bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Hydroxyurea on Elevated Pulmonary Artery Pressures in Children with Sickle Cell Disease
Authors: Pashankar Farzana , Manwani Deepa , Lee Margaret , Green Nancy S.
Published: October 2019

Abstract 4841

Background:

Pulmonary hypertension (PHT) is a significant complication of sickle cell disease. Studies in children report a 16–30% prevalence of elevated pulmonary artery pressures, as estimated by measurement of tricuspid regurgitant jet velocity (TRV) on echocardiography. The pathogenesis of elevated pulmonary artery pressures is multifactorial, with hemolysis induced endothelial dysfunction playing a major role. More recent studies highlight the role of inflammation in the pathogenesis. Hydroxyurea is a well established treatment for sickle cell disease. It acts primarily by induction of fetal hemoglobin, thereby reducing hemolysis, with possible additional effects on vascular and endothelial function. The aim of this study was to determine if early detection and treatment with hydroxyurea will decrease elevated pulmonary artery pressures in children with sickle cell disease.

Methods:

The study was conducted at 3 centers. Children with Hb SS and Hb Sb0 thalassemia between the ages of 5–21 years, with a screening echocardiogram showing a TRV ≥ 2.5 m/sec were identified. An echocardiogram was repeated to confirm elevated TRV. Subjects with persistent elevation of TRV ≥ 2.5 m/sec on repeat echocardiogram, were consented and started on hydroxyurea at 20 mg/kg/d with escalation to a maximum tolerated dose or a total daily dose of 30 mg/kg/d. Laboratory data and echocardiograms were repeated at 6 and 12 months to measure effect of hydroxyurea on TRV. Additionally blood and urine samples were also collected pre treatment, at 6 and 12 months post treatment for biomarker analysis, which will be performed later. Baseline and 6 month laboratory and echocardiogram data were compared using paired t test.

Results:

Twelve patients were enrolled. Mean age was 12.25 years (range 6–19 years) with a M:F ratio of 2:1. Average follow up is 11 months. Patients tolerated hydroxyurea well, and in 90% of patients the dose was escalated to 30 mg/kg/d. 1 patient achieved MTD at 20 mg/kg/d. Two patients went off study at 4 and 5 months respectively.

As shown in Table 1, six months after starting hydroxyurea there was a significant increase in mean oxygen saturation, hemoglobin, mean corpuscular volume and fetal hemoglobin. There was a significant decrease in mean reticulocyte count, LDH and white blood cell count. There was no significant change in TRV six months after treatment with hydroxyurea.

Conclusion:

Hydroxyurea significantly decreased measures of hemolysis in children with sickle cell disease. Six months after treatment with hydroxyurea, there was no significant change in estimated pulmonary artery pressures measured on echocardiography. The study is ongoing to see if hydroxyurea affects pulmonary artery pressures with a longer duration of treatment.

Disclosures:

No relevant conflicts of interest to declare.
Cytotoxic Effects of Curcumin At Various Concentrations and Role of Curcumin on Lipid Peroxidation and Activities of Antioxidant Enzymes of the Rat Peripheral Blood Lymphocytes
Authors: Desai Nisarg , Finosh G T , Panicker N G , Ramachandran R , Varghese Alex
Published: October 2019

Abstract 4933

Background:

Curcumin (diferuloylmethane) is an active ingredient of turmeric and has been suggested to have antiproliferative, pro-oxidant and cytotoxic effects (Das R, Apoptosis 2008). Curcumin has also been shown to have antioxidant effects (Rastogi M, Free Radic Res 2008). Our aim was to assess the cytotoxic, pro-oxidant and antioxidant effects of curcumin on rat peripheral blood lymphocytes (RPL) in culture.

Methods:

Different concentrations of curcumin (0.325 μM, 0.65 μM, 1.3 μM) were analyzed for cytotoxic effects on rat peripheral lymphocytes. The cytotoxic effect of curcumin was assessed by measuring cell viability using Trypan blue exclusion assay. DNA damage by curcumin (at 0.65 μM) was confirmed by the COMET assay. The effect of curcumin (at 0.65 μM) on oxidation was assessed by measuring the activity of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and lipid peroxidation (LP) by spectrophotometry. RPLs were cultured for 72 hr in vitro. The pro-oxidant effect of curcumin was determined by comparing LP induction in control (PBS treated) vs curcumin treated RPLs. Parameters were measured at 0, 24 and 48 hrs of curcumin exposure. Reactive oxygen species (ROS) generated by treatment with FeCl3 (10 mM) and H2O2 (5 mM) (Fe/H) were used to demonstrate the ROS scavenging effect of curcumin. Lymphocyte cultures were treated with either Fe/H alone or in combination with Curcumin. Oxidation levels were assessed as described above at 0, 24, 48 hrs following exposure.

Results:

Cytotoxic effects were seen at different curcumin concentrations (Table 1). Viability of RPLs were 42% at 0.64 μM concentration vs 72% at 0.325 μM. 30% decrease in cell viability of RPLs were seen at 0.65 μM concentration compared to 0.325 μM. Hence, 0.65 μM concentration was used for further experiments. There was a significant increase in DNA damage in curcumin exposed lymphocytes at 48hr (table 2). There was a significant increase in LP in curcumin exposed RPLs compared to control lymphocytes. By contrast, there was no significant change in LP values in presence of Fe/H at all time points compared to control. There was a significant decrease in LP when RPL cultures were incubated with curcumin for 48 hr compared to 0 hr of incubation. There was significant increase in activities of SOD, catalase and GPx in presence of curcumin and Fe/H (table 3 and 4).

Conclusions:

Dose dependent cytotoxic and DNA damage effects of curcumin are seen in RPLs. Treatment with curcumin alone increases lipid peroxidation, However in the presence of oxidative stress, curcumin may act as an antioxidant (ROS scavenging effect) by stimulating activities of antioxidant enzyme and preventing lipid peroxidation.

Disclosures:

No relevant conflicts of interest to declare.
Gemcitabine Is An Active Agent In Acquired Human Immunodeficency HIV Related Lymphomas
Authors: Jajeh Ahmad
Published: October 2019

Abstract 4951

Gemcitabine is a cell cycle phase specific cytotoxic agent. It's phosphorylated products inhibit ribonucleotide reductase an enzyme responsible for DNA synthesis. It also competes with the incorporation of deoxycytosine triphospate (dCTP) into the DNA strands. After Gemcitabine is incorporated into the DNA the growing DNA strands will stop. Gemcitabine has proven to be an active cytotoxic agent in the treatment of lymphomas, either as a single agent or in combination with platinum and dexamethsone. Gemcitabine efficacy has not been investigated in the treatment of HIV patients with Lymphomas. The purpose of this abstract is to show our experience with this agent in HIV related lymphomas. This summarize the experience with three patients. First patient is 42 years old white male with diffuse large B cell type Non-Hodgkin's Lymphoma involving the right psoas muscle and invading the pelvic nerve roarts causing intractable pain that has not been resonsive to CHOP (four cycles), the third patient is a 55 years old hispanic female with stage IV Hodgkin's Lymphoma (mixed cellularity) that has failed ABVD × 6 cycles and progressed in less than two months from completion of treatment. This patient presented with B symptoms and cachexia at relapse. The third patient is 50 years old black male with stage III Immunoblastic B Cell Lymphoma who failed EPOCH. All patients were on HAART medication and viral load were undetectable in two of these patient (2,3). The first patient received Gemcitabine alone. With two weekly doses of 1000mg/M2 the pain resolved and he stopped narcotics and was refered to other institution for autologus bone marrow transplant. The second patient received two cycles of Gemcitabine, Cisplatin and Decadron within one dose the B symptoms and the sever bony pain have disappeared completely. The third patient completed six cycles of treatment; each cycle is of twenty eight days duration; Gemcitabine 1000mg/M2 given IV on week one and two(day1) and Cisplatinum 80/M2 mg given IV on day one with decadron 40 mg po with each chemotherapy treatment. This patient has been in complete remisson and is alive after three years. In conlusion: Gemcitabine is an active agent that can be used safely in HIV/AIDS patients with lymphomas with good results.

Disclosures:

No relevant conflicts of interest to declare.
Ethnic Differences in the Incidence of Leukemia
Authors: MACMAHON BRIAN , KOLLER ERNEST K.
Published: October 2019

A series of 1636 patients with leukemia, representing the white residents of Brooklyn diagnosed in the period 1943-52, is compared with selected census data for the population. After standardization for age differences in the two populations, the incidences of leukemia in the native-born and foreign-born populations of the borough are 45.3 and 61.0 per million per annum, respectively. The data suggest that a high incidence of leukemia in the Russian-born population accounts for the higher rate in the foreign-born group.

Using affiliation of cemetery of burial as an index of religion, a series of 1368 deaths from leukemia is compared with a systematic one in 200 samples of all deaths in the same area. Leukemia is recorded as cause of death twice as frequently among Jews, as among others. This relationship is seen in both native-born and foreign-born groups, in males and females, at all ages and in all the common pathologic varieties of leukemia. No difference in leukemia incidence is seen between predominantly Catholic and predominantly Protestant groups.

Evidence from United States vital statistics is used to show that the commonly observed difference between whites and Negroes imi leukemia death rates is closely associated with social differences between the two groups. Little or no difference in leukemia death rate exists between these two groups in Brooklyn, when allowance is made for the apparently high rate in the white Jewish population.
Immunologic Features of Erythrocyte Sensitization
Authors: VAUGHAN JOHN H. , WALLER MARION V.
Published: October 2019

It has been shown that red cells sensitized with anti-Rh antibody are agglutinated by anti-γ-globulin sera, regardless of the "order" of Rh antibody activity. Red cells sensitized with anti-Lewis antibody, on the other hand, are agglutinated by anti-sera to non-γ-globulin substances. Of six other blood group antibodies studied, five provided γ-globulin red cell coating materials and one, an anti-Kidd, a non-γ-globulin material. The participation of complement, or complement-like, substances was shown in some instances.

Various pitfalls in the technics employed in absorbing Coombs sera so as to render them immunochemically specific are discussed.
The Behavior, as Regards Shape and Volume, of Human Red Cell Ghosts in Fresh and in Stored Blood
Authors: PONDER ERIC , BARRETO DELIA
Published: October 2019

(1) When human red cells are hemolyzed in very hypotonic media (NaCl of a tonicity of 0.167) and when the tonicity is restored, by adding appropriate amounts of NaCl, to tonicities such as 0.3, 0.5, 1.0, and 1.7, the mean volume of the ghosts appears to be linear with the reciprocal of the tonicity. This might lead one to conclude that the ghosts are osmometers and that their volume is governed by simple osmotic considerations such as those expressed by a modified van’t Hoff-Mariotte law. Examination of the shapes and volumes of individual ghosts by a technic which combines phase optics, electronic flash (exposure time 0.001 second) and photography shows that three distinct populations of ghost coexist in any tonicity. These are spherical ghosts with a mean volume of 150 µ3, discoidal biconcave ghosts with a mean volume of 85 µ3, and crenated ghosts with a smaller volume which can be calculated. The most likely reason for this complexity is that the shape and volume of the ghost depends partly on its structure, and not altogether on the tonicity of the surrounding medium. Simple osmotic laws have no real application to systems of this kind.

(2) The changes in ghost volume and shape, as they depend on the duration of storage of the blood, at 4 C, from which the ghosts are prepared, and as they reflect changes in ghost structure, can be expressed simply. Crenated and discoidal ghosts certainly have some of the elements of red cell structure; the spherical ghost, which soon fragments and gives rise to myelin forms, may also retain some of the original elements of structure, but the fragment and the myelin form have certainly lost them. The latter objects are so small and light that they are not thrown down into the ghost column in the hematocrit tube, and so, as ghost structure disappears with increasing time of storage of the red cells from which they are prepared, a discrepancy appears between the volume of the ghost column as measured by the hematocrit and the volume which one would expect. This discrepancy can be used as a measure of the extent to which ghost structure is lost, and there comes a time, as the duration of red cell storage is increased, when the ghosts prepared from these red cells begin to be replaced by breakdown products such as myelin forms, etc.

(3) The less the efficiency of the conditions of red cell preservation, the shorter is this time. In human blood rendered incoagulable with heparin, structural breakdown in the preceding sense and measured by a simple expression which changes sign when the loss of structure has reached a certain point, occurs after about 26 days. In human blood preserved in ACD, structural breakdown measured in an identical manner occurs after about 55 to 60 days. In human blood preserved in ACD-inosine, structural breakdown does not occur until about 75 to 80 days.

These results are based on a large amount of preliminary work of an exploratory nature and then on three runs with heparin, five runs with ACD and five runs with ACD-Inosine.
Studies of Plasma Erythropoietic Factor in Anemic Human Patients
Authors: PRENTICE T. C. , MIRAND E. A.
Published: October 2019

Nine patients with anemia of varied origin have been studied. The relationship between erythron response and the content of erythropoietic factor in the plasma has been observed in these patients. Wide discrepancies between these variables were seen to exist. Further investigation of this aspect of the problem is needed.
The Effect of Irradiation on the Plasma Erythropoietic Stimulating Factor
Authors: LINMAN JAMES W. , BETHELL FRANK H. , TASCOTT HELENA K.
Published: October 2019

1) Protein-free plasma extracts from rabbits made anemic by the administration of phenylhydrazine immediately following total body x-irradiation are capable of stimulating erythropoiesis in the normal rat as demonstrated by erythrocytosis, reticulocytosis, and increased marrow erythropoietic activity.

2) Plasma extracts from rabbits made anemic by total body x-irradiation alone contain an erythropoietic stimulating factor.

3) These data would indicate that the stimulating factor is not produced by hemopoietic or other radiosensitive tissue and its formation is not dependent upon a regenerative marrow.
Combinations of Hemoglobin G, Hemoglobin S and Thalassemia Occurring in One Family
Authors: SCHWARTZ H. C. , SPAET T. H. , ZUELZER W. W. , NEEL J. V. , ROBINSON A. R. , KAUFMAN S. F.
Published: October 2019

1. A Caucasian family is described in which, on the basis of clinical, hematologic and biochemical findings, it is postulated that the genes responsible for hemoglobins S and G and for the thalassemia defect are present.

2. On the basis of the study of this family, it is concluded that:

a. The genes responsible for hemoglobins G and S cannot be alleles.

b. The genes responsible for hemoglobin G and thalassemia cannot be alleles.

c. The absence of hemoglobin A in individuals heterozygous for two "hemoglobin genes" does not provide critical evidence concerning the allelic relations of such genes.

d. In this family, heterozygosity for the gene responsible for hemoglobin G results in an asymptomatic trait condition, in which some 40% of the hemoglobin is abnormal. When the gene responsible for G is combined with a hemoglobin S gene or a thalassemia gene, or both, the presence of hemoglobin G does not significantly alter the expression of these genes on their combinations. For example, an individual of the phenotype SG, whose hemoglobin contained no demonstrable A, was clinically a sickle cell trait, in that he showed no evidence of physiologic handicap.

e. Individuals heterozygous for both the G and thalassemia genes may show on electrophoresis only hemoglobin G. This illustrates the unreliability in some cases of diagnosing genotype on the basis of electrophoretic findings.

f. On the basis of these findings, hemoglobin G should probably be regarded as a normal variant of hemoglobin rather than as an abnormal type of hemoglobin.
The Bone Marrow in Pregnancy and the Puerperium
Authors: LOWENSTEIN LOUIS , BRAMLAGE CATHERINA A. , LALONDE MADELEINE
Published: October 2019

1. The morphologic findings of sternal bone marrow aspirates obtained from thirty healthy women during each of the three trimesters of normal pregnancy, from ten healthy women on each of the first eight puerperal days and from thirty healthy women at six weeks postpartum were compared with the findings of marrow aspirates obtained from 30 non-pregnant normal women of childbearing age.

2. Statistical analysis of the results of differential counts of these aspirates confirmed the impressions obtained from examination of the bone marrow aspirates and their differential smears.

3. Significant quantitative changes occurred in the cellular components of the bone marrow during normal pregnancy, which were maximal in the third trimester. There was a significant increase of normoblastic erythropoiesis and, to a lesser extent, of granulopoiesis during pregnancy, which was associated with an increase of immature cells (shift to the left) of both erythropoietic and of granulopoietic tissues.

4. During the first eight puerperal days the increase of erythropoiesis and of granulopoiesis diminished but did not return to normal non-pregnant normal values. At six weeks postpartum both erythropoiesis and granulopoiesis were more active than in the early puerperium and were significantly increased as compared with the findings in the marrows of normal non-pregnant women.

5. There was some increase of plasma cells and of reticulum cells in the bone marrow during pregnancy and the early puerperium.

6. No megaloblastic or macrogranulocytic changes were found in the two hundred bone marrow aspirates taken from pregnant and puerperal women or in the thirty aspirates taken from non-pregnant normal women.

7. The marrow findings of three women in the early puerperium were reported separately from the above groups. An occasional intermediate macrometamyelocyte was found in the marrows of these patients and in one a few nucleated red cells showed the earliest transitional changes toward megaloblasts. It is believed that these changes represent the earliest morphologic evidence of a deficiency of one or more of the growth factors necessary for normal hemopoiesis.
Electron Microscopic Investigations on Sections from Lymph Nodes and Bone Marrow in Malignant Blood Diseases
Authors: BRAUNSTEINER H. , FELLINGER K. , PAKESCH F.
Published: October 2019

Thin sections from nodes and bone marrow of patients with malignant blood diseases have been investigated with the electron microscope. "Granular bodies" and "vesicular bodies" are found in the cytoplasm of Sternberg cells, and "vesicular bodies" in leukemic cells. An organized ergastoplasm is only to be seen in malignant plasmocytes from myeloma, in isolated plasmocytoma and in lympoid cells from macroglobulinemia. Its development and its relation to protein production and secretion are demonstrated. The submicroscopic organization of cells is described and commented upon.
The Submicroscopic Morphology of Gaucher Cells
Authors: DEMARSH QUIN B. , KAUTZ JEAN
Published: October 2019

The large cells characteristically found in the bone marrow and other organs in Gaucher’s disease have been reinvestigated with the electron microscope, as well as with phase contrast and polarizing microscopes and by standard staining methods. The cytoplasm was filled with a number of dense elongate or crescent shaped bodies. Each of these fibrils was in turn seen with the electron microscope to be bounded by a single dense limiting membrane, and to contain a homogeneous appearing matrix in which were embedded numerous tubular-appearing subunits. These tubular elements measured approximately 130 Å in diameter, and were of very great length. It is suggested that these submicroscopic tubular structures could represent the molecular kerasin or lipoprotein units known from biochemical and histochemical evidence to be present in Gaucher cells.

Some details about the appearance of the ectoplasm in spread cells, and the area near the cell border in cells fixed in situ, are reported. Electron micrographs revealed a complex cell border which was extended into many small pseudopodia and ridge-like projections. Numerous microvesicles about 600 Å in diameter were found in the area near the cell membrane. This is taken as possible evidence of transport of some substance across an active cell membrane. An attempt is made to correlate these morphologic findings with the clinical and biochemical findings of ourselves and others on Gaucher’s disease.
Extrusion of Nuclei of Transfused Avian Erythrocytes in the Mammalian Spleen
Authors: VORHAUS ELAINE F. , TURNER JOSEPH C.
Published: October 2019

1. Duck erythrocytes have been injected intravenously into rabbits, mice, and guinea pigs and at intervals thereafter microscopic examination of certain organs has been made.

2. In the guinea pig spleen extrusion of the nuclei of the duck cells has been observed with great regularity.

3. Attempts to reproduce this phenomenon in vitro have failed.

4. It is suggested that nuclear extrusion of this kind is conditioned by (a) the species into which the cells are transfused and (b) an intact spleen.
Studies on the Serum Haptoglobin Level in Hemoglobinemia and Its Influence on Renal Excretion of Hemoglobin
Authors: LAURELL CARL-BERTIL , NYMAN MARGARETA
Published: October 2019

A short survey is given of the literature on haptoglobin, the hemoglobin-binding serum protein, its properties and biologic variations. The principles of an electrophoretic method for quantitative determination of the serum haptoglobin are described.

Electrophoretic studies showed that haptoglobin has a high affinity for hemoglobin at physiologic pH and that every haptoglobin molecule can bind at least 2 hemoglobin molecules.

Observations made following the intravenous injection of hemoglobin showed:

that hemoglobin administered intravenously is bound by the haptoglobin;

that free hemoglobin is not demonstrable until more hemoglobin has been injected than can be bound by the haptoglobin;

that the complex hemoglobin-haptoglobin is eliminated from the plasma after intravascular hemolysis or intravenous administration of hemoglobin without being excreted in the urine;

that the hemoglobin-haptoglobin complex is removed from the plasma at a constant rate during the major part of the elimination period;

that the haptoglobin level will fall to nil within 24 hours, if the amount of hemoglobin injected is sufficient to bind all the haptoglobin available. During the following days the rate of formation of haptoglobin can be studied.

From the data available it can be concluded that hemoglobinuria cannot appear until the amount of hemoglobin administered intravenously or the amount liberated intravascularly exceeds the binding power of the haptoglobin and the reabsorption capacity of the tubules. The variation observed by earlier authors in the so-called renal threshold for hemoglobin on intravenous injection of hemoglobin can be explained among other things by the variation in the haptoglobin content in one and the same subject, i.e., if the haptoglobin level is low, the threshold value will also be low, and vice versa.
The Combined Use of Typhoid Vaccine and P32 Labeling to Assess Myelopoiesis
Authors: PERRY S. , WEINSTEIN I. M. , CRADDOCK C. G. , LAWRENCE J. S. , PAUL GLORIA , BAKER MARY H. , GREEN JEANINE , CIANKO GEORGE
Published: October 2019

1. The intravenous administration of killed typhoid organisms to dogs results in the development of a severe leukopenia which is followed by a marked leukocytosis.

2. The myelopoietic response of dogs to typhoid injections is quite similar to the response following leukopheresis.

3. Leukocyte DNA-P32 studies indicate that in response to typhoid stimulation, as with leukopheresis, the bone marrow constitutes the main reservoir for granulocytes contributing to peripheral blood leukocytosis.

4. Evidence is presented that, under these experimental conditions, leukocytes, after once having left the vascular tree, are unable to re-enter in any significant numbers.

5. The use of typhoid vaccine to stimulate leukocytosis combined with the use of P32 to measure the fate of cells released from the marrow is presented as an accurate and reasonably simple method to measure certain aspects of myelopoiesis in the experimental animal.
Studies on Erythrocytes in Cases with Past History of Favism and Drug-Induced Acute Hemolytic Anemia
Authors: SZEINBERG A. , SHEBA C. , HIRSHORN NINA , BODONYI EVA
Published: October 2019

Studies have been carried out on erythrocytes of cases with past history of acute hemolysis due to Favism, sulpha drugs or PAS.

No indication was found of abnormal osmotic or mechanical fragility, electrophoretic mobility of hemoglobin or its amino acid content. Similarly no abnormal glycolytic or catalase activities were detected.

Glutathione (GSH) deficiency of erythrocytes was discovered in all cases studied. Similarly, glutathione (GSH) deficiency was a frequent finding among the relatives of affected cases.

No such glutathione deficiency could be established in the cases of thalassemia major so far studied, though both patient-groups without any exception belong to the same ethnographic denomination (non-Ashkenazic).

This investigation was aided by a grant from the Israel Research Council.
Studies on the Hemolysin and Agglutinin of Leukemic Leukocytes
Authors: PIROFSKY BERNARD
Published: October 2019

1. Leukocytes from thirteen cases of leukemia were isolated, homogenized, and extracted in 0.88 M sucrose, 0.14 M NaCl, and 2.0 M NaCl solutions. All thirteen 0.88 M sucrose, twelve of thirteen 2.0 M NaCl, and seven of the thirteen 0.14 M NaCl extracts, were found to contain a hemolysin and agglutinin active against normal and trypsinized erythrocytes.

2. The hemolysin and agglutinin of leukemic leukocyte extracts was found to differ from coating antibody eluted from erythrocytes of patients with acquired hemolytic anemia, from β-glucuronidase, and from anti-H agglutinin. In spite of great similarities, the hemolysin and agglutinin differed in certain respects from the lysin-inhibitor complexes that have been extracted from tissues.

3. An inhibitor of the hemolysin and agglutinin of leukemic leukocyte extracts was found in normal serum. This inhibitor is stable at 56° C. for 30 minutes, is not dialyzable, is not exhausted from leukemic sera, and apparently does not permanently change the hemolysin or agglutinin.

4. The possible role of the leukemic leukocyte hemolysin and agglutinin in instituting premature senescene of the erythrocyte resulting in a "hemopathic" type of hemolytic anemia is discussed.
Platelet Functional Defects in Thrombocytopenic Purpura
Authors: BONNIN J. A. , JENNER MISS MAUREEN
Published: October 2019

A defect in a serum thromboplastic component was found in six patients with severe thrombocytopenic purpura in addition to the platelet thromboplastic defect.

The defective sera returned to normal spontaneously upon storage at -20 C. The addition of washed normal platelets during coagulation completely corrected the serum defect in the one instance in which it was possible to carry out the experiment.

It was postulated that a platelet factor was primarily at fault, resulting in the lack of immediate activation of a serum thromboplastic component.

The two thromboplastic defects are cumulative and are possibly dependent upon separate platelet factors.
Leukemia or Lymphoma and Coexistent Primary Malignant Lesions: A Review of the Literature and a Study of 120 Cases
Authors: MOERTEL CHARLES G. , HAGEDORN ALBERT B.
Published: October 2019

A total of 194 cases of leukemia or lymphoma with coexistent primary malignant disease was found reported in the literature. To this we have added 120 new cases from the files of the Mayo Clinic in which the diagnosis was made in the 10 years from 1944 through 1953.

The following observations were made:

1. The presence of leukemia or lymphoma does not seem to predispose to the development of any other specific type of primary malignancy.

2. The incidence of another primary malignant lesion in patients with leukemia or lymphoma is probably comparable to, and perhaps exceeds, that in any segment of the general population of similar age.

3. The coexistence of Kaposi’s sarcoma and leukemia or lymphoma in the same patient probably represents a morphologic variation of the same basic malignant disease of the reticuloendothelial system.

4. Any patient with leukemia or lymphoma who presents signs or symptoms of a focal malignant lesion should be regarded as having another primary lesion until proved otherwise on pathologic examination.
The Role of the Mature Neutrophil in Bacterial Infections in Acute Leukemia
Authors: SILVER RICHARD T. , BEAL GRACE A. , SCHNEIDERMAN MARVIN A. , MCCULLOUGH NORMAN B.
Published: October 2019

Its acute leukemia, phagocytosis of Brucella organisms by mature neutrophils was not impaired. Phagocytosis of these organisms was not altered by antimetabolite therapy or during periods of bacterial infection. The absolute mature neutrophil count when infection developed was found to vary from patient to patient. The onset of bacterial infection in patients with acute leukemia was preceded by dynamic decreases in the number of mature neutrophils in the peripheral blood. Seventeen of eighteen patients had lower mature neutrophil counts immediately prior to the onset of infection as compared to the period free of infection. The median decrease was 54 per cent. As infection did not always follow decreases in mature neutrophils it is suggested that other factors also play a role in the development of bacterial infections in patients with acute leukemia.
The Osmotic Resistance of Leukocytes in Peripheral Blood and Splenic Circulation of the Rabbit
Authors: STORTI E. , BELLESIA L. , LUSVARGHI E.
Published: October 2019

By studying the behavior of the resistance of leukocytes to hypotonic saline solution we have found that leukocytes leaving the spleen are much less resistant than those entering it.

It is felt that the change in resistance affecting leukocytes on their passage through the spleen is in all probability the manifestation of one of the mechanisms by which the spleen regulates the life span of leukocytes.
Serum 2-Hydroxyglutarate Level Is a Prognostic Marker in Acute Myeloid Leukemia
Authors: Wang Jing-Han , Chen Wen-Lian , Chen Tian-Lu , Zhu Yong-Mei , Zhang Wei-Na , Li Yang , Wu Song-Fang , Qiu Yun-Ping , Zhao Ai-Hua , Li Jun-Min , Mi Jian-qing , Jin Jie , Wang Yun-Gui , Huang He , Wu De-Pei , Li Yan , Yan Xiao-Jing , Yan Jin-Song , Li Jian-Yong , Wang Shuai , Huang Xiao-Jun , Jia Wei , Chen Zhu , Chen Sai-Juan
Published: October 2019

Abstract 1433

BACKGROUND

High levels of 2-HG in serum were found among AML cases with IDH1/IDH2 mutations. However, the impact of 2-HG levels on biological characteristics and clinical outcomes has not been systematically evaluated in a large cohort of AML patients.

METHODS

Serum 2-HG levels in 699 patients with hematologic malignancies, including 405 AML patients, were measured by GC-TOFMS. Clinical prognostic power of 2-HG and genetic risk factors associated with 2-HG were also evaluated in AML patients.

RESULTS

Among all patients with hematopoietic malignancies investigated, high serum 2-HG levels were observed only in AML group. 64 out of 405 (15.8%) AML patients displayed aberrantly higher levels (7.14±2.11μg/ml) of 2-HG. Compared to cases with normal 2-HG (3.65±1.02μg/ml), these patients showed a higher prevalence in AML-M0/M1 subtypes, a closer correlation with mutated IDH1/2 genes, a distinct gene expression profile and an aberrant DNA methylation status in bone marrow blasts. Additionally, the patients with high 2-HG were associated with higher serum levels of α-ketoglutarate (α-KG) and glutamate, suggesting presence of impaired metabolic pathway involved in the biosynthesis of 2-HG. Univariate and multivariate analyses indicate that high level of 2-HG is among the most significant negative indicators for complete remission (CR) rate, overall survival (OS) and event-free survival (EFS) either in all AML cases or in cases with cytogenetically normal AML (CN-AML).

CONCLUSIONS

Serum 2-HG is an independent prognostic marker in AML. Patients with high 2-HG had significantly higher frequency of IDH1/IDH2 gene mutations and unfavorable prognosis compared to those with normal 2-HG.

Disclosures:

No relevant conflicts of interest to declare.
Incidence and Outcomes of a Rare Translocation t(3,5) in Patients (pts) with Acute Myeloid Leukemia (AML) and Myelodysplastic Syndrome (MDS)
Authors: Alattar Mona Lisa , Kantarjian Hagop M. , Cortes Jorge E. , Kadia Tapan M. , Borthakur Gautam , Ohanian Maro , Ravandi Farhad , Garcia-Manero Guillermo , Quintas-Cardamas Alfonso , Jabbour Elias , Daver Naval , Pierce Sherry A. , Brandt Mark , Faderl Stefan , Pemmaraju Naveen
Published: October 2019

Abstract 1456

Background:

Cytogenetic analysis of large pt cohorts allows us to evaluate the prognostic impact of rare and unique translocations. Little is known about the clinical outcomes of pts with t(3,5), which generally results in fusion of NPM and MLF1 in pts with MDS and AML.

Methods:

We retrospectively reviewed the charts of 8,215 pts with a diagnosis of MDS or AML evaluated at our institution from 1985–2011.

Results:

A total of 17 pts with a t(3,5) either at diagnosis or post-treatment were identified (10 pts, as the sole cytogenetic abnormality, and 7 pts, as part of other/complex karyotype). Among evaluable cases (15/17), frequently occurring breakpoints included q25;q34 (n=4), p21;q15 (n=2), and p21;q13 (n=2).10 pts had MDS with IPSS of Int-1(n=5) and Int-2(n=5), and 7 pts had AML. Four pts had therapy-related MDS (3 pts with prior lymphoma, 1 pt small cell lung cancer) and one pt with MDS had PNH. Median age was 56 years (range, 20–78) at diagnosis. Four pts (24%) had a FLT3-ITD mutation (3 with AML and 1 with MDS), 1 of these FLT3-mutated pts had additional mutations in c-KIT and NRAS mutation, 9 pts were tested and negative for molecular abnormalities, and 4 pts did not have molecular analysis available. Therapies were diverse, with two most common: cytarabine-based regimens (n=6) and hypomethylating agent-based therapy (n=6). Overall, median number of therapies for all pts was 1 (range, 0–5), including 2 pts treated with upfront stem cell transplant (SCT) and 1 pt treated with only growth factors/supportive care. Six pts (35%) (MDS 3 pts, AML 3 pts) underwent SCT during their course of therapy (including 1 cord blood, 1 SCT with NK cells, and 4 allogeneic MUD SCT). Overall median survival for pts with MDS was 8.1 months (range, 1–57) and for pts with AML, 21 months (range, 2–53). CR was achieved overall in 12 pts (71%) with median CR duration of 3.2 months (range, 1–60 months). Three (18%) pts were refractory to all chemotherapy and one pt died during induction chemotherapy (infection and diffuse alveolar hemorrhage). One pt never received treatment.

Conclusions:

Survival is particularly poor among patients with MDS and t(3;5) while those with AML have survival comparable to normal karyotype (NK) AML. Further investigation with novel treatment approaches is warranted in this subpopulation of MDS/AML pts.

Disclosures:

No relevant conflicts of interest to declare.
Long-Term Efficacy of Low-Dose All-Trans Retinoic Acid Plus Individually Adapted Chemotherapy Induction Followed by Arsenic Trioxide Based Post-Remission Therapy in Adults with Newly Diagnosed Acute Promyelocytic Leukemia
Authors: Lou Yinjun , Jin Jie
Published: October 2019

Abstract 1480

Acute promyelocytic leukemia (APL) is a distinct subtype of acute myelogenous leukemia (AML), which usually presents with pancytopenia, coagulopathies and bleeding. Molecular studies have revealed that it was caused by leukemogenic PML-RARA fusion gene resulting from a specific chromosomal translocation t(15;17). The administration of target agent all-trans-retinoic acid (ATRA) combined with anthracycline-based chemotherapy for induction and consolidation followed by ATRA plus low-dose chemotherapy maintenance is the standard strategies for patients with newly diagnosed APL. However, despite the high cure rate, early death and leukemia relapse are the two main important obstacles. We evaluated the efficacy of low-dose All-trans-retinoic acid (ATRA) plus individually adapted chemotherapy for induction followed by arsenic trioxide (ATO) based post-remission therapy in newly diagnosed acute promyelocytic leukemia (APL). From January 2004 to September 2011, 109 patients with APL were enrolled the study. The complete remission (CR) rate was 96.3%. The early death rate was 0.9%. Two arms were assigned according to post-remission protocols: ATO group cases were treated with standard chemotherapy, ATO, and ATRA. Without ATO group cases were subsequently treated with chemotherapy and ATRA only. Patients were monitored by reverse transcriptase-polymerase chain reaction (RT-PCR) during and after treatment. The six-year relapse-free survival (RFS) was significantly better for patients in ATO group than in without ATO group, 94.4% versus 50.6% (P < 0.0001) and the six-year overall survival (OS) rate was 95.7% versus 64.1%, in two groups (P = 0.003). This study shows that low-dose ATRA plus tailored chemotherapy is effective in induction therapy, and the addition of ATO to post-remission therapy significantly improves the long-term outcome.

Disclosures:

No relevant conflicts of interest to declare.
Tumor Associated Macrophage Expression, CD163, Is Associated with Poorer Outcome in Diffuse Large B-Cell Lymphoma
Authors: Bohn Olga L. , Maragulia Jocelyn C. , Crispin Hilda , Gonzalez Camille , Miething Cornelius , Zelenetz Andrew D. , Teruya-Feldstein Julie
Published: October 2019

Abstract 1583

Background:

Diffuse large B cell lymphoma (DLBCL) is the most common lymphoma with various subtypes. Recent data of Tumor Associated Macrophages (TAM) in DLBCL have shown association with poor overall survival. We were interested in assessing these markers, in particular, for practical daily clinical use at a high volume cancer center. We therefore performed a retrospective study to evaluate expression of macrophage markers (CD68 and CD163) in DLBCL and determine overall survival (OS) and event free survival (EFS).

Design:

A total of 128 DLBCL cases were reviewed. Tissue microarrays (TMA) were constructed with cores in triplicate. CD163 and CD68 expression was assessed by immunohistochemistry (IHC) using anti-CD68 (KP-1, Ventana) and anti-CD163 (Vector Laboratories) performed on the automated Ventana XT platform according to manufacturer protocols (Tucson, AZ). The assessment of intratumoral CD163 infiltration was performed at objective magnification 40X by three observers (OB, HC and JTF) and a consensus was obtained. The number of CD163 macrophages was counted in three cores. The mean was calculated and the results were scored as follows: 0 (0 to 25 positive cells); 1 (between 26 to 50 positive cells); 2 (51 to 75 positive cells); 3 (76 to 100 positive cells). CD68 expression was assessed by IHC and the degree of intratumoral infiltration was scored as follows: 0 (less than 5% positive cells); 1 (between 5% to less than 50% of positive cells); 2 (more than 50% positive cells). Of 128 patients, 13 had limited clinical data, 23 were biopsies at relapse, and 92 were evaluable for OS and EFS from initial therapy. EFS was defined as progression of disease, secondary malignancy, or death due to disease or any other reason. OS was calculated from date of death or last follow-up date to date of diagnostic biopsy. EFS was calculated from event date or date of last follow-up to end of treatment date. SPSS software was used to calculate Kaplan Meier survival curves and statistics.

Results:

There were 51 (55%) males and 41 (45%) females (M:F ratio 1.2). The median age was 60 years (range from 10 to 90). A high number of tumor infiltrating CD163 expression (scores 0 [n=19], 1 [n=6], 2 [n=9] vs 3 [n=58]) was associated with significantly poorer OS (p=0.028) and EFS (p=0.005). A statistically significant difference was observed in patients with tumors that had abundant tumor associated macrophages with CD163 (score 3 >75 positive cells) on IHC. CD68 expression did not correlate with OS and EFS.

Conclusion:

High CD163 expression is statistically significantly associated with decreased OS and EFS in DLBCL. Because of cleaner staining, scoring on a daily practical basis in a high volume center is useful. Our data supports initial few prior reports of this marker in DLBCL.

Disclosures:

No relevant conflicts of interest to declare.
Salvage Treatment with Ofatumumab and ESHAP (O-ESHAP) for Patients with Relapsed or Refractory Classical Hodgkin's Lymphoma After First-Line Chemotherapy: Interim Analysis of a Phase II Trial of the Spanish Group of Lymphoma and Bone Marrow Transplantation (GELTAMO)
Authors: Martinez Carmen , Rodriguez-Calvillo Mercedes , Terol María José , Xicoy Blanca , García-Sanz Ramón , Perez-Ceballos Elena , Cantalapiedra Alberto , Domingo-Domènech Eva , Hernández Miguel , Sampol Antonia , Espeso Manuel , Briones Javier , Lopez Javier , Sureda Anna
Published: October 2019

Abstract 1630

The management of recurrent or refractory Hodgkin's lymphoma (HL) remains challenging. Previous published data have suggested that infiltrating normal B lymphocytes in classic HL lesions may contribute to the survival of Hodgkin and Reed-Sternberg cells in vivo. The objective of this prospective, multicenter, phase II trial was to investigate the activity of an anti-CD20 monoclonal antibody, ofatumumab, in combination to a standard platinum-based salvage regimen, ESHAP (O-ESHAP) followed by high-dose chemotherapy and autologous stem cell transplantation (ASCT) for patients with classical HL failing to first line chemotherapy. Forty- five patients (25 M / 21 F, median age 34 years, range 18–66) were enrolled in the study between June 2010 and June 2012. Treatment consisted on three cycles of ESHAP plus ofatumumab 1,000 mg days 1 and 8 on first cycle and day 1 on second and third cycles. At the time of study entry, 66% of patients had III-IV Ann Arbor stage, 16% bulky disease, 18% B symptoms, 40% extranodal HL and 52% ≥3 involved nodal areas. We respect to response to first-line therapy, 46% patients had achieved a completed response (CR) and then relapsed, 6% had a partial remission (PR), whereas the remaining 48% were primary refractory. Eighty-one percent patients have received 3 cycles of O-ESHAP as scheduled, three patients 2, and five 1 cycle (1 patient due to toxicity, 1 patient's decision, 2 HL progression, and 4 treatments ongoing). Grade 3–4 WHO hematological toxicity was observed in 16%, 19%, and 20% after cycles 1, 2, and 3, respectively. Non-hematological toxicity was reported in 32%, 10%, and 20%, respectively. Overall response (OR) rate was 63% (49% CR and 14% PR). Response to O-ESHAP according to prior response to first-line chemotherapy is shown in table 1. Adequate PBSCs collection was achieved in 94% mobilized patients. Twenty-six out of 33 patients have already proceeded to ASCT. Two patients died of neutropenic sepsis after ASCT and HL progression, respectively. Preliminary results of this ongoing trial suggest that addition of ofatumumab to ESHAP is safe and has a promising clinical activity in patients with relapsed/refractory HL candidates to ASCT.

Table 1. Response to O-ESHAP according to previous response to first-line treatment Response to first-line chemotherapy Relapsed or partial response (n=17) Refractory (n=16) Response after O-ESHAP OR 16 (94%) 7 (44%) CR 14 (82%) 3 (22%) PR 2 (12%) 4 (22%) Refractory 1 (6%) 9 (56%)

Disclosures:

No relevant conflicts of interest to declare.
A Single Nucleotide Polymorphism in the Coding Sequence of BIM Is Associated with Poor Prognostic in Chronic Myeloid Leukaemia Treated by Imatinib
Authors: Mahon Francois-Xavier , Augis Vanessa , Airiau Kelly , Josselin Marina , Turc Beatrice , Belloc Francis
Published: October 2019

Abstract 1683

BIM is a pro-apoptotic protein essential to tyrosine kinase inhibitors (TKI) response in chronic myeloid leukaemia (CML) patients. Thus, mutations in BIM sequence could lead to imatinib resistance independently of BCR-ABL mutations. Recently, a deletion polymorphism in intron 2 of BIM was demonstrated to confer intrinsic TKI resistance in asiatic patients (Ng et al, nature medicine, 2012). However the Caucasian population is not concerned by this intronic deletion polymorphism. So, in the current study we performed BIM coding sequence analysis in 72 CML imatinib-treated patients and among them 40 were resistant to imatinib in our center from French population of patients. For twenty patients, the disease evolved in either acceleration or blastic phases at least once during the study. We did not find any mutation with amino-acid change in the BIM coding sequence. However, we observed a silent single nucleotide polymorphism (SNP) c465C>T (rs724710) in the region coding for the active BH3 domain. A strong statistic link was found between the presence of the T allele of this polymorphism and the Sokal high risk group (p=0.0065). The frequency of the T allele was found higher in non responsive patients than in a local healthy donor reference population (p=0.0049). Similarly, the analysis showed an association of this T allele with the frequency of mutations on the tyrosine kinase domain of BCR-ABL (p<0.001) but not with additional chromosomal alterations. Moreover, the presence of the allele T significantly lengthened the time of achievement of major molecular remission. Finally, the presence of the T allele was found related to a decreased basal expression of the Bim mRNA in the circulating mononuclear cells of healthy donors. These results suggest that analysis of the c465C>T SNP of BIM could be useful for predicting the outcome of CML patients under imatinib treatment.

Disclosures:

Mahon: Novartis Pharma: Consultancy, Honoraria, Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria; Pfizer: Consultancy.