bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

Abstract

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

Abstract
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

Abstract
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

Abstract
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

Abstract
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.
FOLIC ACID IN THE TREATMENT OF PERNICIOUS ANEMIA
Authors: MEYER LEO M. 
Published: October 2019

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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.