variants associated with CNSHA, so the present variant was provisionally called G6PD Barcelona to distinguish it from other G6PD variants previously described. Possible mechanisms for the severe deficiency of G6PD in erythrocytes and granulocytes was investigated by studies on the immunologic specific activity of the mutant enzyme.
Alterations of polypeptide composition of mature granulocytes obtained from patients with chronic myelogenous leukemia
Authors: Yokota J , Asano S , Teshima S , Morishita K , Iwamoto A , Yoshikura H , Miwa S
Published: October 2019

In order to compare the polypeptide composition of the CML mature granulocytes with that of the normal whole cell lysates, mature granulocytes from four healthy volunteers and six CML patients were analyzed by two-dimensional gel electrophoresis. In the normal subjects, 60 major spots were commonly and reproducibly identified. In the CML, there were constant alterations in some of these major spots. Four spots were totally absent in all the CML samples, and another four spots were absent in five of the six samples. In addition, one spot was larger in CML than in normal cells, and another spot, which was only faintly visible or not detectable in the normal samples, was massively present in all the CML samples. Our data suggest specific changes in the polypeptide compositions of CML granulocytes. This method could be clinically applied for the analysis of granulocytic disorders.
Interaction between vaccinia virus and human blood platelets
Authors: Bik T , Sarov I , Livne A
Published: October 2019

The objective of the present study was to characterize the interaction between human platelets and vaccinia virus and to examine possible impairment of platelet functions. The vaccinia virus was selected for our model system because it lacks detectable neuraminidase activity. Platelets were incubated with purified viral particles labeled with 3H- thymidine and binding parameters were analyzed. Binding reached saturation with an average of 5 particles/platelet. It was not affected by the plasma but was sensitive to temperature and to metabolic inhibitors. 3H-thymidine-labeled vaccinia virus and formaldehyde-fixed platelets were used to measure viral adsorption. The adsorption was temperature-independent but was affected by ionic strength, indicating electrostatic interactions. Treatment of the fixed platelets with neuraminidase or with alkaline phosphatase reduced viral adsorption, indicating that sialate and phosphate residues on the platelet surface may be involved in the adsorption. Platelet activities were markedly affected by vaccinia virus. The virus caused a dramatic 14C-serotonin release with no added inducer. The release was inhibited by aspirin, a known inhibitor of serotonin release related to prostaglandin synthesis. Furthermore, the virus inhibited platelet aggregation, induced by either ADP, collagen, or thrombin. This study demonstrates that although vaccinia virus lacks neuraminidase activity, it does bind to platelets and affects their function.
Platelet protein organization: analysis by treatment with membrane- permeable cross-linking reagents
Authors: Davies GE , Palek J
Published: October 2019

We have examined platelet protein organization by treatment of intact resting or thrombin-activated platelets with two cross-linking reagents, diamide or dithiobis(succinimidyl propionate) (DTSP). Cross- linked complexes were separated by polyacrylamide gel electrophoresis in the absence of reducing agent and their composition determined after reductive cleavage and analysis in a second-dimensional gel. The most prominent cross-linked species produced by diamide treatment of of resting platelets are (A) cytoskeletal protein homopolymers, such as myosin heavy chain dimer and actin oligomers, and (B) high molecular weight material consisting of homo- or heteropolymers of cytoskeletal proteins and 230,000, 170,000, 100,000, 55,000, and 52,000 dalton proteins. DTSP treatment forms similar complexes and also cross-links membrane glycoproteins IIb and III into high molecular weight material. Thrombin activation of platelets before treatment with diamide or DTSP results in increased cross-linking of myosin and increased incorporation of several proteins, particularly myosin and glycoproteins IIb and III, into high molecular weight material. The results provide evidence for reorganization of cytoskeletal and membrane proteins during platelet function.
Decreased actin content of lymphocytes from patients with chronic lymphocytic leukemia
Authors: Stark R , Liebes LF , Nevrla D , Conklyn M , Silber R
Published: October 2019

Actin, a major cytoskeletal protein, was quantitated in normal and chronic lymphocytic leukemia lymphocytes. The actin content of normal human blood lymphocytes was 2.2 +/- 0.4 mg/10(9) cells and represented 6.6% +/- 1.8% of the total cellular protein. A significant decrease (p less than 0.001) was noted in chronic lymphocytic leukemia lymphocytes that contained 1.4 +/- 0.3 mg actin/10(9) cells, constituting 4.3% +/- 1.1% of the total protein. Normal T and B cells did not differ in actin content. Reduced actin levels were found in the T as well as in the B lymphocytes of “B-cell” chronic lymphocytic leukemia. The possible importance of the decreased actin level in the anomalous capping response and motility of chronic lymphocytic leukemia lymphocytes is discussed.
Immature megakaryocytes in the mouse: physical characteristics, cell cycle status, and in vitro responsiveness to thrombopoietic stimulatory factor
Authors: Long MW , Williams N , Ebbe S
Published: October 2019

The heterogeneity among immature megakaryocytes has been examined by physical properties, cell cycle status, and responsiveness to thrombopoietic stimulatory factor. Three types of immature megakaryocytes exist that can be recognized by acetylcholinesterase staining, nuclear shape, high nucleus/cytoplasm ratio, and small size (8--18 mu) with respect to mature megakaryocytes (greater than 18 mu). These three acetylcholinesterase-containing cell types are distinguished by their nuclear configuration: a round, indented, and lobed nucleus. The lobed cell type was found to overlap with and enhance detection of megakaryoblasts (stage I megakaryocytes). These cells had a sedimentation velocity range of 3.5--19.0 mm hr-1 and a density range of 1.072--1.095 g cm-3. Separation of these three classes of immature megakaryocytes was achieved by equilibrium density centrifugation with modal buoyant densities of 1.079 g cm-3 (round), 1.084 g cm-3 (indented), and 1.089 g cm-3 (lobed). In the presence of thrombopoietic stimulatory factor, the round nucleated cells, but not the indented or lobed nuclei morphology, were observed to develop into large mature megakaryocytes in 60-hr semisolid cell cultures. Development of two cell groups, or colonies of megakaryocytes, was not observed during this in vitro incubation period. In vivo treatment with hydroxyurea indicated that 57.5% +/- 19% of the round nucleus form were actively synthesizing DNA. No reduction in the numbers of indented or lobed nucleus forms were observed following hydroxyurea treatment. The data in this report strongly support the concept that these three types of immature megakaryocytes reflect the early maturation stages occurring in megakaryocyte differentiation.
Monoclonal antibodies to porcine factor VIII coagulant and their use in the isolation of active coagulant protein
Authors: Fass DN , Knutson GJ , Katzmann JA
Published: October 2019

Partially purified preparations of porcine factor VIII:C were used to immunize mice and spleen cells from the immunized animals were fused to NS-1 mouse myeloma cells. The ability of hybrid culture fluids to bind factor VIII:C was detected with a radiolabelled, affinity-purified, human antihuman VIII:C inhibitor. Three cloned hybrid lines have been obtained that preferentially bind to VIII:C when compared to von Willebrand factor binding. Two of these monoclonal antibodies partially inhibit VIII:C coagulant activity. The third antibody does not inhibit VIII:C, but it can be used as an affinity reagent to absorb dissociated VIII:C out of solution. Active coagulant can be recovered by elution in 50% ethylene glycol. The VIII:C obtained has a specific activity of 6 units/micrograms based on absorbance measurements. When analyzed on SDS gels, the unactivated VIII:C contains 3 bands of apparent molecular weight 166,000, 130,000 and 76,000. Thrombin treatment results in a 40 fold increase in activity and cleavage to products of 76,000, 67,000 an 50,000 and small amounts of lower molecular weight peptides. EDTA inactivation of the factor VIII:C results in the separation of the 166,000 and 130,000 chains from the 76,000 chain, suggesting a Ca++ dependent noncovalent interaction among the chains.
Distribution of plasma fibronectin (cold-insoluble globulin) and components of the factor VIII complex after heparin-induced precipitation of plasma
Authors: Amrani DL , Mosesson MW , Hoyer LW
Published: October 2019

Factor VIII procoagulant (VIII:C) activity, factor VIII coagulant antigen (VIII:CAg), von Willebrand ristocetin cofactor (VIIIR:RC) activity, factor VIII-related antigen (VIIIR:Ag), and plasma fibronectin (CIg; cold-insoluble globulin) were measured in the heparin precipitable fraction (HPF) and heparin supernatant fraction (HS) of normal human plasma. Following heparin induced precipitation, most measurable VIII:C activity (77% +/- 24%) was recovered in the HS. Although there was little VIII:C activity (less than 1%) in the HPF, 20% +/- 6.5% VIII:CAg was present as well as CIg (81% +/- 5.6%). VIIIR:RC activity (72% +/- 12%), and VIIIR:Ag (34 +/- 5.2%). As assessed by Na dodecyl SO4 glyoxyl agarose electrophoresis, the multimeric forms of plasma VIIIR:Ag could be resolved into a series of bands. Larger multimers tended to precipitate with the HPF whereas the smaller multimers tended to remain supernatant. Plasma from a subject with congenital afibrinogenemia was also studied. Although the afibrinogenemic HPF contained CIg, neither VIIIR:RC activity nor VIIIR:Ag was precipitated. However, both were present in the HPF from afibrinogenemic plasma to which fibrinogen had been added, suggesting that they are incorporated in this precipitate because of an affinity for fibrinogen. The ability of heparin to induce precipitation of CIg while leaving most VIII:C activity in the supernatant plasma may be useful in the preparation of procoagulant-rich plasma subfractions, since VIII:C can subsequently be recovered in good yield by cryoprecipitation.
Factors affecting fibrinopeptide-A levels in patients with venous thromboembolism during anticoagulant therapy
Authors: Yudelman I , Greenberg J
Published: October 2019

The prompt reduction of elevated fibrinopeptide A (FPA) levels (normal less than 1.3 pmole/ml) by heparin therapy in patients with thromboembolism suggests that measuring the FPA level may provide a good index of disease activity and be a useful method of monitoring therapy. Sepsis or malignancy may elevate FPA levels and coexist with thromboembolism. FPA levels were surveyed in 51 patients with thromboembolism (including 15 with concurrent sepsis or malignancy) during heparin treatment in an attempt to distinguish the effects of coexistent disease and the progression of thromboembolism. The anticoagulant effect of heparin was within the therapeutic range for 81% of the study period. In patients with thromboembolism alone and marked resolution of emboli on repeat lung scan, the mean daily FPA levels were lower than the values in patients with minimal resolution (p less than 0.005). In patients with marked resolution of pulmonary embolism or venous thrombosis and a concurrent disorder, the mean FPA level remained elevated compared to normal values in patients with thromboembolism alone. These results suggest that FPA levels monitored during heparin therapy of thromboembolism may be useful as an index of disease activity except in the presence of coexisting sepsis or malignancy.
Exercise-induced hemolysis in sickle cell anemia: shear sensitivity and erythrocyte dehydration
Authors: Platt OS
Published: October 2019

We describe a steady-state patient with sickle cell anemia (SS disease) who developed sporadic hemoglobinuria, historically related to vigorous exercise. We studied him and four other patients with SS disease and demonstrated exercise-induced hemoglobinemia. To see if SS erythrocytes were abnormally fragile when exposed to shear forces that could be generated in small vessels of exercising muscles, we exposed them to physiologic shear rates in a cone-plate viscometer. We show that SS erythrocytes are more shear sensitive than normal erythrocytes. This phenomenon is directly related to the presence of dehydrated cells as demonstrated by the increasing shear sensitivity of increasingly dehydrated cells separated on Stractan density gradients. Normal shear sensitivity could be restored to dehydrated layers by restoring normal hydration. Restoration of shear stability was complete in all layers except for the most dense ISC layer. A control group of patients with SC disease exhibited no exercise-induced hemoglobinemia, no abnormal shear sensitivity of whole blood, and only rare dehydrated ISCs. These studies suggest that the exercise-induced hemolysis in SS patients is related to the lysis of dehydrated, shear-sensitive cells. This same process may also contribute to the chronic hemolysis of SS disease--a phenomenon known to correlate with the numbers of dehydrated ISCs.
Studies of the prothrombin activation pathway utilizing radioimmunoassays for the F2/F1 + 2 fragment and thrombin--antithrombin complex
Authors: Teitel JM , Bauer KA , Lau HK , Rosenberg RD
Published: October 2019

We have evaluated the efficacy of utilizing radioimmunoassays (RIAs) for prothrombin activation fragments (F2/F1 + 2) and for thrombin-- antithrombin complex (TAT) in purified systems and in whole blood. During venipuncture, appropriate anticoagulants were employed in order to prevent the generation of thrombin and factor Xa. The RIAs were shown to be specific for F2/F1 + 2 as well as TAT and did not interact with other plasma components. Initially, thrombin generation was studied in a purified human system of prothrombin, antithrombin, factor Xa, and factor V as well as phospholipid and Ca++. Under these conditions, the kinetics of F2/F1 + 2 and TAT generation were virtually superimposable. However, when factor V was omitted from the reaction mixture, a significantly greater amount of F2/F1 + 2 as compared to TAT was observable. Subsequently, prothrombin activation was monitored during the spontaneous coagulation of freshly drawn blood. Throughout the entire course of thrombin generation, the observable rate of formation of F2/F1 + 2 was considerably greater than that of TAT. We have examined the levels of F2/F1 + 2 and TAT in normal individuals. Our studies indicate that the concentrations of F1 + 2 and TAT average 1.97 nM and 2.32 nM, respectively. We have also quantitated the concentrations of F2/F1 + 2 and TAT in patients with disseminated intravascular coagulation. In these individuals, the levels of both components are elevated. However, the ratio of F1 + 2 to TAT ranges from 2.37 to 5.55. Thus, we conclude that under in vivo conditions, prothrombin activation is characterized by the accumulation of a stable precursor, such as prethrombin-2, and that this phenomenon may be related to an alteration of factor V function.
Hemin preferentially stimulates synthesis of alpha-globin in K562 human erythroleukemia cells
Authors: Rowley PT , Ohlsson-Wilhelm BM , Rudolph NS , Farley BA , Kosciolek B , LaBella S
Published: October 2019

K562 human erythroleukemia cells are an established cell line derived from an adult with chronic myelogenous leukemia. Hemin stimulates their synthesis of embryonic and fetal hemoglobins. We have found that their globin synthetic pattern depends on the concentration of added hemin. Clone RA6 was cultured with 0--100 microM hemin and the globin synthetic pattern determined by 3H-leucine incorporation and analysis of 3H-globins by polyacrylamide gel electrophoresis in Triton X acid urea followed by fluorography and densitometry. The higher the hemin concentration, the greater the synthetic rate of each type of globin. However, the relative increase was greatest for alpha-globin. We propose that the differential dependence of alpha synthesis on added hemin is a reflection of translational inefficiency of alpha messenger RNA and that this property is exposed when the translational capacity of the cell is limited by hemin deficiency. We suggest that the differential dependence of alpha-chain synthesis on added hemin in clone RA6 is evidence of an intrinsic deficiency in heme synthesis.
Spectrin tetramer-dimer equilibrium in hereditary elliptocytosis
Authors: Coetzer T , Zail S
Published: October 2019

The proportion of spectrin tetramers and dimers in 4 degrees C low ionic strength extracts of red cell membranes of 9 subjects with 4 different variants of hereditary elliptocytosis (HE) and 2 subjects with hereditary spherocytosis (HS) was determined by nondenaturing gel electrophoresis. Such extracts reflect the native oligomeric state of spectrin in the red cell membrane. In two hemolytic HE variants (an unclassified adult with increased thermal sensitivity of red cells and an infant also showing increased thermal sensitivity of red cells), the proportion of dimers was increased, whereas the remaining subjects had values within the control range. Conversion of spectrin tetramers to dimers under isotonic conditions at 37 degrees C, or spectrin dimers to tetramers at 30 degrees C, resulted in a high proportion of dimers in the above two HE variants, as well as in a third variant with probable mild HE and sporadic hemolysis. The mother of the infant with elliptocytosis and increased thermal sensitivity of red cells, although hematologically normal, had an increased proportion of dimers in 4 degrees C low ionic strength extracts of her red cell membranes. These findings reflect an underlying primary or secondary abnormality of spectrin in these subjects that affects the association state of spectrin in the red cell membrane. Their exact relationship to the pathogenesis of the elliptical shape of the red cell, or to the presence of hemolysis, is at present unclear.
Emergence of B-immunoblastic sarcoma in patients with multiple myeloma: a clinicopathologic study of 10 cases
Authors: Falini B , De Solas I , Levine AM , Parker JW , Lukes RJ , Taylor CR
Published: October 2019

Immunologic and histologic studies were performed in 10 cases of myeloma that showed progression to a more aggressive proliferation, designated as immunoblastic sarcoma of B-cell type (B-IBS). Several patterns of clinical presentation were observed: eight patients showed typical multiple myeloma, four developed B-IBS within the bone marrow, and four developed B-IBS in multiple extramedullary sites; the remaining two patients had relatively localized myeloma, but also showed development of extramedullary B-IBS. The implications of these findings are discussed with regard to their prognostic import and their relationship to current concepts of plasma cell development.
Quantification of the locomotive behavior of polymorphonuclear leukocytes in clot preparations
Authors: Howard TH
Published: October 2019

Time-lapse videotape recordings of polymorphonuclear leukocytes (PMNs) from clot preparations were used to quantify the locomotive behavior of individual PMNs from normal subjects. Tracings derived from the videotapes allow one to quantify multiple parameters of the locomotive behavior of PMNs--direction, distance, rate, and angle of turn. The results obtained are reproducible from subject-to-subject and from preparation-to-preparation. The method allows the investigator to record the locomotive behavior of 100 cells simultaneously within a 5- min period and analyze the recording as time permits. We utilized this technique to compare the locomotive behavior of slow and fast PMNs (arbitrarily defined as cells that move less than or equal to 7.0 micrometer/min and greater than 7.0 micrometer/min mean rate of locomotion, respectively). The studies show that slow and fast PMNs, thus defined, differ not only in mean rate of locomotion but also in their rate of locomotion during periods of active locomotion, in the number of periods of inactivity/PMN/5 min (slow = 1.65 +/- 0.31; fast = 0.36 +/- 0.12), and in their turning behavior as measured by angle of turn (slow = 92 degrees +/- 39 degrees; fast = 39 degrees +/- 35 degrees). These results show that human PMNs from clot preparations are remarkably heterogeneous in their locomotive behavior, and the results suggest this heterogeneity is due to endogenous differences within cells.
Assay of prekallikrein in human plasma: comparison of amidolytic, esterolytic, coagulation, and immunochemical assays
Authors: Fisher CA , Schmaier AH , Addonizio VP , Colman RW
Published: October 2019

Using the substrate H-D-Pro-Phe-Arg-p-nitroanilide-HCl, an amidolytic assay was designed to measure prekallikrein in plasma. At a substrate concentration of 1 mM (Km = 0.2 mM), the amidolysis of purified kallikrein at 1 coagulant unit/ml was observed to be 2.47 mumole/min/ml. Conditions for plasma prekallikrein activation were optimized to approach complete activation when compared to the amidolytic activity of the purified plasma kallikrein. Plasma treated with chloroform to destroy inhibitors of kallikrein was activated with dilute kaolin (final concentration 1 mg/ml) for 1 min at 25 degrees C. Activated plasma prekallikrein had 78% (1.92 mumole/min/ml) of activity of purified kallikrein at plasma concentration. Comparison of this amidolytic assay with immunochemical, esterolytic, and coagulant assays of three subject populations (normals, women on birth control pills, and patients with hepatocellular disease) showed good correlation both in normals and in the patient groups between the amidolytic and esterolytic assays (r = 0.89). Each enzymatic assay correlated with the immunochemical assay (r = 0.72, r = 0.68, respectively). However, comparison of each of these assays with the coagulant assay showed no significant correlation due to the large inherent error of the latter assay. This standardized plasma prekallikrein amidolytic assay should facilitate studies of plasma prekallikrein concentration in physiologic and pathologic conditions and help identify activation of the contact phase of coagulation in disease states.
The ultrastructural localization of immunoglobulins in human b cells of immunoproliferative diseases
Authors: Gourdin MF , Farcet JP , Reyes F
Published: October 2019

The cellular distribution of immunoglobulins in human malignant and normal B cells was investigated by immunoelectron microscopy by direct incubation of fixed cells with electron microscopy by direct incubation of fixed cells with peroxidase-coupled antibody. These conjugates penetrated into the cell, resulting in the simultaneous detection of surface and cytoplasmic immunoglobulins. The latter were seen as specific intracisternal staining of the perinuclear space and endoplasmic reticulum and occasionally of the Golgi complex. Plasma cells were frequently characterized by a heterogeneity of reactivity of the endoplasmic reticulum. Minute amounts of cytoplasmic immunoglobulin were demonstrated in cells without developed secretory organelles, such as lymphoma cells and lymphocytes from chronic lymphocytic leukemia (CLL). The method allowed us to define several subsets of cells according to the expression of surface and cytoplasmic immunoglobulins and thus to determine the stage of maturation of cells involved in monoclonal proliferation.
Factor XI antigen and activity in human platelets
Authors: Tuszynski GP , Bevacqua SJ , Schmaier AH , Colman RW , Walsh PN
Published: October 2019

Washed platelets, contaminated with less than 0.20% plasma factor XI, were examined for the presence of factor XI antigen and activity. These platelets contained a factor-XI-like coagulant activity (0.67 +/- 0.11 U/10(11) platelets) that remained constant after successive washes. By means of indirect immunofluorescence, a monospecific antibody to factor XI showed specific staining of both normal platelets and platelets from patients deficient in plasma factor XI. Radiolabeled Triton extracts of washed platelets and labeled purified factor XI solutions were analyzed for factor XI antigen by Staph A immunoprecipitation analysis using antibody to purified plasma factor XI followed by SDS gel electrophoresis. On unreduced gels, the platelet material ran as a single band having an apparent molecular weight of 220,000 daltons, whereas purified plasma factor XI gave a single band at 160,000 daltons. On reduced gels, the platelet material analyzed as a single band at 52,000 daltons, whereas purified factor XI gave a single band of 80,000 daltons. Analysis of a partially purified factor XI preparation from platelets by immunoelectrophoresis revealed that the platelet preparation displayed a slightly lower cathodal electrophoretic mobility at pH 8.6 than did plasma factor XI and yet appeared to possess complete antigenic identity with plasma factor XI. These results indicate that platelets possess a form of factor XI that exists as a disulfide-linked 52,000-dalton tetramer in contrast to the plasma form that circulates as a 80,000-dalton disulfide-linked dimer.
The relationship between the hemorrhagic and antithrombotic properties of low molecular weight heparin in rabbits
Authors: Carter CJ , Kelton JG , Hirsh J , Cerskus A , Santos AV , Gent M
Published: October 2019

We have compared the hemorrhagic and antithrombotic effects of a low molecular weight (LMW) heparin fraction and standard heparin in rabbits. Similar LMW heparin fractions have antithrombotic effects when tested in animals, but their hemorrhagic effects relative to standard heparin have not been established. Standard porcine mucosal heparin (mol wt 15,000 daltons) was depolymerized by nitrous acid to a low molecular weight fraction (mol wt 4600 daltons). Using equal USP units, the standard and Dep LMW heparin were compared in vitro, ex vivo, and in vivo. In vitro, when diluted in rabbit plasma, the Dep LMW heparin at equivalent anti-Xa activity showed less prolongation of thrombin clotting times or activated partial thromboplastin times. Ex vivo, platelets from rabbits treated with the Dep LMW heparin showed less inhibition of collagen-induced aggregation. The relative hemorrhagic properties of the two heparins were compared in vivo in rabbits using a sensitive blood loss assay, and the antithrombotic properties were compared in a thrombin-induced venous stasis model. By using an optimal threshold heparin dose in each test system, it was possible to demonstrate that equal USP units of Dep LMW heparin caused less blood loss but showed greater antithrombotic activity than standard heparin.
Glycosylated hemoglobins (GHb): an index of red cell survival
Authors: Panzer S , Kronik G , Lechner K , Bettelheim P , Neumann E , Dudczak R
Published: October 2019

Levels of glycosylated hemoglobins (GHb) are significantly (p less than 0.0005) lower in patients with hemolytic anemia (n = 20; mean = 3.9% +/- 0.1% SD GHb of total Hb) compared to patients with nonhemolytic anemia (n = 20; mean = 7.0% +/- 0.7% GHb) and normal controls (n = 30; mean = 6.7% +/- 0.7% GHb). A curvilinear correlation between GHb and red cell survival is demonstrable (n = 20;r2 = 0.88; p less than 0.001). Determination of GHb may be useful as a screening test for hemolytic anemia and for the evaluation of the degree of hemolysis, provided that diabetes mellitus can be excluded.
Pharmacokinetics of continuous intravenous and subcutaneous infusions of cytosine arabinoside
Authors: Weinstein HJ , Griffin TW , Feeney J , Cohen HJ , Propper RD , Sallan SE
Published: October 2019

The pharmacokinetics of continuous subcutaneous cytosine arabinoside (ara-C) infusions were compared with continuous intravenous infusions. Steady-state serum ara-C levels and myelosuppression were similar with both routes of administration. CSF/serum ara-C ratios ranged from 0.14 to 0.91 (mean, 0.58). Continuous subcutaneous ara-C infusions were a convenient and reliable alternative to intravenous infusions.
Quantitative Analysis of the Normal and Four Alternative Degrees of an Inherited Macrocytic Anemia in the House Mouse
Authors: RUSSELL ELIZABETH S. , FONDAL ELIZABETH L.
Published: October 2019

1. A general description is given of the known effects of a series of allelic genes, W, Wv, w,, which are responsible for varying degrees of a macrocytic anemia as well as for differences in viability, gonad development and intensity and location of hair pigment. A summary is given of previous investigations into the basic nature of these anemias.

2. The mean erythrocyte count for each genotype is given for sixteen day embryos, and for zero day, two weeks and four weeks after birth. All determinations are based on comparisons of normal and anemic littermates.

3. A detailed description is given of the methods used to obtain blood samples from embryos. In all three severely affected genotypes the anemia is already well established at the sixteenth day of embryonic life.

4. In each interval thereafter, some increase in erythrocyte level is found in all genotypes. All genotypes, normal and anemic, made approximately the same relative increase during each time interval, indicating the same relative response to the physiologic stimulus of the individual’s growth.

5. Certain genotypes are lethal at particular stages. It is suggested that these deaths are due to a failure of blood formation to reach the absolute level necessary for continued development and resistance to minor noxious stimuli at these critical periods of rapid growth.

6. Certain other factors possibly affecting blood formation or survival were tested. The presence, absence or degree of anemia of the mother were found to have no effect on the erythrocyte level of newborn young of the slightly anemic Wvw genotype. Continuing injections of diluted whole blood into extremely anemic WWs led to survival for more than 100 days. Two WW individuals survived spontaneously to seven months, indicating that there are influences, as yet unanalyzed, which can lift even this genotype to a level where it can produce sufficient erythrocytes to maintain itself for a considerable period.
The Role of the Lungs in the Removal of Transfused Lymphocytes
Authors: WEISBERGER AUSTIN S. , GUYTON RICHARD A. , HEINLE ROBERT W. , STORAASLI JOHN P.
Published: October 2019

In rats, the intravascular injection of large numbers of lymphocytes obtained from the lymphatic system fails to increase the number of circulating lymphocytes. This is the result, chiefly, of a specific selective action of the lungs in removing the cells, after either intravenous or intra-arterial injection. The spleen, and perhaps the liver and kidneys also play a part in removing the transfused lymphocytes, but are not as active as the lungs in this respect. These results are similar to those obtained with the transfusion of granulocytes obtained from the peritoneal cavity of rabbits and raise the possibility that the lungs may act as a homeostatic organ in controlling the level of the white blood count. Under the conditions of this experiment, relatively large doses of ACTH and cortisone had no significant immediate effect on transfused lymphocytes.

These results indicate that inadequate preservation of leukocytes in stored blood does not account for their failure to appear in the circulation after transfusion, and that better preservation will not, in all probability, alter the situation.
Paroxysmal Nocturnal Hemoglobinuria With Two New Case Reports
Authors: MCILVANIE SAMUEL K. , BEARD MARION F.
Published: October 2019

1. Two patients with paroxysmal nocturnal hemoglobinuria are reported. The chronicity of the hemolytic process is stressed.

2. Venous blood pH was found to be generally more alkaline during sleep and we believe that the nocturnal hemolytic process is not due to increased acidity per se.

3. A trial with (1) oral prostigmine bromide for two months, and (2) witin subcutaneous pilocarpine nitrate and pilocarpine hydrochloride showed no evidence of reduced erythrocyte destruction and the anemia was unaffected.

4. There was no effect with hapten upon in vitro hemolysis.

5. The hemoglobin structure of the erythrocyte in this disease is normal as measured by the electrophoretic method.
Feedback regulation of granulopoiesis: polymerization of lactoferrin abrogates its ability to inhibit CSA production
Authors: Bagby GC Jr , Bennett RM
Published: October 2019

Neutrophil extracts were prepared from the peripheral blood of 40 normal volunteers and tested for their ability to inhibit CSA production by mononuclear leukocytes. Highly dilute neutrophil extracts inhibited CSA production/release, while extracts selectively depleted of lactoferrin by antibody affinity chromatography did not. In addition, higher concentrations of neutrophil extracts and higher doses of lactoferrin (10(-9)-10(-6) M) failed to inhibit CSA production/release. We found no evidence of CSA or CSA-enhancing factors in either our lactoferrin or our neutrophil extracts. However, using gel chromatography and rate zonal density sedimentation, we noted that lactoferrin undergoes concentration-dependent polymerization at 10(-9)-10(-10) M in tissue culture medium and that while monomeric lactoferrin effectively inhibits CSA production/release in vitro, the polymeric form does not. Thus, while we have confirmed that lactoferrin is the activity in neutrophil extracts that inhibits CSA production, we have also found that lactoferrin undergoes reversible polymerization at physiologic concentrations and that the polymerized molecule is inactive. The tendency of lactoferrin to polymerize in tissue culture medium and in vivo should be taken into account in any studies on its potential role as a physiologically relevant regulator of granulopoiesis.
Effects of aclacinomycin-A on murine leukemia
Authors: Nara N , Miyamoto T , Hirashima K , Momoi H
Published: October 2019

Effects of aclacinomycin-A (ACM) on leukemic cells and normal hematopoietic stem cells were studied for the purpose of evaluating the usefulness of this agent as an antileukemic agent. ACM suppressed not only leukemic colony-forming unit (LCFU) in the radiation-induced murine erythroleukemia, but also normal hematopoietic stem cells (CFU- S, CFU-C) in mice. However, LCFU was the most sensitive to ACM. After the administration of ACM at a dose of 5 mg/kg. LCFU was suppressed to 0.02 of the control on day 3 and recovered within 5 days. Splenic CFU-S and CFU-C were suppressed to the nadirs on day 2 and day 1, respectively, and they recovered within 5 days. On the basis of the differences in the sensitivities to ACM and recovery patterns between LCFU and normal hematopoietic stem cells. ACM at a single dose of 5 mg/kg was administered every 3 days (total dose of 25 mg/kg) into leukemic mice, and the increase in mean lifespan obtained 113.9%. Effectiveness of ACM in the treatment of acute leukemia was discussed.
Treatment of paroxysmal nocturnal hemoglobinuria
Authors: Rosse WF
Published: October 2019

Patients with PNH may be treated with a number of known agents. As in all patients with a chronic disease, a regimen tolerable over a long period of time must be selected. Knowledge and anticipation of complications and their proper treatment are essential parts in the treatment. When these principals are used, many patients may live reasonable lives for very long periods of time.
A variant of von Willebrand's disease with abnormal expression of factor VIII procoagulant activity
Authors: Montgomery RR , Hathaway WE , Johnson J , Jacobson L , Muntean W
Published: October 2019

Reports on variants of von Willebrand's disease are numerous, but many of these are based on tests that will show marked fluctuations with time and tests that might not be similar in affected family members. This report describes 8 patients with a new variant of von Willebrand';s disease in which there is a normal APTT, slightly reduced one-stage factor VIII:C assay (VIII:C-1), and a drastically reduced two- stage factor VIII:C assay (VIII:C-2). The VIII:C in this variant is more readily adsorbed to AI(OH)3. This variability in VIII:C assays and excessive adsorption to AI(OH)3 are corrected by the addition of either hemophilic plasma or hemophilic factor-VIII-related antigen. This variant is stable with restudy on multiple occasions and is inherited in a stable fashion in three generations of one family. The multimeric structure of the VIIIR:Ag appears normal, although the concentration is moderately reduced. The differences in functional activity, the adsorption to AI(OH)3, and the differences between functional and antigenic (VIII:C Ag) assays of VIII:C support that this is a functional abnormality of type I von Willebrand's disease.
Acid hydrolases as markers of maturation in B-cell chronic lymphocytic leukemia
Authors: Grossi CE , Zicca A , Leprini A , Cadoni A , Pistoia V , Ferrarini M
Published: October 2019

Malignant lymphocytes from 30 B-cell chronic lymphocytic leukemia (B- CLL) patients were studied for the cytochemical localization of two acid hydrolases, alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (AT). The large majority of the cells stained for both ANAE and AP in 7 cases, for AP only in 18 cases, and were negative for both the enzymes in 5 cases. Ultrastructural analysis revealed that the cells that displayed more mature morphological features, such as well developed smooth and rough membrane compartments, were those positive for acid hydrolases. That ANAE and AP are expressed by B cells at late stage of maturation was confirmed by the finding that some lymphocytes and all of the plasmacytoid lymphocytes and plasma cells from Walderstrom's macroglobulinemia, from mixed cryoglobulinemia, and from multiple myeloma patients stained strongly for both ANAE and AP. Using the expression of acid hydrolases and certain ultrastructural features as markers of cell differentiation, it was possible to demonstrate a process of maturation within the single B-CLL clones with accumulation of the cells at stages that differed in the various cases.
Protein C, an antithrombotic protein, is reduced in hospitalized patients with intravascular coagulation
Authors: Griffin JH , Mosher DF , Zimmerman TS , Kleiss AJ
Published: October 2019

Activated protein C is a potent anticoagulant and profibrinolytic enzyme that can be derived from the vitamin-K-dependent serine protease zymogen, protein C, by the action of thrombin. Protein C antigen concentration was determined in plasmas from normals (n = 40) and from 38 patients with intravascular coagulation as evidenced by positive FDP (greater than micrograms/ml). Plasma protein C was 4 micrograms/ml in normals and was significantly depressed (less than 2 SD below the mean of normals) in 19 of the 38 patients. Of 15 patients with suspected intravascular coagulation but normal FDP, protein C was decreased in 5 individuals; 3 of these 5 patients had liver disease. Based on these results, we suggest that extensive activation of the coagulation system in vivo causes a significant consumption of protein C, presumably due to its activation by thrombin and subsequent clearance.
Isolation of functionally different human monocytes by counterflow centrifugation elutriation
Authors: Figdor CG , Bont WS , Touw I , de Roos J , Roosnek EE , de Vries JE
Published: October 2019

Human peripheral blood monocytes were isolated by counterflow centrifugation elutriation (CCE). This technique was modified in such a way that various monocyte fractions (viability greater than 99%) could be elutriated by increasing the density of the CCE-medium in steps of 0.0027 g/ml. All monocytes showed the same size distributions as determined by electronic sizing, which indicated that they differed in their density only. Both cytoplasmic esterase and peroxidase activity increased with the density of the cells. Furthermore, the monocytes with the highest density were 2.3–4 times more active in an antibody- dependent cellular cytotoxicity (ADCC) assay than those with the lowest density. In contrast, the monocyte with the highest density were less capable to induce the proliferation of lymphocytes in mixed leukocyte cultures (MLC) than those with the lowest density. This observation could not be attributed to differences in the expression of HLA-DR determinants, since a monoclonal antibody directed against HLA-DR antigens reacted equally well with the monocytes in different fractions. These results provide evidence for the existence of functionally different subsets of monocytes or different states of differentiation or maturation.
Antithrombin III deficiency: decreased synthesis of a biochemically normal molecule
Authors: Ambruso DR , Leonard BD , Bies RD , Jacobson L , Hathaway WE , Reeve EB
Published: October 2019

A 29-yr-old white female has suffered from recurrent venous thromboses over the last 12 yr. Plasma antithrombin III (AT-III) levels were 48% of normal by immunoelectrophoresis and 56% by chromogenic assay. Three of four siblings and the father had similar AT-III levels without associated venous thromboses. Heparin-Sepharose chromatography demonstrated normal behavior of the patient's AT-III. Her purified AT- III could not be distinguished from AT-III purified from a normal control either by SDS polyacrylamide gel electrophoresis or by crossed immunoelectrophoresis, and the heparin cofactor activity and the progressive antithrombin activity of both AT-III samples were identical. Turnover studies were made in the patient using her own purified AT-III labeled with 131I, (*I). The results did not differ significantly from studies made with autologous *I-AT-III in two normal control women. Her fractional breakdown rate of 0.54 total plasma AT- III per day compared with 0.45 and 0.52 in the controls. These studies indicate that the patient synthesizes a normal AT-III molecule at half normal rates.