The Effect of Various Metabolites on the Growth of Marrow Cells in Vitro

Abstract Marrow was cultured on chick plasma clots, glass surfaces and in suspension and the influence of various metabolites and antimetabolites on the growth and differentiation of red cell precursors was studied. Vitamin B12 appeared to stimulate cell growth in cultures grown on clots and to convert megaloblastic hematopoiesis to normoblastic in cultures grown on glass. In suspension cultures of megaloblastic marrows in pernicious anemia serum significant cell increases were obtained with the addition of folinic acid, but not with vitamin B12. Both vitamin B12 and folinic acid appeared to accelerate the conversion of megaloblasts to normoblasts in differential counts of stained smears of such cultures. No enhancing effect of intrinsic factor upon the activity of B12 in suspension cultures of megaloblasts was observed. The effect of folinic acid could be blocked by colchicine and folinic acid could reverse the inhibitory effects of Amethopterin. These facts indicated that folinic acid acted in suspension cultures by stimulating mitosis. Megaloblasts could be produced from normoblastic marrows by culturing them in pernicious anemia serum, or in normal serum in the presence of Amethopterin.

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On the Nature of Castle’s Hemopoietic Factor

Blood ◽

10.1182/blood.v6.12.1234.1234 ◽

1951 ◽

Vol 6 (12) ◽

pp. 1234-1239 ◽

Cited By ~ 30

Author(s):

SHEILA T. CALLENDER ◽

L. G. LAJTHA

Keyword(s):

Vitamin B12 ◽

Gastric Juice ◽

Intrinsic Factor ◽

Normal Serum ◽

Relationship Of ◽

The Relationship

Abstract 1. Normal gastric juice (intrinsic factor) and vitamin B12 together form a thermolabile hemopoietic factor which ripens megaloblasts in vitro, both gastric juice and B12 alone being inactive. 2. The hemopoietic factor in normal serum which ripens megaloblasts in vitro also appears to be thermolabile, heating to 56 C. for 2 hours destroying some of its activity. 3. The relationship of these factors is discussed and an extra-gastric as well as a gastric source of intrinsic factor is postulated.

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Absorption of Liver-bound Vitamin B12 in Relation to Intrinsic Factor

Blood ◽

10.1182/blood.v32.2.313.313 ◽

1968 ◽

Vol 32 (2) ◽

pp. 313-323 ◽

Cited By ~ 9

Author(s):

KUNIO OKUDA ◽

ISAO TAKARA ◽

TERUMI FUJII

Keyword(s):

Vitamin B12 ◽

Pernicious Anemia ◽

Rat Liver ◽

Intrinsic Factor ◽

In Vitro Digestion ◽

Enhanced Absorption ◽

Biochemical Studies ◽

Peptide Complexes ◽

Liver Powder

Abstract Rat liver containing radioactive native B12 was prepared by repeated injections of 57Co-OH-B12, and absorption of liver B12 was measured in patients with pernicious anemia and in subjects without stomach, using physiologic doses. It was found that absorption of liver B12 was very poor, not superior to that of free OH-B12, and coadministration of IFC markedly enhanced absorption. In vitro digestion of rat liver with several enzymes, as determined from liberation of dialyzable radioactivity, suggested its easy digestibility. Biochemical studies of the dialyzable products of liver containing 57Co-B12 failed to demonstrate any detectable quantities of radioactivity other than free 57Co-OH-B12. A study in which cow liver powder mixed with a small quantity of 57Co-CN-B12 was fed to humans and digestion of liver was estimated from the reduction in absorption of radioactivity, indicated that most of the extractable liver B12 was liberated free in the intestine. Thus, no evidence has been obtained for the production of B12-peptide complexes from liver by digestion that require no IF for absorption.

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Polycythemia Rubra Vera With Pernicious Anemia Some Observations on Vitamin B12 Metabolism

Blood ◽

10.1182/blood.v34.1.14.14 ◽

1969 ◽

Vol 34 (1) ◽

pp. 14-24 ◽

Cited By ~ 4

Author(s):

ROBERT E. SAGE

Keyword(s):

Vitamin B12 ◽

Pernicious Anemia ◽

Parietal Cell ◽

Intrinsic Factor ◽

Serum Levels ◽

High Serum ◽

Normal Serum ◽

Polycythemia Rubra Vera ◽

The One ◽

Symptoms And Signs

Abstract A patient with polycythemia rubra vera developed symptoms and signs of pernicious anemia three years after institution of therapy for the polycythemia. This is the first satisfactorily documented report of this sequence. Strict criteria utilized make both diagnoses unequivocal. Autoantibody studies, the first performed in such a patient, show the presence of parietal cell and intrinsic factor autoantibodies but no antibodies to other tissues. Vitamin B12 requirements have been excessive despite high or high normal serum levels and extreme marrow sensitivity to withdrawal of the Vitamin is evident in the presence of these high serum levels. Studies on Vitamin B12 binding proteins showed an elevation in α binding globulin with reversal of the α to β ratio. It is suggested that this protein is physiologically abnormal. The literature is briefly reviewed regarding the pathogenesis of the two disorders with the conclusion that their appearance in the one patient represents purely a chance phenomenon.

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Oral Treatment of Pernicious Anemia with Small Doses of Vitamin B12 Combined with Mucinous Materials Derived from the Hog Stomach

Blood ◽

10.1182/blood.v8.10.867.867 ◽

1953 ◽

Vol 8 (10) ◽

pp. 867-892 ◽

Cited By ~ 24

Author(s):

GEORGE B. JERZY GLASS ◽

LINN J. BOYD

Keyword(s):

Vitamin B12 ◽

Pernicious Anemia ◽

Ammonium Sulfate ◽

Oral Treatment ◽

Intrinsic Factor ◽

Pyloric Mucosa ◽

Small Doses ◽

Factor Concentrate ◽

Oral Doses

Abstract 1. Twenty patients with pernicious anemia were treated under rigidly controlled conditions by administering small oral doses of vitamin B12 in combination with intrinsic factor containing mucinous materials processed from the hog stomach. This study was undertaken in view of our earlier observation concerning the relationship of one of the mucin fractions of the human stomach (glandular mucoprotein) to Castle’s intrinsic factor. The following sources of intrinsic factor from animal stomach were used: (a) commercial gastric mucin; (b) acetic acid extracts of hog pyloric mucosa; (c) mucinous materials precipitated by acetone or by saturation with ammonium sulfate from hydrochloric acid extracts of the hog pyloric mucosa; (d) mucous fractions obtained by further fractionation in the electroconvection apparatus of the above mucinous materials precipitated with ammonium sulfate; (e) an intrinsic factor concentrate from the hog stomach in combination with small doses of vitamin B12, processed under the trade name of Bifacton. 2. Results obtained in this study indicate the feasibility of attaining complete clinical remission and suboptimal or optimal hematopoietic responses in patients with pernicious anemia in relapse when small daily doses (below 20 µg.) of vitamin B12 are given orally in combination with mucinous materials processed by various technics from the hog stomach and containing intrinsic factor. The provisions are that: (1) the doses of both vitamin B12 and intrinsic factor containing materials be adequate and optimal in regard to their mutual ratio; (2) the process involved in processing intrinsic factor containing materials not impair their activity and yield a product sufficiently concentrated. 3. The hematopoietic responses and clinical remissions obtained in six cases of pernicious anemia with Bifacton (vitamin B12 with intrinsic factor concentrate) in different batches were uniform in our hands. The doses employed were marginal, in order to detect differences in potency, and although the reticulocyte responses were frequently suboptimal and protracted, the increases in red cells were roughly equivalent to those obtained with daily administration of approximately 1 unit of a standard oral antianemia preparation. Bifacton is supplied in strikingly smaller dosages than the standard liver or stomach oral preparations, and is fully active in a total daily oral dose of about 50 mg. of intrinsic factor concentrate with 15 µg. vitamin B12. 4. Preliminary studies on two patients with pernicious anemia tend to indicate that the administration of massive single oral doses of Bifacton repeated at intervals of one week may imitate even more closely the results of parenteral treatment with injectable liver concentrates or vitamin B12 than does the administration of this concentrate in small daily doses. The single dose of Bifacton which was given once a week to one of these patients contained 150 µg. vitamin B12 with approximately 250 mg. of intrinsic factor concentrate. Five consecutive single doses administered at intervals of from seven to ten days induced a complete clinical and hematologic remission. 5. The studies on two patients with pernicious anemia indicate that Bifacton is resistant to boiling and that it preserves its hematopoietic activity after being boiled in water for from thirty to forty-five minutes at 95 to 100°C. This suggests either that the intrinsic factor of the animal stomach becomes thermostable after the interaction in vitro with vitamin B12, or that the product of this binding is thermostable under conditions described. 6. The data reported here indicate that various extraction and precipitation procedures may be applied to hog stomach tissue to obtain materials which will exhibit definite intrinsic factor activity in patients with pernicious anemia. Although at present some such extracts are active orally in such small doses as from 40 to 50 mg. per day, still further refinement appears to be possible.

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Chronic Atrophic Gastritis with Negative Intrinsic Factor and Parietal Cell Antibody Presenting as a Severe Hemolytic Anemia

Case Reports in Hematology ◽

10.1155/2020/8697493 ◽

2020 ◽

Vol 2020 ◽

pp. 1-4

Author(s):

G. F. Cittolin-Santos ◽

S. Khalil ◽

J. K. Bakos ◽

K. Baker

Keyword(s):

Vitamin B12 ◽

Atrophic Gastritis ◽

Pernicious Anemia ◽

Hemolytic Anemia ◽

Parietal Cell ◽

Intrinsic Factor ◽

Peripheral Blood Smear ◽

Chronic Atrophic Gastritis ◽

Cell Antibody ◽

Parietal Cell Antibody

A 28-year-old Caucasian male with Hashimoto’s disease and vitiligo presented with two weeks of dizziness on exertion following pharyngitis which was treated with prednisone 40 mg by mouth once a day for five days. Initial workup revealed anemia, elevated lactate dehydrogenase (LDH), and low haptoglobin. He underwent workup for causes of hemolytic anemia which was remarkable for a peripheral blood smear with hypersegmented neutrophils and low vitamin B12 levels concerning for pernicious anemia. Parietal cell and intrinsic factor antibodies were negative, and he then underwent an esophagogastroduodenoscopy with biopsy. The biopsy was negative for Helicobacter pylori, and the immunohistochemical stains were suggestive of chronic atrophic gastritis. He was started on vitamin B12 1,000 mcg intramuscular injections daily. His hemoglobin, LDH, and haptoglobin normalized. Given the absence of the parietal cell antibody and intrinsic factor antibody, this is a rare case of seronegative pernicious anemia.

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Vitamin B12 Malabsorption Due to Intrinsic Factor Deficiency in Indian Subjects

Blood ◽

10.1182/blood.v40.5.747.747 ◽

1972 ◽

Vol 40 (5) ◽

pp. 747-753 ◽

Cited By ~ 3

Author(s):

H. G. Desai ◽

F. P. Antia

Keyword(s):

Vitamin B12 ◽

Pernicious Anemia ◽

Indian Population ◽

Intrinsic Factor ◽

Subacute Combined Degeneration ◽

Factor Deficiency ◽

Vitamin B12 Absorption ◽

Intrinsic Factor Deficiency ◽

Intrinsic Factor Antibody ◽

Vitamin B12 Malabsorption

Abstract Sixteen patients (from Bombay) with severe vitamin B12 malabsorption due to intrinsic factor deficiency, presenting as subacute combined degeneration of the cord (7), tropical sprue (3), anemia (2), thyrotoxicosis (2), diabetes mellitus (1), and pain in the abdomen (1), are reported. The difficulties of establishing a definite diagnosis of pernicious anemia in Indian population are described. The lower incidence of circulating intrinsic factor antibody (IFA) in Indian patients with histamine-fast achlorhydria and poor vitamin B12 absorption is emphasized. The necessity of separating atrophic gastritis, with severely impaired vitamm B12 absorption, from pernicious anemia on the basis of absence or presence of IFA in serum and/or gastric juice cannot be overemphasized.

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RED CELL STROMA PROTEIN RICH IN VITAMIN B12 DURING ACTIVE REGENERATION

Journal of Experimental Medicine ◽

10.1084/jem.102.6.725 ◽

1955 ◽

Vol 102 (6) ◽

pp. 725-731 ◽

Cited By ~ 1

Author(s):

G. H. Whipple ◽

F. S. Robscheit-Robbins ◽

W. F. Bale

Keyword(s):

Blood Cell ◽

Vitamin B12 ◽

Pernicious Anemia ◽

Red Blood Cell ◽

Red Cells ◽

Cell Regeneration ◽

Red Cell ◽

Low Levels ◽

Active Regeneration ◽

Protein Rich

During active blood regeneration in anemia in dogs an increase occurs in the stroma protein of the red cells. When vitamin B12 with radioactive cobalt is given at the start of this blood regeneration one finds concentration of labeled B12 in the stroma protein but not in the hemoglobin. After the acute phase of red cell regeneration is ended the concentration of B12 in stroma protein falls rapidly to very low levels within 2 weeks. Subsequent episodes of red blood cell regeneration seems not to cause remobilization of radioactive cobalt into red cells from other body stores. It appears that the vitamin B12 is a factor of importance in the first steps of stroma protein formation in the first few days of the life of the red cell in the dog. This response in dogs and the response in pernicious anemia to vitamin B12 may have some points in common. Distribution of the B12-radioactive cobalt in the organs and tissues at autopsy has been recorded. Some very suggestive localizations were noted and some variation 1 week and 7 weeks after B12 injections. Radioactive cobalt escapes in the urine during the weeks following B12 injections.

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