bluuebunny/crossref_metadata_2025 huggingface dataset:
Time-Space Clustering Among Cases of Acute Leukemia in Two Georgia Counties
Authors: Evatt Bruce L. , Chase Gary A. , Heath Clark W.
Published: October 2019

Statistical analyses were made of the time-space configuration of 406 acute leukemia cases in DeKalb and Fulton Counties, Georgia, over the 13-yr period 1956-1968. Separate analyses were performed for each of three age groups, 0-14 yr, 15-49 yr, and 50 yr-plus, cases being assigned to 2- and 3-yr time segments and to census tract units according to date of onset and place of residence at diagnosis. Pearson chi-square statistics and Haldane’s moments were used as a measure of homogeneity in case configurations. All calculations took into account age-specific populations for each census tract unit corrected for shifting population density. Evidence of inhomogeneity of time-space configurations, based on case clustering, was found among cases in the 0-14 age group but not at older ages. Clustering was present in both counties whether examined separately or combined, and it was more clearly seen in case distributions in DeKalb County. Analyses done for various periods of time showed that degree of clustering depended greatly on placement of time boundaries.
Metabolism of Human Prothrombin and Fibrinogen in Patients With Thrombocytosis Secondary to Myeloproliferative States
Authors: Martinez Jose , Shapiro Sandor S. , Holburn Ruth R.
Published: October 2019

Simultaneous studies of the metabolism of 125I-prothrombin and 131I-fibrinogen were performed in ten patients with thrombocytosis secondary to myeloproliferative syndromes. The mean fractional catabolic rate (FCR) of prothrombin was 61.9%, differing significantly from the normal mean of 42.5% (p <0.01). Similarly, the mean FCR of fibrinogen was 34.0%, differing significantly from the normal mean of 23.9% (p <0.01). Excellent correlation existed between the increased FCRs of prothrombin and fibrinogen but not between these FCRs and the platelet level, at least for platelet counts above 1,000,000/cu mm. Therapeutic reduction of platelet counts to the normal range was associated with a normalization of FCRs as well.
Active transport of GSSG from reconstituted erythrocyte ghosts
Authors: Prchal J , Srivastava SK , Beutler E
Published: October 2019

Human erythrocyte ghosts were loaded with 35S-labeled GSSG and with a sucrose marker, and the transport of GSSG to the suspending medium was studied. GSSG transport from ghosts occurred only when ATP was also present in the ghosts, proceeded against a concentration gradient, and was inhibited by fluoride. The rate of transport was dependent upon the intracellular concentration of GSSG. The relationship between GSSG concentration and rate of transport was sigmoidal. Half-maximal transport was observed at a GSSG concentration of approximately 9.6mM. The maximal velocity was estimated to be in the range of 0.27 umole GSSG per ml of ghosts per hr. These data suggest that the rate of GSSG transport a physiologic concentrations of GSSG is not sufficiently rapid to account for the turnover of glutathione by red cells. It seems more likely that the GSSG transport system serves an emergency function of erythrocytes.
Group-specific human granulocyte antigens on a chronic myelogenous leukemia cell line with a Philadelphia chromosome marker
Authors: Drew SI , Terasaki PI , Billing RJ , Bergh OJ , Minowada J , Klein E
Published: October 2019

Group-specific human granulocyte antigens are serologically detectable with granulocytotoxic-positive human alloantisera on a cell line, K562, of chronic myelogenous leukemia origin which bears a Philadelphia chromosomal marker. The same cell line lacks serologically detectable HLA, B2 microglobulin, and B-lymphocyte antigens. Granulocyte antigens are important cell markers for cell lines of suspected myeloid lineage.
A mechanism for the hydroperoxide-mediated inactivation of prostacyclin synthetase
Authors: Weiss SJ , Turk J , Needleman P
Published: October 2019

Hydroxyl radical generation was demonstrated during heme-catalyzed decomposition of 15-hydroperoxy arachidonic acid. The hydroperoxide- mediated inactivation of prostacyclin synthetase seems to be related to the generation of this highly reactive species. A relationship between hydroperoxides and the hydroxyl radical may be important in the modulation of prostacyclin synthesis in diseased states.
Improved survival of increased-risk myeloma patients on combined triple- alkylating-agent therapy: a study of the CALGB
Authors: Harley JB , Pajak TF , McIntyre OR , Kochwa S , Cooper MR , Coleman M , Cuttner J
Published: October 2019

Two hundred fifty-two previously untreated evaluable patients with multiple myeloma were entered into a study testing a regimen of three intravenous alkylating agents, melphalan, cyclophosphamide, and carmustine (BCNU), given in combination (BCMP) against a regimen employing oral melphalan (MP). Both regimens included a tapering course of prednisone. Objective responses based on the Myeloma Task Force criteria were significantly more frequent in the group receiving BCMP. Survival for the entire group of BCMP-treated patients was not significantly better than that for MP-treated patients (p = 0.62). However, when the survival of the poor-risk (high tumor cell load) group of patients treated with BCMP was compared with the survival of the poor-risk (high tumor cell load) group of patients treated with MP, an improvement in survival attributable to BCMP therapy was seen (p = 0.049 and 0.02, respectively). In the good-risk (low and intermediate tumor cell load) group, BCMP treatment resulted in a trend toward poorer survival, but this did not achieve statistical significance (p = 0.080 and 0.23, respectively). These results indicate that optimal therapy in myeloma may be dependent on the extent of disease at the time of first treatment. Additional studies to explore the effects of treatment intensity and duration are needed in order to design improved myeloma treatment based on the patient's extent of disease.
A New Inherited Abnormality of Hemoglobin and Its Interaction with Sickle Cell Hemoglobin
Authors: KAPLAN EUGENE , ZUELZER WOLF W. , NEEL JAMES V.
Published: October 2019

Through clinical, genetic and physicochemical studies a new inherited abnormality of hemoglobin has been recognized in American Negroes. The new hemoglobin, provisionally called hemoglobin III, can be separated from both normal and sickle cell hemoglobin by electrophoretic analysis. The structural anomaly of the hemoglobin molecule is determined by a gene inherited as a simple Mendelian dominant.

A distinct hemolytic syndrome which is intermediate between the benign sickle cell trait and sickle cell anemia results from the combination of the new hemoglobin with sickle cell hemoglobin. A tentative characterization of the syndrome is presented. In contrast to classical sickle cell anemia this form of sickle cell disease is characterized by a mild hemolytic anemia with slowly progressive splenomegaly in the absence of cardiac or musculo-skeletal manifestations. In vitro, the erythrocytes sickle like those of sickle cell anemia. The bone marrow shows erythroid hyperplasia, fecal urobilinogen excretion is increased, and the survival time of the erythrocytes in normal recipients is shortened, but in the patients the red cell count and hemoglobin concentration are only slightly depressed. Reticulocytosis is slight and icterus is not observed. Whereas in sickle cell anemia both parents are expected to have the sickle trait, only one parent of these individuals shows sickling, while the nonsickling parent is a carrier of the hemoglobin III. The new syndrome does not appear to be identical with that resulting from the simultaneous presence of the sickling gene and the thalassemiagene.

The presence of hemoglobin III when combined with structurally normal hemoglobin is expressed as an asymptomatic carrier state. The erythrocytes do not sickle but have a high incidence of target cell deformity and increased resistance to hypotonic saline. Although there is no evidence of hemolysis in such individuals their erythrocytes are eliminated with abnormal rapidity from the circulation of normal recipients.

The homozygous state with respect to this new hemoglobin has not yet been identified but may well be some already recognized atypical form of chronic hemolytic anemia.

Studies are now in progress to determine the incidence of this new molecular abnormality of hemoglobin.
Chronic Hemolytic Anemia with Erythrocyte Fragility to Cold and Acid
Authors: LIU S. H.
Published: October 2019

Two cases previously diagnosed as aplastic anemia were found to have abnormal susceptibility of the red blood cells to hemolysis after chilling and after addition of acid in the presence of their own as well as control serum. The serum of the patients had no such effect on control erythrocytes. This erythrocyte abnormality was accompanied by a severe anemia of the macrocytic type, slight reticulocytosis, depression of granulocytes and platelets, and erythropoietic hyperactivity of the bone marrow in the early stages and hypoactivity in the late stages. Increased blood destruction was evidenced not by jaundice, but by persistent hyperhemoglobinemia in the serum and increased iron excretion in the urine, without nocturnal intensification. Chilling in vivo produced hemoglobinuria in Case 1, but only slight evidence of increased blood destruction in Case 2. Administration of large doses of ammonium chloride failed to produce hemoglobinuria in either case, although evidence of an increase in blood destruction was detectable. Both patients had historical and serologic evidence of syphilis. Autopsy of Case 1 showed, in addition to syphilitic aortitis, marked hemosiderosis of the kidneys, liver, spleen and lymph nodes.

These were clearly not cases of paroxysmal hemoglobinuria é frigore (Donath-Landsteiner). They resembled cases of chronic hemolytic anemia with nocturnal paroxysmal hemoglobinuria (Marchiafava-Micheli), but the erythrocyte fragility to chilling, the absence of nocturnal increase in the hemolytic process and the presence of syphilis rendered the acceptance of such diagnosis difficult.
Growth factor-mediated proliferation in B cell non-Hodgkin's lymphomas
Authors: Ford RJ , Kouttab NM , Sahasrabuddhe CG , Davis FM , Mehta SR
Published: October 2019

The non-Hodgkin's lymphomas (NHLs) are a heterogeneous group of human lymphoid tumors, primarily of B cell lineage, which appear to represent arrested stages in B lymphocyte differentiation. Control of cell proliferation is a fundamentally important but poorly understood area of study in these tumors. We have studied a representative group of B cell NHLs to assess their potential for growth factor-mediated proliferation in vitro. Our results show that purified monoclonal NHL B cells of the small cell (well-differentiated lymphocytic lymphoma, nodular poorly differentiated lymphocytic lymphoma, etc) type, that were positive for the human malignancy-associated nucleolar antigen could be stimulated by human B cell growth factor (BCGF) to proliferate in vitro. Other B cell activators such as insoluble anti-Ig and the mitogen protein A also could stimulate thymidine incorporation in the lymphoma cell populations. In vitro lymphoma cell growth could be maintained in the presence of the growth factor for up to five weeks. The large B cell type NHL, however, appeared to be refractory to in vitro stimulation by BCGF as well as other stimulators of normal B cells. These studies suggest that human B cell lymphoid tumors are not only phenotypically similar to their normal B lymphocyte counterparts, but are also sensitive in some cases, to the same types of immunoregulatory molecules that control normal lymphoid cell growth.
The characterization of protein 4.1 Presles, a shortened variant of RBC membrane protein 4.1
Authors: Morle L , Garbarz M , Alloisio N , Girot R , Chaveroche I , Boivin P , Delaunay J
Published: October 2019

In a previous report (Blood 60:265, 1982), we described a family with an abnormal RBC membrane protein doublet, which we considered a shortened protein 4.1 on the basis of biochemical and genetic data. Using an anti-4.1 monoclonal antibody, we confirm here that the shortened protein derives from protein 4.1. One of the members of the family contemporaneously displayed the 4.1 (-) trait, eg, the heterozygous state of this variety of hereditary elliptocytosis that lacks protein 4.1. The 4.1a/4.1b ratio was low whenever the 4.1 trait was present, regardless of the type of protein 4.1 involved. The RBCs of the compound heterozygote, containing only the shortened species of protein 4.1, made it possible to analyze without interference the contact between shortened protein 4.1 and sialoglycoprotein beta, or glycoconnectin. Shortened protein 4.1 did not alter the amount of glycoconnectin in the ghosts nor did it change its extractability into the Triton shells. Limited proteolysis of shortened polypeptides 4.1a and 4.1b showed that they are sequence related. It is conflicting that the persons carrying the shortened protein 4.1 are devoid of specific clinical and morphological abnormalities, apart from those pertaining to the 4.1- trait, when the latter is present.
Studies on the Radiosensitivity of Bone Marrow
Authors: VALENTINE WILLIAM N. , PEARCE MORTON LEE , LAWRENCE JOHN S.
Published: October 2019

1. Six animals (cats) were subjected to four whole body irradiation exposures over a period of one and one-half years. Irradiation dosage was 200 r to the whole body, an amount producing severe hematopoietic abnormalities in the cat. Irradiation in each case was immediately preceded by a large phlebotomy, so that functional impairment of erythropoiesis resulting from irradiation could be assessed in terms of a defective regeneration of erythrocytes. The effect of multiple radiation exposures could also be evaluated for each animal in terms of its prior response to phlebotomy and irradiation. The effects of repeated exposures on the leukocytes could likewise be assessed by means of the comparative leukocyte regeneration curves.

2. Little evidence of permanent functional damage of erythropoietic tissue by repeated exposures under the conditions of this experiment were observed. Erythrocyte and hemoglobin regeneration patterns were very similar from one exposure to another. However, hemoglobin regeneration was slightly slower after the fourth and last irradiation. Whether this represents encroachment on marrow reserve by multiple irradiations or whether it was an artifact resulting from iron deficiency cannot be said with certainty. Multiple exposures exerted little apparent effect on the leukocyte regeneration curves.

3. After recovery from the fourth irradiation, the percentage of mononuclear leukocytes was somewhat less than that prior to the first.

4. It is concluded that while repeated irradiation insults under the conditions of our experiment probably result in hidden damage even in the presence of apparent complete recovery, this damage is so subtle that it is difficult to detect by the tests of marrow reserve employed in this study.
A Second Example of Anti-Cellano (Anti-K)
Authors: LEVINE PHILIP , KUHMICHEL A. B. , WIGOD MILTON
Published: October 2019

A second example of anti-Cellano along with anti-D is described in a patient immunized by a series of transfusions. For the final analysis of antibodies acting on almost all bloods, it is essential to have on hand a carefully selected list of bloods with known antigenic constitution.
The Relative Biologic Effectiveness of 200 KV and 23.5 MEV (Beatatron) X-Radiation
Authors: TING T. P. , JOHNS H. E. , JAQUES L. B.
Published: October 2019

1. The leukocyte value in the peripheral blood of three different strains of mice was followed after a single dose of whole body x-radiation. The maximum leukocyte depression was used as a basis for comparing the biologic effectiveness of the betatron radiation and the 200 KV x-radiation.

2. The ratio of effectiveness in producing 70 per cent leukocyte reduction, for betatron radiation to 200 KV x-radiation, for the three strains was 0.65.

3. The onset of recovery from leukopenia for equal doses of radiation occurred earlier in animals treated with betatron radiation than in those treated with a similar dose of 200 KV x-radiation.

4. The degree of leukopenia and delay in recovery was found to increase somewhat with age of animals treated with 200 KV x-radiation and to a less extent in animals treated with betatron radiation.

5. At a dose of 600 r, over 90 per cent of the animals treated with 200 KV died within a few days, while none of the animals exposed to the same dose of betatron radiation died in the same period. The leukocyte count immediately before death was about 2 per cent of control values.

6. For a dose of 600 r, the lymphocytes were equally reduced by 200 KV and betatron radiation, while the granulocytes were much less reduced by betatron than by 200 KV radiation.
The Mechanism of Quinidine Purpura
Authors: LARSON ROGER K.
Published: October 2019

A case of thrombocytopenic purpura was studied and evidence of a specific hypersensitivity mechanism was found. The following experimental observations and their conclusions were discussed.

1. A rapid fall in circulating platelets occurred in the patient after a test dose of the drug. This fall was too rapid to be accounted for by inhibition of the bone marrow megakaryocytes alone.

2. Inhibition of clot retraction could be produced in the patient's blood by the addition of minute quantities of quinidine in vitro. This was assumed to demonstrate a specific peripheral action of quinidine on this patient's platelets.

3. No inhibition of clot retraction could be produced by the addition of quinine (an optical isomer of quinidine) to the blood of this patient in vitro. This was further evidence of the marked specificity of the reaction.

4. Complete inhibition of clot retraction could be produced in the blood of a normal individual by the addition of serum from the patient together with quinidine. No inhibition occurred if one or the other was added alone. This was assumed to demonstrate that quinidine did not effect the platelets directly but did so through a combined action with some other element in platelet-free serum. It was felt that this was strong support for an antigen-antibody type of reaction.

5. No significant fall in platelet concentration by the addition of quinidine in vitro could be demonstrated. This was interpreted as evidence against a complete lysis of the platelets as the method of destruction. The disparity between the platelet counts at the end of 2 hours in vivo and in vitro suggested that the platelets were injured in such a manner as to effect their rate of removal by the reticulo-endothelial system and their ability to produce clot retraction.
Red Cell Stroma in Dogs
Authors: TISHKOFF G. H. , ROBSCHEIT-ROBBINS F. S. , WHIPPLE G. H.
Published: October 2019

Red cell stroma has not been prepared free of iron and/or hemoglobin. It is not certain whether the iron is a contaminant or an essential constituent of the stroma. Extensive washing does not eliminate iron from the stroma but does cause loss of the stroma constituents, protein and lipid.

Red cell stroma protein is increased in anemia due to blood loss in the dog, on the average in severe anemia, almost twice the figures recorded in the pooled normal samples of red cell stroma.

Lipid fractions under the same conditions show minor fluctuations: approximately 90 per cent of total lipids in the red cells are recovered in the stroma by these methods.

The technic for isolation and fractionation of dog erythrocyte stroma is described. Analyses made from the same blood sample by 3 different workers give comparable results.

Red cell stroma deserves careful study in controlled experimental conditions.
Endocytosis of fibrinogen into megakaryocyte and platelet alpha- granules is mediated by alpha IIb beta 3 (glycoprotein IIb-IIIa) [published erratum appears in Blood 1993 Nov 1;82(9):2936]
Authors: Handagama P , Scarborough RM , Shuman MA , Bainton DF
Published: October 2019

Recently, we showed that platelet alpha-granule fibrinogen is derived entirely from endocytic uptake and not from megakaryocyte synthesis, as previously thought. In this present report, we identify the receptor that mediates endocytosis. We have found that barbourin, a unique disintegrin that is a specific antagonist of alpha IIb beta 3, inhibits the endocytic uptake of fibrinogen into alpha-granules. Continuous intravenous infusion of barbourin (200 micrograms/h) into guinea pigs blocked collagen-induced platelet aggregation as well as endocytosis of biotinylated fibrinogen into megakaryocytes; however, endocytosis of biotinylated albumin by megakaryocytes was not affected. Thus, we have shown that endocytosis of fibrinogen into megakaryocyte and platelet alpha-granules is receptor-mediated, and that alpha IIb beta 3 is the primary receptor.
Enhancement of cathepsin G-induced platelet activation by leukocyte elastase: consequence for the neutrophil-mediated platelet activation
Authors: Renesto P , Chignard M
Published: October 2019

We have focused our interest on the platelet-activating properties of two polymorphonuclear neutrophil (PMN)-derived proteinases, namely elastase (HLE) and cathepsin G (Cat.G). First of all, we observed that whereas HLE was unable to trigger platelet activation by itself, it enhanced platelet activation induced by Cat.G when both proteinases were added simultaneously. It has been recently described that, upon stimulation, PMN released Cat.G, which in turn activated surrounding platelets. Thus, we looked for a combined effect of Cat.G and HLE during this cell-to-cell interaction. When PMN (5 x 10(6)/mL) were stimulated by 0.5 mumol/L N-formyl-Met-Leu-Phe, they released 237.9 +/- 49.1 nmol/L Cat.G and 381.7 +/- 28.0 nmol/L HLE. Such a concentration of purified Cat.G (240 nmol/L) induced only a moderate platelet activation when added to a PMN-platelet mixture. However, when Cat.G (240 nmol/L) and HLE (380 nmol/L) were added together, the resulting platelet activation was strictly comparable to that corresponding to the addition of N-formyl-Met-Leu-Phe (P > .05) in terms of aggregation, dense and alpha granule secretion, and thromboxane B2 production. In fact, Elafin, a specific HLE inhibitor, when added to the PMN-platelet cooperation system triggered by N-formyl-Met-Leu-Phe, prevented platelet activation within the same range of concentrations as for inhibition of HLE activity. In conclusion, we now show that not only Cat.G, but also HLE is involved in the PMN-mediated platelet activation.
Advancement of multiple myeloma from diagnosis through plateau phase to progression does not involve a new B-cell clone: evidence from the Ig heavy chain gene
Authors: Ralph QM , Brisco MJ , Joshua DE , Brown R , Gibson J , Morley AA
Published: October 2019

The Ig heavy chain (IgH) gene was used as a marker to investigate clonal succession and the origin of the neoplastic cell in multiple myeloma. The polymerase chain reaction (PCR) was used to amplify a section of the rearranged IgH gene at diagnosis and at progression in 21 patients who had exhibited a plateau phase. A monoclonal PCR product was seen for 16 of the patients and the product present at progression was of the same molecular weight as that at diagnosis. This finding suggests that the IgH rearrangement present at diagnosis and progression was the same. This was confirmed by sequencing the IgH gene in 10 patients. The IgH genes were found to be hypermutated at diagnosis, but no further hypermutation occurred during the course of the disease. The results provide evidence that the neoplastic cell in myeloma may originate as a memory B cell, plasmablast, or plasma cell, and suggest that progression beyond the plateau phase is not caused by clonal succession.
Frequent latent Epstein-Barr virus infection of neoplastic T cells and bystander B cells in human immunodeficiency virus-negative European peripheral pleomorphic T-cell lymphomas
Authors: Korbjuhn P , Anagnostopoulos I , Hummel M , Tiemann M , Dallenbach F , Parwaresch MR , Stein H
Published: October 2019

We investigated 81 cases of peripheral pleomorphic T-cell lymphoma (PMTCL) occurring in human immunodeficiency virus-negative Europeans for the presence of Epstein-Barr virus (EBV)-DNA through polymerase chain reaction (PCR) for the presence of EBV-encoded small nuclear RNAs (EBER) and immediate early mRNAs (Bam H-fragment, lower strand frame [BHLF]) by in situ hybridization (ISH) and for EBV-encoded latent membrane protein (LMP) and nuclear antigen 2 (EBNA2) by immunohistology (IH). EBER-ISH, which could be applied on all cases, showed an overall incidence of EBV-infected cells in 38 of 81 cases (47%) of PMTCL. These data could be confirmed by PCR, which produced congruent results in the cases with amplifiable DNA. By EBER-ISH, the virus was located in the tumor cells in 30 of the 38 EBV-positive cases, with the proportion of the infected cells ranging from 1% to 100%. In 18 of these cases and in the 8 cases without EBV-infected tumor cells, the virus was, respectively, either additionally or exclusively detectable in occasional nonmalignant lymphoid bystander cells. An LMP expression was observed in several of the EBER-expressing tumor cells in 18 cases, whereas EBNA2 was detectable only in one case, which also displayed signs of viral replication. Some nonmalignant EBV-infected B immunoblasts also expressed LMP in several cases. Primary cutaneous and enteropathy-associated PMTCL displayed less frequent EBV infection when compared with other extranodal or nodal manifestations.
Bone marrow transplantation in dogs after radio-ablation with a new Ho- 166 amino phosphonic acid bone-seeking agent (DOTMP)
Authors: Parks NJ , Kawakami TG , Avila MJ , White R , Cain GR , Raaka SD , Hornoff W , Fisher P , Moore P , Seibert JA
Published: October 2019

beta-emitting 166Ho (t1/2 = 26.78 hours, E(beta)max = 1.8 MeV) complexed with the phosphonic acid chelator, 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetra(methylene phosphonic acid) (DOTMP) at a ligand-to-metal ratio of 1.5:1 binds to bone. This radioactive complex is a marrow-ablating radiopharmaceutical that appears useful for preparation of bone marrow (BM) transplant recipients without the morbidity usually associated with total body irradiation preparatory regimens. We have found with seven splenectomized young adult beagle dogs that a 166Ho radiopharmaceutical dosage of 370 MBq/kg body weight provides an initial skeletal radioactivity burden of at least 1.5 GBq/kg skeleton and results in complete ablation of hematopoietic marrow cell populations within 7 days. The beta particle flux distribution in BM-forming skeletal tissue is not uniform. Red marrow radiation doses varied from 30 to 115 Gy as estimated by direct radioassay and autoradiographic analyses of both bone biopsies and postmortem samples; the median value of 61 Gy agreed with our theoretical expectations. In vivo radioactivity distribution was evaluated with nuclear imaging methods. Apparently, normal hematopoiesis was restored in three dogs with autologous BM transplants performed 5 to 6 days after administration of the marrow ablative radiopharmaceutical, 166Ho-DOTMP. BM biopsies at 7 to 10 months posttransplantation indicate continued normal hematopoietic activity.
Regulation of granulocyte colony-stimulating factor and granulocyte- macrophage colony-stimulating factor expression by oncostatin M
Authors: Brown TJ , Liu J , Brashem-Stein C , Shoyab M
Published: October 2019

Oncostatin M (OM) is structurally and functionally related to a subclass of hematopoietic cytokines including leukemia-inhibitory factor (LIF), ciliary neurotrophic factor (CNTF), granulocyte colony- stimulating factor (G-CSF), and interleukin-6 (IL-6). Using human endothelial cells (HEC) as a model for cytokine regulation of hematopoietic growth factor expression, we tested OM as an inducer of colony-stimulating activity. Colony-forming cell assays supplemented with culture supernatants from OM-treated HEC contained a threefold increase in colony-forming unit granulocyte-macrophage colonies. Specific immunoassay (enzyme-linked immunosorbent assay) of culture supernatants indicated that OM treatment of HEC resulted in a dose- and time-dependent increase in the accumulation of G-CSF and granulocyte- macrophage CSF (GM-CSF) (> 28-fold). The ED50 for OM induction of G-CSF and GM-CSF protein expression was 17 and 7 pmol/L, respectively. Increased protein expression was associated with a similar increase in steady-state expression of G-CSF and GM-CSF mRNA. Furthermore, a period of 12 to 24 hours elapsed before there were measurable increases in CSF expression, suggesting that OM may stimulate CSF production through a mechanism requiring the synthesis or activation of a secondary mediating factor or pathway. These findings provide the first evidence that OM may regulate myelopoiesis by inducing the cellular expression of hematopoietic growth factors.
Study of prognosis in Waldenstrom's macroglobulinemia: a proposal for a simple binary classification with clinical and investigational utility
Authors: Gobbi PG , Bettini R , Montecucco C , Cavanna L , Morandi S , Pieresca C , Merlini G , Bertoloni D , Grignani G , Pozzetti U
Published: October 2019

Prognostic evaluation of Waldenstrom's macroglobulinemia (WM) is unreliable, few studies considered prognostic factors in WM and only one was derived from a multivariate analysis. One hundred forty-four retrospective, previously untreated patients with clinically overt WM were studied to learn whether overall survival was related to any of the various clinical features presented at diagnosis. Patients were homogeneously treated with intermittent doses of chlorambucil for as long as this showed an effect on the monoclonal component. The population was randomly subdivided into a 90-patient exploratory sample, on whom investigation would be conducted, and in a 54-patient test sample, on whom the results would be validated. In the exploratory sample univariate analysis identified the following parameters as the most important for prognosis: age (< or > or = 70 years), platelet count (< or > or = 120 x 10(9)/L), presence or absence of an abnormal number of red blood cells in the urine, hemoglobin concentration (< or > or = 9 g/dL), erythrocyte sedimentation rate (< or > or = 110 mm at first hour), presence or absence of cryoglobulinemia and of weight loss. Cox multivariate analysis showed that only hemoglobin, age, weight loss, and cryoglobulinemia independently affected survival. These four clinical variables were also shown to be able to discriminate survival significantly in the test sample. Moreover, it was possible to demonstrate (both in the exploratory and the test sample) that clear-cut, albeit dichotomic, survival discrimination can be reached with the presence at diagnosis of either no more than one, or any two or more, of these four prognosticators. These simple clinical criteria could be the basis of an initial binary, prognostic classification of WM, which could help in differentiating therapy according to the severity of the disease, and in properly designing future clinical trials.
Two acquired immunodeficiency syndrome-associated Burkitt's lymphomas produce specific anti-i IgM cold agglutinins using somatically mutated VH4-21 segments
Authors: Riboldi P , Gaidano G , Schettino EW , Steger TG , Knowles DM , Dalla-Favera R , Casali P
Published: October 2019

We analyzed the reactivity and the structure of the VH and VL segments of two IgM monoclonal antibodies (MoAbs) produced by spontaneously in vitro outgrowing cell lines, HBL-2 and HBL-3, established from two acquired immunodeficiency syndrome (AIDS) patients with Epstein-Barr virus (EBV)-negative Burkitt's lymphoma (BL). These B-cell clones were representative of the respective neoplastic parental clones, as determined by immunophenotypic and molecular genetic analysis. The IgM MoAbs were highly specific for the i determinant on red blood cells (cold agglutinins), but bound none of the other eight self and nine foreign antigens (Ags) tested, including those most commonly recognized by natural antibodies or autoantibodies. Structural analysis showed that the IgM MoAb VH segment sequences were 93.5% and 84.2% identical with that of the germline VH4–21 gene, which encodes the vast majority of cold agglutinins that are specific for the i/l carbohydrate Ag and are produced under chronic lymphoproliferative conditions. The HBL-2 MoAb VH4–21 gene segment was juxtaposed with 20P3 and JH6 genes and paired with a V lambda 1 segment, the sequence of which was 95.5% identical to that of the germline Humlv117 gene; the HBL-3 MoAb VH4–21 gene segment was juxtaposed with DXP'1 and JH5 genes and paired with a V lambda 1 segment, the sequence of which was 86.7% identical to that of the germline Humlv1L1 gene. The high degree of conservation of the VH4–21 gene in the human population, the nature of the nucleotide differences in the expressed VH4–21 segments, and the presence of nucleotide substitutions in the HBL-2 and HBL-3 IgM MoAb JH and/or J lambda segments suggested that the MoAb V segments underwent a process of somatic hypermutation. This was formally shown in the HBL-3 MoAb VH segment, by differentially targeted polymerase chain reaction amplification of the HBL-3 MoAb-producing cell genomic DNA. In addition, cloning and sequencing of the genomic DNA from fibroblasts of the same patient whose neoplastic B cells gave rise to the HBL-3 cell line yielded a germline copy of the VH4–21 gene. Thus, the expression of VH4–21 gene products may be involved in a self Ag-driven process of clonal B-cell expansion and selection associated with BL in these AIDS patients.
Expression of interleukin-2 receptor gamma chain on human neutrophils
Authors: Liu JH , Wei S , Ussery D , Epling-Burnette PK , Leonard WJ , Djeu JY
Published: October 2019

The interleukin-2 (IL-2) receptor gamma is an indispensable functional component of IL-2, IL-4, and IL-7 receptors, and thus, is denoted the common gamma chain, gamma c. The present study was undertaken to determine whether human polymorphonuclear neutrophils (PMNs) expressed gamma c chain. Reverse transcription-polymerase chain reaction and Northern blot analysis showed that fresh human PMN constitutively expressed a remarkable level of gamma c mRNA, which is of the size and intensity of that from the peripheral blood mononuclear cells (PBMCs). Granulocyte macrophage-colony stimulating factor, IL-2, and IL-8, which are known to activate PMN functions, failed to regulate the gamma c gene expression. Western blot analysis with a rabbit anti-gamma c polyclonal antibody identified 64-, 58-, and 50-kD gamma c bands in lysates from PMN, but only 64- and 58-kD bands from PBMCs. After the PMNs and PBMCs were treated with tunicamycin to prevent N-linked glycosylation, Western blot analysis detected a single 39-kD band, which is equal to the calculated molecular weight from the cloned cDNA. Thus, our results indicate that PMNs constitutively express high levels of gamma c and the three forms detected are caused by different glycosylation of a protein translated from a single mRNA species.
Regulation of interleukin-11 protein and mRNA expression in neonatal and adult fibroblasts and endothelial cells
Authors: Suen Y , Chang M , Lee SM , Buzby JS , Cairo MS
Published: October 2019

Interleukin-11 (IL-11), a newly-identified cytokine produced by stromal cells, elevates platelet counts in neonatal rats in vivo and synergizes in vitro with IL-3 in supporting murine megakaryocyte colony formation and stimulating hematopoietic stem cells. Megakaryocytopoiesis is also enhanced by other colony-stimulating factors (CSFs), including IL-3, IL- 6, and Steel factor (SLF). Dysregulation of neonatal thrombopoiesis predisposes newborns to develop thrombocytopenia during sepsis, despite increased circulating pools of committed thrombopoietic progenitors in newborn cord blood compared with adult. We previously reported reduced expression of granulocyte-macrophage colony-stimulating factor (GM- CSF), granulocyte-colony-stimulating factor (G-CSF), and IL-3 from stimulated cord mononuclear cells, but increased expression of SLF in human umbilical vein endothelial cells (HUVEC). Therefore, we hypothesized that IL-3, IL-6, and SLF might modulate megakaryocytopoiesis by inducing IL-11 expression, and newborns might express altered levels of IL-11 mRNA expression during activated conditions, contributing to the difference in circulating colony- forming unit-megakaryocyte (CFU-Meg) cord and adult blood. Phorbol myristate acetate (PMA) induced a twofold greater increase in IL-11 mRNA expression in neonatal fibroblasts (NFb) compared with adult fibroblasts (AFb), and a 3.6-fold greater increase in HUVEC than human adult aorta endothelial cells (HAEC) by Northern blot analysis. PMA also induced a threefold greater increase in IL-11 protein production in NFb than AFb. Physiologic agonists IL-1 alpha, transforming growth factor-beta 1 (TGF-beta 1), and TGF-beta 2 triggered upregulation of IL- 11 mRNA expression in both NFb and AFb. However, IL-3, IL-6, PIXY321 (a GM-CSF-IL-3 fusion protein), and SLF failed to upregulate IL-11 mRNA expression from the basal level, while macrophage-colony stimulating factor (M-CSF) mRNA was significantly induced. These data suggest that the hematopoietic effect of IL-6, SLF, and IL-3 on megakaryocytopoiesis is probably not mediated by secondary IL-11 mRNA expression. Similarly, inflammatory agonists IL-1 beta, lipopolysaccharide (LPS), and tumor necrosis factor-alpha (TNF-alpha) alone did not upregulate IL-11 expression from the basal level in endothelial cells, whereas intracellular adhesion molecule-1 (ICAM-1) and endothelial leukocyte adhesion molecule-1 were strongly induced. Minimal basal IL-11 expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in NFb, AFb, HUVEC and HAEC. The quantitative RT-PCR assay also verified that IL-1 beta and TNF-alpha-stimulated HUVEC and HAEC, and IL-3- and IL-6-stimulated NFb and AFb only expressed minimal levels of IL-11 mRNA.(ABSTRACT TRUNCATED AT 400 WORDS)
Regulation of cytokine expression by interferon-alpha in human bone marrow stromal cells: inhibition of hematopoietic growth factors and induction of interleukin-1 receptor antagonist
Authors: Aman MJ , Keller U , Derigs G , Mohamadzadeh M , Huber C , Peschel C
Published: October 2019

We investigated the effects of interferon-alpha (IFN-alpha) on the expression of cytokines by human bone marrow stromal cells. Production of granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte-CSF (G-CSF), and interleukin-1 beta (IL-1 beta) in stromal cell layers was induced by incubation with IL-1 alpha, tumor necrosis factor (TNF), or lipopolysaccharide (LPS). Addition of IFN-alpha to such stimulated cultures resulted in a strong downregulation of mRNA expression of GM-CSF and IL-1 beta. Similarly, the protein levels of GM- CSF and IL-1 beta were significantly reduced by IFN-alpha, whereas G- CSF production was only moderately inhibited. In contrast, IFN-alpha markedly stimulated the production of IL-1 receptor antagonist (IL-1RA) by stromal cells. The inhibition of cytokine expression resulted in a reduced hematopoietic activity of stromal cells, indicated by a reduced proliferation of the factor dependent cell line MO7e on IFN-alpha- treated stromal cells. In the presence of cycloheximide (CHX), IFN- alpha failed to inhibit IL-1 mRNA expression, whereas the regulation of GM-CSF and IL-1RA by IFN-alpha was not affected. Our results indicate that the myelosuppressive effects of IFN-alpha, as observed in therapeutic applications or associated with viral infections, are, in part, indirectly mediated by inhibition of the paracrine production of hematopoietic growth factors.
Cytokine production by primary bone marrow megakaryocytes
Authors: Jiang S , Levine JD , Fu Y , Deng B , London R , Groopman JE , Avraham H
Published: October 2019

Primary human bone marrow megakaryocytes were studied for their ability to express and release cytokines potentially relevant to their proliferation and/or differentiation. The purity of the bone marrow megakaryocytes was assessed by morphologic and immunocytochemical criteria. Unstimulated marrow megakaryocytes constitutively expressed genes for interleukin-1 beta (IL-1 beta), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha), by the polymerase chain reaction (PCR) and Northern blot analysis. At the protein level, megakaryocytes secreted significant amounts of IL-1 beta (53.6 +/- 3.6 pg/mL), IL-6 (57.6 +/- 15.6 pg/mL), and GM-CSF (24 +/- 4 pg/mL) but not TNF-alpha. Exposure of human marrow megakaryocytes to IL-1 beta increased the levels of IL-6 (87.3 +/- 2.3 pg/mL) detected in the culture supernatants. Transforming growth factor- beta was also able to stimulate IL-6, IL-1 beta, and GM-CSF secretion, but was less potent than stimulation with phorbol-12-myristate-13- acetate (PMA). The secreted cytokines acted additively to maintain and increase the number of colony-forming unit-megakaryocytes colonies (approximately 35%). These studies demonstrate the production of multiple cytokines by isolated human bone marrow megakaryocytes constitutively or stimulated in vitro. The capacity of human megakaryocytes to synthesize several cytokines known to modulate hematopoietic cells supports the concept that there may be an autocrine mechanism operative in the regulation of megakaryocytopoiesis.
Expression and function of the human granulocyte-macrophage colony- stimulating factor receptor alpha subunit
Authors: Jubinsky PT , Laurie AS , Nathan DG , Yetz-Aldepe J , Sieff CA
Published: October 2019

To determine the expression and function of the granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor alpha chain (GMR alpha) during hematopoiesis and on leukemic cells, monoclonal antibodies were raised by immunizing mice with cells expressing high levels of human GMR alpha. A pool of five antibodies isolated from three different mice was used to characterize GMR alpha. This antibody pool (anti-GMR alpha) immunoprecipitated a protein with the expected molecular weight of GMR alpha from COS cells transiently transfected with the GMR alpha gene. In factor-dependent cells, GMR alpha existed as a phosphoprotein. However, its phosphorylation was not stimulated by the presence of GM- CSF. Anti-GMR alpha inhibited the GM-CSF-dependent growth of cell lines and normal bone marrow cells and inhibited the binding of iodinated GM- CSF to its receptor. Cell surface expression of GMR alpha was examined using anti-GMR alpha and flow cytometry. GMR alpha was readily detectable on both blood monocytes and neutrophils. In adherence- depleted normal bone marrow, two separate populations expressed GMR alpha. The most positive cells were predominantly macrophages, whereas the cells that expressed less GMR alpha were largely myelocytes and metamyelocytes. A small population of lin-CD34+ or CD34+CD38- cells also expressed GMR alpha, but they were not capable of significant growth in colony-forming assays. In contrast, the majority of lin-CD34+ and CD34+CD38- cells were GMR alpha-, yet they produced large numbers of myeloid and erythroid colonies in the same assay. Malignant cells from patients with leukemia were also tested for GMR alpha expression. All of the myeloid leukemias and only rare lymphoid leukemias surveyed tested positive for GMR alpha. These results show that anti-GMR alpha is useful for the functional characterization of the GMR alpha and for the detection of myeloid leukemia and that GMR alpha is expressed on certain lineages throughout hematopoietic development; however, progenitors that express the receptor may have a reduced capacity to proliferate in response to hematopoietic growth factors.
Immunomorphologic analysis of bone marrow biopsies after treatment with 2-chlorodeoxyadenosine for hairy cell leukemia
Authors: Ellison DJ , Sharpe RW , Robbins BA , Spinosa JC , Leopard JD , Saven A , Piro LD
Published: October 2019

Treatment of hairy cell leukemia with 2-chlorodeoxyadenosine (2-CdA) induces complete remissions in 85% of patients. Complete remission has been defined as the absence of hairy cells in the bone marrow after routine morphologic examination. To determine if hairy cells could be detected in complete remission bone marrows using immunohistochemical techniques with antibodies L26 (CD20) and DBA.44, 154 bone marrow biopsies performed between 3 months and 25 months after therapy were studied. Of the biopsies, 50% exhibited staining with L26 and/or DBA.44 in five or more cells with morphologic features of hairy cells. Minimal residual disease was usually less than 1% of the total cellular population. DBA.44-positive cells were demonstrated in 91% of the biopsies, although in 48% of these the morphologic features of the positive cells were not sufficiently distinctive for hairy cells. The proportion of biopsies with residual hairy cells was similar over the 25 months of follow up, indicating a relatively stable amount of residual disease. Immunomorphologic analysis is a more sensitive method for detecting residual hairy cells than morphology alone. Although further follow up is necessary to determine the clinical significance of the L26/DBA.44-positive staining in cells with and without distinctive morphologic features of hairy cells, we conclude that many patients in a stable clinical remission may have residual hairy cells.
Characterization of three erythropoietin (Epo)-binding proteins in various human Epo-responsive cell lines and in cells transfected with human Epo-receptor cDNA [see comments]
Authors: Takahashi T , Chiba S , Hirano N , Yazaki Y , Hirai H
Published: October 2019

Molecular cloning of a cDNA for a mouse erythropoietin (Epo) receptor (EpoR) has facilitated the understanding of the structure of this receptor. However, there is, as yet, no explanation for the discrepancy between the protein recognized by specific antibodies against mouse EpoR and the unexpectedly larger species that can be cross-linked to labeled Epo. It is unclear whether the product of an unidentified gene is included in the EpoR complex. In the present study, we directly compared the cross-linking patterns for human EpoR that were endogenously expressed in three types of Epo-responsive cell, and that was artificially expressed in nonhematopoietic cells after transfection with cDNA for human EpoR. We observed that 85-kD and 105-kD proteins formed ligand-receptor complexes in all the human Epo-responsive cells and, furthermore, that the formation of a complex derived from the 70- kD protein was dependent on the level of expression of the cloned EpoR mRNA in these cells. By contrast, a prominent cross-linked band derived from the 70-kD protein and a weaker band derived from the 80- to 85-kD protein, but no band derived from the 105-kD protein, could be shown in the case of nonhematopoietic cells transfected with the human EpoR cDNA. These observations suggest that the cloned cDNA for human EpoR alone does not allow generation of the complete EpoR in nonhematopoietic cells and that the 105-kD Epo-binding protein may represent the product of an as yet unidentified gene that is expressed in hematopoietic cells.