bluuebunny/crossref_metadata_2025 huggingface dataset:
Adenosine Deaminase Deficiency in Adults
Authors: Ozsahin Hulya , Arredondo-Vega Francisco X. , Santisteban Ines , Fuhrer Hanspeter , Tuchschmid Peter , Jochum Wolfram , Aguzzi Adriano , Lederman Howard M. , Fleischman Amy , Winkelstein Jerry A. , Seger Reinhard A. , Hershfield Michael S.
Published: October 2019

Adenosine deaminase (ADA) deficiency typically causes severe combined immunodeficiency (SCID) in infants. We report metabolic, immunologic, and genetic findings in two ADA-deficient adults with distinct phenotypes. Patient no. 1 (39 years of age) had combined immunodeficiency. She had frequent infections, lymphopenia, and recurrent hepatitis as a child but did relatively well in her second and third decades. Then she developed chronic sinopulmonary infections, including tuberculosis, and hepatobiliary disease; she died of viral leukoencephalopathy at 40 years of age. Patient no. 2, a healthy 28-year-old man with normal immune function, was identified after his niece died of SCID. Both patients lacked erythrocyte ADA activity but had only modestly elevated deoxyadenosine nucleotides. Both were heteroallelic for missense mutations: patient no. 1, G216R and P126Q (novel); patient no. 2, R101Q and A215T. Three of these mutations eliminated ADA activity, but A215T reduced activity by only 85%. Owing to a single nucleotide change in the middle of exon 7, A215T also appeared to induce exon 7 skipping. ADA deficiency is treatable and should be considered in older patients with unexplained lymphopenia and immune deficiency, who may also manifest autoimmunity or unexplained hepatobiliary disease. Metabolic status and genotype may help in assessing prognosis of more mildly affected patients.
Strict Lymphotropism of Epstein-Barr Virus During Acute Infectious Mononucleosis in Nonimmunocompromised Individuals
Authors: Karajannis M.A. , Hummel M. , Anagnostopoulos I. , Stein H.
Published: October 2019

Previous investigations of exfoliated oropharyngeal cells from individuals suffering from infectious mononucleosis (IM) suggested that the oropharyngeal epithelia are the primary target and also the site of life-long persistence of the Epstein-Barr virus (EBV). This concept was widely accepted. However, the investigation of histological sections with more sensitive EBV detection techniques has drawn this concept into doubt since EBV proved to be constantly absent in normal epithelial cells. To elucidate the discrepancy, throat washings and peripheral mononuclear blood cells from 16 patients suffering from IM were investigated for EBV-DNA and EBV gene products employing highly sensitive in situ hybridization, immunocytochemistry, and polymerase chain reaction. Although all patients exhibited latently infected B lymphocytes in peripheral blood, samples of exfoliated oropharyngeal cells were constantly EBV-negative with the exception of three cases. In these cases, the patients additionally suffered from purulent ulcerating tonsillitis, EBV-infected B cells, but no EBV-infected epithelial cells were detectable. These findings support the view that recirculating lymphocytes of B-cell origin, but not epithelial cells are the initial target of EBV during primary infection and that B cells also represent the site of life-long viral persistence.
CD8+ T Cells Activated During the Course of Murine Acute Myelogenous Leukemia Elicit Therapeutic Responses to Late B7 Vaccines After Cytoreductive Treatment
Authors: Dunussi-Joannopoulos Kyriaki , Krenger Werner , Weinstein Howard J. , Ferrara James L.M. , Croop James M.
Published: October 2019

We have previously shown in a murine acute myelogenous leukemia (AML) model that leukemic mice can be cured with a B7 vaccine if immunized early in the disease and that CD8+ T cells are necessary for tumor rejection. However, when B7 vaccine is administered 2 weeks after leukemia inoculation, the effect is only prolonged survival, ending in death virtually of all the mice. To distinguish between tumor kinetics and tumor-induced immunosuppression as potential mechanisms eliminating the therapeutic potential of late B7 vaccines, we performed in vitro T-cell studies during leukemia progression and in vivo studies on the clinical outcome of late B7 vaccines in combination with prior cytoreductive chemotherapy. Our results show that CD8+ T cells from leukemic mice 1 and 2 weeks after leukemia inoculation proliferate more vigorously in response to in vitro activation than cells from normal mice and produce Th1-type cytokines interleukin-2 and interferon-γ. Cytotoxic T lymphocyte (CTL) assays demonstrate that cells from week-2 vaccinated mice (which succumb to their leukemia), surprisingly develop a stronger CTL activity than cells from week-1 vaccinated mice (which reject their leukemia). Finally, the combination of late chemotherapy and late B7 vaccine administration can cure only 20% of leukemic mice, whereas early chemotherapy and the same late B7 vaccine administration cure 100% of leukemic mice. These results demonstrate that in murine AML tumor growth does not induce T-cell anergy or a Th2 cytokine profile and suggest that tumor growth is most likely to be the limiting factor in the curative potential of late B7 vaccines.
TRA1, a Novel mRNA Highly Expressed in Leukemogenic Mouse Monocytic Sublines But Not in Nonleukemogenic Sublines
Authors: Kasukabe Takashi , Okabe-Kado Junko , Honma Yoshio
Published: October 2019

Mouse monocytic Mm-A, Mm-P, Mm-S1, and Mm-S2 cells are sublines of mouse monocytic and immortalized Mm-1 cells derived from spontaneously differentiated, mouse myeloblastic M1 cells. Although these subline cells retain their monocytic characteristics in vitro, Mm-A and Mm-P cells are highly leukemogenic to syngeneic SL mice and athymic nude mice, whereas Mm-S1 and Mm-S2 cells are not or are only slightly leukemogenic. To better understand the molecular mechanisms of these levels of leukemogenicity, we investigated putative leukemogenesis-associated genes or oncogenes involved in the maintenance of growth, especially in vivo, by means of differential mRNA display. We isolated a fragment clone (15T01) from Mm-P cells. The mRNA probed with 15T01 was expressed at high levels in leukemogenic Mm-P and Mm-A cells but not in nonleukemogenic Mm-S1 and Mm-S2 cells. The gene corresponding to 15T01, named TRA1, was isolated from an Mm-P cDNA library. The longest open reading frame of the TRA1 clone predicts a peptide containing 204 amino acids with a calculated molecular weight of 23,049 D. The predicted TRA1 protein is cysteine-rich and contains multiple cysteine doublets. A putative normal counterpart gene, named NOR1, was also isolated from a normal mouse kidney cDNA library and sequenced. NOR1 cDNA predicts a peptide containing 234 amino acids. The sequence of 201 amino acids from the C-terminal NOR1 was completely identical to that of TRA1, whereas the remaining N-terminal amino acids (33 amino acids) were longer than that (3 amino acids) of TRA1 and the N-terminus of NOR1 protein contained proline-rich sequence. A similarity search against current nucleotide and protein sequence databases indicated that the NOR1/TRA1 gene(s) is conserved in a wide range of eukaryotes, because apparently homologous genes were identified in Caenorhabditis elegans and Saccharomyces cerevisiae genomes. Northern blotting using TRA1-specific and NOR1-specific probes indicated that TRA1 mRNA is exclusively expressed in leukemogenic but not in nonleukemogenic Mm sublines and normal tissues and also indicated that NOR1 mRNA is expressed in normal tissues, especially in kidney, lung, liver, and bone marrow cells but not in any Mm sublines. After leukemogenic Mm-P cells were induced to differentiate into normal macrophages by sodium butyrate, the normal counterpart, NOR1, was expressed, whereas the TRA1 level decreased. Furthermore, transfection of TRA1 converted nonleukemogenic Mm-S1 cells into leukemogenic cells. These results indicate that the TRA1 gene is associated at least in part with the leukemogenesis of monocytic Mm sublines.
Erythropoietin and Interleukin-3 Activate Tyrosine Phosphorylation of CBL and Association With CRK Adaptor Proteins
Authors: Barber Dwayne L. , Mason Jacqueline M. , Fukazawa Toru , Reedquist Kris A. , Druker Brian J. , Band Hamid , D'Andrea Alan D.
Published: October 2019

Transformation of hematopoietic cells by the Bcr-abl oncoprotein leads to constitutive tyrosine phosphorylation of a number of cellular polypeptides that function in normal growth factor-dependent cell proliferation. Recent studies have shown that the CrkL adaptor protein and the Cbl protooncoprotein are constitutively tyrosine phosphorylated and form a preformed complex in cells expressing Bcr-abl. In the current study, we have examined cytokine-dependent tyrosine phosphorylation of Cbl and its association with Crk proteins. Erythropoietin (EPO) and interleukin-3 induced a dose and time-dependent tyrosine phosphorylation of Cbl in both EPO-dependent Ba/F3 and DA-3 transfectants, and the erythroid cell line HCD-57. Furthermore, once phosphorylated, Cbl associated with Crk adaptor proteins. Of the three Crk isoforms expressed in hematopoietic cells (CrkL, CrkII, and CrkI), tyrosine phosphorylated Cbl binds preferentially to CrkL and CrkII. The amount of Cbl associated with CrkL and CrkII exceeded the fraction of Cbl associated with Grb2 indicating that unlike other receptor systems, the Cbl-Crk association represents the dominant complex of Cbl in growth factor-stimulated hematopoietic cells. In factor-dependent hematopoietic cell lines, CrkL constitutively associated with the guanine nucleotide release factor, C3G, which is known to interact via Crk src-homology 3 (SH3) domains. Our data suggest that the inducible Cbl-Crk association is a proximal component of a signaling pathway downstream of multiple cytokine receptors.
Receptors That Induce Erythroid Differentiation of Ba/F3 Cells: Structural Requirements and Effect on STAT5 Binding
Authors: Pless Miklos , Norga Koenraad , Carroll Martin , Heim Markus H. , D'Andrea Alan D. , Mathey-Prevot Bernard
Published: October 2019

Ectopic expression of the erythropoietin receptor (EpoR) in the interleukin-3 (IL-3)–dependent cell line Ba/F3 results in growth and partial erythroid differentiation in Epo. In contrast, introduction and activation of the interleukin-5 receptor (IL-5R) or of the granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR) results in proliferation only. As this effect is specific to the EpoR, the role of its extracellular or cytoplasmic domain in differentiation was tested after construction of two chimeric receptors. One receptor contained the extracellular domain of EpoR fused to the endodomain of IL-3R β-chain (E/β), while the other contained the EpoR cytoplasmic region fused to the extracellular domain of GM-CSFR α-chain (GMER). Surprisingly, both receptors induced differentiation ruling out a strict specificity of the extracellular or cytoplasmic region of EpoR in this process. Instead the ability to signal differentiation correlated with structural features shared by the EpoR, GMER, and E/β receptors. Dimerization of all three receptors results in the pairing of two signal transducing chains in the cytoplasm, in contrast to the mitogenic receptors IL-3R, IL-5R, GM-CSFR, which assemble as αβ heterodimers. Two new chimeric receptors that fulfilled the structural requirement exemplified by EpoR, but lacked any part of EpoR, were designed to consolidate this model. They consisted of the ectodomains of the GMR-α and IL-5Rα, respectively, fused to the endodomain of IL-3R β-chain. Both receptors were as effective as EpoR in signaling differentiation in response to their cognate ligand. Another property of receptors fulfilling these structural requirements is that they cause a marked delay in signal transducers and activators of transcription 5 (STAT5) activation on ligand stimulation. Taken together our studies show that structural assembly of receptors dictates their potential to signal erythroid differentiation in Ba/F3 cells, that differentiation can take place in the absence of Epo and that a delay in STAT5 activation is highly predictive of this process.
Thiol-Disulfide Isomerization in Thrombospondin: Effects of Conformation and Protein Disulfide Isomerase
Authors: Huang Evelyn M. , Detwiler Thomas C. , Milev Youli , Essex David W.
Published: October 2019

Thiol-disulfide isomerization in thrombospondin may affect the function of this adhesive protein. Two assays were developed to analyze the determinants of thiol-disulfide exchange and to correlate this exchange with thrombospondin conformation. (1) A competitive immunoassay for the EDTA-conformation of thrombospondin was developed with monoclonal antibody D4.6. (2) The free thiol(s) in thrombospondin was labeled with [3H]N-ethylmaleimide (NEM) under various conditions (the presence or absence of calcium, temperature, and pH), and thrombin digests of the labeled protein were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Consistent with previous reports, thrombin digest fragments of 150, 120, 20, and 14 kD were observed, each with radioactivity under some condition, plus a 25-kD peptide that was not labeled. Sequence data for these fragments and comparisons of SDS-PAGE analyses under reducing and nonreducing conditions indicated that Cys974 was the free thiol. The appearance of thiol label in the 120-kD fragment was previously shown to be a consequence of thiol-disulfide exchange (J Biol Chem 265:17859,1990) and label was recovered in this peptide only under conditions (absence of calcium, 37°C and pH 8.4) that led to the appearance of the EDTA-conformation of thrombospondin. Additional evidence for the correlation of EDTA-conformation and thiol-disulfide exchange was the enhanced conversion of thrombospondin to its EDTA-conformation in the presence of protein disulfide isomerase and the inability of thrombospondin pretreated with NEM to attain the EDTA-conformation. Flow cytometry with antibody D4.6 revealed platelet-associated thrombospondin in the EDTA-conformation in the presence of calcium, suggesting that the EDTA-conformation is a physiological conformation that does not necessarily require EDTA.
Thymocyte Fas Expression Is Dysregulated in Myasthenia Gravis Patients With Anti-Acetylcholine Receptor Antibody
Authors: Moulian Nathalie , Bidault Jocelyne , Truffault Frédérique , Yamamoto Ana Maria , Levasseur Philippe , Berrih-Aknin Sonia
Published: October 2019

Myasthenia gravis (MG) is a human autoimmune disease mediated by anti-acetylcholine receptor (AChR) antibodies. The thymus is probably the site where the autoimmune response is triggered and maintained. Recent reports have linked various autoimmune disease with defective Fas expression. We thus analyzed Fas expression in thymocytes and peripheral blood lymphocytes (PBL) from MG patients. The proportion of a thymocyte subpopulation with strong Fas expression (Fashi) was markedly enhanced in MG patients with anti-AChR antibodies (P < .0003, compared with controls). In this group of patients, the proportion of CD4+Fashi and CD4+CD8+Fashi thymocytes were significantly increased (P < .002 for both subsets). Fashi thymocytes were enriched in activated cells and showed intermediate CD3 expression. They were preferentially Vβ5.1-expressing cells, previously shown to be enriched in potentially autoreactive cells. The proliferative response of thymocytes from MG patients to peptides from the AChR was abolished after depletion of Fashi cells. Fashi thymocytes were sensitive to an agonistic anti-Fas antibody. In peripheral blood, Fashi lymphocytes proportion was not significantly modified in MG patients whatever their anti-AChR antibody titer, compared with controls. Altogether, these results indicate that Fashi thymocytes, which accumulate in MG patients with anti-AChR antibodies, could be involved in the autoimmune response that targets the AChR.
Abnormal Erythrocyte Endothelial Adherence in Hereditary Stomatocytosis
Authors: Smith Brian D. , Segel George B.
Published: October 2019

Hereditary stomatocytosis is a red cell membrane protein disorder, which results in hemolytic anemia. Some patients with hereditary stomatocytosis experience dyspnea, chest pain, and abdominal pain, particularly after splenectomy. These symptoms may represent vaso-occlusion secondary to adherence of an abnormal erythrocyte membrane to vascular endothelium. We studied three members of a family with varying clinical expression of hereditary stomatocytosis. Adherence of red cells to endothelium was quantified by measuring the shear force required to separate individual cells from endothelial monolayers using a micropipette technique. Two patients with symptoms of in situ thromboses had a higher percentage of adherent cells compared with their asymptomatic sibling and normal controls. Correlation between this in vitro phenomenon and the clinical course suggests that flow abnormalities in the microcirculation attributable to erythrocyte endothelial adherence may play an important pathogenetic role in the illness. When the proportion of adherent red cells was reduced by a chronic transfusion program in one patient and pentoxifyllin therapy in another, the vaso-occlusive complications were eliminated.
The Effect of ACTH in Periodic (Cyclic) Neutropenia
Authors: CHAIKEN BERNARD H.
Published: October 2019

A case of periodic neutropenia of twenty years’ duration is presented. ACTH was administered intravenously in an attempt to abort or prevent the neutropenic cycles. No effect in raising the total leukocyte or neutrophil count was noted, and the neutropenic cycles could not be prevented by the ACTH. A striking absence of fever, stomatitis, or other secondary manifestations occurred during the neutropenic periods treated with ACTH. The background for treating such conditions with hormones is discussed.
Blood Coagulation and the L. E. Cell Phenomenon
Authors: LEE STANLEY L. , SCHWARTZ LAWRENCE I. , PARISER SANFORD
Published: October 2019

A factor derived from blood platelets has been shown to be necessary for the activation of the L. E. cell factor. This L. E. cell cofactor may be identical with the platelet thromboplastic factor. The relation of these findings to occurrence of the L. E. cell phenomenon in vivo and to the occurrence of a coagulation defect in systemic L. E. has been discussed.
The Significance of Proplasmocytes and Atypical Plasma Cells in the Benign Lymphocytic Meningitis Syndrome
Authors: WOLF PETER
Published: October 2019

A nontuberculous, lymphocytic meningitis with a low C.S.F. sugar level may be confused with a tuberculous meningitis. Additional diagnostic help may be obtained from examination of the blood. The blood pictures of twelve patients with tuberculous meningitis were compared with those from ten patients with a nontuberculous, nonbacterial lymphocytic meningitis, four of whom had low C.S.F. sugar levels. These ten patients appeared clinically to be suffering from the same disease and only the serum complement fixation test for lymphocytic choriomeningitis showed them to be a heterogeneous group.

The main differences in the blood were (1) a uniformly low total white cell count in the benign meningitis group as opposed to a widely varying total white cell count in tuberculous meningitis; (2) a lower percentage count of polymorphs in the benign lymphocytic meningitis group; (3) the presence of atypical plasma cells in nine of the ten patients in the benign, lymphocytic meningitis group. Two of the patients with benign meningitis, who had repeatedly given negative Paul-Bunnell tests showed a blood picture indistinguishable from that of glandular fever.

Although these atypical plasma cells are not specific for lymphocytic choriomeningitis or for virus diseases in general, they were found in no case of tuberculous meningitis. The presence of these cells is therefore of considerable aid in excluding a diagnosis of tuberculous meningitis.
An Unidentified Reticuloendothelial Cell in Bone Marrow and Spleen
Authors: SAWITSKY ARTHUR , HYMAN GEORGE A. , HYMAN JULIAN B.
Published: October 2019

1. A cell is described in the bone marrow aspirates of two patients which has not previously been fully described. This is a large reticuloendothelial-like cell filled with blue-staining granules. This cell has been found in successive bone marrow films aspirated from sternum and iliac crest.

2. These cells were also found in a splenic aspiration from one of the patients and were compared to similar cells from a case summarized by Möschlin.

3. The case reports show striking clinical similarities in the two patients, i.e. hepatosplenomegaly in relatively symptom-free individuals. This suggests that a common syndrome is present.

4. Differential histochemical studies of the granular material in the cytoplasm are reported.

5. The significance and function of these unusual cells in the marrow is unknown.
Metabolic Interrelations between Gastric Intrinsic Hematopoietic Factor and Vitamin B12
Authors: GLASS GEORGE B. JERZY , LILLICK LOIS C. , BOYD LINN J. , GUNNIS CARRIE , CORTI LUCY GORMAN
Published: October 2019

The concentration of vitamin B12 in blood serum and its excretion in the urine were assayed by the E. coli mutant technic in two patients after total gastrectomy and by use of the more sensitive Euglena gracilis method in four cases of pernicious anemia in relapse and 1 case of well controlled nutritional macrocytic anemia.

Following parenteral administration of 100 µg. vitamin B12 intramuscularly to two patients with total gastrectomy, much higher blood levels were observed within the first few hours than those reported by others under similar circumstances in normals.

Following oral administration of vitamin B12 together with a potent intrinsic factor concentrate from hog stomach, a rise in blood levels of vitamin B12 was observed in patients with pernicious anemia.

The normal serum levels of vitamin B12 could not be maintained, however, for the entire duration of the oral treatment, in spite of the apparent hematologic and clinical remission. The urinary output of vitamin B12 was in all instances less than 0.5 per cent of the total dose of vitamin B12 ingested during the experimental period, in spite of an optimal or suboptimal hematopoietic response obtained. A similar situation existed in patients with total gastrectomy, as well as in the patient with controlled macrocytic nutritional anemia, when vitamin B12 was ingested alone or with intrinsic factor concentrate.

The intensity of the obtained hematopoietic response in patients with pernicious anemia indicates the absorption of much larger amounts of vitamin B12 from the intestine than is indicated by the total urinary output of vitamin B12; therefore the anchorage and retention of a large part of vitamin B12 in the storage depots of the body after its absorption from the intestine must occur.

A sharp increase in the urinary output of vitamin B12 was observed during or immediately following the hematopoietic response in all patients with pernicious anemia treated orally with vitamin B12 and intrinsic factor concentrate, i.e., coincident with or immediately following the reticulocyte peak in blood. Similar observation was reported by the authors in the previous series of investigations. Of many possible interpretations of this finding, the most plausible appears to be the metabolic release of free vitamin B12 from its complex binding in the body during hyperactivity of the hematopoietic organs and its escape through the kidneys.
Number and Distribution of Human Hemic Cells
Authors: OSGOOD EDWIN E.
Published: October 2019

1. Methods are described which make possible the calculation of the number and distribution of the hemic cells of the body.

2. The numbers of these cells in the various blood-forming organs, in the blood outside these areas, and in the total body are summarized in table 3, together with the ratios of the number in the blood to the number outside the hematopoietic organs for the standard 70 Kg. human male. Dividing these figures by 70 will give the values per Kg.

3. By similar methods the values for other age and sex groups in man and for other species may be calculated.

4. Leukocytes apparently are not primarily blood cells but are merely en route in the blood to the defense of body tissue generally. Only 1/40 to 1/400 of the total leukocytes outside the hematopoietic organs are present in the blood stream at any one time.

5. These somewhat surprising figures account for many previously made observations which were hitherto hard to explain.
Radioactive Sodium Chromate for the Study of Survival of Red Blood Cells
Authors: WEINSTEIN IRWIN M. , SPURLING CARROLL L. , KLEIN HERMAN , NECHELES THOMAS F.
Published: October 2019

Cr51 erythrocyte survival times are reported in a group of patients with a variety of abnormal hemoglobin syndromes. Marked decreases in survival time are demonstrated in pure sickle cell anemia. Shortened survival times are reported in one case each of hemoglobin C disease and sickle cell-hemoglobin C disease with compensated hemolysis. Normal survival times are reported in sickle cell trait and hemoglobin C trait.

Red cell life span as measured by the Cr51 technic agrees well with most published reports of survival times in these disorders in cases performed with the Ashby technic. The Cr51 method appears to be as useful in measuring the survival of erythrocytes containing abnormal hemoglobins as it has been shown to be in other hemolytic disorders as well as in normals. Its decided advantages are its simplicity, adaptability, and reliability.
Intraerythrocytic Crystals in a White Patient with Hemoglobin C in the Absence of Other Types of Hemoglobin
Authors: DIGGS L. W. , KRAUS A. P. , MORRISON D. B. , RUDNICKI R. P. T.
Published: October 2019

Tetragonal crystals of hemoglobin were observed in approximately 2 per cent of erythrocytes of a sixteen-year-old white male of Italian parentage, who had hemoglobin C in the absence of A, S, and F.

The clinical and hematological features of the patient described are similar to those with "pure" hemoglobin C disease reported by others. The essential features are: chronic anemia of a mild degree, congestive splenomegaly, evidences of increased red cell destruction, hyperplastic marrow with increased numbers of immature erythrocytes in the peripheral blood, target cells and increased erythrocyte resistance to hyptonic saline solutions.

Electrophoretic analysis of hemolysates of both parents, one brother and a maternal uncle revealed hemoglobin A and C. The hemolysate of one sister revealed only hemoglobin A. None of the relatives examined had hemoglobin S or F and none showed hematologic abnormalities.
The Effect of Pyridoxine Deficiency Induced by Desoxypyridoxine on Acute Lymphatic Leukemia of Adults
Authors: WEIR DAVID R. , MORNINGSTAR WILLIAM A.
Published: October 2019

1. Pyridoxine deficiency can be induced in human beings by giving large doses of desoxypyridoxine while maintaining the patients on a normal diet.

2. In four cases of acute lymphoid disease of adults some modification of the disease appears to have been accomplished by the induction of pyridoxine deficiency. In one case a brief clinical remission occurred which seemed almost complete.

3. Possible other effects of the deficiency were adrenal cortical degeneration and deposition of crystals in the convoluted tubules of the kidneys. The adrenal changes could well be due to postmortem degeneration.

4. Other toxic effects of the desoxypyridoxine therapy were not observed.

5. Further trials of this type of therapy appear to be justified.
Symposium: What Is Hemophilia?: On the Nature and Diagnosis of Hemophilia
Authors: QUICK ARMAND J.
Published: October 2019

The primary cause of the defective hemostasis in hemophilia is the lack of thromboplastinogen in the blood. This anomaly is congenital and hereditary. The mechanism whereby the concentration of thromboplastinogen is depressed in the blood is not known and no satisfactory explanation can be given why the defect manifests itself only in the male. The degree of the hypothromboplastinogenemia determines the intrinsic severity of the disease. The diagnosis is based on establishing the existence of a bleeding tendency from birth, in demonstrating a deficiency of thromboplastinogen in the blood, and in finding a typical hereditary pattern.
The Effects of Tyrosinase on Certain Components of the Blood Clotting System
Authors: KRUGELIS EDITH J. , SIZER IRWIN W.
Published: October 2019

The clotting time of human and rat plasma was found to increase markedly after exposure to mushroom tyrosinase. The main, but not exclusive, effect of tyrosinase on the clotting system seems to be on fibrinogen, since the inactivating effect of tyrosinase can be greatly reduced by the subsequent addition of excess fibrinogen just before the clotting time of the plasma is measured. No evidence was found for any major effect of tyrosinase on any of the other components of plasma considered with the possible exception of prothrombin. Purified prothrombin is susceptible while prothrombin in plasma or serum is refractory to tyrosinase.
Leukemia in Atomic Bomb Survivors
Authors: MOLONEY WILLIAM C. , LANGE ROBERT D.
Published: October 2019

1. The ten cases of leukemia occurring among 3480 atomic bomb survivors in this study present convincing evidence of the high incidence of leukemia in atomic bomb survivors.

2. In all but one case blood studies were carried out prior to the development of pathognomonic features of leukemia. One case of reticulum cell leukemia was encountered on routine examination in a preclinical stage but with primitive or blast cells in the peripheral blood smears.

3. Of the ten cases, seven were acute or subacute leukemia and three were chronic myelogenous leukemia.

4. Observations on the acute cases of leukemia were of limited value because of the long interval in most cases between routine blood studies and the development of leukemia. However, in one case, examination ten weeks before clinical onset revealed no hematologic or physical findings suggestive of leukemia.

5. In contrast to acute leukemia, the long preclinical stage in two cases of chronic myelogenous leukemia allowed repeated hematologic and biochemical studies which were of considerable interest.

6. In these cases the long latent period and prolonged preclinical course were similar to those reported in cases of chronic myelogenous leukemia occurring spontaneously and following exposure to x-ray and other forms of radioactivity.

7. In the preclinical stage, disturbances in the red cells and platelets as well as leukocytosis with immature neutrophilic granulocytes were noted. The consistent presence of increased numbers of basophils, many of them atypical, was an outstanding feature of the peripheral blood smears in early myelogenous leukemia.

8. In two cases of preclinical myelogenous leukemia, studies on the separated leukocytes of the peripheral blood gave extremely low values for alkaline phosphatase.

9. Certain features of chronic myelogenous leukemia following irradiation suggest that the disorder may be due to a loss of growth-promoting or regulating factors.
Cytochemical Changes in Lymph Nodes and Spleen of Rats after Total Body X-Radiation
Authors: ACKERMAN G. ADOLPH , BELLIOS NICHOLAS C. , KNOUFF RALPH A. , FRAJOLA WALTER J.
Published: October 2019

Marked cytochemical changes were observed in the cells of the lymph nodes and spleen of one hundred and twenty-seven albino rats observed over periods of two hours to thirty-two days following a single dose of 600 r total body x-radiations. The intensity of the cytochemical reactions for pentosenucleoprotein, protein-bound sulfhydryl groups, and alkaline phosphatase decreased within two hours after x-radiation. This decrease in the cytochemical reactions occurred very rapidly and preceded any discernible morphologic alteration. Lymphocytes that were extremely sensitive to x-ray exhibited in most instances irreversible cytochemical changes. The cells that survived the lethal effects of x-radiations exhibited an increase in intensity of the reactions for pentosenucleoprotein, protein-bound sulfhydryl groups, and alkaline phosphatase between twelve hours and four days following irradiation. The reactions for desoxypentosenucleoprotein and mucopolysaccharides were unaltered in the cells of the lymph nodes and spleen following x-radiation.
Centrifugal Subfractionation of Polymorphonuclear Leukocytes, Lymphocytes and Erythrocytes
Authors: AGRANOFF BERNARD W. , VALLEE BERT L. , WAUGH DAVID F.
Published: October 2019

The separations of polymorphonuclear leukocytes from lymphocytes and erythrocytes has been accomplished by employing a method which takes advantage of their density differentials. During a single centrifugations the lymphocytes accumulate at a plasma-albumin interface (albumin I, density = 1.065 to 1.085), the polymorphonuclear leukocytes accumulate at an albumin I-albumin II interface (albumin II, density = 1.094) and the erythrocytes pass to the base of the Wintrobe tube. The yields are over 90 per cent pure cell types. All layers are isosmotic with blood.
IRC011, a New Synthetic Chelator With Selective Interaction With Catabolic Red Blood Cell Iron: Evaluation in Hypertransfused Rats With Hepatocellular and Reticuloendothelial Radioiron Probes and in Iron-Loaded Rat Heart Cells in Culture
Authors: Rivkin Gurion , Link Gabriela , Simhon Elliot , Cyjon Rosa L. , Klein Joseph Y. , Hershko Chaim
Published: October 2019

A major consideration in the selection of new and improved iron chelators for clinical use is preferential interaction with the most toxic iron compartment. We describe the biologic properties of a new synthetic hexadentate iron chelator (IRC011) that is a substituted polyaza compound. Unlike deferoxamine (DF ), the polyaza structure of IRC011 does not contain any readily hydrolyzable covalent bonds and is anticipated to resist in vivo biotransformation. In the present studies, the ability of IRC011 to remove radioiron from iron-loaded heart cells in vitro was similar to DF, with a decrease to 20.0 ± 0.4% and 19.7 ± 0.5% of initial values after 24 hours of incubation with 0.3 mmol/L of DF or IRC011, respectively. The in vivo interaction of IRC011 with specific iron stores was studied in hypertransfused rats using selective labeling of reticuloendothelial (RE) iron stores with 59Fe-heat-denatured red blood cells (DRBCs) and of hepatocellular stores with 59Fe-ferritin. The pattern of radioiron excretion with IRC011 was quite different from that with DF. Although with both compounds, hepatocellular iron excretion was through the bile, whereas RE iron excretion was mainly in the urine, the magnitude of these effects was quite different. After the administration of a single parenteral dose of 200 mg/kg representing a 53% higher iron-binding capacity for IRC011 compared with DF, 48-hour urinary excretion of RE iron with IRC011 was 22.8% ± 1.1% (% of total body 59Fe), but only 6.0% ± 3.6% with DF. By contrast, the corresponding biliary excretion of hepatocellular radioiron was 14.2% ± 3.2% with DF, but only 0.7% ± 0.3% with IRC011. Thus, the new iron chelator IRC011 is distinguished from DF by the following features: (1) a higher affinity to Fe(III), (2) anticipated resistance to in vivo catabolism, (3) preferential interaction with RE iron derived from RBC breakdown, and (4) selective renal excretion. Because RBC breakdown is the most likely source of the toxic nontranferrin plasma iron, IRC011 may be a useful iron chelator for protecting vital organs from peroxidative damage.
Exclusion of Three Candidate Genes as Determinants of Congenital Dyserythropoietic Anemia Type II (CDA-II)
Authors: Iolascon Achille , Miraglia del Giudice Emanuele , Perrotta Silverio , Granatiero Matteo , Zelante Leopoldo , Gasparini Paolo
Published: October 2019

Congenital dyserythropoietic anemia type II (CDA-II) is the most common form of inherited dyserythropoiesis. Previous studies have shown that the anion transporter (band 3) is narrower and it migrates faster on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); this aspect was related to insufficient glycosylation. Biochemical data support the hypothesis that this disease is due to a deficiency of N-acetylglucosaminyltransferase II (GnT II) or α-Mannosidase II (α-Man II), which represent the key to glycosylation. In addition, a third candidate gene is α-Man IIx, which shows a strong homology with α-Man II. The knowledge of the chromosomal localization of these putative genes allowed us to perform a linkage study using three sets of microsatellite markers flanking the candidate genes. Six families with two or more affected children were enrolled in this study. The data obtained exclude linkage to all three candidate genes. In consideration of the biochemical data (reduction of enzymatic activity) of the same enzymes, our results suggest the hypothesis that a defect in an unknown transcriptional factor is involved in CDA-II.
Phenotypic and Functional Analysis of Fas (CD95) Expression in Primary Central Nervous System Lymphoma of Patients With Acquired Immunodeficiency Syndrome
Authors: Baiocchi Robert A. , Khatri Vijay P. , Lindemann Matthew J. , Ross Mary E. , Papoff Giuliana , Caprio Anthony J. , Caprio Thomas V. , Fenstermaker Robert , Ruberti Giovina , Bernstein Zale P. , Caligiuri Michael A.
Published: October 2019

The poor prognosis associated with patients afflicted with the acquired immunodeficiency syndrome and primary central nervous system lymphoma (AIDS-PCNSL) is due in part to the intrinsic resistance of this Epstein-Barr virus (EBV)-associated tumor to conventional antineoplastic therapy. Fas (CD95) is a transmembrane protein receptor that transmits an intracellular signal leading to rapid programmed cell death following ligation with its natural ligand or anti-Fas antibodies. Fas expression and function were assessed in AIDS-PCNSL biopsy samples and in EBV+ human B-cell tumors that spontaneously developed in severe combined immune deficient (SCID) mice engrafted with human lymphocytes (hu-PBL-SCID mice). All tumors samples showed high-density surface expression of Fas by flow cytometry or immunohistochemical staining. Cells from two AIDS-PCNSL biopsy samples that did not express pan B-cell markers did not express Fas antigen. All tumors examined were susceptible to Fas-mediated apoptosis, as measured by standard assays for endonucleolytic cleavage of DNA. The response to Fas-mediated apoptosis was dependent on log-fold increases in the concentration of immobilized anti-Fas antibody, but could also be induced with a mobilized anti-Fas antibody. No evidence for intrinsic resistance to Fas-mediated apoptosis (ie, secreted or truncated forms of Fas) could be shown. Radiation-induced apoptosis of neoplastic EBV+ B cells was enhanced by activation of Fas, and prolonged exposure to interleukin-2 increased both Fas expression and Fas-induced apoptosis. As the normal brain parenchyma appears to have either low-density or absent expression of Fas, and antineoplastic therapy can be selectively delivered to the CNS with little systemic toxicity, local delivery of Fas-activating molecules could prove to be a useful component in the multimodal treatment of AIDS-PCNSL.
Infection of Human Marrow Stroma by Human Immunodeficiency Virus-1 (HIV-1) Is Both Required and Sufficient for HIV-1–Induced Hematopoietic Suppression In Vitro: Demonstration by Gene Modification of Primary Human Stroma
Authors: Bahner Ingrid , Kearns Karen , Coutinho Sunita , Leonard Earl H. , Kohn Donald B.
Published: October 2019

Patients with human immunodeficiency virus-1 (HIV-1) infection often present with bone marrow (BM) failure that may affect all hematopoietic lineages. It is presently unclear whether this failure reflects a direct viral impairment of the CD34+ hematopoietic progenitor cells or whether the virus affects the BM microenvironment. To study the effects of HIV-1 on the BM microenvironment, we examined the stromal cell monolayers in long-term BM culture (LTBMC), which are the in vitro equivalent of the hematopoietic microenvironment. We assessed the hematopoietic support function (HSF ) of human stromal layers by determining the cellular proliferation and colony-forming ability of hematopoietic progenitors from BM cells grown on the stromal layers. We show that the HSF is reduced by in vitro infection of the human stromal cell layer by a monocytotropic isolate of HIV-1 (JR-FL). There is no loss of HSF when the stromal cell layer is resistant to HIV-1 replication, either using murine stromal cell layers that are innately resistant to HIV-1 infection or using human stromal cells genetically modified to express a gene that inhibits HIV-1 replication (an RRE decoy). Decreased HSF was seen using either human or murine hematopoietic cells, if the stromal cells were human cells that were susceptible to HIV-1 infection. These in vitro studies implicate HIV-1 replication in the stroma as the essential component causing decreased hematopoietic cell production in HIV-1 infection.
Loss of Tolerance to Exogenous and Endogenous Factor VIII in a Mild Hemophilia A Patient With an Arg593 to Cys Mutation
Authors: Thompson Arthur R. , Murphy Michael E.P. , Liu MiaoLiang , Saenko Evgueni L. , Healey John F. , Lollar Pete , Scandella Dorothea
Published: October 2019

A 42-year-old patient with mild hemophilia A developed spontaneous muscle hematomas 1 month after intense therapy with factor VIII concentrates. Factor VIII clotting activity was less than 1% and his factor VIII inhibitor was 10 Bethesda units (BU)/mL. The titer peaked at 128 BU despite daily infusions of factor VIII; 1 year later, the titer was 13 BU with no spontaneous bleeding for 4 months. The plasma inhibitor was 95% neutralized by factor VIII A2 domain but less than 15% neutralized by light-chain or C2 domain. His inhibitor did not cross-react with porcine factor VIII and was at least 10-fold less reactive to a series of hybrid factor VIII proteins in which human residues 484-508 are replaced by the homologous porcine sequence (Healey et al, J Biol Chem 270:14505, 1995). The inhibitor patient's DNA encoding his A2 domain and flanking sequences showed a C-T transition predicting Arg593 to Cys. Thirteen patients from 5 unrelated families with Cys593 have not developed inhibitors. Factor VIII clotting activity from one of them was inhibited similarly to diluted normal plasma by inhibitor patient plasma. In an homologous structure, ceruloplasmin (Zaitseva et al, J Biol Inorgan Chem 1:15,1996), the residue equivalent to Arg593, is in a loop distinct from residues 484-508. On solution phase immunoprecipitation with labeled factor VIII fragments, A2, light chain, and C2 domains bound. In contrast to typical immune responses to factor VIII in patients with severe hemophilia A, this patient's inhibitor was almost entirely reactive with common epitopes within the A2 domain whereas by more sensitive immunoprecipitation testing antibodies to light chain epitopes were also present. Accordingly, immune responsiveness to exogenous factor VIII (antigen burden) appears to be more critical than his endogenous, hemophilic factor VIII to his developing high-titer anti–factor VIII antibodies and loss of tolerance to both native and hemophilic factor VIII proteins.
Altered Tumor Growth and Metastasis of a T-Cell Lymphoma in Timp-1 Transgenic Mice
Authors: Krüger Achim , Fata Jimmie E. , Khokha Rama
Published: October 2019

The concept of tumor suppression by tissue inhibitor of metalloproteinases (TIMPs) has evolved primarily from studies of genetically modulated tumor cells. The next step is to focus on the host and assess the protective potential of host TIMP-1 on primary tumor growth and metastasis. We generated two transgenic mouse lines with altered Timp-1 expression in skin and liver: one overexpressed Timp-1 (Timp-1high), and the other had antisense RNA–mediated Timp-1 reduction (Timp-1low). ESbL-lacZ T-lymphoma cells provided the tumor challenge, as they form primary tumors upon intradermal injection with spontaneous metastasis to liver. Metastases were examined in X-Gal–stained whole-organ mounts. Timp-1 overexpression inhibited intradermal tumor growth and spontaneous metastasis, leading to prolonged survival of the mice. The opposite effects occurred in Timp-1low mice, leading to shorter host survival. Experimental metastasis assays showed that Timp-1–compromised livers in Timp-1low mice showed at least a doubling of metastatic foci and numerous additional micrometastases, indicative of increased host susceptibility. However, Timp-1high mouse livers showed an unaltered metastatic load in the experimental metastasis assay. In conclusion, these data demonstrate that Timp-1 levels within a tissue predetermine the development and progression of T-cell lymphoma.
Growth of Human T-Cell Lineage Acute Leukemia in Severe Combined Immunodeficiency (SCID) Mice and Non-obese Diabetic SCID Mice
Authors: Steele Jeremy P.C. , Clutterbuck Robyn D. , Powles Ray L. , Mitchell Paul L.R. , Horton Clive , Morilla Ricardo , Catovsky Daniel , Millar John L.
Published: October 2019

Primary leukemic cells from patients with acute lymphoblastic leukemia (ALL) can be injected intravenously into mice with severe combined immunodeficiency (SCID) to create a model of human leukemia. Leukemic cells disseminate to murine tissues in a clinicopathologic pattern similar to that seen in humans. Thus far, reports of engraftment of lymphoid leukemia in SCID mice have mainly been from patients with B-cell lineage ALL, for which engraftment occurs more frequently with cells from high-risk patients. There are few data on the engraftment of T-cell lineage ALL in SCID mice. Leukemic cells from 19 patients (16 adult and three pediatric) with T-cell lineage ALL were injected into SCID mice, with overt engraftment of 12 cases (63%). Engraftment of leukemia in SCID mice was associated with earlier death due to leukemia of the patient donors (P < .01, log-rank test). The recently developed non-obese diabetic (NOD)/SCID mouse may expand the uses of the SCID model. Cells from the seven patients with T-cell lineage ALL that failed to cause leukemia in SCID mice were injected into NOD/SCID mice. Overt leukemia engraftment was observed in all seven cases. Thus, growth of human T-cell lineage ALL cells in SCID mice was associated with a high-risk patient group. However, this association was not observed when NOD/SCID mice were used, suggesting that this model would no longer predict patients likely to die early of leukemia, but may provide a more realistic system for studying the biology and treatment of the disease.