bluuebunny/crossref_metadata_2025 huggingface dataset:
HSC Hierarchy in Primary Myelofibrosis
Authors: Triviai Ioanna N , Zeschke Silke , Panagiota Victoria , Heuser Michael , Stocking Carol , Kroeger Nicolaus
Published: October 2019

Primary Myelofibrosis (PMF) is a Myeloproliferating Neoplasm (MPN) of hematopoietic stem cell origin, characterized by expansion of aberrant myeloid progenitors in the chronic phase that can lead to bone marrow fibrosis development and/or osteosclerosis. In 20-25% of PMF cases transformation to acute myeloid leukemia (AML) is observed. Identification of multiple molecular lesions suggests complex clone dynamics that indicates exceeding sub-clone dominance as PMF progresses. Our aim is to determine the HSC hierarchy orchestrating initiation and development of PMF.

In our previous studies we reported a CD133+ HSC population in PMF peripheral blood that represents the aberrant long-term stem cell fraction responsible for PMF reproduction of chronic and acute phases in vivo. Molecular analysis of PMF xenografts indicates sustenance of genetically different HSC clones exhibiting variation in both their engraftment potential and reproduction of PMF parameters in the first mouse model of the disease.

To further characterize the succession of molecular lesions determining HSC clone propagation we performed targeted exon sequencing of PMF HSC from 100 PMF patients for 54 genes. Mutations in the epigenetic regulators ASXL1 and EZH2 were detected in 38% and 15% of PMF patients respectively. HSC clonal evolution was determined by single cell molecular and phenotypic analysis in vitro and graft analysis in vivo.

Mutations detected in the epigenetic regulators ASXL1 and EZH2 represent founding molecular lesions at the top of PMF HSC hierarchy. ASXL1 mutations precede and are connected with sustenance of clonal hematopoiesis without any significant influence on the HSC differentiation potential. EZH2 mutations are connected with impaired erythropoiesis in vitro and anemia, high engraftment and expansion of pre-leukemic clones in vivo. Occurrence of JAK2 and CALR mutations in the background of mutated epigenetic regulator genes is connected with expansion of HSC subclones and reproduction of chronic phase PMF as atypical myelopoiesis, splenomegaly and induction of fibrosis.

Our results indicate genetic heterogeneity of PMF neoplastic HSC is comprised of three different mutational clusters. Mutations in epigenetic regulator genes (Group 1) precede and shape the epigenetic landscape conferring genetic instability to sustain the expansion of pre-leukemic clones (Group 3 mutations). JAK2 and CALR mutations (Group 2) occur later on and are connected with aberrant myelopoiesis of chronic phase disease. HSC clonal dynamics reflect genotype related phenotypes as determinants of chronic and acute phases of PMF.

Disclosures

No relevant conflicts of interest to declare.
KIR Gene Haplotype: An Independent Predictor of Clinical Outcome in MDS Patients
Authors: Daher May , Marin David , Sobieski Catherine , Shaim Hila , Basar Rafet , Mendt Vilchez Mayela Carolina , Gokdemir Elif , Li Li , Imahashi Nobuhiko , Muftuoglu Muharrem , Champlin Richard E. , Shpall Elizabeth J. , Kantarjian Hagop M. , Garcia-Manero Guillermo , Rezvani Katayoun
Published: October 2019

Myelodysplastic syndromes (MDS) are a group of hematopoietic disorders affecting the myeloid lineage, characterized by cytopenias and clonal evolution to AML. Immune responses against MDS, partly mediated by NK cells, have the potential to affect disease progression. However, immune evasion remains an important barrier. NK cells express both activating and inhibitory killer immunoglobulin-like receptors (KIRs) that interact to regulate NK effector function. Based on the number and distribution of inherited KIR genes, individuals can be classified in two broad haplotypes. Haplotype A comprises a single activating KIR (aKIR) gene (KIR2DS4), while haplotype B incorporates various combinations of aKIR genes (up to 6). We hypothesized that the aKIR gene repertoire may be useful in refining predictions of clinical outcomes in MDS. Thus, we studied the variations in aKIR gene content and haplotype in MDS and their relationship to the risk of AML transformation and patient outcomes.

We first compared the number of aKIR genes in 108 MDS patients treated at MDACC with that in 139 HSC donors. The median number of aKIR genes was significantly lower in MDS than controls: 2 (range 0-6) vs 3 (range 0-6), p=0.001. Functional studies revealed that compared to healthy controls, NK cells from MDS patients demonstrate less Interferon gamma production (p<0.0001) and less degranulation (p=0.0028) in response to K562 cell line, and have lower killing ability evidenced by chromium release assay (p=0.04).

We next examined the influence of KIR haplotype on the risk of AML transformation and outcomes in the cohort of 108 MDS patients (28 KIR haplotype A and 80 KIR haplotype B patients). On multivariate analysis, cytogenetic risk group and KIR haplotype were identified as independent predictors of MDS progression to AML. The relative risk of MDS-AML transformation in patients with haplotype A vs. haplotype B was 2.67 (1.13-6.31) (p=0.02). Similarly, cytogenetic risk group, IPSS and KIR haplotype independently predicted survival. MDS patients with KIR haplotype A had worse adjusted PFS (RR with 95%CI 2.96 [1.59-5.52], p=0.001) and OS (2.25 [1.17-4.31], p=0.02), compared to patients with haplotype B.

These novel findings may help identify a subgroup of MDS patients with a high risk of disease progression and poor outcomes, who would likely benefit from adoptive NK cell therapy.

Disclosures

Kantarjian: ARIAD: Research Funding; Bristol-Myers Squibb: Research Funding; Amgen: Research Funding; Pfizer Inc: Research Funding; Delta-Fly Pharma: Research Funding; Novartis: Research Funding.
TP53 Mutations and Outcome in Patients with Myelodysplastic Syndromes (MDS)
Authors: Al-Issa Karam , Sekeres Mikkael A. , Nielsen Alek d , Jha Babal , Przychodzen Bartlomiej P , Aly Mai , Carraway Hetty E. , Advani Anjali S. , Patel Bhumika , Clemente Michael J. , Mukherjee Sudipto , Gerds Aaron T. , Maciejewski Jaroslaw P , Nazha Aziz
Published: October 2019

Objectives:

TP53, a tumor suppressor gene, is frequently mutated in myeloid malignancies. The negative impact of TP53 mutations in myelodysplastic syndromes (MDS) has been described previously but there is controversy regarding the prognostic impact of the mutation's characteristics (location, type, passenger vs. driver, and others).

Methods:

We sequenced DNA samples from 732 patients (pts) with MDS and related myeloid malignancies for the presence of TP53 mutations and 61 other genes that have been described as recurrently mutated in MDS. Overall survival (OS) was measured from the time of diagnosis to time of death or last follows up. Variant allele frequencies (VAFs) adjusted by zygosity were used to define clonal architecture of driver clones.

Results:

Of 80 mutations detected in 73 (10%) pts, 66 (88%) were missense, 7 (9%) were nonsense, and 7 (9%) were frame shift deletions/insertions. Pts with TP53 mutations had a higher WBC (4.6 vs. 3.9 X 109/L, p = .04), higher bone marrow blast % (median 9 vs. 3, p =0.1), and a higher risk category by revised International prognostic Scoring System (56% vs. 27%, p =.01) compared to pts with TP53 wild type. TP53 mutations were commonly occurred with TET2 (16%), PRPF8 (13%), ASXL1 (11%), DDX54 (8%), DNMT3A (8%), and IDH2 (8%). The mean VAF for TP53 was 41.9 (5-100). TP53 mutations were defined as drivers in 20% of samples, passengers in 40%, and mosaic in 40%. Mutation positions included: 19 (24%) in the DNA binding domain, 2 (3%) in the transactivation domain, 1 (1%) in the tetramerization domain and 58 (72%) other. With a median follow up of 16.4 months, the median OS for the entire group was 8.24 months. Patients with TP53 as driver mutations had a worse OS compared with patients with TP53 as passenger mutations (median, 2.2 vs. 13.0 months, respectively, p =.02). Similarly, OS by TP53 VAF categorized as low (<25%), intermediate (25-50%), and high (>50%) was 12.4, 8.5, and 3.4 months, respectively. Among patients with available treatment data, 29 patients were treated with the hypomethylating agents, azacitidine or decitabine, 17% responded (60% CR, and 40% PR), and OS was similar compared to patients treated with other therapies. Patients treated with Hematopoietic stem cell transplant (HCT) had superior OS compared to pts not receiving HCT (median, 14.9 vs. 8.9 months, respectively, p =.05).

Conclusions:

TP53 mutations are associated with poor outcome in MDS, but the outcome varies depending on the type of mutation and VAF. Treatment with HCT remains a valid treatment option in a subset of patients but novel treatment strategies are desperately needed.

Disclosures

Mukherjee: Novartis: Consultancy, Honoraria, Research Funding; Ariad: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria, Research Funding.
Congenital Hemolytic Anemia with Abnormal Pigment Metabolism and Red Cell Inclusion Bodies: a New Clinical Syndrome
Authors: LANGE ROBERT D. , AKEROYD JOSEPH H.
Published: October 2019

1. A case report and special studies of a 14-year-old girl with a congenital hemolytic anemia are reported.

2. Fourteen per cent of her erythrocytes contained unusual inclusion bodies.

3. In addition, the child has been known to pass dark urine since the age of 2½ years. The pigment probably belongs to the bilifuscin and mesobilifuscin group.

4. It is believed that the syndrome is probably caused by an inborn error in erythrocytic metabolism.

5. It has been proposed that the syndrome be named "congenital hemolytic anemia with abnormal pigment metabolism and red cell inclusion bodies."
Rate of Production of P32-Labeled Lymphocytes
Authors: PERRY SEYMOUR , CRADDOCK CHARLES G. , VENTZKE LUTZ , CREPALDI GAETANO , LAWRENCE JOHN S.
Published: October 2019

1. Lymphocytes in dogs have been labeled by means of the DNA-P32 incorporation technic.

2. In the thoracic duct, a high level of lymphocyte DNA-P32 specific activity is reached within two hours. After 24 to 30 hours, the activity falls to a lower level which is then maintained for at least 160 hours.

3. Lymphocyte-granulocyte separations on the blood revealed that DNA-P32 specific activity reached a peak on the fourth day for both lymphocytes and granulocytes.

4. These findings are consistent with a short maturation time for the lymphocyte and a short intravascular time.
Multiple Hemangiomata Associated with Thrombocytopenia: Remarks on the Pathogenesis of the Thrombocytopenia in this Syndrome
Authors: GILON ELIAHU , RAMOT BRACHA , SHEBA CHAIM
Published: October 2019

1. A 13-year-old girl with multiple hemangiomata and thrombocytopenia is described.

2. Complications of surgical treatment included severe bleeding, septicemia and a fibrinolytic event.

3. Simultaneous platelet counts from the tumor and peripheral blood were performed and showed a significantly higher concentration of platelets in the tumor vessels.

4. With the rise in the platelet count, a decreased platelet fragility as well as a persistently abnormal prothrombin consumption could be demonstrated.

5. The possible role of the tumor in the pathogenesis of the thrombocytopenia is discussed.
Some Factors Involved in the Effect of X-Irradiation on the Phosphatase Activity of Hematopoietic Tissues
Authors: UYEKI EDWIN M. , SALERNO PAUL R.
Published: October 2019

Factors which modify lymphoid distribution of tissues were found to modify the adenosine triphosphatase activity of these tissues. Starvation or cortisone injection, which produces destructive changes in lymphoid tissues, was found to increase the enzyme activity of spleen and thymus tissues. The greater increment of enzyme activity of the thymus as compared to that of the spleen was correlated with its normally higher content of lymphoid tissue.

The increase in adenosine triphosphatase activity of hematopoietic tissues appears to be associated with the type of cells present in the assay medium. With respect to peripheral blood leukocytes of the rat, the cell type is confined largely to lymphocytes and granulocytes. The increase in adenosine triphosphatase activity of the leukocytes after total-body x-ray was seen to parallel the increase in granulocytes present in the assay medium. The ratio of granulocytes to lymphocytes is not appreciably altered in dog peripheral blood after exposure to total-body x-ray; the adenosine triphosphatase activity similarly was not significantly altered. After total-body x-ray (390 r and 780 r), cells isolated from the rat bone marrow displayed a fivefold increase in adenosine triphosphatase activity. This increase was seen to correspond with an increase in the ratio of segmented leukocytes and reticuloendothelial cells and a decrease in the immature forms of the erythroid and myeloid cells.

The heterogeneous cell mixtures used for our assay procedures permit the observation that total-body x-irradiation results in an increased enzyme activity of the isolated cells of the peripheral blood, bone marrow and spleen tissue of the rat. The increased enzyme activity was associated with the increased ratio of cells with high enzyme activity present in the assay medium.
Hematologic Changes in Rat Following Administration of Desoxypyridoxine, Aminopterin, Oxythiamine and 6,7-Dimethyl-9-(2'-acetoxyethyl)isoalloxazine
Authors: DOCTOR V. M.
Published: October 2019

1. Administration of aminopterin to rats at a level of 100 γ/Kg./day for a 10 day period produced leukopenia which was rapid in onset and led quickly to death, whereas the leukopenia produced by the administration of desoxypyridoxine at the level of 40 mg./Kg./day for a 2 week period or at 100 mg./Kg./day for a 4 week period was not accompanied by toxic effects such as loss of appetite, diarrhea and hemorrhage and was characterized by lymphocytopenia and granulocytosis.

2. The results of combining the administration of aminopterin (70 γ/Kg./ day) with desoxypyridoxine (40 mg./Kg./day) for a 2 week period indicated an additive effect of the two in the production of lymphocytopenia; the granulocytosis resulting from desoxypyridoxine administration was counter-acted by the granulocytopenic effect of aminopterin. Also, the leukopenia produced by the combined administration of aminopterin and desoxypyridoxine was not accompanied by the toxic effects which follow aminopterin administration.

3. Oxythiamine alone or in combination with desoxypyridoxine or aminopterin did not influence the leukocyte count.

4. Administration of aminopterin with 6, 7-dimethyl-9-(2'-acetoxyethyl)isoalloxazine (U-2112) produced marked lymphocytopenia and granulocytopenia accompanied by toxic effects such as loss of appetite, diarrhea and hemorrhage.

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Thrombocytopenia with a Circulating Platelet Agglutinin, Platelet Lysin and Clot Retraction Inhibitor
Authors: ZUCKER MARJORIE B. , LEY ALLYN B. , BORRELLI JENNIE , MAYER KLAUS , FIRMAT JORGE
Published: October 2019

A case is reported of thrombocytopenia developing suddenly in a patient with metastatic carcinoma. Studies of this patient’s serum revealed a platelet agglutinin presumed to be related to the development of the thrombocytopenia in spite of the failure to demonstrate autoagglutination. In addition, a technique is described for the detection of a platelet lysin and clot retraction inhibitor, both of which could be demonstrated in this patient’s serum. The identity of the lysin and retraction inhibitor is suggested.

Studies on the serums of other patients showed a dissociation between the occurrence of platelet agglutinin on the one hand and of platelet lysin and retraction inhibitor on the other.
Behavior of Total Serum Complement in Hodgkin’s Disease and Other Malignant Lymphomas
Authors: ROTTINO ANTONIO , LEVY ARTHUR L.
Published: October 2019

Using the method of Mayer and collaborators for the determination of complement in the blood serum, 67 normal persons, 72 patients with Hodgkin’s disease, 28 with other types of lymphoma and 65 with cancer involvement of various organs were studied.

Fifty-eight of 72 Hodgkin’s disease patients were found to have an elevated level of complement, 12 a normal level and 2 a below-normal level. Elevated levels were found also in patients with other lymphomas and cancer.

An association between elevated complement, increased sedimentation level, decreased serum properdin, positive C-reactive protein and elevated beta globulin in the serum was noted.

The conclusion reached is that an elevation of complement in the serum indicates that Hodgkin’s disease is in an active phase. As to the nature of the disease, the statistics assembled seem to give no clue.
Distribution of Abnormal Hemoglobins in Puerto Rico and Survival Studies of Red Blood Cells Using Cr51
Authors: SUAREZ RAMON M. , BUSO ROBERTO , MEYER LEO M. , OLAVARRIETA S. T.
Published: October 2019

1. A survey of the incidence of abnormal hemoglobins in different racial groups distributed over the island of Puerto Rico was performed. The relation of the rate of destruction of red cells to the presence of abnormal hemoglobin patterns in the affected population was also studied.

2. The abnormal hemoglobins were classified by the difference in paper electrophoretic mobility. Fetal hemoglobin was measured by its resistance to alkaline denaturation. The red cell life span was determined by measuring the survival of erythrocytes labelled with radioactive sodium chromate.

3. A total of 2,089 inhabitants were studied. There were 1,487 white subjects and 602 Negroes. Forty-two individuals were found to harbor abnormal hemoglobins. All but one were Negroes or Negroid of African descent, and their relative numbers agreed closely with the geographical distribution of ethnic groups in the island. Abnormal hemoglobins were found in 2.01 per cent of the entire series of 2,089 persons, but in those considered Negroes or Negroids the incidence of abnormal hemoglobins was 6.8 per cent.

Of the 42 persons showing abnormal hemoglobins, thirty-four or 81 per cent, had hemoglobin S; only two of these had sickle cell anemia. The incidence of the sickle cell trait among the Puerto Rican Negro population was 5.2 per cent; and the incidence of hemoglobin S disease among those harboring the trait was 5.9 per cent.

Eight, or 19 per cent of the abnormal cases had hemoglobin C; only one of these had hemoglobin C disease with clinical hemolytic anemia. The incidence of the hemoglobin C trait among the Puerto Rican Negro population is 1.3 per cent; and the incidence of hemoglobin C disease among those harboring the trait may reach 12.5 per cent.

4. All 42 cases harboring abnormal hemoglobins showed very small quantities of fetal or alkali-resistant hemoglobin ranging from 0.45 per cent to 3.25 per cent, averaging 1.12 per cent.

5. The "apparent" half-life of the red blood cells was found to be 10 days in sickle cell anemia, 7 days in SC disease, 20.6 days in cases of hemoglobin SA, 18.5 days in one case of hemoglobin C disease and 21.5 days in 2 cases of the combination CA. The normal "apparent" half-life in our laboratory is 24.5 days.
Leukokinetic Studies. I. A Method for Labeling Leukocytes with Diisopropylfluorophosphate (DFP32)
Authors: ATHENS J. W. , MAUER A. M. , ASHENBRUCKER HELEN , CARTWRIGHT G. E. , WINTROBE M. M.
Published: October 2019

1. A method has been presented by which granulocytes can be labeled in vivo with diisopropylfluorophosphate containing radioactive phosphorus. The leukocytes are isolated from blood by dextran sedimentation of erythrocytes and are then treated with gramicidin and lysolecithin to remove remaining red cells. Platelets are removed by differential centrifugation. The isolated leukocytes are placed between two squares of scintillating plastic and counted with a scintillation counter.

2. Leukocytes essentially free of erythrocytes and platelets can be obtained by the method outlined. The efficiency of the plastic scintillation counting method for radioactive phosphorus is about 74 per cent and leukocyte samples obtained from 20 ml. samples of normal blood can be counted with a reproducibility of ±10 per cent.

3. The administration of 2 mg. of diisopropylfluorophosphate either intramuscularly or intravenously is without significant toxic side effects.

4. No evidence has been obtained that the label damages the leukocytes.

5. No evidence has been obtained that the label elutes from leukocytes under the conditions of these studies.

6. Diisopropylfluorophosphate labels granulocytes for a brief period of time following injection. The label is not reutilized after death of the cells.
What Are Normal Blood Values for Leukemia in Remission?
Authors: HYMAN CAROL B. , BORDA C. EDUARDO , DIGUMARTHI GOPALASWAMI
Published: October 2019

1. The values for hemoglobin concentration, total white blood cells, granulocytes, lymphocytes and monocytes at the midpoint of 78 drug-induced remissions in a group of children with acute leukemia have been presented. These values have been compared with those from a group of normal children, and the differences analyzed statistically.

2. Statistical analysis shows that, in general, blood values in remission are reduced by the particular agent employed. White blood cells are more markedly affected than hemoglobin.

3. The effects are essentially the same with 6-M.P. given alone or in combination with azaserine or DON.

4. The reduction in values is greatest with 6-M.P., alone or in combination, less with amethopterin and least with prednisone.

5. The data presented indicate that criteria for complete blood remission should make allowances for the influence of the drug employed.
Congenital Hypoplastic Thrombocytopenia with Skeletal Deformities in Siblings
Authors: SHAW SIDNEY , OLIVER R. A. M.
Published: October 2019

The occurrence of congenital hypoplastic thrombocytopenia with absent radii is described in siblings.

Both presented with hemorrhagic manifestations within a few weeks of birth and the first died at the age of six and a half months.

Reference is made to two other examples in siblings.

These abnormalities are considered as due to a genetic defect and the cases are probably variants of Fanconi’s syndrome.
Total Cell Counts of the Bone Marrow of Normal Albino Rats from 1 to 50 Weeks of Age
Authors: BURKE WILLIAM T. , HARRIS CHARLES
Published: October 2019

A method is described by which the total nucleated cell count of femoral bone marrow of the rat can be estimated and cell population expressed in terms of differential counts. Normal values of total nucleated cell counts and the cellular distributions are given for seven age groups. These data indicate considerable change in bone marrow total cell population in rats one to 10 weeks of age.
Ferritin and Ferruginous Micelles in Normal Erythroblasts and Hypochromic Hypersideremic Anemias
Authors: BESSIS MARCEL C. , BRETON-GORIUS JANINE
Published: October 2019

1. All normal erythroblasts contain some iron in the ferritin form. It may be present in either a dispersed state or in compact clusters. When large enough, these clusters may be observed in the optical microscope: they are the granular particles of the siderocytes.

2. Iron can be found in the mitochondria. It may exist either in the form of ferritin granules or as ferruginous micelles.

3. In thalassemia, large quantities of iron accumulate in the erythroblasts and are even found in the erythrocytes as ferritin, in cluster formation or dispersed. Occasionally, iron is present in great quantities in the mitochondria as ferritin or micelles. It seems that the various disorders encountered in thalassemia may thus be ascertained; the disturbance in hemoglobin synthesis results in the accumulation of the unused iron in the hypochromic erythrocytes.

4. In diseases very similar to thalassemia and in which no fetal hemoglobin is found, i.e., the hypochromic-hypersideremic anemias (sidero-achestic anemia, hypochromic hypersideremic anemia, lead-poisoning ), similar findings are observed.

5. Normally, it is probable that iron metabolism occurs in the mitochondria. In thalassemia and hypochromic hypersideremic anemias, on the other hand, iron metabolism often appears to be "blocked" in the same areas.
Studies on the Biosynthesis of Heme from Iron and Protoporphyrin
Authors: SCHWARTZ H. C. , CARTWRIGHT G. E. , SMITH EMIL L. , WINTROBE M. M.
Published: October 2019

1. Additional evidence has been presented that the formation of heme in vitro from iron and protoporphyrin is enzyme dependent.

2. An enzyme solution was prepared from the active particulate matter of a chicken erythrocyte hemolysate. The enzyme is soluble in 0.15 M KCl and optimally active at pH 7.9.

3. The rate of heme synthesis was constant over the first 30 minutes and approached maximal values at 3 to 4 hours. At optimal concentrations of protoporphyrin and iron the rate of heme synthesis over the first 30 minutes was proportional to enzyme concentration.

4. The enzyme solution was 89 per cent inactivated after heating at 56 C. for 30 minutes. Optimal stability for 3 hours was at pH 7.4. It was unstable when lyophilized or on storage at 5 C. or -30 C.

5. The enzyme was inhibited by 1 x 10-2 M p-chloromercuriphenylsulfonate and 1 x 10-2 M iodoacetamide.

6. There was a 70 per cent loss of activity after dialysis of the enzyme solution for 22 hours against water. The dialyzed enzyme solution was reactivated with either reduced glutathione or cysteine.

7. The enzyme solution augmented heme synthesis in vitro from δ-amino-levulinic acid, porphobilinogen and protoporphyrin, but not from glycine.
The Proportion of Reticulocytes in the Erythrocytes of the Spleen as Compared with Those of Circulating Blood, with Special Reference to Hemolytic States
Authors: BERENDES MARGRET
Published: October 2019

1. Comparative studies of reticulocyte percentages have been made on the spleens of normal dogs, dogs with phenylhydrazine hemolytic disease, and human cases of hemolytic jaundice. Splenic artery and vein blood has been compared in some of these, both before and after adrenalin.

2. No significant differences were found, as between peripheral, splenic arterial and venous blood, with or without the use of adrenalin.

3. Imprint and "squeeze" preparations of splenic parenchyma consistently gave higher percentages ranging from 1.5 to 3.5 times the peripheral values, the most marked differences being noted in the normal dog (11.5 to 27 x).

4. The possible significance of these findings is discussed.
Pseudotumor of Hemophilia in a Patient with PTC Deficiency
Authors: SILBER R. , CHRISTENSEN W. R.
Published: October 2019

1. A case of pseudotumor of the ilium in a PTC deficient patient is reported.

2. The natural history and treatment of this serious complication of hemophilia are reviewed.
Studies on Erythropoiesis. XI. Reticulocyte Response of Transfusion-Induced Polycythemic Mice to Anemic Plasma from Nephrectomized Mice and to Plasma from Nephrectomized Rats Exposed to Low Oxygen
Authors: JACOBSON LEON O. , MARKS EDNA K. , GASTON EVELYN O. , GOLDWASSER EUGENE
Published: October 2019

Mice with transfusion-induced polycythemia have been used to assay erythropoietic activity in plasma derived from mice and rats subjected to various stimuli and experimental procedures.

1. Anemic, anoxic, or cobalt mouse plasma increased erythropoiesis from the zero baseline to about the normal range after four 0.5 cc. injections. No significant effect is observed from the same number of injections of normal rat or mouse plasma or normal saline. Similarly, plasma from rats starved for 4 days had no erythropoietic activity, but erythropoietic activity was present in the plasma of pregnant mice during the last trimester of pregnancy.

2. Plasma from mice made anemic by phenylhydrazine and then subjected to bilateral nephrectomy contained slight erythropoietic activity (less than 0.2 per cent reticulocytes). The plasma of sham-operated anemic animals had considerably more erythropoietic activity (2.0 per cent). The plasma was collected for assay 10 hours after the operative procedures.

3. Plasma from nephrectomized rats exposed to hypoxic anoxia for 8, 16, or 24 hours, had slight erythropoietic activity when measured by the peripheral reticulocyte response of the recipients (0.18, 0.11, and 0.00 per cent reticulocutes, respectively). The same plasma had no erythropoietic activity when judged by the Fe59 red cell incorporation response in starved rats.

Plasma from rats subjected to bilateral ureter ligation and similarly exposed had an erythropoietic activity (0.7, 1.38, and 1.26 per cent reticulocytes, respectively), comparable with that of the plasma from normal rats exposed to hypoxic anoxia (0.9, 1.71, and 1.78 per cent, respectively). The low and inconstant erythropoietic activity in the plasma of nephrectomized mice and rats subjected to anemic or hypoxic anoxia is more or less comparable with the response occasionally produced by normal plasma. We are, therefore, disinclined to consider the data as evidence against the renal origin of erythropoietins, but the alternative possibilities are discussed.
Hereditary Spherocytosis-Sicklemia in the Negro
Authors: MARTIN WILLIAM W. , KOUGH ROBERT H. , BRANCHE GEORGE C.
Published: October 2019

1. A case of coexistent hereditary spherocytosis and sicklemia in a Negro female is reported. Hematologic and clinical amelioration followed splenectomy.

2. Study of three generations of the family of the propositus revealed one additional case of hereditary spherocytosis-sicklemia and 10 cases of hereditary spherocytosis in the Negro.

3. Further evidence is adduced that the combination of this heterozygous hemoglobinopathy with the red blood cell defect of hereditary spherocytosis produces an illness which is not grossly different from hereditary spherocytosis associated with normal hemoglobin.
The Mechanism of the Anemia Associated with Hodgkin’s Disease
Authors: GIANNOPOULOS P. PIKOULI , BERGSAGEL DANIEL E.
Published: October 2019

1. The survival of autotransfused Cr51-tagged erythrocytes was shortened in a group of five patients with the anemia of Hodgkin’s disease, but since the normal marrow is believed to be capable of compensating for destruction rates of six to eight times greater than normal, it is felt that the increased rate of hemolysis was not sufficient to account for the development of anemia in any of these patients.

2. There was evidence of an increased rate of erythropoiesis in all of the patients with Hodgkin’s disease. This was manifested by normoblastic hyperplasia of the marrow, a moderate reticulocytosis and increased plasma iron clearance rates. The shortened red cell survival times were associated with the increased plasma iron clearance rates. However, in view of the fact that all of the patients were anemic, it is evident that these patients were incapable of increasing the rate of red cell production sufficiently to compensate for the increased rate of destruction.

3. Intravenously injected Fe59 citrate was incorporated more rapidly into the circulating red cell mass in the patients with Hodgkin’s disease than in the normal subjects.

4. The tissue iron stores of the liver and spleen were greatly increased, but no iron was demonstrated in the marrow of a group of seven patients with Hodgkin’s disease.

5. The plasma iron was low and the UIBC normal in a group of 12 patients with active Hodgkin’s disease.

6. It is suggested that one of the factors which limits the ability of the marrow to produce hemoglobin in the anemia of Hodgkin’s disease may be a relative hypoferremia caused by a defect in the mobilization of iron from tissue stores.
A Review of Lesions in the Optic Fundus in Various Diseases of the Blood
Authors: MARSHALL RICHARD A.
Published: October 2019

1. The frequency of retinopathy as seen on an active hematologic service in a two year period is reported. The report is based on the findings in 50 patients who had retinopathy associated with a blood disorder.

2. The occurrence of retinopathy in the course of leukemia did not significantly alter the mortality or duration of illness.

3. Anemia, thrombocytopenia, septicemia and immaturity of the leukocytes appear to be the most important factors in the production of retinopathy in hematologic diseases.

4. The various retinal lesions are illustrated in photographs. Perivascular sheathing was seen only in chronic granulocytic leukemia. Otherwise, retinopathy was not specific for any particular blood disorder.
A Mobile Retroclavicular Lymph Node of Some Clinical Significance
Authors: TERRY R. BARRATT
Published: October 2019

The only clinically enlarged peripheral lymph node in three patients was one lying hidden deep behind the medial end of the clavicle. Its presence was established by instructing the patient to cough and strain; this dislocated the node up into the supraclavicular fossa, thus allowing biopsy.
Congenital Factor VII Deficiency with Normal Stuart Activity: Clinical, Genetic and Experimental Observations
Authors: KUPFER HENRY G. , HANNA BERTRAM L. , KINNE DIXIE R.
Published: October 2019

The case of a four year old white girl with a bleeding tendency characterized by the spontaneous occurrence of petechiae, ecchymoses and hemarthroses is presented. Laboratory studies indicate a deficiency of factor VII, with normal levels of all other clotting components, and a vascular deficiency possibly related to frequent upper respiratory infection.

Serum from the patient and from four close relatives having reduced levels of factor VII is shown to correct the abnormal thromboplastin generation of Stuart factor-deficient serum. The rate of conversion of prothrombin into thrombin is markedly delayed in the patient’s plasma but essentially normal in that of other family members. The patient’s prothrombin conversion is corrected by the addition of normal or Stuart-deficient plasma. The partial thromboplastin time of the patient is somewhat prolonged.

It is concluded that factor VII deficiency is inherited in this family as an incompletely recessive autosomal characteristic. The penetrance of the deficiency allele in the heterozygous individual appears to be variable. Analyses of the distributions of clotting factor levels in the family and in samples from the normal population indicate that an individual may vary greatly in his clotting factor levels, the average variation being nearly as great as the variation among individuals. The magnitude of the variation among siblings suggests that the penetrance of the factor VII deficiency allele is affected by extrinsic factors and that the observed population variation does not result from the occurrence of multiple alleles. It is suggested that sex may be a factor determining clotting factor levels.

The bleeding of the patient is controlled by transfusion of plasma and by injection of a plasma preparation called ACC-76 (Behringwerke, Germany), the latter followed by a transient elevation of blood factor VII.
The Duffy Blood Group System in Transfusion Reactions: A Review of the Literature and Report of Four Cases
Authors: COMPTON ALLAN , HABER JANE M.
Published: October 2019

1. The literature on the incidence and inheritance of the "Duffy" blood group factors and their significance in blood transfusion and erythroblastosis fetalis is briefly reviewed.

2. Four new cases in which anti-Fya was detected are reported.

3. The necessity for the use of the Coombs’ compatibility test with a potent Coombs’ serum to prevent reactions due to anti-Fya is re-emphasized.

4. The inadequacy of a 15 minute incubation period for the Coombs’ compatibility test in certain cases is noted.

5. The desirability of specific identification of an antibody detected in a patient’s serum prior to transfusion whenever possible is suggested.
Platelet Life Span in Normal, Splenectomized and Hypersplenic Rats
Authors: HJORT PETER F. , PAPUTCHIS HELEN
Published: October 2019

1. A method is described for the isolation of rat platelets. The method gives high yields of platelets with negligible white and red cell contamination.

2. Rat platelets were labeled in vivo with radioactive diisopropylfluorophosphate (DFP32). The platelet life span was the same in normal, splenectomized and hypersplenic rats.

3. Some problems involved in the use of DFP32 as a blood cell label are discussed.
Cyanocobalamin, Ascorbic Acid and Pteroylglutamates in Normal and Megaloblastic Bone Marrow
Authors: COX E. V. , MATTHEWS D. M. , MEYNELL M. J. , COOKE W. T. , GADDIE R.
Published: October 2019

The B12 activity as estimated by Lactobacillus leichmannii, the folic-acid-like activity by Streptococcus faecalis (F.A.A.) and the ascorbic acid concentration have been determined in the blood and buffy coat of bone marrow of normal subjects, 10 patients with pernicious anemia in relapse, a group of patients with non-Addisonian megaloblastic anemia and some patients with iron deficiency.

A correlation between the serum B12 and the plasma ascorbic acid and their respective levels in bone marrow was observed. The marrow and serum B12 levels in prenicious anemia were abnormally low, but they did not differ from a group of 5 patients with hypochromic normoblastic anemia who had both low serum and marrow levels. The concentration of F.A.A. in the marrow of patients with pernicious anemia was reduced, but it was felt that this was more likely a manifestation of the megaloblastic anemia rather than a causative factor.

One of six patients with megaloblastic anemia of pregnancy had no detectable deficiency, while the other five had reduced B12, folic acid and ascorbic acid concentrations. The possible therapeutic implications are discussed.

There was a significant reduction in the bone marrow concentration of ascorbic acid in all patients with megaloblastic anemia.
Platelet and Fibrinogen Sequestration
Authors: ADELSON EDWARD , RHEINGOLD JACK J. , PARKER OLIVE
Published: October 2019

1. Small amounts of thromboplastin intravenously cause platelet sequestration. This is followed by fibrinogen sequestration. Large doses of intravenously administered thromboplastin lead to irreversible destruction or utilization of platelets and fibrinogen.

2. The following theories are advanced: (a) This platelet sequestration may be due to platelet clumping caused by coagulation occurring on the platelet surface. Surface coagulation may be initiated by the action of thromboplastin or thrombin on the plasma proteins and calcium adsorbed on the platelet surface. The platelet clumps may be filtered out in the sinusoidal spaces of the body, especially in liver and spleen. (b) The coagulation on the platelet surface may lead to partial polymerization of the fibrinogen molecule. (c) As long as this polymerization is reversible, the entire process remains reversible. When polymerization proceeds far enough so as to become irreversible, the platelets and fibrinogen can no longer return to the circulation. (d) This same mechanism may explain other examples of platelet sequestration.
The Maturation Rate of Reticulocytes
Authors: BALDINI MARIO , PANNACCIULLI IVO
Published: October 2019

An in vitro culture technic for the study of reticulocyte maturation was described. The method gave reproducible results and proved to be of value in the comparative study of reticulocyte maturation in blood disorders. By this method it was shown that variations in the reticulocyte maturation in vitro paralleled similar variations present in vivo.

The maturation of reticulocytes from patients with different types of anemia was investigated. In some anemias the in vitro maturation of reticulocytes was prolonged, not only because younger reticulocytes were present in the blood, but also because the rate at which the reticulum substance disappeared was delayed. This was particularly evident in the anemia of chronic uremia, in Cooley’s anemia and in pernicious anemia in relapse. In only occasional cases of hereditary spherocytosis and of autoimmune hemolytic anemia was the rate of reticulocyte maturation found to be moderately delayed. In patients with iron deficiency anemia or bleeding anemia it was always normal.

From the above findings the following conclusions were derived:

1. The reticulocyte number in the circulating blood is the resultant of three variables: (a) the rate of output of new reticulocytes from the bone marrow; (b) the stage of maturation at which reticulocytes are delivered into the peripheral circulation; (c) the rate of disappearance of the reticulum substance.

2. The number of reticulocytes in the circulating blood cannot be indiscriminately used as a precise index of red cell production in erythrokinetics.

3. There is good reason to believe that a defect in the rate at which the reticulocytes mature in the circulating blood is an index of a similar defect in the process of erythroblastic differentiation in the bone marrow.
Hereditary Spherocytosis: Observations on Hemolytic Mechanisms and Iron Metabolism
Authors: CROSBY WILLIAM H. , CONRAD MARCEL E.
Published: October 2019

1. Two healthy patients with hereditary spherocytosis were phlebotomized until they developed iron deficiency and the erythrocytes became hypochromic. The hereditary spherocytes were no longer spheroidal: they became thin, and the fragility tests improved. However, the life span of the cells in the circulation was not improved. Later, splenectomy corrected the hemolytic disease.

2. In both patients, prior to the experiment, the hemolytic disease was compensated. There was no anemia despite the rapid turnover of red cells. In one of the patients, whose average red cell life span was only five days, the output of hemoglobin must have been exceedingly high. It was computed to be 135 Gm. per day, or 20 times the normal rate.

3. Some aspects of iron metabolism in hereditary spherocytosis are discussed.

4. The shape of the red cell in HS does not appear to be responsible for its premature destruction by the spleen. Iron deficiency corrects the spherocytosis, but it does not correct the hemolytic disease. Splenectomy corrects the hemolytic disease, but it does not correct the spherocytosis.
Congenital Vascular Defect Associated with Platelet Abnormality and Antihemophilic Factor Deficiency
Authors: RACCUGLIA GIOVANNI , NEEL JAMES V. , Davidson Ruth T. , Ussery Mary Jane
Published: October 2019

1. A kindred of 311 individuals, many members of which are affected by a hemorrhagic diathesis, has been described.

2. The variability in the manifestations of this diathesis is extreme. In its fullest expression the disease is characterized by a prolonged bleeding time with evidence of a morphologic defect in the platelets, and a deficiency in antihemophilic globulin. Some possibly affected individuals exhibit only a prolonged bleeding time, while, on the other hand, the clinically most severely affected individual, with AHF levels on several occasions of 5 to 10 per cent, has not been observed by us to have a prolonged bleeding time, although his platelets are morphologically abnormal.

3. Genetic analysis suggests that the hemorrhagic tendency is determined by a single dominant gene of variable penetrance and expressivity.

4. No satisfactory explanation can be developed on the basis of these studies for the association between platelet abnormality and AHF deficiency. More specifically, it is impossible to conclude whether the platelet defect is precursor to the AHF deficiency, or whether—as on a priori grounds seems less likely—this is an example of true genetic pleiotropy.

5. The terminologic chaos which afflicts the literature on hemorrhagic diatheses characterized by a prolonged bleeding time is discussed in the light of the findings in this one large kindred, and suggestions are advanced for minimizing confusion based on terminology alone.
Identification of Blood Group Antigens and Minor Cell Populations by the Fluorescent Antibody Method
Authors: COHEN FLOSSIE , ZUELZER WOLF W. , EVANS MARGARET M.
Published: October 2019

It is possible to produce fluorescence of erythrocytes as the result of specific antigen-antibody reactions of various blood group antigens; thus far, the factors A and B, a variety of Rh antigens and Kidd, have been successfully demonstrated with this method. It is important to realize that in the presence of adequate negative controls, low intensity fluorescence like that obtained with Rh antigens is nevertheless specific in systems involving erythrocytes.

The method discriminates between A1 and A2 cells. More antibody must be attached to the red cell for fluorescence than for agglutination. The relative paucity of antigenic sites of Rh substance as compared to A and B antigens is reflected by the difference in intensity of fluorescence.

The fluorescent antibody technic has been used successfully for the demonstration, and, to some extent, quantitation of minor cell populations in in vitro mixtures and in vivo. Injected Rh-positive erythrocytes were demonstrated in the blood of an Rh-negative volunteer during a period of 20 days. Fetal Rh-positive erythrocytes were demonstrated in several Rh-negative women, both with and without antibody, in the last trimester of gestation.
Brief Note: The Use of a Special Preservation Medium for the Maintenance of Platelet Viability at 4 C
Authors: BALDINI MARIO , EBBE SHIRLEY , DAMESHEK WILLIAM , Donovan Janet
Published: October 2019

Viability of rabbit platelets stored for 1 to 6 days in different media at 4 C. was studied using the Cr51-platelet survival technic.

Platelets stored for 24 hours in Triton-saline or DAS-gelatin had practically no viability.

When platelets were stored for 24 hours in diluted plasma, significant degrees of viability were found, but this decreased rapidly in the successive 48 hours.

The viability of platelets stored in diluted plasma containing inosine, adenine, glucose and phosphate buffer was maintained at significant levels over a six day period.

The data presented here suggest that viability and, consequently, the hemostatic properties of platelets stored at 4 C. can be maintained over relatively long periods of time provided that loss of the ATP reserve of the cells can be prevented.
The Application of Fluorescent Antibody Methods to the Study of Platelets
Authors: SILBER ROBERT , BENITEZ ROBERTO , EVELAND WARREN C. , AKEROYD JOSEPH H. , DUNNE CHARLES J.
Published: October 2019

1. Through the use of fluorescent antibody technic, it is possible to detect the antigen-antibody reaction between platelets and heterospecific antiserum, but no antigen-antibody reaction between normal human platelets and sera from patients with idiopathic thrombocytopenic purpura could be demonstrated.

2. The method permits the histologic localization of platelets in splenic tissue.

3. Whereas serum proteins can be easily removed from platelets by washing, there is an intimate association with fibrinogen so that this protein is still not removed from the platelet after 10 washings in saline.
Immunocytochemical Studies on Platelets. The Demonstration of a Common Antigen in Human Platelets and Megakaryocytes
Authors: VAZQUEZ JACINTO J. , LEWIS JESSICA H.
Published: October 2019

By means of the fluorescent antibody technic of Coons and Kaplan it was possible to demonstrate a common antigenic structure in human platelets and megakaryocytes, both in nonthrombocytopenic cases and in cases with idiopathic thrombocytopenic purpura. Direct evidence for a marked increase in the number of platelets in the spleens of two cases of idiopathic thrombocytopenia is given. The pathogenetic significance of this finding is discussed. It is concluded that the fluorescent antibody technic is a valuable tool for the chemical and morphologic study of platelets and megakaryocytes both in tissues and smears.