bluuebunny/crossref_metadata_2025 huggingface dataset:
Small-molecule inhibitors in myeloproliferative neoplasms: are we aiming for the right targets?
Authors: Constantinescu Stefan N. , Vainchenker William
Published: October 2019

The ATP-binding pocket of the kinase domain of JAK2 is the major target of the present treatment of myeloproliferative neoplasms. Several inhibitors of JAK2 that are ATP competitive have been developed, but they do not discriminate between wild-type and mutant JAK2. These inhibitors have been used in myelofibrosis and, for the first time, treatment induced a reduction in spleen size and in constitutional symptoms. However, no dramatic effects on BM fibrosis, allele burden, or peripheral blast numbers were observed. These data indicate that other avenues should be explored that would either target mutant molecules (JAKs or receptors) more specifically and spare wild-type JAK2 or that would address other pathways that contribute to the malignant proliferation. Future success in treating myeloproliferative neoplasms will depend on advances of the understanding of JAK-STAT signaling and also on a better understanding of the disease pathogenesis, especially the role that mutants in spliceosome factors and epigenetic regulators play in the phenotype of the disease and the precise mechanism of fibrosis development.
Interpreting new molecular genetics in myelodysplastic syndromes
Authors: Abdel-Wahab Omar , Figueroa Maria E.
Published: October 2019

The myelodysplastic syndromes (MDS) are a clinically and cytogenetically heterogeneous group of clonal diseases characterized by ineffective hematopoiesis, peripheral blood cytopenias, and an increased risk of progression to acute myeloid leukemia. The precise molecular mechanisms behind the development of MDS have remained elusive; however, the distinct sensitivity of this disease to DNA methyltransferase inhibitors and the presence of markedly abnormal epigenetic profiles suggested the existence of an epigenetic mechanism underlying the disease. Recently, the advent of new technologies for the detection of genetic abnormalities has led to the description of a set of novel recurrent mutations in patients with this disease. The majority of these novel mutations have been described in genes encoding different components of the epigenetic machinery, many of which are associated with distinct clinical outcomes. Finally, mutations in mRNA splicing genes have also been described recently in MDS, underscoring the molecular complexity that underlies the development of this heterogeneous disease.
Are you sure this is Waldenström macroglobulinemia?
Authors: Ghobrial Irene M.
Published: October 2019

Waldenström macroglobulinemia (WM) is a rare lymphoproliferative disorder characterized by the presence of lymphoplasmacytic cells in the BM and IgM monoclonal protein in the serum. The origin of the malignant clone is thought to be a B cell arrested after somatic hypermutation in the germinal center and before terminal differentiation to plasma cells. In this review, recent advances in the genetic and epigenetic regulators of tumor progression are discussed. Risk factors include IgM-monoclonal gammopathy of undermined significance, familial disease, and immunological factors. The clinical manifestations of the disease include those related to clonal infiltration of the BM, lymph nodes, and, rarely, other sites such as pulmonary or CNS infiltration (Bing-Neel syndrome). Other manifestations are related to the IgM monoclonal protein, including hyperviscosity, cryoglobulinemia, protein-protein interactions, Ab-mediated disorders such as neuropathy, hemolytic anemia, and Schnitzler syndrome. IgM deposition in organs can lead to amyloidogenic manifestations in WM. The diagnostic workup for a patient with WM and rare presentations of WM are described herein. Prognosis of WM depends on 5 major factors in the International Staging System, including age, anemia, thrombocytopenia, β-2 microglobulin, and IgM level. The differential diagnosis of WM includes IgM-multiple myeloma, marginal zone lymphoma, mantle cell lymphoma, and follicular lymphoma.
New insights into genotype and phenotype of VWD
Authors: Flood Veronica H.
Published: October 2019

Recent advances in VWD research have improved our understanding of the genotype and phenotype of VWD. The VWF gene is highly polymorphic, with a large number of sequence variations reported in healthy individuals. This can lead to some difficulty when attempting to discern genotype–phenotype correlations because sequence variations may not represent disease. In type 1 VWD, mutations can be found throughout the VWF gene, but likely pathogenic sequence variations are found in only ∼2/3 of type 1 VWD patients. Sequence variations in type 2 VWD are located in the region corresponding to the defect in the VWF protein found in each type 2 variant. In type 3 VWD, sequence variations are not confined to a specific region of the VWF gene and also include large deletions that may not be picked up using conventional sequencing techniques. Use of genetic testing may be most helpful in diagnosis of type 2 VWD, in which a larger number of known, well characterized mutations are present and demonstration of one of these may help to confirm the diagnosis. Bleeding symptoms in general are more severe with decreasing VWF levels and more severe in type 2 and type 3 VWD compared with type 1 VWD. Prediction of phenotype for an individual patient, however, is still difficult, and the addition of genetic data will be most helpful in ascertaining the correct diagnosis for VWD patients.
Advances in therapies for non-Hodgkin lymphoma in children
Authors: Kobos Rachel , Terry William
Published: October 2019

Pediatric patients with newly diagnosed, non-Hodgkin Lymphoma (NHL) have an excellent overall survival. However, therapy regimens are associated with acute toxicity and late effects. Furthermore, patients with relapsed or refractory disease have relatively few options with proven clinical benefit. Both histologic and molecular differences exist between adult and pediatric NHL preventing simple translation of adult NHL successes into improvements in pediatric NHL treatment. This review summarizes the introduction of targeted therapies into frontline treatments for patients with anaplastic large-cell lymphoma and CD20–positive tumors, with the goal of improving overall survival while limiting both short- and long-term toxicities. In addition, newer approaches that have limited data in children but may have a significant role in how we treat pediatric NHL in the future are reviewed, which include CD19 directed therapy, Notch inhibition, the tri-functional antibody, FBTA05, and EZH2 inhibition.
Treatment of Langerhans cell histiocytosis: role of BRAF/MAPK inhibition
Authors: Abla Oussama , Weitzman Sheila
Published: October 2019

Langerhans cell histiocytosis (LCH) is a clonally derived neoplasm with a highly variable clinical course. Although LCH was once considered a disorder of immune regulation, the identification of activating mutations in the proto-oncogene BRAF-V600E in ∼50%-60% of cases and MEK and ERK phosphorylation in 100% of examined cases, has changed the definition of LCH to a dendritic cell neoplasm with a strong inflammatory component. Current international LCH trials are focused on further improving the outcome of high-risk multisystem LCH patients, by decreasing the reactivation rate, optimizing early salvage regimens, and preventing late sequelae. Anecdotal responses to vemurafenib, a BRAF-V600E inhibitor, have been reported in a few cases of LCH and Erdheim–Chester disease. However, the development of resistance, as well as the potential risks of cutaneous and pancreatic cancers in patients with BRAF-V600E-mutated melanoma treated with single inhibitors, suggest the need for prospective trials with BRAF inhibitors, alone or in combination with other inhibitors of this pathway, for patients with refractory or multiply-relapsed LCH. The recent discovery of somatic mutations in ARAF and in MAP2K1, which lead to activation of the RAS-RAF-MEK –ERK pathway in the setting of wild-type BRAF, as well as the finding that activating mutation in MAP2K1 are relatively insensitive to MEK inhibitors, suggest that a more detailed understanding of this pathway in LCH may be necessary for the development of more effective targeted therapies.
Therapy-related myeloid neoplasms: does knowing the origin help to guide treatment?
Authors: Heuser Michael
Published: October 2019

Therapy-related myeloid neoplasms (t-MN) combine t-MDS and therapy related acute myeloid leukemia (t-AML) patients in one entity because of their similar pathogenesis, rapid progression from t-MDS to t-AML, and their equally poor prognosis. Treatment with epipodophyllotoxins like etoposide has been associated with a short interval between treatment and development of t-AML, with fusion oncogenes like KMT2A/MLL-MLLT3 and a better prognosis. In contrast, treatment with alkylating agents has been associated with a longer latency, an initial MDS phase, adverse cytogenetics, and a poor prognosis. The pathogenesis of t-MN can be explained by direct induction of an oncogene through chromosomal translocations, induction of genetic instability, or selection of a preexisting treatment-resistant hematopoietic stem cell clone. Recent evidence has highlighted the importance of the last mechanism and explains the high frequency of TP53 mutations in patients with t-MN. After previous cytotoxic therapy, patients present with specific vulnerabilities, especially evident from the high nonrelapse mortality in patients with t-MN after allogeneic hematopoietic cell transplantation. Here, the prognostic impact of currently known risk factors and the therapeutic options in different patient subgroups will be discussed.
Prolonged survival with the early use of a novel extracorporeal photopheresis regimen in patients with Sézary syndrome
Authors: Gao Crystal , McCormack Christopher , van der Weyden Carrie , Goh Michelle S. , Campbell Belinda A. , Twigger Robert , Buelens Odette , Harrison Simon J. , Khoo Christine , Lade Stephen , Prince H. Miles
Published: October 2019

Gao et al report that early use of extracorporeal photopheresis improves time to next treatment and survival in patients with Sézary syndrome.
PRIMARY CONGENITAL AND SECONDARY ACQUIRED SPLENIC PANHEMATOPENIA
Authors: DOAN CHARLES A. , WRIGHT CLAUDE-STARR
Published: October 2019

1. The spleen is an organ of multiple structures and many functions, but in the interests of human health and disease, it is probably far more important pathologically than physiologically.

2. It has been abundantly proved that instability in splenic functional balance toward any one of the essential elements of the blood passing through this organ may be an inherited trait, as in congenital hemolytic icterus. Recognition is now made of a syndrome in which, despite intensive compensatory panmyeloid hyperplasia, indiscriminate elimination of all circulating elements occurs, actually simulating panmyeloid hypoplasia. Splenectomy in such a syndrome is often dramatically curative. "Primary splenic panhematopenia" is suggested as an appropriate descriptive designation.

3. The potentially important role which may be played by the spleen, secondarily involved in a wide variety of syndromes, with the precipitation of varying degrees of peripheral cellular disequilibria, demands careful diagnostic discrimination. A dependable experience in the specific technics by which bone marrow and splenic functions are appraised is essential to sound judgment and clinical acumen.

4. The normal spleen is apparently not essential to life and health at any age and, therefore, may be surgically removed without prejudice to future hemolytopoietic equilibria and longevity. The pathologic spleen may at times constitute a very real hazard to health and an actual threat to survival; in the more acute syndromes, prompt surgical intervention may be lifesaving.
PROGNOSIS IN THE NEUROLOGIC MANIFESTATIONS OF PERNICIOUS ANEMIA
Authors: RUNDLES R. WAYNE
Published: October 2019

Four types of disturbed neurologic function occur in pernicious anemia: (1) cerebral, (2) olfactory, (3) peripheral nerve and posterior columns of the spinal cord, and (4) lateral columns of the spinal cord. The neurologic prognosis depends upon the specific type of disability present, its severity, duration, and the adequacy of treatment. With intensive use of highly potent liver extracts intramuscularly the prognosis is far better than generally recognized. Serious neurologic residuals can be avoided if the diagnosis is made reasonably early.
THE EXCRETION OF UROBILINOGEN IN THE STOOLS AND URINE DURING MALARIAL INFECTION
Authors: DE VRIES A. , Schiffer F.
Published: October 2019

In 10 cases of malaria (6 benign tertian, 4 malignant tertian), the excretion of urobilinogen in the stools and in the urine was studied. In all 10 cases the amount of urobilinogen excreted in the stools was found to be increased. After defervescence and disappearance of parasites from the blood the excretion gradually declined. The increased excretion of urobilinogen in the stools was the constant and sometimes the only evidence of increased blood destruction occurring at times in the complete absence of jaundice and reticulocytosis. Increased excretion of urobilinogen in the urine was not a constant feature.

It is suggested that the development of jaundice and of urobiligenuria is due not only to the liberation of pigments by the hemolysis, but to a disturbance in the liver function.

This study lends further confirmation to the concept that the only unequivocal evidence of increased blood destruction is shown in an increased output of urobilinogen in the feces.
PRESERVATION OF NORMAL HUMAN PLASMA IN THE LIQUID STATE
Authors: LOZNER EUGENE L. , LEMISH SONIA , CAMPBELL A. SUE , NEWHOUSER LLOYD R.
Published: October 2019

1. During preservation of human plasma in the liquid state at room temperature for three years, the alpha-amino nitrogen and the nitrogen content of the tungstic acid filtrate rise slowly until about two years have elapsed, after which time these levels do not increase significantly.

2. During the entire period of storage the nitrogen contents of the trichlor-acetic acid filtrate and the "polypeptide index" increase progressively. The actual increase represents hydrolysis to nonprotein size of 3 to 4 per cent of the original protein.

3. The colloid osmotic pressure of stored plasma is slightly but significantly greater than that of fresh plasma.

4. The viscosity of stored plasma is slightly but significantly greater than that of fresh plasma.

5. The electrophoretic patterns of stored plasma show increases of alpha globulin and albumin concentration, complete disappearance of gamma globulin (containing immune properties) and fibrinogen, and some reduction of beta globulin concentration as compared to fresh plasma.

6. Analysis of 3,384 questionnaires completed after administration of liquid plasma more than a year old indicates that the transfusion of such plasma continues to be safe and beneficial up to at least three years of storage. The untoward reaction rate following these administrations was significantly less than that following a comparable series of 1500 administrations of commercially prepared dried plasma.
A-B Hemolytic Disease of the Newborn
Authors: ROSENFIELD RICHARD E. , EISINGER FLORENCE
Published: October 2019

A study was made of oxalated umbilical vein blood of nearly every infant born at The Mount Sinai Hospital in a nine month period. A specimen of maternal blood was available for intragroup antibody screening and six cases of Rh-Hr hemolytic disease were eliminated from the data. The umbilical vein blood was tested, where possible, for: (1) group and Rh, (2) direct antiglobulin test, (3) hemoglobin, (4) reticulocyte count and examination of red cell morphology, (5) plasma bilirubin, and (6) osmotic fragility in 0.52 per cent NaCl. From the mothers’ blood groups, the infants were classified into group compatible and group incompatible, and the arithmetic means of the hemoglobin, reticulocyte count, and plasma bilirubin obtained for each class. A third class of infants, those with positive direct antiglobulin test, were analysed separately for comparison.

1. A weakly positive direct antiglobulin test was obtained on the umbilical vein blood of over 11 per cent of group incompatible infants but in none of the group compatible infants.

2. It appears that the weakly positive direct antiglobulin test detects an abnormal class of group incompatible infants, since their mean hemoglobin is low, their mean reticulocyte count is high, and their mean bilirubin is high, when these means are compared with those of the other group incompatible infants.

3. Thirty-eight of thirty-nine mothers of incompatible infants with positive direct antiglobulin test were group O. In comparison with the distribution of the blood groups of the mothers of other incompatible infants, this disproportion is of significance.

4. The mean reticulocyte count of incompatible infants with negative direct antiglobulin test is slightly (but with statistical significance) higher than the mean reticulocyte count of compatible infants. This difference was found to be associated almost entirely with group O mothers.

5. Thirty-one out of thirty-eight infants with positive direct antiglobulin test had increased osmotic fragility in hypotonic NaCl. Two of the negative cases appeared to have slight spherocytosis on blood smear.
Acute Intestinal Iron Intoxication
Authors: REISSMANN KURT R. , COLEMAN THOMAS J.
Published: October 2019

Following oral or rectal administration of toxic doses of dissociable iron salts in dogs and rabbits, the rapidly and excessively absorbed iron produced a profound metabolic acidosis with blood pH values as low as 6.7. The acidosis was mainly due to the hydrolysing effect of ferric ions and partly due to an increase in lactic and citric acid. The latter findings suggest a possible interference of the absorbed iron with enzymes in the Krebs cycle.

The respiratory changes were those seen in metabolic acidosis: greatly increased respiratory rate and minute volume, lowering of the blood CO2, excessive CO2 output. The cardiac output decreased progressively due to diminished venous return, but a normal blood pressure was maintained by arteriolar constriction until the final collapse occurred, which was preceded by respiratory failure.

A marked capillary congestion and increased capillary permeability were noted, the latter possibly being the result of a direct action of the high non-protein bound serum iron upon the capillary wall. The increased capillary permeability caused a reduction in plasma volume and hemoconcentration.

No abnormal hemoglobin derivatives were found.
Thrombocytopenic Purpura without Anemia and Leukopenia in Gaucher’s Disease
Authors: DE VRIES A. , IZAK G.
Published: October 2019

A patient with Gaucher’s disease is described who had thrombocytopenic purpura, but no anemia or leukopenia and only slight enlargement of the spleen. The hemorrhagic diathesis and platelet count responded well to splenectomy.
Studies on Human Myoglobin
Authors: SINGER KARL , ANGELOPOULOS BASIL , RAMOT BRACHA
Published: October 2019

Hemoglobin-free met-myoglobin preparations from premature infants and normal newborns contain a fetal type of muscle pigment (Mb F) which differs spectroscopically and electrophoretically from adult myoglobin (Mb A). Acid met-Mb F can not be distinguished from fetal acid met-Hb (Hb F) by these technics. Alkaline met-Mb F has, however, a characteristic absorption spectrum which is dissimilar from that of alkaline met-Hb F, and, electrophoretically, it moves faster than the fetal met-Hb compound. Mb F is gradually replaced by the adult muscle pigment within the first 6 months of life. Thus, the period of disappearance of the fetal red cell pigment from the circulation approximates the period of disappearance of the fetal pigment from the muscle tissue.
Some Characteristic Properties of Hemoglobin C
Authors: HUISMAN T. H. J. , VAN DER SCHAAF P. C. , VAN DER SAR A.
Published: October 2019

1. The present paper describes an apparently white family in which a few members suffered from homozygous and heterozygous hemoglobin C disease. Since the paternal great grandfather and the maternal grandfather of the patients examined were in a family relation of the first order, it is surmised that the hemoglobin C was derived from the same homozygous negroid (?) ancestor. Besides its occurrence in this family Hb-C is also found in a colored family from the same island (Curaçao).

2. Hemoglobin C shows, in addition to abnormal electrophoretic mobility, three other more or less specific properties:

a. During chromatographic examination carried out with the aid of the cation exchanger Amberlite IRC 50 (XE 64), a citrate citric acid buffer at pH 6 and sodium ion concentration of 0.15, COHb-C displaces itself noticeably slower than the three other forms of hemoglobins (S, A and F).

b. The solubility of the COHb-C in a phosphate buffer pH 6.5 of various concentrations is significantly less than that of COHb-S, COHb-A and COHb-F. The reduced form of hemoglobin C does not show this property; the solubility of this is practically the same as that of reduced adult hemoglobin.

c. A comparison of the amino acid composition of hemoglobin C and hemoglobin A, S and F, shows that COHb-C contains a higher number of lysine and perhaps histidine residues. This increase amounts to four residues per mol. for lysine and one residue per mol. for histidine. The possible connection between the more basic character of the proteins and the abnormal behavior in the electrophoretic and chromatographic experiments is discussed.

3. The results obtained with five experimental methods (electrophoresis, alkali denaturation, chromatography, salting-out experiments, and amino acid composition determination) all seem to indicate that in the homozygous form of this disease only the abnormal Hb-C is present. None of the other three hemoglobins (A, S and F) were shown to be present in determinable amounts.

4. In addition to the electrophoretic abnormalities, the above three properties may be of importance in the characterization of this abnormal hemoglobin.
Histochemical and Biochemical Studies on Leukocyte Alkaline Phosphatase Activity
Authors: WILTSHAW E. , MOLONEY W. C.
Published: October 2019

(1) In a large series of individuals alkaline phosphatase activity in the leukocyte was studied by biochemical and histochemical methods.

(2) The value of combined histochemical and biochemical measurements is demonstrated in this report.

(3) In chronic myelogenous leukemia the neutrophil contain very little alkaline phosphatase activity while in myeloid metaplasia and pyogenic infections morphologically similar cells exhibit a great increase in this enzyme.

(4) Various factors which might influence alkaline phosphatase activity in

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leukocytes were investigated and it was found that on incubation in serum or saline the cell alkaline phosphatase activity increased. However, this in vitro rise was unrelated to the source of the serum menstrum.
The Clinical Significance of Erythrocyte Cholinesterase Titers
Authors: SABINE JEAN CAPTAIN
Published: October 2019

A method for determining erythrocyte cholinesterase titers on small samples of whole blood is described in detail. The method is one which requires little special equipment and can be used by any competent technician.

Data for erythrocyte cholinesterase are presented on a group of 101 normal adults, and it is shown that the observed data fit the normal distribution. Eighteen children, aged three months to ten years, all had values within the adult distribution, and the mean value was the same as that for adults. Seven newborn babies all had values at or below the lower limit for adults.
The Rare Blood Factor rh('') or Eu
Authors: SUSSMAN LEON N.
Published: October 2019

1) A family study including the identification and transmission of the variant of time rh'' factor known as Eu is presented.

2) The need for caution in interpretation of unusual variations in reactions to standard sera, especially in Negroids is emphasized.

3) The value of special technics such as use of serum containing only a single antibody, anti-human globulin test, enzyme treated cells and elution methods for clarification of atypical findings is demonstrated.
Thalassemia in Jews from Kurdistan
Authors: MATOTH Y. , SHAMIR Z. , FREUNDLICH E.
Published: October 2019

1. Seven cases of Cooley’s anemia in Jews from Kurdistan residing in the Jerusalem area are presented with family studies. Four homozygous cases were severe and progressive. One ended fatally and the remaining 3 were kept alive by frequent blood transfusions. Three heterozygous cases showed a milder course.

2. It is suggested on the basis of these and other observations that the Jews of Kurdistan constitute a hitherto unknown reservoir of the genetic abnormality responsible for Cooley’s anemia.
The Survival Time of Canine Erythrocytes Modified by Influenza Virus
Authors: STEWART W. B. , PETENYI C. W. , ROSE H. M.
Published: October 2019

Canine erythrocytes labeled with radioactive iron were modified by the action of influenza virus. Modified cells were transfused to compatible recipients and the survival of the donated cells was followed by means of their radioactivity. A rapid fall of radioactivity occurred in the first half hour, followed by a rise which reached a maximum in 2 to 3 hours. The donated cells were then destroyed and had all disappeared by 3-4 days. During the time modified erythrocytes were in the circulation, panagglutinins to them were undetectable. However, if the initial concentration of panagglutinin was unusually high, the virus-treated red cells were permanently removed from the circulation within one-half hour after transfusion.

Hence, it is concluded that virus-modified canine erythrocytes have a greatly shortened survival-time. In addition, it is suggested that the panagglutinins to virus-modified erythrocytes may be responsible for the rapid destruction of the modified cells.
Cold Hemagglutination in Acute and Chronic Hemolytic Syndromes
Authors: PISCIOTTA ANTHONY V. , DOWNER ESTELLE , HINZ JEAN
Published: October 2019

1. Three cases have been described of hemolytic anemia associated with autoagglutination of erythrocytes in the cold. These cases were characterized clinically by pallor, slight icterus and lack of splenomegaly. Raynaud’s phenomenon was present in two of the three patients. The first patient had an acute self-limited course. The second and possibly the third were chronic and apparently persistent.

2. A hemolysin was demonstrated in the chronic variety of hemolytic anemia. This antibody was active at body temperature, required complement and appeared to be independent of the cold agglutinin. It disappeared following therapy with cortisone. In the third case, hemolysin was demonstrated directly attached to the surface of the patient’s erythrocytes by the addition of complement to the washed red cells. The hemolytic reaction was enhanced by the use of trypsinized test erythrocytes.

3. "Complete and incomplete" cold agglutinins were demonstrated by the use of saline, albumin and trypsin technics as well as by the Coombs’ antiglobulin reactions. Erythrocyte coating antibodies were not neutralized with human gamma globulin. Cold agglutinins and erythrocyte coating antibodies were unaffected by cortisone therapy.

4. Survival of transfused erythrocytes was increased in one patient during the period of cortisone treatment, despite the fact that cold agglutinins persisted.

5. Cold agglutinated erythrocytes have increased mechanical fragility when the cold agglutinin is present in high titer. Incubation at 37 C. for 24 hours causes a marked shift to the left in the osmotic fragility curve.

6. Study of the serum in one patient for antibodies to Newcastle virus disease and influenza failed to disclose these antibodies in significant titer.
Particulate Component in the Blood Plasma of Mice with Transplantable Leukemia
Authors: DE ROBERTIS E. , CANZANI R. , GASIC G. , EPSTEIN B.
Published: October 2019

Blood and plasma smears of mice inoculated with two leukemic lines were studied under the electron microscope.

In 15 mice of the AKR strain and in two C58 mice inoculated with the Hch leukemia line of the Instituto de Biología of Chile (372 to 382 transfers), dense submicroscopic particles were found in the blood plasma. In some cases isolated particles or clumps of them were found adsorbed on the red cell membranes. The particle size ranged between 19 and 58 mµ, with a large peak at 38 mµ. The survival time of the inoculated animals was of 5 days and the submicroscopic particles were constantly found between the 3rd and 5th day.

Similar dense submicroscopic particles were found in blood plasma smears of three C58 mice at the 16th day after inoculation with the Ich leukemia line (86th transfer).

The probable significance of these particles is discussed on the basis of the literature regarding the etiology of mouse leukemia.
The Influence of Anoxia of Lactating Rats and Mice on Blood of Their Normal Offspring
Authors: GRANT WILSON C.
Published: October 2019

1) Lactating rats and mice 3 to 4 days post partum were discontinuously exposed daily to an ambient pressure of 400 to 300 mm. Hg for 6 hours while their litters remained at sea level pressure. At the same time, control rats and mice were separated from their litters but were kept at sea level pressure.

2) When baby rats were sampled from 10 to 25 days of age, the hematocrits, erythrocyte counts and O2 capacity values of those nursed by anoxic mothers were greater than those nursed by the normal mothers. The total hemoglobin content of baby mice nursed by anoxic mothers generally exceeded those obtained from mice nursed by normal mothers.

3) The evidence indicates that a substance is present in the milk of anoxic rats and mice which produces an increased hemoglobin content of the nursing young.
The Chemical Composition of Normal Human Red Blood Cells, including Variability among Centrifuged Cells
Authors: KEITEL HANS G. , BERMAN H. , JONES H. , MACLACHLAN E.
Published: October 2019

1. Red cells from different layers of centrifuged cells vary in composition. Cells obtained from the upper layer, which is relatively richer in reticulocytes, contain more water, sodium, potassium, chloride and phosphorus than the remaining cells.

2. The direct method of analysis of red blood cells using a constricted type centrifuge tube to separate the entire red cells sample from buffy layer cells and from plasma avoids the errors in direct analysis caused by different cell population in upper and lower layers of centrifuged cells and the cumulative errors inherent in indirect analysis.

3. Using the direct method and a constricted type centrifuge tube, the means and standard deviations of the water and mineral content of the erythrocytes and plasma of 11 normal males and 11 normal females were determined. Males were found to have a higher sodium content of red cells and plasma.

4. The sum of the molal concentrations of sodium, potassium, chloride and phosphorus in red cells is not always equal to the sum of the molal concentrations of these minerals in the plasma.
In Vitro Studies of Erythropoiesis
Authors: THOMAS E. D.
Published: October 2019

The effect of various oxygen tensions on the in vitro synthesis of heme by rabbit bone marrow was measured. At levels above 4 per cent oxygen there was no effect of oxygen tension on heme synthesis. Total anoxia stopped heme synthesis completely. No level of oxygen tension was found to stimulate heme synthesis.
Erythropoietic Function in Dilution Anemia
Authors: ERSLEV ALLAN J.
Published: October 2019

For a period of four days, four rabbits received intravenously at 12-hour intervals large amounts (25 ml./Kg.) of a 6 per cent solution of dextran. This caused a pronounced dilution anemia. However, no increase in erythropoietic activity was observed despite the reduced oxygen content of each unit of arterial blood.

Since hypervolemia and dextran per se were found not to interfere with red cell production it was concluded that the increased cardiac output in dilution anemia compensates for the reduction in the oxygen content of each unit of arterial blood.

Consequently, the stimulus to erythropoietic function must depend on the tissue tension of oxygen rather than on the oxygen content of arterial blood.
The Influence of Chemotherapy on Survival in Acute Leukemia
Authors: HAUT ARTHUR , ALTMAN STANLEY J. , CARTWRIGHT G. E. , WINTROBE M. M.
Published: October 2019

One measure of the effectiveness of therapy in the treatment of acute leukemia is its influence on longevity. Median survival was calculated for 103 of our own cases and 782 cases published in the literature. These were divided into treated and untreated groups. Separate analyses were made for several modes of therapy.

It was not possible to demonstrate that the newer therapies (folic acid antagonists, ACTH, cortisone and 6-mercaptopurine) significantly increased the total survival in these representative groups. However, in those who achieved at least one complete remission, the median survival had been enhanced from four and a half to eight months, a difference which was found to be statistically significant.

Among the reasons for this rather surprising finding is the possibility that the use of chemotherapeutic agents may fail to alter the course of acute leukemia in some instances while prolonging it in others; the fact that of the series suitable for analysis which were found in the literature, only one was of a more recent date than 1952, although publications up to May 1954 were reviewed; and finally, that the use of 6-mercaptopurine was not adequately represented in the survey.
Bortezomib Demonstrates Superior Efficacy to High-Dose Dexamethasone in Relapsed Multiple Myeloma: Final Report of the APEX Study.
Authors: Richardson Paul , Sonneveld P. , Schuster M. , Irwin D. , Stadtmauer E. , Facon T. , Harousseau J. , Ben-Yehuda D. , Lonial S. , Goldschmidt H. , Reece D. , Miguel J. San , Blade J. , Boccadoro M. , Cavenagh J. , Dalton W. , Boral A. , Schenkein D. , Anderson K.
Published: October 2019

Introduction/Methods: This international phase 3 study conducted at 93 sites is the largest randomized study to date in relapsed multiple myeloma (MM). Patients (pts) had to have received 1–3 prior therapies and those with dexamethasone (Dex) refractory disease were excluded. Pts were randomized to bortezomib (VELCADE®) 1.3 mg/m2 IV d 1, 4, 8, 11 q3wk for 8 cycles followed by 3 cycles on d 1, 8, 15, 22 q5wk, or Dex 40 mg PO d 1–4, 9–12, 17–20 q5wk for 4 cycles followed by 5 cycles on d 1–4 every 28 d. The 1° endpoint was time to progression (TTP); 2° endpoints included survival, response rate (CR+PR) using EBMT criteria, duration of response, time to response (TTR), time to skeletal events, infections ≥gr 3 and safety; exploratory endpoints included pharmacogenomics and quality of life. Pts with progressive disease on Dex were eligible to crossover to bortezomib in a companion study.

Results: 669 pts were randomized and baseline characteristics were balanced in both arms. Based on a median follow-up of 8.3 mo, bortezomib demonstrated a 78% improvement in median TTP (hazard ratio 0.55, P<0.0001) and a significant 1-y and overall survival benefit over Dex (hazard ratio 0.53, P=0.0005; hazard ratio 0.57, P=0.0013, respectively). The response rate (CR+PR) was significantly higher (P<0.0001) with CR/nCR rates of 13% for bortezomib vs 2% for Dex. 132 (40%) and 119 (35%) pts received bortezomib or Dex 2nd line, respectively. Importantly, in 2nd-line bortezomib vs Dex, median TTP was 7.0 v 5.6 mo, 1-y survival was 89% vs 72%, and CR+PR rate was 45% vs 26%, respectively. The incidence of ≥ gr 3 adverse events and the discontinuation rate were similar across treatments in all pts.

Conclusion: This is the first and largest randomized study demonstrating a survival advantage in relapsed MM, with single-agent bortezomib proving superior to Dex based on TTP, survival, and response. Superiority was also observed in 2nd line and later salvage therapy. The safety profiles of bortezomib and Dex were predictable and toxicities were manageable.

Efficacy Bortezomib Dex P (n=333) (n=336) NC = not calculated. Median TTP, mo 6.2 3.5 <0.0001 1 prior line 7.0 (n=132) 5.6 (n=119) 0.0021 > 1 prior line 4.9 (n=200) 2.9 (n=217) <0.0001 1-year survival, % 80 66 0.0005 1 prior line 89 72 0.0098 > 1 prior line 73 62 0.0109 CR, % (n/N) 6 (20/315) 1 (2/312) 0.0001 1 prior line 6 (8/128) 2 (2/110) 0.0842 > 1 prior line 6 (12/187) 0 (0/202) 0.0002 CR/PR, % (n/N) 38 (121/315) 18 (56/312) <0.0001 1 prior line 45 (57/128) 26 (29/110) 0.0035 > 1 prior line 34 (64/187) 13 (27/202) <0.0001 Median TTR (CR/PR), d 43 43 NC
Treatment with Pegylated Interferon α (PegIntron) for High-Risk Essential Thrombocythemia: Results of a Phase II Study.
Authors: Langer Christian , Lengfelder Eva , Thiele Jurgen , Kvasnika Hans M. , Pahl Heike , Matzdorff Axel , Stade Barbara , Schauer Stefanie , Gisslinger Heinz , Dohner Hartmut , Griesshammer Martin
Published: October 2019

In high risk Essential Thrombocythemia (ET) there is a indication for cytoreductive therapy. However, especially in younger patients long-term use of alkylating agents or hydroxyurea is a matter of concern. Modification of the pharmacokinetic profile of interferon α, a non-leukemogenic agent, through addition of a polyethylenglicol (pegylation) has resulted in slower absorption and lower elimination rate, thus enabling a weekly application with potentially increased compliance compared to conventional interferon α.

In this phase II study we evaluate the safety, tolerability and efficacy of pegylated Interferon α (PegIntron, Essex Pharma) in high risk essential thrombocythemia. PegIntron is administered subcutaneously at a starting dose of 50 μg weekly. In patients not achieving a platelet count < 500 G/l after 8–12 weeks, the dosages can be increased by 25 μg/week up to 150 μg/week. Between 12/01 and 09/03 a total of 36 high risk ET patients (either platelet > 1500 G/l, or age> 60 years, or previous ET related complications) were enrolled by a total of 16 centers (16 male and 20 female patients, median platelet count at diagnosis 900 G/l, median age at diagnosis 54 years [range: 24–72 years]). Treatment was indicated because of at least one of the following reasons: Age > 60 years (38%), platelet count > 1500 G/l (36%) or previous ET related complications (50%). 10 patients (28%) were previously treated (hydroxyurea: 8 patients, anagrelide: 2 patients).

In this analysis there are 35 of 36 patients evaluable, 1 patient was lost to follow-up. Of these 35 patients, 24 (69%) are still on therapy with PegIntron after a median observation time of 18 months. The individual dosages of PegIntron are shown in table 1. A platelet count < 500 G/l was achieved in 57% of the patients after 3 months and in 75% and 89% after 6 and 12 months, respectively.

In 9 patients (26%) therapy was stopped because of drug related side effects (alopecia: 2 patients, arthralgia: 2 patients, flu-like symptoms: 2 patients, psoriasis: 1 patient, depression: 1 patient, diarrhea: 1 patient). In one patient an alternative cytoreductive therapy was initiated because of an insufficient platelet control. One patient suffered from a cerebral stroke and subsequently was taken off PegIntron application. After a median observation time of 18 months, we observed one potentially ET related complication (stroke) during PegIntron therapy so far.

These preliminary data indicate the efficacy of PegIntron in the treatment of high risk ET at relatively low doses and reasonable toxicity and safety compared to conventional interferon α.
A New Model to Evaluate Signaling of Raf in Hematopoietic Cells.
Authors: Okuda Keiko , Sonoda Yoshiaki , Griffin James D.
Published: October 2019

The Raf/MEK/ERK pathway is thought to be critical in mediating cell survival and proliferation by cytokine receptors. However, the exact contribution of Raf is complex and not well understood. A better understanding of Raf signaling is important because of the recent observation that B-Raf is frequently mutated in various human cancers. We have generated a new model system that activates Raf directly by linking the extracytoplasmic and transmembrane domains of the erythropoietin receptor (EPOR) with the catalytic domain of Raf (CR3). This synthetic oncogene in which dimerization can be controlled by an exogenous ligand, is fixed at the cellular membrane, while the endogenous Raf is normally activated by binding with Ras. The chimeric receptor (EPOR/CR3) was stably expressed in Ba/F3 cells which lack EPO receptors, and eight independent sublines were established. Although the lines remained dependent on IL-3 for proliferation, EPO treatment reduced the rate of cell death in the absence of IL-3. Also, EPO was synergistic with sub-optimal concentrations of IL-3 in inducing long-term cell proliferation, but did not augment proliferation of cells cultured with full concentrations of IL-3. EPO induced a rapid activation of ERK and also phosphorylation of endogenous Raf. It also induced tyrosine phosphorylation of several cellular proteins, estimated molecular masses of 150, 130, 110, 95, 60 and 42 kD. Many of the prominent targets of the wild type IL-3 or EPO receptors, such as Shc, Stat1, Stat3 and Stat5 were not tyrosine phosphorylated by the EPOR/CR3 receptor. The MEK1 inhibitor PD98059 reduced EPO-induced tyrosine phosphorylation, suggesting these substrates are downstream of MEK kinase. Interestingly, PD98059 also reduced the phosphorylation of endogenous Raf, indicating there is a positive feedback mechanism in Raf activation. We conclude that Raf can be activated by a mechanism that induces clustering at the cell membrane, and that this leads directly to activation of MEK and ERK. This is insufficient to induce proliferation by itself, but can add to sub-optimal growth signals from another receptor. This EPOR/CR3 system may serve as a useful model to evaluate the effects of signal transduction inhibitors for Raf as a target.
COAT Platelet Formation Is P2Y12-Dependent.
Authors: Kempton Christine L. , Hoffman Maureane , Lenkowski Angie , Roberts Harold R. , Monroe Dougald M.
Published: October 2019

Inhibition of the platelet ADP receptor, P2Y12, has been shown to reduce thrombin-stimulated exposure of platelet phosphatidylserine and tissue factor (TF)-dependent thrombin generation in human platelet rich plasma (ATVB2003;23(10):1941–1947). Platelets stimulated by the combination of convulxin (Cvx) and thrombin develop two populations of platelets, one with low coagulation factor binding and a second with high levels of coagulation factor binding (Blood2000;95(5):1694–1702). Platelets in the population with increased coagulation factor binding have been termed COAT platelets. Previously we have demonstrated that the percentage of COAT platelets correlates with increased thrombin generation on Cvx plus thrombin-stimulated platelets compared to platelets stimulated with thrombin alone (Blood2003; 102(11): 293a). In this study we investigated whether inhibition of the platelet P2Y12 receptor reduced the development of the COAT platelets in addition to reducing thrombin generation.

METHOD: A well established cell-based model of hemostasis was used. Components included: LPS-stimulated monocytes expressing TF, coagulation factors (II, V, VII, VIII, IX, X, XI) at plasma concentrations, coagulation inhibitors (AT, TFPI) at plasma concentrations, platelets from normal donors, and calcium chloride. Thrombin generation in the system was measured by cleavage of a chromogenic substrate and compared to a standard curve. Model systems without the addition of Cvx (20 ng/ml) were compared to model system with the addition of Cvx with and without a 2-methylthiol AMP (2MeSAMP, an inhibitor of the P2Y12 receptor). To determine the percentage of COAT platelets formed, samples were removed at 9.5 minutes after the model system was initiated, fixed in 1% paraformaldehyde, and analyzed on FACScan flow cytometer for binding of factor V using a polyclonal primary and secondary.

RESULTS: In the absence of 2MeSAMP approximately 30% of Cvx plus thrombin-stimulated platelets were COAT platelets. In the presence of 2MeSAMP, none of the platelets were COAT platelets. 2MeSAMP led to a significant reduction in the peak and rate of thrombin generation in the presence of Cvx but no change in the total amount of thrombin generated as measure by the area under the curve.

CONCLUSIONS: Inhibition of the platelet P2Y12 receptor prevents COAT platelet formation. The absence of COAT platelet formation correlates with a reduction in both the rate of thrombin generation and the peak of thrombin generated. COAT platelet formation and up-regulated thrombin generation induced by the combination thrombin plus moderate concentrations of Cvx are P2Y12-dependent.
The Addition of Rituximab to Combination Chemotherapy with CHOP Has a Long Lasting Impact on Subsequent Treatment in Remission in Follicular Lymhoma but Not in Mantle Cell Lymphoma: Results of Two Prospective Randomized Studies of the German Low Grade Lymphoma Study Group (GLSG).
Authors: Hiddemann Wolfgang , Forstpointner Roswitha , Kneba Michael , Schmitz Norbert , Schmits Rudolf , Metzner Bernd , Reiser Marcel , Lengfelder Eva , Woermann Bernhard , Harder Hariett , Hegewisch-Becker Susanna , Berdel Wolfgang , Hess Georg , Eimermacher Hartmut , Aldaoud Ali , Planker Manfred , Parwaresch Reza , Dreyling Martin , Unterhalt Michael
Published: October 2019

The addition of Rituximab (R) to combination chemotherapy has been shown to increase the remission rate (RR) and to prolong the time to treatment failure (TTF) both in follicular lymphoma (FL) and mantle cell lymphoma (MCL). The impact of the R-chemo combination on subsequent therapy in remission, however, remains unclear. The GLSG embarked on two parallel studies in FL and MCL comprizing a prospective randomized comparison of R-CHOP versus CHOP alone followed by a second randomization in remission for Interferon alpha maintenance (IFN) versus myeloablative radio-chemotherapy with subsequent stem cell transplantation (PBCT) in patients < 60 yrs. of age while all older pts. received IFN maintenance. In 428 pts. with FL R-CHOP revealed a significantly higher RR (96% vs. 90%, p = 0.011) and a longer TTF (median not reached vs. 2.6 yrs, p<0.0001). From 347 patients evaluable for subsequent treatment in remission 79 younger patients (35 after CHOP and 44 after R-CHOP) received PBCT. 52 younger patients after CHOP and 69 after R-CHOP received IFN maintenance. 122 older cases received IFN (59 after CHOP and 63 after R-CHOP). For IFN treated pts. a significantly longer progression free survival (PFS) was observed after initial therapy with R-CHOP (estimated 2 yrs - PFS: 63% vs. 84%, p = 0.0004) while no differences were encountered at the time of this analysis after PBCT (estimated 2 yrs - PFS for the whole group: 86%). Hence, in FL initial therapy with R-CHOP appears to have a long lasting beneficial effect on PFS which may be in the range that has previously only been achieved by chemotherapy followed by PBCT. Similar to FL a significantly higher RR (94% vs. 75%, p = 0.005) and a longer TTF (median 14 months vs. 21 months, p = 0.0131) was observed in 122 pts. with MCL undergoing the same randomized comparison. In contrast to FL, however, no differences were revealed for the PFS after R-CHOP versus CHOP and subsequent therapy with IFN or PBCT. These data show a differential effect of adding R to CHOP for initial therapy in FL and MCL. In FL the addition of R to CHOP has a long lasting beneficial effect with a substantial impact on subsequent treatment in remission while in MCL the benefit of R seems restricted to the remission induction period only.