bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationships among Hematopoiesis, Blood Coagulation, Hemorrhage and Mortality in the Guinea Pig after Total Body X-Irradiation
Authors: ROSENTHAL ROBERT L. , GOLDSCHMIDT L. , PICKERING B. I.
Published: October 2019

Studies have been presented on blood, bone marrow, coagulation and their relation to mortality in guinea pigs exposed to 200, 400 and 600 r total body x-irradiation.

The initial responses of the blood and bone marrow were remarkably similar for the 3 doses of x-irradiation (ranging from LD10 to LD100). Marked differences among the dosages were noted in regeneration which occurred earliest after 200 r, later after 400 r and not at all after 600 r. Once started, regeneration was equally active after 200 and 400 r.

A coagulation defect, closely related to thrombocytopenia, was observed in the 6 to 13 day period following all doses of irradiation. An additional clotting defect, characterized by increases in the prothrombin time and heparin clotting time, was noted at 4 and 24 hours following all 3 doses of irradiation. The etiology of this defect requires further evaluation.

The pattern of hemorrhage, evaluated in various tissues by a semiquantitative scoring method, revealed that hemorrhage was most prominent in the 9 to 13 day period after exposure, and was closely related to thrombocytopenia. As the dose of x-ray increased, hemorrhage tended to become more protracted and severe and involved more organs. Hemorrhage disappeared rapidly as platelet formation returned in the recovery period. Hemorrhage was the chief factor in the severity and incidence of anemia after irradiation.
Metastatic Acropachy in Lymphatic Leukemia
Authors: CALVERT ROWLAND J. , SMITH ERIC
Published: October 2019

A patient with chronic lymphatic leukemia exhibiting the unique combination of bulbous digits from metastatic invasion and rhinophyma is described. The associated dermal and skeletal changes are discussed. Although two weeks of urethane administration produced a prompt hematological response, the peripheral lesions were unaffected by this very short therapeutic course.
New Data on Microdrepanocytic Disease
Authors: SILVESTRONI E. , BIANCO I.
Published: October 2019

1. Geographic, clinical and genetic data of 35 new cases of microdrepanocytic disease are presented.

2. These data suggest that sickle cell trait and microdrepanocytic disease are not infrequent in the eastern district of Sicily, perhaps with some small foci of sicklemic subjects.

3. The clinical history of adult patients and the personal examination of some cases of microdrepanocytic disease in children show the existence of mild forms of the disease and of forms appearing late in life.

4. From the clinical viewpoint, this series shows several interesting findings in the patients with microdrepanocytic disease: painful osteoarticular crises are inconstantly present; splenomegaly may undergo a spontaneous lessening during the course of the disease, analogous to that which occurs in sickle cell anemia; in women with microdrepanocytic disease miscarriages are frequent and successful pregnancies and births are extremely rare.

5. Hematologic studies reveal hypochromic anemia, leukocytosis, nucleated erythrocytes in the circulating blood, marked abnormalities in the morphology of the erythrocytes, and decreased cell fragility to hypotonic saline.

6. Genetic studies of the new cases again show the simultaneous presence in the patients with microdrepanocytic disease of the genes responsible for microcythemia and sickle cell trait.
Relationships of Adrenals, Gonads, and Thyroid to Thymus and Lymph Nodes, and to Blood and Thoracic Duct Leukocytes
Authors: SHREWSBURY MARVIN M. , REINHARDT WM. O.
Published: October 2019

The effects of gonadectomy, adrenalectomy, and thyroidectomy, and of thyroxine administration on the weight of lymphatic tissues and on the numbers of blood and thoracic duct leukocytes have been studied over a 40 day period in young male rats. The results of the present experiment are interpreted to indicate that:

1. Removal of the gonads caused a marked increase in the weight of the thymus, whereas but slight effect was noted on the weights of the lymph nodes. Removal of the adrenal glands, however, resulted in a marked increase in the weight of all lymphatic tissues.

2. Thyroxine administration was demonstrated to exert a hyperplastic effect on the lymph nodes, an effect which was accentuated by the removal of the adrenals and gonads. The lymph nodes were more sensitive than the thymus to the stimulating influence of thyroxine administration in the intact animal.

3. A comparison of the weights of the thymus and lymph nodes to control values was more indicative of the hypoactivity or hyperactivity of the thyroid, adrenals and gonads, than were the weights of the thymus or lymph nodes considered separately.

4. The increase in the level of blood lymphocytes following removal of the adrenals appeared to result from a failure of normal removal of the lymphocytes from the bloodstream, rather than from an increased delivery of the lymphocytes to the bloodstream via the thoracic duct. Administration of thyroxine to normal animals did not produce a blood lymphocytosis, lending further weight to the suggestion of Marine and others that the blood changes seen its toxic hyperthyroidism are the result of adrenocortical insufficiency.

5. Changes in the weight of the lymphatic tissues were not necessarily reflected by a proportional change in the number of thoracic duct lymphocytes, nor did the number of blood lymphocytes necessarily reflect the number of lymphocytes delivered to the bloodstream via the thoracic duct lymph.

6. The marked increase in the volume and cell content of thoracic duct lymph noted its thyroxine treated adrenalectomized-gonadectomized animals appears to be directly related to an increased sensitivity to the effects of thyroxine administration in the absence of these glands.

7. Removal of the thyroid gland vitiated the increases in weight of thymus and lymph nodes produced after castration and/or adrenalectomy.

8. The eosinophilic leukocytosis consequent on adrenalectomy was markedly augmented by removal of the thyroid and gonads.
The Measurement of the Survival of Human Erythrocytes by In Vivo Tagging with Cr51
Authors: SUTHERLAND DONALD A. , MCCALL MARY SUE
Published: October 2019

1. The deviation from a rectilinear form of the complex curvilinear curves of erythrocyte decay produced by the in vitro tagging of the cells with Na2Cr51O4 is not a result of the in vitro tagging procedure per se.

2. Human erythrocytes can be tagged its vivo with Na2Cr51O4. One can determine the in vivo survival of such cells if the gamma detecting unit is sufficiently sensitive and the amount of radiochromium used is sufficient. Such decay curves are similar to those produced by the in vitro Cr51 tagging method.

3. The in vivo method permits a study of the life span of erythrocytes in patients who have a very active cold agglutinin or in whom the cells are easily hemolyzed by mechanical trauma.
The Mechanics of the "L.E." Cell Phenomenon, Studied with a Simplified Test
Authors: SNAPPER I. , NATHAN DANIEL J.
Published: October 2019

1. A simple method is described for the formation of large numbers of lupus cells by positioning one drop of finger or ear blood of a lupus patient upon an accumulation of dried, normal polynuclear leukocytes. Thick leukemic smears and perhaps even imprints of various carcinomas or Hodgkin’s tissue can also be used as substrates.

2. No false positive results were obtained in a group of control patients.

3. Striking results were obtained with this method itt 20 of 21 known cases of disseminated lupus erythematosus. The remaining case in whom a single "L.E." cell was found by a conventional method, to date has yielded no lupus cells by our method.

4. The difference in appearance of the inclusion body of some of these cells from the characteristic Hargraves cell is highlighted.

5. Theoretic considerations of the mechanism of lupus cell formation are discussed and reassessed in the light of our experience with this method. For the formation of "L.E." cells it seems necessary that living polynuclear leukocytes, dead cells and lupus serum be brought together. The lupus serum can depolymerize the desoxyribose nucleic acid of the nuclei of the dead cells, but not of living cells. The depolymerized material is then phagocytized by living polynuclear cells and formation of lupus cells results.
Alkaline Phosphatase Activity of Leukocytes in Shock
Authors: VACCARI FRANCESCO , SABOTTO BENITO , MANZINI ENRICO
Published: October 2019

In animals and patients subjected to stress procedures and shock, there occurs simultaneously with the increase in general metabolism a rise of the cytoplasmic localizations of the alkaline phosphatases in granulocytes.
Megaloblastic Anemia Associated with Anatomic Lesions in the Small Intestine
Authors: THOMPSON R. B. , UNGLEY C. C.
Published: October 2019

This paper describes the development of anemia in six patients with strictures and anastomoses in the small intestine. The marrow proved to be megaloblastic in three instances, and megaloblastic change is presumed in the other three because of the clinical and laboratory findings, and the characteristic hemopoietic response to liver therapy. Responses to crude and refined liver extracts and vitamin B12 compared unfavorably with those to be expected in Addisonian pernicious anemia; they were similar to the poorer responses often observed in megaloblastic anemia associated with idiopathic steatorrhea. The relationship of the intestinal lesion to the development of megaloblastic anemia is briefly discussed.
Case Report: Infectious Mononucleosis and Acute Hemolytic Anemia; Report of Two Cases and Review of the Literature
Authors: THURM RICHARD H. , BASSEN FRANK
Published: October 2019

1. Two cases of infectious mononucleosis complicated by acute hemolytic anemia are presented. One of these patients had, in addition, an underlying Mediterranean anemia.

2. Hemolytic anemia is a rare complication of infectious mononucleosis.

3. Thirteen similar cases have been collected from the literature, and the cumulative data summarized.

4. Recovery occurred in all cases. Splenectomy was considered necessary in only two.

5. The etiology of hemolytic anemia in infectious mononucleosis is obscure but there is good evidence to indicate it may be different in different cases. Hypersplenism initiated by infectious mononucleosis could best explain the authors’ cases whereas the elaboration of hemolytic antibodies would best explain some of the reported cases.
Effect of Repetitive Exposures to Gamma Rays on the Hematopoietic System of the Rat
Authors: BAUM SIEGMUND J. , KIMELDORF DONALD J. , JACOBSEN ELLY M.
Published: October 2019

Rats were subjected to a series of one or more doses of 75 r gamma irradiation, at seven day intervals between exposures. Hematologic examinations were made following the last exposure in each series of irradiations. The maximum changes observed were, to a great extent, independent of the number of previous exposures, and appeared to reflect principally the effects of the last irradiation. These results were ascribed to the regenerative capacity of the hematopoietic system to recover during the seven day interval between radiation exposures.
A Second Example of Anti-Fyb
Authors: LEVINE PHILIP , SNEATH JOAN S. , ROBINSON ELIZABETH A. , HUNTINGTON P. W.
Published: October 2019

A second example of anti-Fyb was found in the serum of a patient immunized by transfusions and pregnancies. In addition to anti-Fyb, anti-c and anti-E were also present. The identification of the antibody as anti-Fyb was made in part with the aid of a 2 x 3 table in parallel tests with anti-Fya.
Phase I Study Using Alternate Week Administration of the Farnesyl Transferase Inhibitor R115777 (Zarnestra™) in Patients with Myelodysplastic Syndrome.
Authors: Kurzrock Razelle , Verstovsek Srdan , Wright John J. , Pilat Susan R. , Cortes Jorge E. , Estey Elihu H. , Giles Francis J. , Beran Miloslav , Kantarjian Hagop M.
Published: October 2019

R115777 is an oral selective inhibitor of farnesyltransferase with potent antiangiogenic, antiproliferative, and apoptotic effects. The current study assesses the tolerance and response rate in patients (pts) with myelodysplastic syndrome (MDS) receiving R115777 on a one-week on/one-week off schedule. Starting dose was 100 mg bid with a standard 3+3 design for a total initial treatment period of eight weeks. Escalation was 100 mg po bid until grade 2 non-hematologic toxicity was observed, then 100 mg total dose per day escalation until MTD. Fifty-three pts were treated (median age 68.5 yrs; range 35–83) with RA (n=12), RARS (n=4), RAEB (n=22), RAEBt (n=7), and CMMoL (n=8). Three pts (5%) had IPSS score low; 24 (45%), INT-1; 19 (36%), INT-2; and 7 (13%) high score. Three patients had an N-RAS mutation and one had a K-RAS mutation. Fifty-three patients are evaluable for toxicity and 51 are evaluable for response (1 non-compliant, 1 ineligible). This regimen was well tolerated. The most common toxicity was myelosuppression (60% of the patients). Twenty percent reported no side effects. Non-hematologic toxicities included mild skin rash (15%), diarrhea (13%), increase in SGPT (13%), increase in bilirubin (10%), and nausea (8%). DLTs of ataxia (n=1) and fatigue (n=1) occurred at doses of 1300 mg/day. The MTD appears to be 1200 mg/day. Fifteen of 51 evaluable patients responded (3 CRs and 12 hematologic improvements). Interestingly, 3 of 6 pts at the lowest dose level (100 mg bid) responded, including a CR. Major platelet responses were most common; 11 of the 15 responders had a platelet response. Only one patient among the responders (hematologic improvement) had a RAS (N-RAS) mutation. Median time to response is eight weeks (range: 4–32 weeks). In conclusion, alternate week therapy with R115777 is well tolerated and a response rate of 29% is achieved in the Phase I setting. This intermittent schedule would lend itself well to combination or maintenance regimens.
Global Gene Expression in Leukemic Blasts as a Function of Germline Genetic Variation.
Authors: French Deborah , Wilkinson Mark R. , Yang Wenjian , de Chaisemartin Luc , Cook Edwin H. , Das Soma , Ratain Mark J. , Evans William E. , Downing James R. , Pui Ching-Hon , Relling Mary V.
Published: October 2019

The possible impact of common, functional, germline polymorphisms on clinical outcome in patients with leukemia is under active investigation. However, little attention has been paid to the possible consequences of germline genetic variation on genome-wide phenotypic variation in human tissues. We determined the germline status of 16 well-characterized functional genetic polymorphisms (using normal leukocyte cell DNA) in unrelated children with newly diagnosed acute lymphoblastic leukemia (ALL). We assessed whether global gene expression (using oligonucleotide arrays that interrogated approximately 10,000 genes) of diagnostic ALL blasts from the same patients differed by germline polymorphic genotypes. Gene expression values were adjusted for ALL-subtype-specific patterns. In a supervised analysis that divided patients based on their germline genotypes, we identified two polymorphisms that were significant predictors of global gene expression: the UGT1A1 promoter repeat polymorphism [A(TA)nTAA] (181 probe sets, p=0.01, false discovery rate [FDR] = 35.6%) and the GSTM1 deletion (112 probe sets, p=0.008, FDR = 42.5%). Genes whose expression differed significantly in ALL cells from patients with and without the UGT1A1 (TA) 7/7 genotype included ATM and ABL1; genes whose expression differed significantly between patients with the GSTM1 null genotype and the non-null genotype included PCNA, PRIM1, CDC6 and SKP1A. In an unsupervised analysis, patients were separated into two clusters; only the UGT1A1 promoter repeat polymorphism differed in frequency between the two clusters (p=0.001, Fisher’s exact test). These findings illustrate that polymorphisms in genes whose direct products are restricted to other tissues (e.g. UGT1A1 is primarily expressed hepatically) may affect gene expression elsewhere (e.g. ALL blasts). Because UGT1A1 is involved in the conjugation (and thus the transport and excretion) of numerous endogenous steroids and hormones, it is plausible that genotypic variation in its activity could have consequences for gene expression in distant tissues, such as ALL blasts.
Valuation of Cytogenetics and FISH in Evaluation of Myelodysplastic Syndromes: A Cancer Cytogenetics Reference Lab’s Experience.
Authors: Sun Guoxian , Lorenzo Michael , Tripodi Joseph , Chrysostomou Chrysostomos , Stephenson Christine F. , McGill John R.
Published: October 2019

Cytogenetics has proven an essential tool not only for confirming a diagnosis/classification, but for providing prognostic value as well in myelodysplastic syndromes (MDS). However, approximately 50% of primary MDS do not show discernible chromosome changes. In recent years, the fluorescence in situ hybridization (FISH) technique using gene or chromosome locus/region specific probes has emerged as a promising test in various hematopoietic and lymphoid neoplasms. To evaluate the application of FISH panels and cytogenetic studies in MDS, we retrospectively analyzed 1,885 consecutive bone marrow results from patients with suspected MDS due to cytopenia(s). In particular, we assessed the additional information a FISH reflex testing might have given in cytogenetically normal cases. The probes used in the panel included the EGR1 at 5q31, the D7S522 at 7q31, the D8Z2 for the centromere of chromosome 8, the MLL at 11q23 and the D20S108 at 20q12 (Vysis, Inc.). Among all patients, 190 (10%) had clonal chromosome abnormalities, mostly as reported in the literatures, eg, -5/5q- accounted for 34.7% of abnormalities, trisomy 8 29.5%, -7/7q- 14.2%, 20q- 13.7%. Of 345 cases with a FISH reflex test ordered and performed, only 3 (0.87%) showed positive results: a deletion of 7q31, a deletion of 20q12 and a deletion of 5q31 in 9.6%, 8.2% and 71.5% of interphase cells respectively. For the case with 5q- detected by FISH, only 12 metaphases were available for cytogenetic analysis. From our data and experience, at present, interphase FISH panel testing seems not to be an efficient and cost-effective method used as a screening test for cytopenia(s) in the diagnosis of MDS, different from its applications in B-cell chronic lymphoid leukemias, non-Hodgkin lymphomas and plasma cell neoplasms where neoplastic cells inherited not to divide easily in culture for metaphase analysis. Rather, it should be used for suspected MDS cases as a technique of choice for problematic specimens compromised for cytogenetic analysis such as cellular insufficiency, extended transit time and extremely low mitotic index or poor chromosome morphology. Until more genetic defect targeted probes become available with a better understanding of the stem cell biology and pathogenesis in MDS, cytogenetics is still the best and a “must” technique for detecting genomic aberrations in MDS and nearly all other myeloid hematopoietic neoplastic disorders.
Administration of an HE2100 Depot Preparation after 60Co Total Body Irradiation (400 cGy) Reduces the Duration of Severe Neutropenia, Thrombocytopenia and Anemia in Rhesus Macaques.
Authors: Dowding Charles , Frincke James , Garsd Armando , Stickney Dwight R. , Reading Christopher L.
Published: October 2019

HE2100 (androst-5-ene-3β,17β-diol) is a naturally occuring adrenal steroid hormone that exhibits a relatively weak interaction with the androgen receptor, estrogen receptor-α and estrogen receptor-β but not the glucocorticoid receptor. We previously reported (Blood [2003] 1002;20) that subcutaneous HE2100 administration (daily for 10 days) eliminated severe neutropenia in rhesus macaques exposed to 400 cGy 60Co total body irradiation (TBI). The current study utilized the same model of myelosuppression in rhesus macaques and evaluated the activity of HE2100 when administered as a depot-type preparation on duration of Grade 4 neutropenia (<500/μL, G4 Neut), at least Grade 3 thrombocytopenia (<50,000/μL, G3 Thromb) and Grade 4 anemia (<6.5 g/dL, G4 Anemia). Blood was analysed daily during the cytopenic phase and approximately every three days at other times through Day 35 (irradiation Day 1) to minimize the effect of phlebotomy. HE2100 (15 mg/kg) was administered by daily intramuscular injection for 5 days (Regimen 5d-IM), by four once-weekly intramuscular injections (Regimen 1w-IM) or by four once-weekly subcutaneous injections (Regimen 1w-SQ). The first dose for all regimens was administered 2 to 4 hours after TBI. All groups comprised of 2 male and 1 female young adult rhesus macaques with the exception of the vehicle group (2 males and 2 females). Vehicle-treated animals experienced on average G4 Neut, G3 Thromb and G4 Anemia for 11.8, 3.5 and 4.0 days, respectively. Animals treated by 5d-IM experienced on average 2.7 days of G4 Neut, 0 days of G3 Thromb and 0 days of G4 Anemia. The values for Regimens 1w-IM and 1w-SQ were 6.0 and 4.5 days (G4 Neut), 0.3 and 0.3 days (G3 Thromb) 0 and 0 days (G4 Anemia), respectively. These results indicate that HE2100 depot-type preparations delivered by either 4 or 5 SQ or IM injections substantially reduced the duration of severe neutropenia following 400 cGy TBI. Major morbidity or mortality associated with acute Hematopoietic Syndrome is believed to result from sepsis associated with severe neutropenia. Hence it is possible that HE2100 would protect against acute Hematopoietic Syndrome-related major morbidy or mortality. The contribution of thrombocytopenia or severe anemia to Hematopoietic Syndrome-related major morbidity or mortality is unknown but it is possible that HE2100-mediated protection of these blood elements might be beneficial by, for example, reducing tissue hypoxia. In conclusion, HE2100 affords significant protection as a countermeasure to TBI in rhesus macaques. There is an umet medical need for a cost-effective countermeasure for acute Hematopoietic Syndrome that may result from exposure to a nuclear event. It is possible that the protection seen in rhesus macaques may extend to man.
Cholinergic Stress Responses Facilitate the Molecular Mechanisms Underlying Granulocytosis.
Authors: Deutsch Varda R. , Pick Marjorie , Grisaru Dan , Perry Chava , Naparstek Elizabeth , Lapidot Tzvee , Soreq Hermona
Published: October 2019

Glucocorticoid-initiated granulocytosis following trauma persists long after cortisol levels decrease, through yet unknown modulators of hematopoiesis. An intriguing candidate for such a modulatory role is the AChE-R splice variant of acetylcholinesterase. In brain and muscle a functional glucocorticoid response element (GRE) in the distal promoter of the ACHE gene induces overexpression of AChE-R following trauma. To explore the possibility that circulating and/or blood cells-associated AChE-R regulates sustained granulocytosis following stress, we employed labor and childbirth as the paradigm for acute, transient stress. In intra-partum women, increased AChE-R expression in granulocytes was observed by flow cytometry, which correlated significantly with their high cortisol levels and elevated AChE activity in plasma. In post-partum women, leukocyte counts, AChE-R positive granulocytes and AChE activity in plasma remained high for several days after cortisol levels declined. Furthermore, exposure to ARP, a synthetic peptide with the C-terminal sequence of AChE-R, promoted AChE gene expression in human CD34+ progenitors and facilitated their efficient proliferation in liquid cultures (1) in an antisense suppressible manner, supporting auto-regulatory prolongation of AChE-R expression. The intracellular interaction of the C-terminal domain of AChE-R with RACK1 and protein kinase Cε (2) supports such a mechanism. In 2 weeks cultures of CD34+ HPC, 2nM ARP induced granulocyte expansion and maturation. Additionally, exposure to ARP for 24 hours facilitated the production of pro-inflammatory cytokines interleukin (IL)-6, IL-10 and TNFα in adult PB MNC and enhanced the engraftment of CD34+ cells in NOD/SCID mice. We conclude that stress-induced AChE-R accumulation and consequent inflammatory reactions facilitate prolonged granulocytosis.
The Transcriptional Program of Terminal Granulocytic Differentiation in Man.
Authors: Borregaard Niels , Jacobsen Lars C. , Borup Rehanah , Rasmussen Thomas , Bjerregaard Malene D. , Nielsen Finn C. , Cowland Jack B. , Theilgaard-Moench Kim
Published: October 2019

To characterize the transcriptional program that governs terminal granulocytic differentiation in vivo, we performed comprehensive microarray analyses of human bone marrow (BM) populations highly enriched in promyelocytes (PMs), myelocytes/metamyelocytes (MYs), and neutrophils (bm-PMNs). These analyses identified 11,310 genes involved in differentiation, of which 6,700 were differentially regulated including granule proteins and surface receptors previously not identified in neutrophils. Functional clustering demonstrated that differentially expressed genes were assigened to all of the major gene categories including apoptosis, cell cycle, chaperones, enzymes, immunity proteins, kinases, motor proteins, signal transducers, structural proteins, and transcription factors. In an attempt to define the developmental distance between PM, MY, and bm-PMN populations, we assessed the Pearsons correlation coefficient (γ) by correlating the transcriptomes (=all 44.760 probe sets, Affymetrix) of BM populations. The correlation coefficients among replicates of BM populations were in the range of 0.97–0.99. On the other hand the correlation coefficients between BM populations were 0.81 (γPM - MY), 0.79 (γMY - bm-PMN), and 0.52 (γPM - bm-PMN), and thus, provided a useful quantitative measure reflecting the hierarchical relationship between the three BM populations. Additional correlation of bm-PMN and peripheral blood-PMN transcriptomes revealed a high similarity of both populations (γbm-PMN - pb-PMN=0.95) indicating a termination of granulocytic differentiation in the BM microenvironment during steady state hematopoesis. Differentiation of PMs towards MYs was accompanied by a marked decline of proliferative and general cellular activity as defined by downregulation of E2F target genes, cyclin dependent kinases 2/4/6, and various metabolic, proteasomal, and mitochondrial genes. Expression patterns of apoptosis genes indicated death control by the p53-pathway in PMs and by death receptor pathways in bm-PMNs. Effector proteins critical for host defense were expressed successively throughout granulocytic differentiation, whereas receptors and receptor ligands essential for the activation of the host defense program were terminally upregulated in bm-PMNs. The upregulation of ligand-receptor pairs, which are defined inducers as well as target genes of NF-kB, suggests a constitutive activation of NF-kB in bm-PMNs by autocrine loops. Overall, these results define a granulocytic differentiation model governed by a highly coordinated fail-safe program, which promotes completion of differentiation before cells gain responsiveness towards activating stimuli that accompany infections.
2-Chlorodeoxyadenosine (2-CdA) and Cyclophosphamide (Cy) Alone or in Combination with Rituximab (Rit) for Previously Untreated Waldenstrom’s Macroglobulinemia (WM).
Authors: Weber Donna M. , Wang Michael , Delasalle Kay B. , Gavino Maria , Alexanian Raymond
Published: October 2019

Between 3/96-11/03 we treated 62 consecutive, newly diagnosed pts with symptomatic WM with either 2-CdA 1.5 mg/m2 sc tid x 7d + Cy 40 mg/m2 po bid x 7d (35 pts, 3/96-2/99) or with identical doses of 2-CdA and cyclophosphamide + Rit 375 mg/m2 IV q wk x 4 wk (27 pts, 11/98-11/03). For both regimens a second and final course was repeated after 6 weeks. Partial response (PR), defined by at least 50% reduction of monoclonal IgM, adenopathy, and organomegaly, was noted in 83% (29 pts) of pts treated with 2-CdA-Cy and 81% (22 pts) with 2-CdA-Cy-Rit; complete disappearance of clonal evidence of disease (CR) by negative serum and urine immunofixation, bone marrow, and CT scan was noted in 6% (2 pts) treated with 2-CdA-Cy and 11% (3 pts) with 2-CdA-Cy-Rit. Overall response rate was 89% and 93% for 2-CdA-Cy and 2-CdA-Cy-Rit, respectively. Responding patients were followed without further treatment until relapse, defined by a 25% increase in M-protein from nadir, or recurrence of adenopathy or organomegaly. Median time to remission was similar at 2.5 mos for 2-CdA-Cy and 2.4 months for 2-CdA-Cy-Rit (p.42), but time to treatment failure (TTF) appeared longer for 2-Cda-Cy-Rit (med. 64.2 mos) than for 2-CdA-Cy (med. 31.4 mos) (p.20). Since many pts have not required retreatment for symptomatic disease at the time of relapse, we also evaluated time to retreatment (TTRT) as a separate endpoint. The median time to retreatment (TTRT) has not been reached with 2-CdA-Cy-Rit, in comparison with a median TTRT of 51.2 mos with 2-CdA-Cy (p<.01), illustrating the prolonged period of stability after limited treatment with either 2-CdA combination and the superiority of added rituximab. Median overall survival has not been reached with either regimen since only 27% of all pts have died. Both treatment regimens were associated with minimal toxicity, high response rate, long duration of unmaintained remission, even longer time to the need for retreatment, and long survival. Thus treatment with a limited 2-CdA regimen represents the therapy of choice, in our opinion, for previously untreated, symptomatic WM.
PAD Combination Therapy (PS-341/Bortezomib, Adriamycin and Dexamethasone) for Previously Untreated Patients with Multiple Myeloma.
Authors: Cavenagh James D. , Popat Rakesh , Curry Nikki , Stec Jim , Morris Treen C. , Drake Mary , Agrawal Samir , Smith Patricia , Schenkein David P. , Esseltine Dixie-Lee , Oakervee Heather
Published: October 2019

Background: The most favourable outcomes for patients with MM are seen in those achieving a complete or near complete response following initial therapy. Therefore, it is logical to explore strategies aimed at improving initial response rates. Bortezomib has shown significant activity in patients with advanced MM and has been shown to be superior to pulsed dexamethasone in this setting. Additional efficacy is seen when dexamethasone is combined with bortezomib and in vitro synergy is observed with cytotoxic agents such as adriamycin. On this basis, the PAD regimen was investigated as front line therapy for patients with MM.

Aims: The primary objective of this Phase I/II study was to assess the feasibility of harvesting PBSCs after PAD, with secondary objectives being assessments of safety, toxicity, response rate (RR), PFS and OS.

Methods: Patients with previously untreated MM were eligible. Patients were treated with 4x21 day cycles of PAD comprising bortezomib 1.3 mg/m2 on days 1,4,8 & 11 along with 40 mg dexamethasone on days 1–4, 8–11 & 15–18 during cycle 1 and days 1–4 during cycles 2–4. During days 1–4 of each cycle, patients also received 0 mg/m2, 4.5 mg/m2 or 9 mg/m2 of adriamycin at levels 1,2 & 3 respectively. Following harvesting, patients received high dose melphalan (MEL200) with stem cell (PBSC) rescue. Gene expression profiling was performed on purified plasma cells from diagnostic samples for pharmacogenomic analysis.

Results: Thus far, 21 patients have been enrolled (19 male 2 female, median age 55 years [range 36–66]). All 21 have completed PAD with a 95% PR/CR rate. 20/21 patients mobilised PBSC successfully (median collection 3.8 x 106 CD34+ cells/kg, range 1.6–10.4). 16 patients have received MEL200 with median neutrophil (>0.5 x109/L) and platelet engraftment (>20 x109/L) of 15 (1–24) and 13 (10–33) days respectively. Of patients who are assessable at 3 months post MEL200, 11/12 have achieved at least a PR (CR 4, nCR 1, VGPR 4 & PR 2). Toxicities have generally been acceptable with 15 grade 3 events: 6 infections, 4 episodes of shingles, 2 nausea and vomiting, 2 neuropathy and 1 postural hypotension. Overall, 48% of patients have experienced sensory or painful neuropathy which is of Grade 1 severity in 43% of cases. Of note, neuropathic symptoms are improving in all patients after completion of therapy.

Summary: These preliminary data show that PAD is well tolerated in the majority of patients, is highly effective and does not prejudice subsequent PBSC collection. A further cohort of patients will receive PAD with bortezomib at 1.0 mg/m2 in order to determine whether the frequency of neuropathy can be reduced whilst preserving efficacy. PAD should be further evaluated as first line therapy in prospective randomised trials.
Thalidomide and Rituximab in Waldenström’s Macroglobulinemia.
Authors: Branagan Andrew , Hunter Zachary , Santos Daniel Ditzel , Treon Steven P.
Published: October 2019

Rituximab and thalidomide are active agents in Waldenstrom’s macroglobulinemia (WM) producing major response rates of 25–50%. Moreover, as previously shown by us, thalidomide enhances rituximab mediated ADCC killing of lymphoplasmacytic cells (Blood 100:314b). As such, we conducted this phase II study using combination rituximab and thalidomide in WM patients who previously had not received rituximab or thalidomide. Intended therapy was as follows:

Weeks 1–52 Thalidomide (200 mg po qhs for 2 weeks, then 400 mg po qhs)

Weeks 2–5 Rituximab (375 mg/m2/week)

Weeks 13–16 Rituximab (375 mg/m2/week)

Patients were evaluated at week 12, and if they had at least stable disease (SD) were eligible for further therapy and for response evaluation. Dose modifications, delay, and/or discontinuation of thalidomide were permitted for toxicities. Twenty-five patients have been enrolled, 20 of whom were previously untreated. Six patients are off study, 4 because of no response and 2 because of deaths unrelated to protocol therapy. Grade 3/4 toxicities to thalidomide included neuropathy (n=15); somnolence or confusion (n=12); rash (n=7); tremors (n=2); bradycardia (n=2); other (n=3); and led to its eventual discontinuation in 11/19 patients. All evaluable patients received the intended rituximab therapy. Paradoxical IgM spikes following rituximab occurred during both courses of therapy and were observed in 12/25 (48%) patients similar to those spikes which we reported with rituximab monotherapy (ASH2003; 102:690a). Responses to date among the 23 evaluable patients are as follows: CR (n=1); PR (n=12); MR (n=2); SD (n=4). Four patients had no response to therapy. No patients with stable disease or better have progressed with a median follow-up of 10 months (range 6–13 months). Response to therapy is associated with cumulative dose of thalidomide. Patients who responded to therapy received a median thalidomide cumulative dose of 37,600 (range 5700–74,500) mg as compared to 5,650 (range 2100–21,000) mg among non-responding or stable disease patients. These studies demonstrate that thalidomide in combination with rituximab is active in WM, and associated with the cumulative dose of thalidomide. In view of the encouraging response rates obtained with this combination and the potential for improved toxicity profile, a phase II study examining the thalidomide analogue CC-5013 (Revlimid) plus rituximab is planned.
Prognostic Significance of Magnetic Resonance Imaging (MRI) of Bone Marrow in Previously Untreated Patients with Multiple Myeloma.
Authors: Moulopoulos Lia Angela , Gika Dimitra , Dellasale Kay , Weber Donna , Anagnostopoulos Athanasios , Alexanian Raymond , Dimopoulos Meletios A.
Published: October 2019

Purpose: To determine the prognostic value of spinal bone MRI in symptomatic patients with multiple myeloma requiring treatment.

Materials and methods: Between January 1990 and Decmber 2001, 142 patients with symptomatic multiple myeloma (MM) underwent MRI of the spine before initiation of treatment. All patients received primary treatment based on high dose pulse dexamethasone (D) such as VAD or Melphalan + D. High-dose therapy with autologous stem cell transplantation was administered to 61 patients. MRI patterns of involvement were correlated with known prognostic variables of myeloma (including the International Staging System (ISS)), with response to treatment and with survival.

Results: Focal marrow lesions with intervening normal marrow were identified in 50% of patients, diffuse marrow replacement in 28%, a variegated pattern consisting of innumerable small foci of marrow replacement on a background of normal marrow in 14% and normal MRI pattern in 8% of patients. When patients with diffuse pattern were compared to patients with other MRI patterns, they had features of more advanced disease such as higher ISS, anemia, hypercalcemia, extensive bone marrow plasmacytosis, elevated serum LDH and impaired renal function. Patients and disease features were similar among patients with normal, focal or variegated MRI patterns. The frequency of response to primary treatment was similar among patients with different MRI patterns. Median survival was 24 months for patients with a diffuse pattern, 51 months for those with a focal pattern, 52 months for variegated and 56 months for patients with normal pattern (p=0.001). The presence or absence of diffuse MRI pattern separated the patients with ISS 1 and 2 into two subgroups with significantly different survival times of 28 months and 61 months respectively (p=0.01). Among patients with ISS 3, those with a diffuse pattern had a median survival of 18 months, whereas the remaining patients survived for a median of 30 months (p=0.5). Furthermore, a diffuse pattern predicted an inferior outcome both in patients who did or did not receive high dose therapy with autologous stem cell transplantation.

Conclusion: MRI of the spine before treatment provides prognostic information for symptomatic patients with myeloma, especially for those with ISS 1 or 2. Diffuse marrow replacement on MRI of the spine identifies patients with advanced MM who have a poor prognosis. Such patients are candidates for innovative treatments.
Long-Term Id Vaccination Combined with IL-12 or IL-12 and GM-CSF in Early Stage Multiple Myeloma Patients.
Authors: Hansson Lotta , Abdalla Amir O. , Rabbani Hodjattallah , Osterborg Anders , Mellstedt Hakan
Published: October 2019

The idiotypic structures expressed on the myeloma Ig protein may serve as tumor specific antigens and can be used as target structures for Id (Id) vaccination. In this study, 28 patients with IgG myeloma stage I-II were repeatedly immunized intracutaneously during a 110 week period with the autologous M-protein, either in combination with interleukin-12 (IL-12) (n=15), or with IL-12 and granulocyte-macrophage colony-stimulating factor (GM-CSF) (n=13). An Id-specific T cell response, as assessed by [3H]-thymidine incorporation, ELISPOT assay and delayed type hypersensitivity (DTH) reaction and strictly defined by 3 specified criteria, developed in 4/15 patients (27%) in the IL-12 group and 11/13 patients (85%) in the GM-CSF/IL-12 group (p=0.003). Three of 15 patients (20%) showed a gradually increasing Id-specific T cell response during the study whereas 10/15 patients (67%) showed an initial response which then disappeared rapidly; the latter pattern was frequently associated with subsequent progressive disease. In two patients (both in the IL-12 group) a reduction of the M-protein concentration by > 50% and >25%, respectively, was observed 18 and 21 months after the last immunization. These results indicate that Id immunization of myeloma patients using IL-12 and GM-CSF may more frequently induce a specific T cell response than with IL-12 alone, but also that repeated immunizations may result in a non-reversible state of tolerogenic T cells.
Pediatric Essential Thrombocythemia (ET): Evaluation of Clonality Status.
Authors: Pacquola Enrica , Scapin Margherita , Putti Maria C. , Tucci Fabio , Randi Maria L. , Fabris Fabrizio
Published: October 2019

ET is a very rare disease in pediatric age and some evidence suggest that ET in childhood may be a possible variant of the ET as described in adults. Clonal assays using X chromosome inactivation patterns (XCIPs) have been used to demonstrate the clonal origin of ET. However, about half women with ET seem to have a polyclonal XCIPs and the occurrence of thrombosis seems to be higher in patients with monoclonal XCIPs. We report 7 females in pediatric age affected with ET (mean age at diagnosis 9 years). The diagnosis of ET was made in agreement with the Polycythemia Vera Study Group Criteria. The main clinical data of our children are summarized in the table. None of the children had heavy haemorrhages nor splenomegaly. HUMARA trinucleotide repeated polymorphism in granulocytes DNA amplified using two primers flanking the STR, was used to assess myeloid clonality of our patients. The PCR products will be quantified after electrophoresis with density analysis. T lymphocytes DNA of the patients were also tested for the HUMARA polymorphism as control. Monoclonal granulocytes XCIPs was demonstrated in only 1 asymptomatic child who had polyclonal T-lymphocytes, while 5 other children had polyclonal XCIPs of both granulocytes and T-lymphocytes. Omozygous XCIP of 1 chlid did not permit clonality evaluation.

In conclusion, as observed in adults, also children with ET seem to be heterogeneous concerning clonality status. The percentage of polyclonal cases seem to be higher in pediatric age if compared with adults (data of the literature). The sole patient with a major thrombotic complication is polyclonal. Again pediatric ET seems to be different than adults ET: more studies are needed to clarify the biological characteristics of pediatric ET

age plts x 10to9/L clinical symptoms TPO pg/ml clonality granulocytes/T lymphocytes CV 9 m 945 Budd Chiari syndrome 30 polyclonal/polyclonal GM 12 y 1611 headache 30 omozygous TF 10 y 1038 headache 9 polyclonal/polyclonal FE 6 y 3266 headache + purpura 15 polyclonal/polyclonal BC 14 y 1308 asymptomatic 25 monoclonal/polyclonal GI 9 y 2718 asymptomatic 22 polyclonal/polyclonal 8 y 1550 asymptomatic 64 polyclonal/polyclonal
Cooperation between MYC and BCL2 to Induce Lymphoma Is Uncovered in an Adult Context.
Authors: Fan Alice C. , Giuriato Sylvie , Feng Charles , Padua Rose Ann , Felsher Dean
Published: October 2019

MYC overexpression is thought to induce tumorigenesis through a variety of different mechanisms including the induction of proliferation, inhibition of differentiation and disruption of genomic stability. Conventional transgenic systems that have been used to study the role of oncogenes in tumorigenesis continuously overexpress transgenes and hence preclude the investigation of the initial and age specific consequences of oncogene activation. To investigate the developmental specific consequences of MYC overexpression in the pathogenesis of lymphoma in vivo, we used transgenic mice in which the MYC proto-oncogene is conditionally regulated via the Tetracycline Regulatory System (Tet system). The ability of MYC to induce lymphomagenesis was found to be inversely correlated with the age of the host at the time of MYC activation. When MYC was activated constitutively, the mean time until tumor development was 13 weeks. When MYC was activated at increasing developmental ages of 3 and 5 weeks, identical tumors arose, but with an increased mean tumor latency. When MYC was activated at 7 weeks, an age equivalent to an adult mouse, no tumors developed even after 40 weeks of observation. However, we found that in adult mice, if both MYC and BCL2 are overexpressed at 7 weeks of age, mice succumbed to lymphoma with a mean latency of 27 weeks. Surprisingly, we could not find evidence that MYC overexpression induces apoptosis or that BCL2 overexpression reduced apoptosis. We conclude that the ability of MYC to induce lymphomagenesis is highly dependent on the developmental context. MYC and BCL2 cooperate to permit tumorigenesis in adult mice. Since lymphoma occurs generally as a disease in adults, this suggests that previous reports may have greatly overestimated the ability of MYC activation to induce tumorigenesis and underestimated the potential cooperation between MYC and BCL2 oncogenes. Our results also suggest that BCL2 may cooperate with MYC to induce lymphomagenesis through additional mechanisms other than preventing MYC from inducing apoptosis.
Disruption of the Hexb/Enc-1 Antisense Gene and Identification of Elevated Enc-1 Expression in Patients with Hairy Cell Leukemia.
Authors: Lerner Mikael , Hammarsund Marianne , Sangfelt Olle , Zhu Chaoyong , Merup Mats , Jansson Monika , Gahrton Goesta , Kluin-Nelemans Hanneke , Einhorn Stefan , Grander Dan , Corcoran Martin
Published: October 2019

Chromosomal aberrations on chromosome 5 and in particular band 5q13 is frequently seen in Hairy Cell Leukemia (HCL). Detailed characterization of a constitutional inv (5)(p13.1q13.3) breakpoint in one HCL patient yielded a number of candidate genes that might be important for the development of this malignancy. One of these was beta-hexosaminodase, HEXB, commonly mutated in the lysosomal storage disorder Sandhoff disease. The 5q13.3 breakpoint disrupts a novel evolutionarily conserved alternative isoform of HEXB. This isoform directly overlaps, in a cis-antisense fashion, the first exon of the gene for ectodermal neuronal cortex 1, ENC-1, and was thus named ENC-1AS. The ENC-1 gene has previously been shown to be overexpressed in a variety of malignancies and has been described as a potential oncogene. Analysis of ENC-1 in purified primary HCL tumor cells showed marked upregulation of ENC-1 expression in all patients examined, compared to normal peripheral blood lymphocytes from healthy donors. Although initially found as a p53-induced gene, the upregulation of ENC-1 in HCL follows a p53-independent pathway since activation of the p53 pathway did not lead to further increase in ENC-1 expression in HCL cells. Examining the ENC-1/ENC-1AS locus in more detail, we identified a complex regulatory mechanism involving an inverse expression of the ENC-1 sense and the ENC-1AS transcripts in several tissues. Thus, it appears as if upregulation of ENC-1 is a common phenomenon in HCL and that the regulation of this gene may be dependent on its novel antisense partner ENC-1AS.
Human Platelet Glycoprotein VI: Identification of Residues Involved in the Binding to Collagen.
Authors: Jandrot-Perrus Martine , Lecut Christelle , Arocas Veronique , Ulrichts Hans , Villeval Jean-Luc , Lacapere Jean-Jacques , Deckmyn Hans
Published: October 2019

Glycoprotein VI (GPVI), an essential platelet collagen receptor, belongs to the immunoglobulin (Ig) superfamily and presents two Ig-like loops in its extracellular domain (EC1, EC2). Little is known about the structures involved in GPVI binding to collagen and to its specific ligands, convulxin and collagen related peptides (CRP). Our aim was to characterize these sites using different approaches: competition assays, monoclonal antibodies (MoAb) directed against the GPVI extracellular par, phage display, molecular modelling and site-directed mutagenesis. In binding assays using soluble or cell-expressed human GPVI, we observed that: (i) collagen, CRP and convulxin, competed with one another for the binding to GPVI; (ii) GPVI interaction with either ligand is inhibited by at least two MoAb directed to different independent epitopes, and (iii) MoAb displayed selective inhibitory properties on GPVI interaction with each ligand. These observations indicate that the binding sites on GPVI for the diverse ligands seem to be distinct, exhibiting specific features but at the same time sharing common elements. MoAb 9O12 is a potent inhibitor of GPVI binding to collagen. Its epitope was mapped by screening a 15 mer linear peptide library. This allowed the identification of a motif of 7 residues of which 5 are conserved and that presented a relative homology with the GPVI 30–40 sequence. In a computer model, these residues appeared in close vicinity on a surface loop exposed at the surface of the molecule and accessible to ligands. Site-directed mutagenesis revealed that Val34 and Leu36 are critical for GPVI interaction with collagen and CRP. This study allowed the identification of residues within a loop on GPVI EC1 that might be part of a collagen/CRP-binding site.