bluuebunny/crossref_metadata_2025 huggingface dataset:
Epidemiology Data of Patients with Chronic Myeloid Leukemia in West Central Mexico
Authors: Campos-Cabrera Virginia , Campos-Villagomez Jose-Luis , Campos-Cabrera Salvador , Campos-Cabrera Gregorio
Published: October 2019

Background:Chronic myeloid leukemia (CML) treatment in Mexican mestizos since the use of TKIs has been well described. Most of the studies only refers to the response rather than epidemiology. There are two large cohorts reported this years (Gad Med Mex 2016;152:208-212 and Clin Lymphoma Myeloma Leuk 2016;16:57-62), but all national information is not available.

Methods:Epidemiological data from samples referred to Laboratorios Fatima de Michoacan for identification of BCR-ABL translocation by chromosome or molecular analysis in patients with suspected chronic myeloid leukemia.

Results:77 positive samples for identification of BCR-ABL were analyzed. By physician indication 46 karyotype and 31 PCR. Male 40 and females 37, ratio 1.08/1. Age range from 15 to 87 years, mean 44.8. Three less than 19 years, thirteen from 20 to 29, twenty from 30 to 39 years, twelve from 40 to 49, twelve from 50 to 59, seven from 60 to 69, nine from 70 to 79 and one more than 80 years.

Conclusions: Compared with data from other Mexican CML studies (cited above) the age at diagnosis was similar: 44.8, 45.8 and 47.0 years; important to mention that in the three studies there are patients less than 15 years old; in the ratio M/F there was a difference: 1.08, 1.37 and 1.32. Compared to CLL epidemiology data that we presented last year in the 57thASH Annual Meeting (Blood 2015;126:5269), CML presented more than 20 years earlier, 66.0 to 44.8 years; M/F ratio is elevated in CLL 1.6 a 1; and CML is more frequent, 77 to 52 cases in CLL.

Disclosures

No relevant conflicts of interest to declare.
Emerging Potential Roles of Lysyl-Oxidase-like 2 Stimulates Cancer Associated Fibroblasts Promoting Myelofibrosis of Myeloproliferative Neoplasms
Authors: Liu Xiaoli , Xu Na , Pan Chengyun , Zhou Xuan , Liu Qifa , Feng Ru , Sun Jing , Zhou Hongsheng
Published: October 2019

Backgroud and objective: Myeloproliferative neoplasms (MPNs) are a group of clonal haematological disorders that characteristic with a multipotent haematopoietic stem cell transformation, often associating with the progression of myelofibrosis in the evolution course of disease. Progressive myelofibrosis finally turned out a high risk factor of transformation to leukemia and bone marrow failure. Cancer Associated Fibroblasts (CAFs) are recently thought to be a critical mediator in several hematological malignancies tumor microenvironment and associate with fibrosis. Lysyl-oxidase-like 2 (LOXL2) is a member of the lysyl oxidase family that promote the crosslinking of collagens or elastins in the extracellular matrix and mediate epithelial-mesenchymal transition (EMT). Here, we want to verify CAFs mediating myelofibrosis and explore the potential role of LOXL2 association with CAFs in simulated vivo microenvironment.

Patients and methods:For bone marrow specimens, normal samples (n=19) and patients with polycythemia vera (PV) (n=21), essential thrombocythemia (ET) (n=32), and primary myelofibrosis (PMF) (n=9) were contrasted. Markers of CAFs including α-smooth actin(α-SMA), fibroblast activation protein(FAP), transforming growth factor-β1(TGF-β1) and LOXL2 were detected by quantitative reverse transcription-PCR(RT-PCR). we also detected α-SMA, FAP, LOXL2 and reticulin protein by western blot and immunohistochemical staining. For cell lines, α-SMA and FAP were measured after cocultured mesenchymal stem cell(MSCs) with recombinant human lysyl oxidase homolog 2 Protein(rhLOXL2) in hypoxic niche for 24, 48, 72 and 96 hours, respectively.

Results: Markers of CAFs displayed a differential pattern of expression in MPNs especially in PMF(P<0.010) compared with normal samples(P=0.023) according to RT-PCR. Among these markers, the expression of α-SMA is the highest. For western blot and immunohistochemical staining, we discovered the level of α-SMA, FAP and LOXL2 expression was associated with the grading of reticulin fibrosis in bone marrows. we also founded that α-SMA and FAP were significantly expressed after cocultured MSCs with rhLOXL2 in hypoxia for 96 hours, but there were no significant increase in gene and protein level expression with the prolonged exposure time.

Conclusion: Over expression of CAFs played a critical role in promoting bone marrow fibrosis. LOXL2 may be a key factor in stimulating MSC to CAFs and further promoting disease progression, which may provide a new target spot for further excavation of the pathogenesis of MPNs and looking for more effective targeted therapies.

Disclosures

No relevant conflicts of interest to declare.
Early Bortezomib Failure Predicts Shorter PFS: a Retrospective Analysis from 4 Victorian Centres
Authors: Bergin Krystal , Lew Thomas E , Chow Annie W.S. , Kopanidis Paul , Lim Hui Yin , Chan Kah Lok , Chai Khai-Li , Chua Chong Chyn , Reynolds John , Spencer Andrew
Published: October 2019

Aim: To compare patients with newly diagnosed multiple myeloma (NDMM) withbortezomib responsive and resistant disease (defined as <PR after 2 cycles) and assess the impact on clinical outcome particularly progression-free survival (PFS) and overall survival (OS).

Method: Retrospective review of all patients identified with NDMM treated with upfront bortezomib in four tertiary centres in Victoria, Australia between 1st of January 2009 and 1st of March, 2016. Bortezomib responsiveness was defined as achievement of PR or better after completing 2 cycles of any bortezomib containing therapy (total eight doses of bortezomib). Prior radiotherapy was allowed. Data were analysed via log rank tests for PFS and OS using SAS (V9.4) software. Predictors of bortezomib resistance were assessed by chi-square analysis.

Result: A total of 187 eligible patients were included (147bortezomib responsive and 40bortezomib resistant patients). Bortezomib resistance was associated with shorter median PFS, 27.4 (95%CI: 16.3, 30.8) vs 30.4 (95%CI: 23.4, 37.1) months (log-rank p= 0.047) with a median potential follow up of 27.4 vs 29.7 months in the resistant and responsive groups respectively. Estimated PFS at 12 months was 68.0% (95%CI: 50.3%, 80.5%) vs 88.3% (95%CI: 81.2%, 92.8%) and, at 24 months, 60.4% (95%CI: 41.7%, 74.7%) vs 60.0% (95%CI: 49.9%, 68.6%) in the resistant and responsive groups respectively, indicating non-constant hazard in the resistant group (see figure). Bortezomib resistance was not associated with higher ISS (ISS3 24% vs 15%, p=0.17), high riskcytogenetics (17p del ort(4;14)) (23% vs 23%, p= 0.99), MM sub-type (p= 0.63) or length ofbortezomib cycle (21 vs 35 day p= 0.56)). In responders (n=147), maintenance therapy was associated with improved PFS (p=0.021), and PFS at 24 months was, 70.7% (95%CI: 57.3%, 80.5%) vs 46.9% (32.0%, 60.5%) in those who received maintenance therapy and those who did not respectively. In non-responders (n=40) PFS at 24 months was 68.8% (95%CI: 35.7%, 87.3%) vs 54.9% (31.6%, 73.2%) in those patients who received maintenance therapy and those patients who did not respectively (p=0.84). Median OS has not been reached in either thebortezomib resistant or responsive groups however estimated OS at 24 months is 82.8% (95%CI: 62.8%, 92.7%) vs 89.5% (95%CI: 81.5%, 94.1%) in the resistant and responsive groups respectively.

Conclusion: Bortezomib resistance is a poor prognostic factor and is associated with a shorter PFS. Consideration of early therapy change, maintenance therapy and/or referral for assessment for allograft in eligible patients appears to be warranted.

Figure Figure.

Disclosures

No relevant conflicts of interest to declare.
VD Versus VTD Induction: Similar Efficacy in Controlling Disease in Transplant Eligible Multiple Myeloma Patients Outside Clinical Trials
Authors: Cocito Federica , Mangiacavalli Silvia , Ferretti Virginia Valeria , Cartia Claudio Salvatore , Ganzetti Maya , Cazzola Mario , Corso Alessandro
Published: October 2019

Autologous stem cell transplant (ASCT) remains the standard of care for Multiple Myeloma (MM) patients younger than 70 years old. The role of induction therapy is crucial within a program of high-dose therapy since deeper is the response before, higher is the outcome of transplant.

In this study, we analyzed a real life setting of patients treated with three different induction approaches: VAD (Vincristine-Adriamycin-Dexamethasone), VD (Bortezomib - Dexamethasone), and VTD (Bortezomib-Thalidomide-Dexamethasone) in terms of depth of response, 2 years therapy-free rate and toxicity.

One hundred and sixty-three MM patients (pts) were included in the analysis: 62 pts treated with VAD (38%), 44 with VD (27%) and 57 with VTD (35%). In VTD group 49 pts (86%) received Bortezomib subcutaneously. As shown in Table 1, patients of the three groups were similar for D&S stage (p 0.59), a higher rate of ISS stage 3 was observed in VAD group (p=0.019), patients in VTD group were significantly older (p=0.024), median follow-up was significantly lower in VTD pts (p<0.001). The overall response rates after induction were similar in all three groups (p=0.156), with higher rate of responses of good quality (CR+VGPR) for patients treated with Bortezomib-based combinations: VAD 24.2%, VD 52.3%, VTD 63.2% (p<0.001). No difference was observed between VTD and VD (p=0.258). A different pattern of responses was observed after transplant, VTD, in fact, was superior to VAD (p<0.001) and VD (p=0.012), while no difference was highlighted between VAD and VD (p=0.352). As a matter of fact, 2 years therapy-free rate were: 48% for patients treated with VAD vs 73% with VD vs 74% with VTD (p=0.189). Of note, however, bortezomib base therapy maintained its superiority with respect to VAD (p=0.03). No differences were observed between VD and VTD regimens in terms of toxicity of any grade and type (52.3% VD vs 52.6% VTD, p> 0.9), and of discontinuation rate (14% in VTD and 18% in VD, p=0.395). The incidence of all grades peripheral neuropathy (PN) was similar between VD and VTD (28.2% and 31.4% p=0.835), but grade 3-4 PN was significantly higher in VD group (no patients in VTD vs 18% in VD pts, p=0.078), probably due to different way of Bortezomib administration in VTD group (86% of pts received subcutaneous Bortezomib).

In this study, we confirm that Bortezomib-based regimens are better than VAD in terms of overall response rate, good quality responses and long-term disease control. VTD is superior to VD in terms of good quality responses after transplant but disease control at 2 years is similar.

Disclosures

Corso: Janssen-Cilag: Honoraria.
Robust Production of Cytomegalovirus pp65-Specific T Cells By a Fully Automated IFN-γ Cytokine Capture System
Authors: Kim Nayoun , Nam Young-Sun , Im Keon-Il , Lim Jung-Yeon , Jeon Young-Woo , Song YuneJin , Lee Jong Wook , Cho Seok-Goo
Published: October 2019

Cytomegalovirus (CMV) related diseases are a serious cause of morbidity and mortality following hematopoietic stem cell transplantation (HSCT). As an alternative to antiviral drugs, CMV-specific cytotoxic lymphocytes (CMV-CTLs) can provide long-term CMV-specific immunity without major side effects. In this study, we apply the IFN-γ cytokine capture system (CCS) using the fully automated CliniMACS Prodigy device to rapidly produce CMV-CTLs. Five validation runs were performed using apheresis samples from randomly selected CMV-seropositive healthy blood donors. CliniMACS Prodigy automatically performed successive processes including antigen stimulation, anti-IFN-γ labelling, magnetic enrichment, and elution which took approximately 13 hours. The original apheresis samples consisted of 0.3% CD3+IFN-γ+ T cells which were mainly CD45RA+CD62L+ naïve T cells. Following IFN-γ enrichment, the target fraction contained 51.3% CD3+IFN-γ+ cells with reduction in naïve T cells and the selection of CD45RA-CD62L- and CD45RA+CD62L- memory T cells. Interestingly, the frequency of CMV pp65 specific CD3+ T cells was also increased by 1.7-fold. Furthermore, extended culture of isolated cells revealed efficient proliferation with sustained antigen-specific IFN-γ secretion and cytotoxicity against pp65 pulsed target cells. Therefore, we suggest IFN-γ CCS by CliniMACS Prodigy as a simple and robust approach to produce CMV-CTLs, which may be highly feasible and applicable for clinical use.

Disclosures

No relevant conflicts of interest to declare.
New Therapeutic Options: Alternates to Gene Therapy for Treating Hemoglobinopathies
Authors: De Franceschi Lucia
Published: October 2019

Hemoglobinopathies are chronic, invalidating globally distributed hereditary red cell disorders, which impact patient survival and quality of life. Sickle cell disease (SCD) and β-thalassemias are the more frequent hemoglobinopathies worldwide. In both SCD and β-thalassemias, novel therapeutic strategies might be divided into (1) pathophysiology related therapies and (2) innovative curative therapeuties such as hematopoietic stem cell transplantation and gene therapy, the latter discussed in another presentation in this session. In the last two decades, studies have highlighted the biocomplexity of SCD and β-thalassemias and the need for the development of new therapeutic options targeting one or more of the mechanisms involved in their pathogenesis. In SCD, the pathophysiology related novel therapies might be divided into: (i)Molecules targeting sickling and red cell dehydration; (ii) Molecules targeting SCD vasculopathy and sickle cell-endothelial adhesive events; (iii) Molecules modulating SCD related oxidative stress. Since inflammatory vasculopathy is a key factor in SCD related organ damage, major efforts have been made to develop agents that interfere with SCD vasculopathy and adhesion events, involving sickle red cells, neutrophils and endothelial interplay. β-thalassemic syndromes are characterized by ineffective erythropoiesis, which plays a crucial role in anemia and in the development of extramedullar erythropoiesis. In β-thalassemic syndromes, pathophysiology related novel therapies might be divided into: (i) Agents reducing the ineffective erythropoiesis such the TRAP ligands, targeting the GDF11 pathway (sotatercept or luspatercept); (ii) Agents affecting Jak2 activation to reduce splenomegaly in β-thalassemia and (iii) Agents modulating hepcidin - erythropoiesis axis such as minihepcidins. The main goal of these molecules is to ameliorate the ineffective erythropoiesis resulting in increased hemoglobin levels with reduction of transfusion requirement, iron overload, and facilitating iron chelation therapy. This review highlights new therapeutic strategies in SCD and β-thalassemia and discusses future development, research implications, and possible directions toward future clinical trials.

Disclosures

De Franceschi: F. Hoffmann-La Roche Ltd, Basel, Switzerland: Research Funding.
The Effect of Homologous Marrow Transplantation on the Survival of Monkeys Following Sublethal Who1e-Body Radiation
Authors: NEWSOME FRANCES E. , OVERMAN R. R.
Published: October 2019

Homologous bone marrow given to the rhesus monkey 30 hours after LD 10 radiation dose of 550r increased the mortality rate expected from the radiation alone. The symptoms produced resembled those obtained at higher doses of radiation. The effects of marrow given 48 hours after radiation differed little from those produced by the radiation alone. It is suggested that the 30-hour injection elicited a more violent reaction or that the reaction occurred during a more decisive time in the process of recovery.
ON HOOKWORM ANEMIA
Authors: STRANSKY EUGENE , QUINTOS FLORENCIO N.
Published: October 2019

We have discussed three types or stages of blood changes in hookworm disease. Attention is directed to a severe irreversible aplastic anemia which occurs not infrequently in hookworm disease. The pathogenesis is obscure, but we believe that it is due to bone marrow exhaustion following continuous blood loss in the presence of various factors, including dietary deficiency.

The symptomatology, hematologic and bone marrow findings are described.
THE RELATIONSHIP BETWEEN HEPARIN DOSAGE AND CLOTTING TIME
Authors: JAQUES L. B. , RICKER ANN G.
Published: October 2019

1. The relationship between clotting time and heparin dosage has been studied in the dog.

2. On the addition of heparin to blood in vitro, a linear relation is found between heparin dosage and the logarithm of the clotting time obtained. The sensitivity of the blood sample to the action of added heparin is influenced both by the individual (coagulability of the blood before withdrawal) and by the technics of withdrawal and of determination of the clotting time. It is indicated that alterations in the latter may be used to extend the range of measurable hypocoagulability due to heparin. Incubation of heparin with blood for ten minutes increases its anticoagulant effect.

3. When moderate doses of heparin are injected intravenously, five to fifteen minutes are required for the clotting time to reach a maximum. No evidence of a biphasic response was obtained. The maximum clotting time obtained is greater than it is with the same amount of heparin added to the blood in vitro, due to the effect of incubation of heparin with blood on its anticoagulant activity. The in- terval required for the clotting time to return to normal is quite short, and with a given dosage is constant with different animals. Factors influencing the relation between duration of hypocoagulability and dosage are discussed.

4. A test has been devised to determine the sensitivity of the animal to the anticoagulant action of heparin. The clotting time response to certain concentrations of heparin added to the blood in vitro is determined. A fixed dose of heparin is then injected intravenously and the clotting time response is again determined. The response in vitro measures the sensitivity of the clotting system to heparin, while the in vivo response, when interpreted in the light of the in vitro response, measures the ability of the body to remove heparin from the circulation.

5. By means of this test, it has been determined that anesthesia with pentobarbital decreased the coagulability of the blood, urethane had no effect on coagulability, while the effect of ether was variable. The injection of india ink and evisceration caused a hypercoagulability, while removal of the kidneys had little effect.

6. When the sensitivity of the blood to the anticoagulant action of heparin was tested during these procedures, pentobarbital and nephrectomy had no effect, ether caused an increase in sensitivity, urethane a decrease. The injection of india ink and also evisceration markedly decreased the sensitivity of the blood to the anticoagulant action of heparin.

7. Anesthesia with pentobarbital, ether or urethane, the injection of india ink, removal of the kidneys, or removal of the gastrointestinal tract, had no effect on the duration of heparin action in the body.
RESPONSE OF TROPICAL SPRUE TO VITAMIN B12
Authors: SPIES TOM D. , SUAREZ RAMON M.
Published: October 2019

These findings show that the administration of vitamin B12 to patients with tropical sprue was followed by general clinical and hematologic improvement provided the dosage was adequate. A single dose of 4 micrograms administered in case 4 produced little or no change. The larger dosage of 10-25 micrograms administered in the other cases was accompanied by striking increase in strength and vigor and a decrease in the diarrhea; however, in no instance was a maximal dose given and these patients quickly tended to relapse clinically and hematologically. They could be relieved promptly again either by another injection of vitamin B12 or by a compound of folic acid. (The conjugated compounds of folic acid used in these cases were used for experimental purposes, and they produced the same hematologic response as that of folic acid per se.) Case 3, who had an excellent hematologic response after eating one serving of 400 grams of liver, is regarded as especially significant in that it suggests that, as powerful as vitamin B12 is as a therapeutic agent, it is more effective when given with liver. It is especially noteworthy that cases 1 and 2, who had three injections of vitamin B12, have had steady clinical and hematologic improvement. The reader should have in mind that a single injection of approximately 100 micrograms of vitamin B12 probably would be needed to produce a full hematologic response in persons so ill. This tentative appraisal would suggest that this therapeutic compound, per unit of weight, is more effective in treating human disease than any compound that yet has been used.
STUDIES CONCERNING THE PATHOGENESIS OF GAUCHER’S DISEASE
Authors: OTTENSTEIN BERTHA , SCHMIDT GERHARD , THANNHAUSER S. J.
Published: October 2019

1. The serum of normal individuals and of patients with Gaucher’s disease does not contain cerebrosides in measurable amounts. Cerebrosides in Gaucher’s disease are increased only in those organs containing abundant numbers of cells characteristic of the disease.

2. Normal red blood cells contain approximately 0.19 per cent cerebrosides. The cerebroside in red blood cells is a galactosidocerebroside.

3. Red blood cells in Gaucher’s disease do not differ quantitatively and qualitatively in their cerebroside content from the red blood cells of normal individuals.

4. In four different cases of Gaucher’s disease separate determinations of splenic galactosido- and glucosidocerebrosides were made. The spleen of two adults showed mainly glucosidocerebrosides and only traces of galactosidocerebrosides, while the analysis of the spleen of two other adults showed that galactosido- as well as glucosidocerebrosides may be accumulated simultaneously in Gaucher cells. These findings are of importance, since it is demonstrated that the deviation of the cerebroside metabolism in Gaucher cells not only results in the formation of an abnormal glucosidocerebroside, but also may lead to the increased formation of the normal galactosidocerebroside, kerasin. It is demonstrated that the relative proportions of galactosido- and glucosidocerebrosides in Gaucher cells may differ considerably in individual cases.

5. The organs of infantile siblings with "generalized infantile Gaucher’s" disease were analyzed. The organs of one infant showed mainly galactosidocerebrosides while in the organs of the other sibling both kinds of cerebrosides, glucosido- as well as galactosidocerebrosides, were present.

6. The findings reported in this paper lend support to the theory that Gaucher’s disease is the result of a deviation of the intracellular metabolism of reticulum cells and histiocytes. The cerebrosides are not transported by the serum or by the red blood cells and secondarily deposited in the cells involved, but are formed and stored in the reticulum cells and histiocytes were they are found.
STUDIES OF PHOSPHORUS METABOLISM IN MAN: II. A STUDY OF THE PERMEABILITY OF THE HUMAN ERYTHROCYTE TO INORGANIC PHOSPHATE IN VITRO BY THE USE OF RADIOACTIVE PHOSPHATE (P32)
Authors: TAYLOR F. H. L. , LEVENSON S. M. , ADAMS M. A. , KENDRICK MARY
Published: October 2019

1. Phosphate exchange in red cells and plasma was studied in vitro using P32 in the form of sodium phosphate as a tracer.

2. No phosphate was added other than the isotopic preparation which was of high specific activity.

3. Inorganic phosphate exchanged freely between the plasma and the erythrocytes at 37.5 C. in a period of four hours. Minimal transfer occurred at 7 C.

4. Most of the added P32 which passed into the erythrocytes during this time remained in the inorganic fraction, less than 15 per cent being found in the organic acid soluble fraction.

5. The specific activity of the inorganic phosphate of the erythrocytes was equal to or greater than that obtaining for the inorganic phosphate of the plasma at the end of the four hour incubation period at 37.5 C.
STUDIES IN IRON TRANSPORTATION AND METABOLISM
Authors: DUBACH REUBENIA , CALLENDER SHEILA T. E. , MOORE CARL V.
Published: October 2019

1. The isotope technic for studying iron absorption has been extended to measure the unabsorbed isotope in feces as well as the amount synthesized into hemoglobin. The recovery of radioiron from feces was shown to be accurate within 10 per cent.

2. There was suggestive evidence to indicate that, with the 1 mg. per kilogram dose employed, normal subjects may sometimes absorb more iron than is converted within a two week period into hemoglobin.

3. Patients with fever, untreated pernicious anemia, and refractory anemia were shown to absorb more iron than they use for hemoglobin.

4. Patients with hemolytic anemia may absorb more iron than can be recovered in the peripheral blood at any one time because isotopic hemoglobin is removed from the circulation at a rapid rate.

5. Except in afebrile patients with hypochromic anemia, acceptance of the per cent of a given dose of radioiron which appears in circulating hemoglobin as a measure of iron absorption must be made with caution.

6. The theory that mucosal cells accept iron for absorption or block its assimilation provides the best known explanation for iron absorption; but patients with adequate iron stores may assimilate considerable quantities of the metal and the block must be regarded as relative.
THE EFFECT OF RADIATION ON HEMOPOIESIS. IS THERE AN INDIRECT EFFECT?
Authors: LAWRENCE JOHN S. , VALENTINE WILLIAM N. , DOWDY ANDREW H.
Published: October 2019

1. The general aim of the investigations here reported has been to obtain evidence for or against "indirect" radiation effects.

2. To this end, twenty-six successful cross circulation experiments (carotid to carotid anastomoses) have been performed between normal cats and radiated cats.

3. Cross circulation was established in most instances at some specified time interval after the radiation of one partner. All intervals up to eighty-two hours after radiation of one partner were covered.

4. In seven experiments cross circulation was established and then one animal radiated while the other was shielded. These were considered the most critical experiments of the group.

5. Detailed data on leukocyte and lymphocyte counts in the normal animals obtained during an approximately twenty-eight day period of follow-up are presented.

6. These data are not considered to support the thesis of indirect effects peculiar to radiation. A trend toward slightly lowered absolute lymphocyte counts in normal animals after cross circulation was not considered significant, and in no instance did leukopenia develop in the normal animal.

7. The literature is reviewed and discussed.
CLOTTING OF PLASMA AND SILICONE SURFACES
Authors: PATTON THOMAS B. , WARE ARNOLD G. , SEEGERS WALTER H.
Published: October 2019

Blood was collected with special care and centrifuged in silicone treated glass tubes at 22-23,000 R.P.M. for various periods of time. Even after prolonged centrifugation, incoagulable plasma could not always be obtained. When clots formed, only a small portion of the fibrinogen was represented as fibrin. Minute amounts of thrombin cause this fibrin formation. No evidence was found to support the view that plasma may contain a soluble factor which can initiate clotting independent of platelet action.
CARBAMATES IN THE CHEMOTHERAPY OF LEUKEMIA
Authors: SKIPPER HOWARD E. , RISER WILLIAM H. , STELZENMULLER ANNE , HOLT HELEN
Published: October 2019

A simple procedure for screening compounds for leukotoxic action is described. Results obtained on assaying known leukopenic agents such as urethan, nitrogen mustard, x-ray, potassium arsenite, and benzene indicate a reasonable measure of reliability.
A METHOD FOR OBTAINING LIVING LEUKOCYTES FROM HUMAN PERIPHERAL BLOOD BY ACCELERATION OF ERYTHROCYTE SEDIMENTATION
Authors: MINOR ALLEN H. , BURNETT LEE
Published: October 2019

A simple method for obtaining living leukocytes from human peripheral blood is presented. It consists of the addition of fibrinogen to heparinized blood. Essentially all the leukocytes remain in plasma suspension and essentially all the erythrocytes are sedimented out at the end of one hour.
NUCLEAR STRUCTURE VERSUS NUCLEAR PATTERN
Authors: JONES OLIVER P.
Published: October 2019

1. Lymphoid cells from rat and human fetal livers and adult rat lymph nodes were studied by means of various cytologic, cytochemical and optical technics.

2. Freshly dried smears preserve cytoplasmic detail with an extraordinary degree of accuracy.

3. Mitochondria, both numerous and pleomorphic, are preserved in the cytoplasm which lies beyond the nuclear membrane as well as in the thin layers of cytoplasm above and beneath the nucleus.

4. In dry-fixed preparations stained with Romanowsky dyes, mitochondria appear as negative images (light areas) in basophilic cytoplasm. These negative images within the nuclear area contribute to the formation of what hematologists in the past have described as parachromatin.

5. The dry smear method of studying lymphoid blood cells produces nuclear patterns formed by the sum total of nuclear and cytoplasmic alterations which reflect functional activity.

6. The meaning of the terms nuclear structure and nuclear pattern have been contrasted.
DIAGNOSIS AND MECHANISM OF HEMOLYSIS IN CHRONIC HEMOLYTIC ANEMIA WITH NOCTURNAL HEMOGLOBINURIA
Authors: DACIE J. V.
Published: October 2019

1. Studies in vitro and transfusion experiments indicate that the "cause" of nocturnal hemoglobinuria is an abnormality of the erythrocytes. In vitro, patient’s corpuscles undergo hemolysis in fresh human sera, but only within a pH range of 6 to 8, a range more restricted on the alkaline side than the limits within which isohemolysis will take place.

2. The nonspecific nature of the acid-serum test is emphasized. In addition to nocturnal hemoglobinuria, positive tests for hemolysis may be obtained in this way with certain "warm" and "cold" hemolysins, and in the presence of marked spherocytosis. The control observations necessary for the diagnosis of nocturnal hemoglobinuria are described.

3. The erythrocytes in nocturnal hemoglobinuria are remarkably sensitive to hemolysis by anti-A (or anti-B) but are not hemolyzed by anti-Rh.

4. It is suggested as a hypothesis that the same abnormality, presumably at the corpuscular surface, which is the cause of the increased sensitivity to hemolysis by anti-A results in the erythrocytes being fatally sensitive in vivo to a hemolytic factor distinct from complement and normally present in serum.
HEMOLYTIC REACTIONS PRODUCED IN DOGS BY TRANSFUSION OF INCOMPATIBLE DOG BLOOD AND PLASMA
Authors: YOUNG LAWRENCE E. , ERVIN DONALD M. , YUILE CHARLES L.
Published: October 2019

1. Dogs injected intravenously with dog erythrocytes containing one or more antigenic factors lacking in their own red cells developed iso-hemagglutinins and hemolysins exhibiting characteristics of immune antibodies.

2. Transfusions of incompatible whole dog blood and plasma were carried out under controlled conditions. Pretransfusion observations were made and followed by closely spaced post-transfusion measurements of serologic and hematologic alterations.

3. The rate of destruction of incompatible donated corpuscles was determined by tagging the cells with radioactive iron and also by employing the technique of differential agglutination of erythrocytes. It was thereby shown that all of the incompatible donated cells disappeared from the recipient’s circulation within the first thirty to ninety minutes following transfusion. The probable mechanisms and relative importance of intra- and extravascular destruction of erythrocytes are briefly discussed.

4. Destruction of recipient dogs’ corpuscles by donated immune plasma was relatively slow, and spherocytosis and increased osmotic fragility of the recipients’ cells were evident for periods as long as twenty days. These observations are compared with those made in human beings after transfusions of plasma and of blood from dangerous universal donors.

5. The titer of complement in the sera of recipient dogs was sharply reduced for at least five hours after all transfusions of incompatible whole blood, but isoagglutinin titers were less regularly reduced after such transfusions.

6. Other notations of interest included estimates of the concentrations of serum bilirubin, sodium and potassium, determinations of clotting time, prothrombin concentration, and observations on red cell morphology, intravascular erythrophagocytosis, and shifts in distribution of leukocytes and in the electrophoretic patterns of plasma.
EFFECTS OF THE INTRAMUSCULAR ADMINISTRATION OF BAL (2,3 DIMERCAPTOPROPANOL) IN A SUBJECT WITH THE SICKLE CELL TRAIT: CASE REPORT
Authors: KUHNS WILLIAM J.
Published: October 2019

The effects of the intramuscular administration of BAL in a Negro harboring the sickle cell trait have been presented. It was observed that the rate of sickling was accelerated and the erythrocyte sedimentation rate was retarded in the presence of BAL both in vitro and apparently in vivo. However, the administration of BAL produced none of the pathologic sequelae characteristic of sickle cell disease.
THE PATHOGENESIS OF ERYTHROBLASTOSIS FETALIS
Authors: KLINE B. S.
Published: October 2019

The mechanism of transfer, in cases of erythroblastosis fetalis, of incompatible fetal red blood cells to the mother and of maternal blood with antibody to the fetus, was observed especially well in 2 cases in which the infants were born alive.

The two placentas showed occlusion of peripheral blood vessels of many villi and trunks by agglutinated red blood cells and fibrin. Associated with the vascular thromboses, there were, in places, necrosis of the walls and of regional tissues with rupture and hemorrhage of fetal blood, containing numerous intact nucleated red blood cells, into regional intervillous spaces. Through the broken surfaces, adjacent maternal blood was in contact with the fetal circulation.

A more accurate designation for "erythroblastosis fetalis" would be "transplacental erythrocytotoxic anemia."
OBSERVATIONS ON THE INFLUENCE OF THE HYPOPHYSIS AND THE ADRENAL CORTEX ON BLOOD PLATELET LEVELS
Authors: ADAMS ELIJAH
Published: October 2019

Observations were made to investigate possible endocrine influences on blood platelets. Adrenal cortex extract failed to influence the platelet counts of mice, rats, or rabbits. Adrenalectomy and sham-adrenalectomy were followed by almost identical platelet increases in mice and rats. Administration of adrenal cortex extract, or physiologic saline, to adrenalectomized rats was followed by a consistent fall in platelets not observed in sham-adrenalectomized rats, or after administering distilled water to adrenalectomized rats. Platelet levels in hypophysectomized rats were significantly lower than in unoperated controls. Splenectomy in hypophysectomized rats was followed by a maximum rise in platelets markedly lower than following splenectomy in intact rats. Bone-marrow megakaryocytes in hypophysectomized rats were significantly fewer than in intact rats. No changes in megakaryocyte number or morphology appeared following splenectomy either in intact or hypophysectomized rats.
The relationship between anemia, fecal stercobilinogen, erythrocyte survival, and globin synthesis in heterozygotes for beta-thalassemia
Authors: Gallo E , Pich P , Ricco G , Saglio G , Camaschella C , Mazza U
Published: October 2019

Erythrocyte survival, fecal stercobilinogen, and the alpha/beta-chain synthesis ratio in recticulocytes and bone marrow (six cases only) were determined and related to the hemoglobin levels in ten heterozygous carriers of beta-thalassemia with different degrees of anemia. Erythrocyte survival values were low, though not related to the degree of anemia, whereas stercobilinogen values and changes in the peripheral blood alpha/beta-chain ratios were so correlated. In the case of bone marrow, on the other hand, the ratio between alpha-chain and beta-chain synthesis was virtually 1, irrespective of the degree of anemia. It is suggested that the severity of anemia in heterozygous carriers is independent of peripheral hemolysis and entirely attributable to destruction of erythroblasts within the marrow.
Thymidylate synthetase activity in bone marrow cells in pernicious anemia
Authors: Sakamoto S , Niina M , Takaku F
Published: October 2019

The tritium release assay for the demonstration of thymidylate synthetase activity has been applied to the measurement of enzyme activity in the bone marrow of four patients with pernicious anemia and nine normal subjects. On the average, an approximately ninefold increase in enzyme activity was observed in patients with pernicious anemia. In the absence of 5, 10-methylene-tetrahydrofolate, enzyme activity was reduced in both normal and in pernicious anemia cells. Addition of 5, 10-methylene-tetrahydrofolate to the assay medium resulted in a far greater activation of thymidylate synthetase activity in megaloblastic bone marrow cells than in the cells of control subjects.
In vivo release and turnover of secreted platelet antiheparin proteins in rhesus monkey (Macaca mulatta)
Authors: Musial J , Niewiarowski S , Edmunds LH Jr , Addonizio VP Jr , Nicolaou KC , Colman RW
Published: October 2019

Human and rhesus monkey platelets secrete at least two antiheparin proteins: platelet factor 4 (PF4) and low affinity platelet factor 4 (LA-PF4). Neither of these proteins showed species-related antigenic differences. As determined by radioimmunoassay, the levels of PF4 and LA-PF4 antigen per 10(9) monkey platelets amounted to 10.7 and 20.3 microgram, respectively. One milliliter of monkey plasma prepared from blood collected into an anticoagulant composed of EDTA, prostaglandin E1, and theophylline solution contained 22.4 ng LA-PF4 and 8.0 ng PF4. Concentrations of these two platelet-specific proteins in monkeys closely resembled levels found in human platelets and plasma. Infusion of prostacyclin (PGI2) (100 or 300 ng/kg/min) into monkeys for 15 min resulted in a significant decrease of plasma levels of LA-PF4 antigen and of PF4 by 40%--60% (p < 0.0001). This decrease was related to the inhibitory effect of PGI2 on the secretion of platelets stimulated by a catheter or by venipuncture. Longer infusion of PGI2 did not produce further significant change. The supernate obtained after aggregation of human platelets stimulated by thrombin was injected into monkeys receiving PGI2 infusion. The disappearance of LA-PF4 antigen in monkey plasma followed a biphasic exponential curve with half-lives for the fast and slow components of 8.4 and 63 min. PF4 disappeared faster but followed the same pattern (half-lives for the fast and slow component of 2.1 and 70 min). Analysis of the experimental data suggests that the low levels of secreted platelet proteins in monkey plasma are related to their minimal in vivo release and to their rapid clearance.
Detection of factor IX antibodies by radioimmunoassay: effect of calcium on antibody-factor IX interaction
Authors: Lewis RM , Reisner HM , Chung KS , Roberts HR
Published: October 2019

A radioimmunoassay for alloantibodies (inhibitors) and heteroantibodies to human factor IX has been developed using radioiodinated human factor IX and formalin-fixed, heat-killed Staphylococcus aureus cells (Staph A). Staph A was used as a solid-phase adsorbent for immune complexes. The assay is specific, shows excellent correlation with factor IX coagulant neutralization assays in detecting alloantibodies (r = 0.98), and is 60 times more sensitive. The Staph A method allows rapid separation of immune complexes (within 10 min of addition) and binds immunoglobulin with equivalent efficiency in the presence and absence of calcium. These characteristics have made the Staph A binding method useful for observing the Ca++ effect on the antigenicity of factor IX as detected by hetero- and alloantibodies. All antisera investigated showed some increase in antibody titer when measured in the presence of Ca++. One particular alloantibody showed the greatest increase in titer (> twofold). The reversibility of the calcium effect by excess EDTA indicates that it was not caused by Ca++-dependent proteolysis of the factor IX molecule. The Staph A procedure can also be used as a sensitive competitive radioimmunoassay for human factor IX using alloantibody. The assay could detect factor IX antigen to a dilution of 1:1280 of normal plasma and correctly classified hemophilia B plasmas as cross-reacting material positive or negative.
Artificial microvascular graft thrombosis: the consequences of platelet membrane glycoprotein Ib inhibition and thrombin inhibition
Authors: Dascombe WH , Hong C , Garrett KO , White JG , Lyle VA , Miller JL , Johnson PC
Published: October 2019

Early thrombosis of artificial microvascular grafts (AMG, grafts < or = 2 mm internal diameter) prevents their reliable clinical use. The present studies were undertaken to examine the effect of hirudin, a thrombin-specific inhibitor, and of the F(ab')2 fragment of PG-1, a monoclonal antibody (MoAb) directed against guinea pig platelet membrane glycoprotein Ib (GPIb), on AMG patency in an animal model. One- centimeter long segments of expanded polytetrafluoroethylene (ePTFE), 0.88 mm internal diameter, were serially implanted as interposition grafts in the guinea pig femoral arterial systems bilaterally. A control group was treated with 0.5 mL saline intravenously (IV) 30 minutes before limb 1 and limb 2 graft implantation. Three experimental groups were treated with 0.5 mL saline IV before limb 1 graft implantation as an animal control and with either 0.5 mL saline containing 1.25 mg/kg IV PG-1 F(ab')2, (which inhibits ristocetin- induced platelet agglutination and von Willebrand factor binding), hirudin 1 mg/kg IV, or a combination of both agents before limb 2 graft implantation. GPIb inhibition, thrombin inhibition, and the combination resulted in a significant prolongation of AMG patency (P < .005). Whereas thrombin inhibition with hirudin prolonged AMG patency similar to that observed with GPIb inhibition, the combination of GPIb and thrombin inhibition provided the overall longest prolongation of AMG patency. These results indicate that both platelet membrane GPIb and thrombin play a role in AMG thrombosis.
Localization of ligands for L-selectin in mouse peripheral lymph node high endothelial cells by colloidal gold conjugates
Authors: Kikuta A , Rosen SD
Published: October 2019

L-selectin, a Ca(2+)-dependent lectin-like receptor, mediates lymphocyte attachment to high endothelial venules (HEV) of peripheral lymph nodes (PLN) during the process of lymphocyte homing. Two endothelial-derived ligands for L-selectin, known as GlyCAM-1 (Sgp50) and CD34 (Sgp90), have been identified by affinity precipitation of lymph node extracts with a chimeric molecule that combines the extracellular domains of L-selectin with the human IgG1 Fc region (L- selectin-IgG) (J Cell Biol 110:2221, 1990). Here, using a histologic probe based on colloidal gold conjugated to L-selectin-IgG (LS-Ig), we performed morphologic mapping of the HEV ligands in PLN at both the light and electron microscopic levels. With a postembedding labeling method, intense LS-Ig-gold staining of PLN HEV was observed, while the HEV of Peyer's patches (PP) were negative. The specificity of LS-Ig- gold staining was established by pretreatment of sections with sialidase and coincubation of sections with EGTA, fucoidin, or L- selectin-IgG itself. In ultrastructural studies of high endothelial cells(HEC), gold particles were bound to the trans-Golgi network(TGN) and to peripheral vesicles in the cytoplasm. Gold labeling was also detected in a patchy distribution on the entire luminal vascular surface of HEC. Although the perivascular fibroreticular sheath of HEV was frequently labeled limited labeling was observed on the basolateral surfaces of the HEC. In most cases, the HEC membrane surrounding migrating lymphocytes was negative. These results show that L-selectin ligands pass through the Golgi apparatus during their biosynthesis, are stored in secretory granules, and are expressed on the vascular luminal surface of the HEC. A polyclonal antiserum to GlyCAM-1 intensely stained intracellular organelles in the biosynthetic pathway including cytoplasmic vesicles, but failed to stain the cell surface of HEC. Given its presence in serum as a soluble factor, GlyCAM-1 is likely to be a secretory product.
Early mortality and the retinoic acid syndrome in acute promyelocytic leukemia: impact of leukocytosis, low-dose chemotherapy, PMN/RAR-alpha isoform, and CD13 expression in patients treated with all-trans retinoic acid
Authors: Vahdat L , Maslak P , Miller WH Jr , Eardley A , Heller G , Scheinberg DA , Warrell RP Jr
Published: October 2019

All-trans retinoic acid (RA) has proven a major advance in the treatment of acute promyelocytic leukemia (APL). However, the proper management of patients who present with or develop leukocytosis during remission induction with all-trans RA is not established, nor is there a clear relation between leukocytosis and the development of the retinoic acid syndrome. We reviewed the course of our patients who underwent induction with all-trans RA to identify potential factors that might predict for the development of this syndrome and to identify which patients, if any, might specifically benefit from additional treatment with cytotoxic chemotherapy. Seventy-eight courses of all- trans RA therapy were administered to patients with a molecular diagnosis of APL. Initial and peak leukocyte counts, their rate of rise, leukocyte count criteria developed in Europe, and cell surface marker expression were all analyzed relative to subsequent development of both the RA syndrome as well as all causes of early mortality. The outcome of patients who received specific treatment for retinoid- induced leukocytosis was also examined. No factor was found to consistently predict for the development of the RA syndrome. Although the occurrence of the syndrome was positively associated with the peak value of the peripheral blood leukocyte count (P = .001), neither the initial leukocyte count nor the rate of rise in leukocyte counts on days preceding onset of the syndrome were sufficiently well-correlated to be clinically useful (P = .21). The leukocyte count criteria developed in Europe had a sensitivity of 62%, a specificity of 69%, and a positive predictive value that ranged from only 44% to 72%. However, we unexpectedly found that basal expression of CD13 (aminopeptidase N), a cell surface enzyme previously linked to tumor cell invasion and an inferior outcome in patients with acute myeloid leukemia, was highly associated with both development of the syndrome (P < .05) as well as an elevated leukocyte count (P = .006). Neither low-dose chemotherapy nor leukapheresis prevented development of the syndrome nor ameliorated its effects. In fact, 9 of 11 patients who received these interventions sustained fatal or near-fatal events, most of which were due to hemorrhage. However, early treatment with a short-course of high-dose corticosteroids halted progression of the syndrome in most cases. Finally, we found that expression of the type “A” isoform of PML/RAR- alpha (also known as bcr3 or “short”) was associated with a significantly shorter duration of relapse-free and overall survival (P = .005).(ABSTRACT TRUNCATED AT 400 WORDS)
Natural protection against severe Plasmodium falciparum malaria due to impaired rosette formation
Authors: Carlson J , Nash GB , Gabutti V , al-Yaman F , Wahlgren M
Published: October 2019

Genes for two lethal diseases, thalassemia and sickle cell anemia, are favored by evolution because, in their heterozygous form, they protect against cerebral malaria. Rosette formation, the binding of uninfected red cells (RBCs) to Plasmodium falciparum-infected RBCs (PRBCs), has previously been found to be associated with cerebral malaria, the most important severe manifestation of P falciparum malaria. We show here that thalassemic RBCs and, under certain conditions, even hemoglobin S (HbS)-containing RBCs possess an impaired ability to bind to PRBCs, forming small and weak erythrocyte rosettes compared with rosettes formed by normal RBCs. This decreased rosetting ability is associated with the small size of the thalassemic RBCs and with distortion of the mechanical properties of HbS-containing RBCs. The impairment of rosette formation may hinder the development of cerebral malaria by abatement of sequestration.