bluuebunny/crossref_metadata_2025 huggingface dataset:
Incidence of Cytomegalovirus (CMV) Reactivation after Autologous Peripheral Blood Stem Cell Transplantation (ASCT) in 324 Patients with Multiple Myeloma and Lymphoma: A Single-Center Study at the American University of Beirut Medical Center (AUBMC)
Authors: Massoud Radwan , Assi Rita , Fares Elie , Kreidieh Nabila , Mahfouz Rami , Kanj Souha , El-Zakhem Aline , Kharfan-Dabaja Mohamed A. , Bazarbachi Ali , El Cheikh Jean
Published: October 2019

CONTEXT: Cytomegalovirus reactivation (R-CMV) is often diagnosed in allogeneic hematopoietic cell transplant recipients and could determine a CMV-related disease in these immunocompromised patients, involving any organ. R-CMV and end-organ disease after ASCT has not been studied thoroughly. Autograft recipients are generally considered to have low risk of R-CMV or end-organ disease.

OBJECTIVE: Evaluate the incidence, risk factors, and outcome of R-CMV in adult patients with hematologic malignancies undergoing ASCT.

DESIGN: Retrospective single center study.

SETTING: This study was approved by the institutional review board of AUBMC and conducted at our institution

PATIENTS OR OTHER PARTICIPANTS: A total of 324 consecutive ASCT were performed at AUBMC between January 2005 and March 2016. All patients and transplant-related characteristics are listed on Table 1. CMV DNA load in blood was measured by quantitative polymerase chain reaction (PCR) weekly as a routine monitoring strategy in all patients. Irrespective of PCR results, some patients suspected to have gastrointestinal involvement were biopsied. Also, in the presence of symptomatic CMV reactivation, appropriate anti-CMV therapy was instituted.

MAIN OUTCOMES MEASURES: The primary outcome is understanding the potential relationship between R-CMV and Overall Survival (OS). Secondary outcomes included the effect of CMV reactivation on transplant related mortality (TRM), and progression free survival (PFS).

RESULTS: Overall, 53 (16%) patients had R-CMV and 38 (72%) required anti-CMV treatment. Five (1,5%) had CMV disease with positive PCR on colon biopsy, yet two had PCR negative in blood. After a median follow up of 21.5 months (range: 1 to 125 months), there was no significant difference in OS or PFS between patients with or without R-CMV. TRM has increased from 1.1% in patients with no R-CMV to 13% in patients with R-CMV (P=0.003). We didn't observe any impact for age, sex, type of disease, pre-transplant treatment types/lines on the incidence R-CMV following ASCT.

CONCLUSIONS: Our data suggest that R-CMV is not uncommon in ASCT recipients and may contribute to increased TRM. Biopsy is recommended in case of high suspicion of R-CMV irrespective of PCR results.

Disclosures

No relevant conflicts of interest to declare.
Increased Mir-155 Expression Is Associated with FLT3-ITD Mutation and Poor Therapy Response in Chinese AML and Allo-HSCT Could Improve Survival of These Patients
Authors: Yang Xiaofei , Sun Aining , Hu Xingxing , Ma Wenjuan , Wu Depei
Published: October 2019

The aim of this study was to evaluate the diagnostic and prognostic value of miR-155 in Chinese acute myeloid leukemia (AML) patients. The miR-155 levels of 142 de novo AML patients and 28 healthy donors were detected by real-time quantitative PCR (RQ-PCR). The miR-155 expression was significantly up-regulated in AML compared with control (p<0.001). A receiver operating characteristic (ROC) curve revealed that miR-155 expression could differentiate patients with AML from control subjects (AUC=0.907, 95%CI: 0.853-0.960, P<0.001). We also monitored the miR-155 level in 22 de novo AML patients and found that the miR-155 level decreased significantly from the initial diagnosis to post-CR (P<0.001). FLT3 internal tandem duplication (FLT3-ITD) mutation was significantly more frequent in miR-155 high-expressed group versus low-expressed group (p=0.002). Considering all patients, miR-155 high-expressed cases had significantly lower complete remission (CR) and part remission (PR) rate than miR-155 low-expressed cases (p=0.04). Survival analysis was performed in non-M3-AML patients who received allogeneic hematopoietic stem cell transplantation (allo-HSCT) as consolidation therapy. Kaplan-Meier demonstrated that there was no difference in leukemia free survival (LFS) and overall survival (OS) between miR-155 high-expressed and low-expressed patients (p>0.05). Collectively, Increased miR-155 expression is associated with FLT3-ITD mutation and poor therapy response in Chinese AML and allo-HSCT could improve survival of these patients.

Disclosures

No relevant conflicts of interest to declare.
Use of Computed Tomography Pulmonary Angiography for Suspected Pulmonary Embolism in Patients with Malignancy- a Single Center Experience
Authors: Ravulapati Sravanthi , Leung Cerena K , Mehta Mudresh R , Christopher Kara M , Woelich Susan K. , Poddar Nishant
Published: October 2019

Background: Pulmonary embolism (PE) is a potentially lethal condition commonly suspected in patients with malignancy. Computed Tomography Pulmonary Angiography (CTPA) is increasingly used in the diagnosis of PE, and guidelines have incorporated various screening tools including the Modified Geneva and Wells criteria to facilitate exclusion of pulmonary embolism. There is an increased risk of venous thromboembolism in patients with active malignancy and therefore an increased suspicion in patients who present to the emergency department (ED) with concerning symptoms.

Methods: This is a retrospective analysis at a single tertiary care institution. All patients initially diagnosed with an active malignancy since 2005 and underwent a CTPA between January 2010 and October 2015 were reviewed. Patients were excluded if the CTPA was performed in the setting of trauma, a history of benign malignancy, or if the diagnosis of malignancy was made subsequent to the CTPA. Data collected included patient demographics, clinical presentation, type of malignancy and treatment regimen received. The modified Geneva and Wells criteria were applied to all patients independent from the initial ED risk assessment for a PE.

Results: There were 796 patient records reviewed, of which 162 patients met inclusion criteria. Out of these 162 patients, only 8 (4.9%) were found to have a pulmonary embolism. All patients with a positive CTPA had an intermediate risk per the Geneva criteria while only 62.5% had an intermediate risk per the Wells criteria. Of the 154 patients with a negative CTPA, 71.5% and 78.7% had an intermediate risk; 22.5% and 18.7% were classified as low risk based on Wells and Geneva criteria, respectively. The median age of patients was 59 years old, and the majority were male (58%). The most common malignancies in which a CTPA was ordered were lung cancer (27.7%) followed by breast cancer (14.9%) and prostate cancer (6.8%). Despite a negative CTPA, 82 out of 154 patients (53%) were admitted to the hospital.

Conclusion: Pulmonary embolism is commonly associated with and frequently suspected in patients with active malignancy. The incidence of PE over a 5-year period in oncology patients was 5% in our emergency department. In total, 18.7% to 22.5% of patients could have avoided a CTPA if scoring was based on the Wells or Geneva criteria. Based on the review at our institution, the modified Geneva and Wells criteria are not adequate, and a new tool needs to be developed for risk stratification for the diagnosis of PE specifically in patients with active malignancy.

Disclosures

No relevant conflicts of interest to declare.
Test-Retest Reliability Analysis of the Patient Reported Outcomes Burdens and Experiences (PROBE)
Authors: Chai-Adisaksopha Chatree , Iorio Alfonso , Curtis Randall , Frick Neil , Nichol Michael , Noone Declan , O'Mahony Brian , Page David , Stonebraker Jeffrey , Skinner Mark
Published: October 2019

Background: Patient Reported Outcomes Burdens and Experiences (PROBE) questionnaire was developed for assessing health status in patients with bleeding disorders, specifically, hemophilia. Phase I of PROBE study confirmed robustness of methodology and feasibility of the questionnaire.

Objective: To investigate the test-retest reliability of PROBE questionnaire

Methods: Patients and non-hemophilia individuals who attended an hemophilia related workshops were invited to participate in this study. All participants were asked to fill out a PROBE questionnaire for 3 times. The paper-based questionnaires were collected immediately after being filled out on day 1 and day 2. Participants were instructed to log into the website within 2 weeks and fill out the web-based questionnaire. Test-retest reliability was analyzed.

Results: A total of 63 participants were enrolled in this study with a median age of 50 (range 14-76) years. Of these, 30 (47.6%) were hemophilia patients or carriers, 33 (52.5%) were participants with no known bleeding disorders. On general health domain, Kappa coefficients ranged from 0.69 to 0.90, indicating substantial to almost perfect agreement for all items. Reliability of web-based questionnaire showed moderate to substantial agreement for all except 1 item. For hemophilia related domain, Kappa coefficients ranged from 0.5-1.0. Of these, 5 of 11 items were in perfect agreement (Kappa=1.0). Reliability of web-based questionnaire were in substantial to almost perfect agreement. For overall health related quality of life, EQ-5D had Kappa coefficients of 0.62 to 0.89. Intraclass correlation coefficient of visual analog scale (VAS) was 0.90 (95% confidence interval; 0.83-0.94). Test-retest reliability was comparable between hemophilia patients and participants with no known bleeding disorders.

Conclusions: The results suggest that PROBE is a reliable tool to assess patients reported outcomes for hemophiliacs. Test-retest reliability analysis showed that the agreement was comparable between two participant populations. This result suggests that PROBE tool may be used to collect and compare not only patient reported outcome in hemophilia, but to generate norm values in appropriate control population. The Web-based questionnaire has an acceptable agreement with the standard paper-based version.

Disclosures

No relevant conflicts of interest to declare.
A Dominant Mutation in Mitochondrial Unfoldase CLPX Results in Erythropoietic Protoporphyria
Authors: Yien Yvette Y , Nicolas Gael , van der Vorm Lisa , Gouya Laurent , Begonia Hector , Kafina Martin , Deybach Jean-Charles , Baker Tania , Kardon Julia , Phillips John D. , Puy Hervé , Paw Barry H.
Published: October 2019

Red cells synthesize large quantities of heme during terminal differentiation. Central to erythropoiesis is heme synthesis, which requires tight coordination between mitochondrial iron import and synthesis of protoporphyrin IX (PPIX).

Most individuals with erythropoietic porphyria carry loss of function mutations in FECH, or gain of function mutations in ALAS2, resulting in protoporphyrin IX accumulation. We performed whole exome sequencing to identify novel mutations in an individual exhibiting symptoms in whom FECH and ALAS2 mutations were absent (Fig. A, asterisk). We identified a novel CLPX point mutation in this individual (III.2), her father (II.4) and her paternal uncle (II.2), who also exhibited increased porphyrin levels relative to healthy individuals. The individual's mother was healthy and had a wild-type CLPX genotype (Fig. A). CLPX encodes a mitochondrial protein unfoldase that partially unfolds ALA synthase (ALAS) to allow efficient incorporation of its cofactor, pyridoxal phosphate (Kardon et al. 2015 Cell). This greatly stimulates the synthesis of d-aminolevulinic acid (ALA), the first step in heme biosynthesis.

To determine if the CLPX mutation was causative for porphyria, we expressed mutant CLPX in HEK293T embryonic kidney cells and Friend mouse erythroleukemia (MEL) cells. Mutant CLPX expression (MUT) caused a significant increase in ALAS1 (non-erythroid isoform) (Fig. B) and ALAS2 (erythroid isoform) (Fig. C) activity relative to control and wild-type CLPX expressing samples (WT). This increase in ALAS enzymatic activity translated to an increase in PPIX levels (Fig. D, E), consistent with the porphyria phenotype observed in the individuals in this study.

We observed that MUT-expressing samples had increased levels of ALAS1 and ALAS2. To determine if mutant CLPX altered ALAS protein stability, we transfected WT or MUT CLPX into HEK293T and MEL cells. Cells were treated with cycloheximide to block translation. We quantitated degradation rate of ALAS by western blot analysis of cell lysates obtained at several time points after cycloheximide treatment (chase). Expression of MUT CLPX stabilized both ALAS1 and ALAS2, accounting for the increase in ALAS protein levels, activity and downstream production of PPIX.

The control of ALAS enzymatic activity and protein stability by CLPX unveils a novel cause of protoporphyria and insights revealing the ways in which mitochondrial physiology and heme synthesis are interdependent. Our results reveal an important regulatory node where the mitochondrial protein quality control machinery intersects with a key step in heme synthesis and provides an important genetic tool for understanding the pathology of porphyrias.

Figure 1. Figure 1.

Disclosures

No relevant conflicts of interest to declare.
A Chemical Modifier Screen in Zebrafish Mutants for the Spliceosomal Component Sf3b1reveals Genotype-Selective Vulnerabilities
Authors: Cameron Rosannah C. , McKinstry Mia , De La Garza Adriana , Nik Sara , Smith Peter G , Buonamici Silvia , Bradley Robert K. , Bowman Teresa V.
Published: October 2019

Myelodysplastic syndrome (MDS) is a disorder arising from hematopoietic stem and progenitor cell (HSPC) dysfunction resulting in ineffective hematopoiesis. Mutations in the spliceosomal component SF3B1 (Splicing Factor 3B, subunit 1) are prevalent in MDS, but how this leads to aberrant hematopoiesis is unclear. We elucidated the in vivo consequences of sf3b1 misregulation on hematopoiesis and splicing using a zebrafish sf3b1 (sf3b1hi3394) loss-of-function mutant. Primitive erythropoiesis initiates normally in sf3b1 mutants, as evidenced by normal expression of scl (stem cell leukemia), gata1 and beta-globin, but maturation is defective as shown by decreased hemoglobinization and an increase in immature erythroblasts at 48 hours post fertilization (hpf). Similarly, primitive myelopoiesis initiates normally as early expression of scl and pu.1 are normal, but expression of the differentiation markers l-plastin and myeloperoxidase are diminished in mutants at 24 and 28 hpf. Hemogenic endothelial and HSPC markers runx1 and gata2b are also diminished, while the pan-endothelial marker kdrl (kinase insert domain receptor like) and aorta-specific markers notch1b and notch3 are normally expressed at 24 hpf, indicating hematopoietic induction is more sensitive to sf3b1 loss than vascular formation. RNA-seq analysis of sf3b1 mutants revealed large-scale splicing defects and alterations, particularly affecting introns with suboptimal splice sites. Comparison of RNA-seq data from zebrafish sf3b1 mutants and MDS patients harboring SF3B1 mutations revealed a significant overlap in misspliced genes and in the corresponding affected pathways, including many pathways implicated in MDS pathology such as TGFβ (Transforming Growth Factor-Beta) and immune signaling. Together, these results show conservation between zebrafish mutants and human cells with SF3B1 mutations. We next took a chemical genetic approach and discovered sf3b1 mutants were more sensitive than wild type siblings to treatment with E7107, a small molecule inhibitor of the spliceosome, indicating a potential therapeutic window for spliceosome modulators in MDS. Motivated by this finding, we performed a large-scale chemical modifier screen and identified many new lead compounds (6 out of 562 screened) that also show synthetic lethality in sf3b1 mutants. These small molecules are enriched in those with anti-inflammatory properties, suggesting aberrant immune signaling could underlie some of the defects in sf3b1 mutants. Together, our data demonstrate the molecular and cellular consequences of sf3b1 loss in vivo and establish sf3b1 zebrafish mutants as a novel model for discovery of therapeutics for MDS.

Disclosures

Smith: H3 Biomedicine: Employment. Buonamici:H3 Biomedicine: Employment.
Megaloblastic Anemia Associated with Inhibition of Thyrmine Synthesis (Observations During 5-fluorouracil Treatment)
Authors: BRENNAN M. J. , VAITKEVICIUS V. K. , REBUCK J. W.
Published: October 2019

Megaloblastic changes in the erythrocytic series and cellular giantism in the myeloid line characterize the bone marrow response to 5-fluorouracil in man.

Cytoplasmic vacuolizations occur in reticulo-endothelial cells, megakaryocytes, and myeloid elements.

During development of these morpholigic changes in marrow, associated cellular giantism, polynucleism, and nuclear pattern changes appear in the oral, vaginal, colonic mucosa, and in some tumors resembling those encountered in the bone marrow.

The myeloid-erythroid ratio was found to be useful for the purpose of identifying dosage end-points.

The physiological implications Of these changes as related to nucleic acid metabolism were discussed.
Acanthrocytosis, Pigmentary Degeneration of the Retina and Ataxic Neuropathy: A Genetically Determined Syndrome with Associated Metabolic Disorder
Authors: MIER MANUEL , SCHWARTZ STEVEN O. , BOSHES BENJAMIN
Published: October 2019

Clinical and laboratory observations are reported in an 18 year old white male who had a celiac syndrome during childhood and subsequently developed a pigmentary degeneration of the retina and a progressive ataxic neuropathy. The red cells show a peculiar malformation, described as acanthrocytosis. The clinical manifestations are similar to those of four previously reported cases.

Laboratory studies demonstrate many biochemical abnormalities. These are characterized by increased tolerance to carbohydrates, defective intestinal absorption of lipids, hypocholesterolemia, increased urinary excretion of copper, and elevation of certain enzymes in the serum.

Although many vital facts remain unknown, it is suggested that a relationship exists between the clinical manifestations and the metabolic features of this syndrome.
The Viability of Stored Human Platelets
Authors: BALDINI MARIO , COSTEA NICHOLAS , DAMESHEK WILLIAM , LIMAURO ALVERA
Published: October 2019

Viability of normal human platelets preserved for short intervals at 4 C. was studied by in vitro labelling with Na2Cr51O4. As a criterion of viability two parameters were used: (1) the survival time of the infused platelets, and (2) the maximum percentage of platelet radioactivity which could be recovered in the circulation after infusion. From these two parameters the platelet viability index was calculated, and for stored platelets this was expressed in per cent of the value obtained with freshly prepared platelets.

After 3 hours of storage at 4 C. the platelet viability index was reduced to 62 per cent. With longer periods of storage the viability of the platelets fell rapidly with viability indices of 12 per cent after 24 hours and 2 per cent after 48 hours. No significant difference was seen whether the platelets were stored as whole blood, as platelet-rich plasma, or as platelet concentrates suspended in a plasma medium. When stored in saline the platelets lost their viability more rapidly and the viability index was less than 5 per cent after only 24 hours. When the platelets were stored for 24 hours in a DAS-gelatin medium, their viability fell to insignificant levels within 24 hours.

Platelets frozen in glycerol-plasma and stored for 24 hours at —75 C. showed reduction of viability to one-fifth the value obtained with fresh, unfrozen platelets. Even without storage the frozen platelets showed similar values of viability.

From these results the following conclusions may be drawn:

1. Conventional methods of storage at 4 C. result in rapid loss of platelet viability. An inverse, almost logarithmic, relationship exists between time of storage at 4 C. and platelet viability. The glycerol-freezing technique, although better than most available methods, induces a great loss in platelet viability.

2. At present no method can be advised by which platelets can be preserved in viable form even for relatively short periods of time. However, it is important to know that in the absence of plasma, platelet viability is lost more rapidly than in a plasma medium.

3. From the data obtained, it seems advisable for the practice of platelet transfusion to infuse platelets within less than 3 hours after collection.

4. Methods of improving the viability of stored blood platelets based upon their metabolic needs are strongly warranted and are now under study.
Brief Note: Hemoglobin J in an American Caucasian Family of Swedish Ancestry
Authors: WASI PRAWASE , GITHENS JOHN , HATHAWAY WILLIAM
Published: October 2019

Two instances of the heterozygous state of hemoglobin J are described in the male members of an American Caucasian family of Swedish ancestry.
FOLIC ACID IN THE TREATMENT OF PERNICIOUS ANEMIA
Authors: MEYER LEO M.
Published: October 2019

1. Folic acid in daily doses of 15 to 50 mg., orally, or 20 mg. intramuscularly, usually produced a submaximal reticulocytosis in patients with pernicious anemia.

2. In 3 patients the hemoglobin and red cells rose to a level of about 12.0 Gm. and 4.3 million respectively without further rise after 3 months of therapy.

3. Folic acid in the above doses failed to prevent the development or progression of neurological symptoms indicative of subacute combined sclerosis.

4. In 5 patients folic acid in doses of 5 or 10 mg. orally daily combined with ½ unit of liver extract injected intramuscularly daily produced a reticulocytosis greater than that anticipated from adequate liver extract therapy alone.

5. With combined liver extract and folic acid therapy there was evidence of improvement in the symptoms and signs of subacute combined sclerosis in 3 patients.

6. Folic acid, combined with ½ unit of liver extract, was found to produce a complete hematological remission.

7. Folic acid, alone or in combination with small doses of liver extract, produced an improvement in appetite and general well-being in patients with pernicious anemia.

8. The possible enhancing effect of liver extract when combined with folic acid cannot be due to the folic acid content of the former since 1 unit of liver extract contains only 0.38 micrograms of folic acid.31

9. Folic acid administered to a patient with macrocytic anemia due to faulty postoperative intestinal digestion and absorption, produced a complete remission in the blood picture and a marked improvement in signs and symptoms.
PERNICIOUS ANEMIA FROM ADDISON TO FOLIC ACID
Authors: HADEN RUSSELL L.
Published: October 2019

I have tried to review and clarify steps leading to our present knowledge of pernicious anemia as a clinical and etiologic entity. The early history is most illuminating. The development of the present concept of this complicated disease is a triumph of medical research. Many great names both in clinical and research fields are associated with the advance in knowledge of pernicious anemia. Further research will almost certainly clarify problems still unsolved.
PLATELET ADHESIVENESS
Authors: WEINER MURRAY , ZELTMACHER KURT , REICH CARL , SHAPIRO SHEPARD
Published: October 2019

The method described by Helen Wright for estimation of the adhesiveness of platelets has been found to be reproducible. Adhesiveness of platelets is reduced in the presence of hypocoagulable blood. It may or may not be enhanced when the blood is hypercoagulable. In hemophilia, the adhesiveness of platelets has been found to become increased beyond the premedication level when the coagulation defect is corrected by Fraction 1 derived from human plasma. Two cases exhibiting bleeding are described in which deficient adhesiveness of the thrombocytes was the only demonstrable defect noted in the coagulation mechanism.
JAUNDICE AND THE SULFONAMIDE DRUGS
Authors: RAMMELKAMP CHARLES H.
Published: October 2019

It is apparent from a review of the reported cases of hepatitis associated with sulfonamide therapy that it is usually possible to recognize this toxic manifestation. This is of considerable practical importance since, in every instance in which the sulfonamide is responsible for the jaundice, treatment with the drug should be discontinued and some other form of therapy, such as the antibiotics, instituted. If the jaundice is not secondary to the sulfonamide, therapy may be continued even in the presence of hepatitis secondary to the infection.

Jaundice which appears during the first week of chemotherapy is usually associated with a previous history of ingestion of sulfonamides and accompanying signs of toxicity (immediate sulfonamide jaundice) or with acute hemolytic anemia and jaundice (intermediate jaundice). In either instance the diagnosis is not difficult because the clinical and laboratory abnormalities are characteristic. Finally, jaundice which occurs after ten days of chemotherapy is usually associated with other toxic manifestations, especially fever and various forms of rashes (delayed jaundice). Occasionally jaundice may be the only toxic manifestation of sulfonamide therapy and, in such patients, a definite diagnosis may be difficult.
CONGENITAL HEMOLYTIC JAUNDICE. THE PATHOGENESIS OF THE "HEMOLYTIC CRISIS"
Authors: OWREN PAUL A.
Published: October 2019

Six cases of congenital hemolytic jaundice with "hemolytic" crises are reported. It is demonstrated that during the development of the anemia an acute aplastic condition is present in the erythropoietic tissue of the bone marrow with complete cessation of the formation of red cells. The reticulocytes disappear from the blood; jaundice, serum bilirubin and urobilinuria decrease to normal values; and the serum iron increases. This period is further characterized by leukopenia and thrombocytopenia.

The spontaneous recovery is caused by a rapid regeneration of the erythropoietic tissue resulting in a marked reticulocytosis in the peripheral blood, and there is also leukocytosis, an increase in thrombocytes and a rapid fall of serum iron.

During the period of severe anemia an increase in the blood urea and uric acid occurs.

Transfusion experiments revealed an average lifetime of approximately fifteen days for the red cells in congenital hemolytic jaundice, a fact which fully explains the development and symptoms of the crisis as a result of cessation in the formation of red cells.

The findings definitely contradict the theory that an acute increase in the hemolytic process is the reason for the crisis.

The crisis should be called aplastic and not hemolytic.
THE EFFECTS OF ESTROGENS ON THE BONE MARROW OF ADULT FEMALE DOGS
Authors: CRAFTS ROGER C.
Published: October 2019

Large doses of estrogens have a profound effect on the bone marrow of adult dogs. The initial reaction is a great increase in the number of neutrophilic elements in the bone marrow. These neutrophils are released into the blood stream causing a marked rise in the total white cell count. This is followed by congestion of the bone marrow and a destruction of the white cell elements in these marrows. Congested areas and locations formerly occupied by white cell elements are replaced by edema, leaving erythroid elements intact. This accounts for the marked drop in the total white cell count in the blood stream. This is followed by destruction of remaining erythroid elements in the bone marrow and replacement by "edema" until a stage is reached where practically no cells can be found in the marrow.
PERNICIOUS ANEMIA FROM ADDISON TO FOLIC ACID
Authors: HADEN RUSSELL L.
Published: October 2019

The editorial footnote in "Pernicious Anemia from Addision to Folic Acid," by Dr. Russell L. Haden, in the January Blood (3:-24, 1948) should have read: "Wilkinson and Israëls, Waldenström and others report that free hydrochloric acid occurs in approximately 1 case of 100." The term "achlorhydria" was used in error.
THE BONE MARROW ON STERNAL ASPIRATION IN MULTIPLE MYELOMA
Authors: BAYRD EDWIN D.
Published: October 2019

Generalizing, it can be said that the pathologic cells seen in smears of the bone marrow in multiple myeloma resemble the plasma cell and vary from the very anaplastic and immature cell to the well-differentiated and almost characteristic plasma cell.

The feature which the "myeloma" cell shares with the plasma cell is the abundant, granular, basophilic cytoplasm which tends to be fragile and undergo the same degenerative changes in each; namely, the formation of Russell bodies and vacuolization. Fairly frequently a perinuclear clear area or Hof is present and the nucleus tends to be eccentrically placed. Cytoplasmic extensions or pseudopodia may also be seen in either case, but they occur more often and more dramatically in instances of multiple myeloma. Multinucleated cells are commonly seen.

In addition, myeloma-plasma cells will often have a large clear nucleolus and a leptochromatic nucleus and will exhibit a tendency to the formation of isolated areas of condensed chromatin. Cytoplasmic extrusions, free cytoplasmic bodies, occasionally complete with Russell bodies and vacuoles are almost universally present.

All cases were of the plasma cell type; there was no exception. In these cases, the myeloma-plasma cell constituted from 2.5 to 96 per cent of the leukocytic elements present. The opinion was expressed that all so-called types of multiple myeloma are merely variations in differentiation of this same cell.

It was noted that anaplasia, hypernucleation and lack of plasma cell predominance in certain cases were diagnostic pitfalls. Additional evidence was adduced to confirm the reticulo-endothelial origin of the myeloma-plasma cell. It was further observed that certain prognostically valuable information could be gleaned from a careful review of the cytologic characteristics in these cases.
THE A AND B FACTORS AS A POSSIBLE CAUSE OF ERYTHROBLASTOSIS
Authors: OROZCO ALFONSO C. VÉLEZ
Published: October 2019

The author establishes that there is no reason to think that the isoimmunization must be limited to the Rh factor, especially in pregnancy cases, because such a condition brings the ideal means for its presence.

A brief historical review is presented, establishing some evidence for the antigenic capacity of the A and B blood factors, such as the experimental production of serum in animals, the increase of agglutinin titre following an incompatible transfusion or intramuscular blood injection and heterospecific pregnancies. Some fetal antigens are analyzed, as well as the possible pathogenic process of sensitization. Some factors of the antigen-antibody conflict in the fetal organism are studied. The pregnancies are also analyzed from the viewpoint of the fetal maternal incompatibility explaining the low frequency of the observed cases, due to nonpermeable placentas, the anergy of the mother, and the fetal characteristic of being secretor or nonsecretor, considering that the clinical form of the fetal alteration depends upon the presence or absence of A and B substances in fetal tissues. The writer gives as a basis to consider a true isoimmunization the presence in the blood stream of erythroblasts with nuclear alterations, and a positive "developing test."

A clinical case of twins in which the sick twin was of incompatible blood type in respect to the mother is presented.
Hereditary stomatocytosis: membrane and metabolism studies
Authors: Mentzer WC Jr , Smith WB , Goldstone J , Shohet SB
Published: October 2019

A defect in the protein kinase-mediated phosphorylation of erythrocyte membrane proteins, previously unrecognized in stomatocytosis, was discovered in a boy with hereditary stomatocytosis and severe hemolytic anemia. The high-sodium, low-potassium erythrocytes of this patient were remarkably permeable to both sodium and potassium. The rate of ouabain-inhibitable active cation transport was more than ten times normal and was sustained by an increase of similar magnitude in glycolysis. The deformability in vitro of fresh stomatocytes was reduced and deteriorated further after a brief period of incubation with glucose. Ferrokinetic studies showed that these rigid cells were sequestered by the spleen. When stomatocytes were deprived of glucose in vitro, ATP depletion and ATPase cation pump failure rapidly ensued. Because of their permeability defect, such depleted cells rapidly became swollen and lysed. Prolonged entrapment in acidic, hypoglycemic regions of the spleen would recapitulate these unfavorable events in vivo. In this regard, splenectomy was followed by an improvement in erythrocyte survival, although evidence of continuing hemolysis was obtained.
Human megakaryocytes express clusterin and package it without apolipoprotein A-1 into alpha-granules
Authors: Tschopp J , Jenne DE , Hertig S , Preissner KT , Morgenstern H , Sapino AP , French L
Published: October 2019

Clusterin, a 70-Kd disulfide-linked two-chain plasma glycoprotein circulates in blood as a high-density lipoprotein particle and is highly induced after tissue injury and tissue remodeling. In this study, peripheral blood leukocytes were assayed for clusterin expression. The protein was predominantly detectable in human platelets by immune cytochemistry. The content of clusterin was determined and amounts to 2.5 +/- 1.3 micrograms/10(9) platelets, thus representing about 2% of the blood pool. Clusterin purified from human platelets had the same molecular weight as plasma clusterin under nonreducing conditions and was composed of two disulfide-linked nonidentical subunits of the same size. Both preparations were sensitive to reduction yielding the two subunits of 35 Kd. In contrast to plasma clusterin, the platelet form was not complexed to apolipoprotein A-I. By immunogold labeling, alpha-granule localization of clusterin was observed. Complete release of platelet clusterin occurred at optimal doses of A23187, phorbol myristate acetate (PMA), and thrombin. Because clusterin mRNA was detected by hybridization in situ in bone marrow- derived megakaryocytes, platelet clusterin is most likely produced and packaged into alpha-granules during megakaryocyte development.
New variant of von Willebrand disease type II with markedly increased levels of von Willebrand factor antigen and dominant mode of inheritance: von Willebrand disease type IIC Miami
Authors: Ledford MR , Rabinowitz I , Sadler JE , Kent JW , Civantos F
Published: October 2019

A variant of von Willebrand disease (vWD) was identified in six members of a kindred spanning four generations. The proband was a 46-year-old woman with a lifelong history of bleeding, a prolonged bleeding time (> 15 minutes), markedly elevated von Willebrand factor (vWF) antigen (vWF:Ag = 2.09 U/mL), slightly reduced ristocetin cofactor activity, and a plasma vWF multimer pattern similar to that of vWD type IIC. Similar findings were observed in her three children, mother, and brother. In affected family members, platelet and plasma vWF multimer patterns were discrepant with higher molecular weight multimers observed in platelet vWF. Following a 1-Des-amino-8-D-arginine vasopressin (DDAVP) challenge, the proband failed to normalize her bleeding time even though vWF: Ag rose by 70% and higher molecular weight multimers were increased slightly. Genetic studies were consistent with autosomal dominant inheritance of a mutation within the vWF gene. By sequencing of cloned genomic DNA, mutations were excluded in exons 4, 5, 14, and 15, which encode regions of the vWF propeptide proposed to be important in multimer biosynthesis. Mutations also were excluded in exons 28 to 31, which encompass the known mutations that cause vWD types IIA, IIB, and B. This new variant of vWD, characterized by autosomal dominant inheritance, a qualitative defect that resembles vWD type IIC, and increased plasma vWF:Ag, was tentatively designated vWD type IIC Miami.
Sera from patients with heparin-induced thrombocytopenia generate platelet-derived microparticles with procoagulant activity: an explanation for the thrombotic complications of heparin-induced thrombocytopenia
Authors: Warkentin TE , Hayward CP , Boshkov LK , Santos AV , Sheppard JA , Bode AP , Kelton JG
Published: October 2019

Heparin-induced thrombocytopenia is characterized by moderate thrombocytopenia and thrombotic complications, whereas quinine/quinidine-induced thrombocytopenia usually presents with severe thrombocytopenia and bleeding. Using flow cytometry and assays of procoagulant activity, we investigated whether sera from patients with these immune drug reactions could stimulate normal platelets to generate platelet-derived microparticles with procoagulant activity. Sera or purified IgG from patients with heparin-induced thrombocytopenia stimulated the formation of platelet-derived microparticles in a heparin-dependent fashion. Further studies showed that heparin-induced thrombocytopenia sera also produced a marked increase in procoagulant activity. In contrast, sera from patients with quinine- or quinidine-induced thrombocytopenia did not generate platelet-derived microparticles nor generate increased procoagulant activity. However, quinine/quinidine-induced thrombocytopenia sera produced a significant increase in the binding of IgG to platelets in a drug-dependent fashion, whereas sera from patients with heparin-induced thrombocytopenia demonstrated no drug-dependent binding of IgG to platelets. We also observed increased levels of circulating microparticles in patients with acute heparin-induced thrombocytopenia compared with control patients. Our observations indicate that the generation of procoagulant platelet-derived microparticles in vivo is a plausible explanation for the thrombotic complications observed in some patients with heparin-induced thrombocytopenia.
Platelet adhesion to fibronectin in flow: dependence on surface concentration and shear rate, role of platelet membrane glycoproteins GP IIb/IIIa and VLA-5, and inhibition by heparin
Authors: Beumer S , IJsseldijk MJ , de Groot PG , Sixma JJ
Published: October 2019

Platelet adhesion to purified surface-immobilized fibronectin under flow conditions was investigated. Fibronectin was found to support attachment and spreading of platelets. The extent of platelet spreading depended on the amount of immobilized fibronectin. An antiglycoprotein (anti-GP) IIb/IIIa antibody and an Arg-Gly-Asp (RGD)-containing peptide inhibited adhesion almost completely, whereas antibodies directed against platelet GP Ic/IIa (very late antigen 5) inhibited by 50%. Similar results with the antibodies and the peptide were found in a static system. A comparison of different anticoagulants showed no difference in adhesion using citrate or hirudin. However, unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH) as the only anticoagulant or in combination with citrate maximally inhibited adhesion by 80% and 60%, respectively. Preincubation of the immobilized fibronectin with UFH resulted in a maximal inhibition of 90%, whereas preincubation with LMWH had no effect. When we preincubated the surface with heparins of different size, we observed 40% inhibition of adhesion with heparins with an average MW of up to 18 kD, whereas a heparin with an average MW of 21 kD almost completely blocked adhesion. These results indicate that platelet adhesion to fibronectin in flow involves several receptors, is highly RGD-mediated, does not require physiologic levels of divalent cations, and can be inhibited by direct binding of heparin to the fibronectin surface.
Absence of WAF1 mutations in a variety of human malignancies
Authors: Shiohara M , el-Deiry WS , Wada M , Nakamaki T , Takeuchi S , Yang R , Chen DL , Vogelstein B , Koeffler HP
Published: October 2019

A newly cloned gene named wild-type p53-activated fragment 1 (WAF1; also known as p21, Pic-1, Cip-1, or SDI1) is directly regulated by p53 and can itself suppress tumor cell growth in culture. Induction of expression of WAF1 may be an important means by which cells with DNA injury arrest their growth to repair DNA or undergo apoptosis. Based on the hypothesis that mutations of this gene may play a role in carcinogenesis, we have studied 351 DNAs from 14 kinds of malignancies, as well as 36 human transformed cell lines, for alterations of WAF1 gene by single-strand conformation polymorphism analysis of polymerase chain reaction amplification of the DNA coding region of the WAF1 gene. No abnormal band shifts of WAF1 were noted in any of the samples or cell lines, but three major variants in exons 2 and 3 of the gene were found that are consistent with the existence of two different DNA polymorphisms. Sequence analysis of the amplified products producing these three variants in each exon from normal DNAs confirmed the presence of the polymorphisms in the WAF1 gene. Of 290 selected tumor samples previously evaluated for p53 mutations by single-strand conformation polymorphism, 90% had no detectable p53 alterations. In summary, mutations within the coding portion of the WAF1 gene were undetectable in a large series of human tumors, many of which had a normal p53 gene. This suggests that WAF1 alterations are generally caused indirectly, through p53 mutations rather than through intragenic mutation of the WAF1 itself.
Regulation of the human cellular glutathione peroxidase gene during in vitro myeloid and monocytic differentiation
Authors: Shen Q , Chada S , Whitney C , Newburger PE
Published: October 2019

We have used the HL-60 and PLB-985 myeloid leukemia cell lines to examine the regulation of expression of the important intracellular antioxidant enzyme, glutathione peroxidase (GSH-Px), during phagocytic cell differentiation in vitro. Induction of differentiation along the monocytic pathway by phorbol ester results in an approximately twofold rise in enzyme activity and a parallel increase in the rate of 75Se incorporation into immunoprecipitable GSH-Px protein. Induction along the granulocytic pathway by dimethyl formamide (DMF) results in similar changes in steady-state enzyme levels and rates of GSH-Px protein synthesis. Steady-state levels of GSH-Px gene transcripts also increase more than twofold, approximately in parallel with the enzyme levels. Nuclear run-on transcription assays of GSH-Px mRNA synthesis show ratios of induced to uninduced transcript levels of 2.24 and 1.59 with phorbol myristate acetate (PMA) induction and DMF, respectively, in HL- 60 cells, and ratios of 1.34 and 3.46 with PMA and DMF, respectively, in PLB-985 cells. Half lives of GSH-Px mRNA are unchanged or slightly shorter after differentiation of HL-60 cells, and slightly longer after induction of PLB-985. Overall, the present studies show that GSH-Px activity rises during in vitro-induced monocytic or granulocytic differentiation of myeloid cell lines and that the increased expression of the cellular GSH-Px gene occurs through complex mechanisms that include transcriptional up-regulation. This pattern contrasts with the nearly complete cotranslational regulation of GSH-Px expression by exogenous selenium.
Transfection of wild-type deoxycytidine kinase (dck) cDNA into an AraC- and DAC-resistant rat leukemic cell line of clonal origin fully restores drug sensitivity
Authors: Stegmann AP , Honders WH , Willemze R , Ruiz van Haperen VW , Landegent JE
Published: October 2019

The AraC-resistant rat leukemic cell line RO/1-A has been shown to have a typical deoxycytidine kinase (DCK)-deficient phenotype and cannot metabolize the antileukemic drugs cytarabine (AraC) and decitabine (DAC). To investigate the relative contribution of mutations in the dck gene to the development of in vitro-induced AraC-resistance, a neomycin selectable plasmid construct harboring the wild-type dck coding region was transfected into RO/1-A. Polymerase chain reaction analysis confirmed the presence of vector DNA in the target cells (RO/1-ADCK) that were stably transfected and monitored over a period of 14 weeks. Northern and Western blot analysis showed restoration of dck mRNA and protein expression. Initial rate measurements of DCK activity showed that Km values for dck were only slightly altered as a result of transfection, whereas strongly increased Vmax values were observed, resulting in a 12-fold increased phosphorylation efficiency for both dC and AraC, compared with the AraC-sensitive parental cell line RO/1 from which the RO/1-A was originally derived. In vitro sensitivity to AraC- and DAC-mediated cytotoxicity was fully restored in RO/1-ADCK. The data pinpoint acquired DCK deficiency caused by mutations of the dck gene as the major cause of AraC resistance in this model.
Myelosuppressive conditioning improves autologous engraftment of genetically marked hematopoietic repopulating cells in dogs
Authors: Barquinero J , Kiem HP , von Kalle C , Darovsky B , Goehle S , Graham T , Seidel K , Storb R , Schuening FG
Published: October 2019

We have studied the role of different conditioning regimens for engraftment of genetically marked hematopoietic repopulating cells in dogs. Peripheral blood (PB) and/or marrow cells collected after treatment with recombinant canine stem cell factor (rcSCF) or cyclophosphamide were transduced in a vector-containing long-term culture system. Three different vector-producing cell lines with similar viral titers were used. In two of them, the neo-containing LN vector was packaged either in the PA317 cell line with an amphotropic murine retrovirus envelope or the PG13 cell line with the gibbon ape leukemia virus (GALV) envelope. The MFG/GC vector produced in PA317 cells contained the human glucocerebrosidase gene. Nineteen dogs received either no conditioning (group A, n = 5), irradiation to both humeri with 1,000 cGy (group B, n = 5), a sublethal dose of cyclophosphamide 40 mg/kg (group C, n = 4), a sublethal dose of 200 or 300 cGy total body irradiation (TBI) (group D, n = 3), or an otherwise lethal dose of 920 cGy TBI (group E, n = 3) before intravenous (groups A, C, D, E) or intramedullary (group B) infusion of the transduced autologous hematopoietic cells. Transduction efficiency of hematopoietic cells at the time of infusion into the animals was similar among the different conditioning groups. Dogs were observed for at least 6 months. PB granulocytes were obtained at least every 3 weeks after transplant and analyzed by polymerase chain reaction for the presence of the transduced genes. The percentages of positive results in dogs more than 4 weeks after transplantation were 0% without conditioning, 5% with local irradiation, 18% with sublethal cyclophosphamide, 33% with sublethal TBI, and 17% with otherwise lethal TBI. Analyzing the influence of conditioning regimens by a generalized estimating equation (GEE) technique, which considered the use of different retrovirus vectors and the number of mononuclear cells infused as potential confounding variables, we found that engraftment of genetically marked repopulating cells was significantly improved (P < .001) in dogs receiving systemic conditioning with either otherwise lethal TBI, sublethal TBI, or sublethal cyclophosphamide compared to dogs with local irradiation only or no conditioning. Within the limitation of the experimental design, these data suggest that myeloablative or myelosuppressive conditioning improves engraftment of genetically marked hematopoietic repopulating cells.