The Effect of Chronic Iron Deficiency on Some Biochemical Functions of the Human Hemopoietic Tissue

In view of the erythroid hyperplasiafound in the bone marrow of subjectswith chronic iron deficiency anemia, thedelayed appearance of reticulocytosis following iron therapy is unexplained. Profound disturbances were observed insome biochemical functions of the bonemarrow cells isolated from such patients.There was a substantial decrease in thecellular nucleic acid content, associatedwith a marked drop in the rate of ³H-thymidine incorporation into DNA. Theutilization of ⁵⁹iron and of glycine-2-¹⁴Cfor heme production, and of the lattercompound for protein synthesis was alsoreduced, as compared to the findings inbone marrow cells from normal subjects.These metabolic alterations returned tothe normal pattern in the bone marrowcell suspensions obtained from the patients following recovery after irontherapy.

The possible implications of these findings are discussed in the light of available information.

Submitted on September 15, 1969 Revised on October 12, 1969 Accepted on March 12, 1970     

The Bone Marrow Colony Forming Cell in Megaloblastic Anaemia and Iron Deficiency Anaemia

Summary: The bone marrow colony forming cell in megaloblastic anaemia and iron deficiency anaemia. K. A. Rickard, R. D. Brown, L. Dunleavy, H. Robin and H. Kronenberg, Aust. N.Z. J. Med. , 1975, 5, pp. 424–430.     

Leukocytosis, Bone Marrow Hyperplasia and Leukemia in Chronic Magnesium Deficiency in the Rat

Hematologic studies, including bone marrow examinations, have been doneon a series of rats which were on a magnesium-deficient diet, with or without2-AAF supplementation. Leukocytosis and granulocytosis could be maintainedin these deficient rats by decreasing their intake of magnesium and could bereversed by the addition of magnesium. The bone marrows of the magnesium-deficient group showed granulocytic hyperplasia; the bone marrows ofthe deficient 2-AAF supplemented rats showed erythrocytic hyperplasia, as wellas granulocytic hyperplasia. Chronic granulocytic leukemia developed in twomagnesium deficient rats and persisted after magnesium supplements weregiven. This leukemia has been successfully transplanted into newborn rats.

Submitted on August 25, 1966 Accepted on December 20, 1966     

Anemia in Chronic Obstructive Pulmonary Disease and the Potential Role of Iron Deficiency

The purpose of this review is to evaluate the role of anemia on patient outcomes in chronic obstructive pulmonary disease (COPD), the potential contribution that low iron stores may play in this process, and possible treatment considerations. A review of research studies found that anemia is associated with declining functional outcomes, increased health care utilization and costs, and increased mortality in COPD. Associations exist between reduced iron intake and progression of COPD and in reduction of iron status with declining lung function. Currently data are limited on the effects of either treating anemia or utilizing iron supplementation in anemic COPD patients. If iron supplementation might therefore reverse some of the declines that patients experience, then routine screening and treatment may turn out to be an effective, simple and inexpensive intervention. Iron supplementation models utilized in other inflammatory-related disease states were reviewed as a possible starting point to evaluate treatment options in COPD. Future research can be directed to establish best practice standards for the use of iron supplementation in COPD.     

Effects of Ethanol on Tissue Folate Incorporation and Recovery from Folate Deficiency in Rats

Studies in folate-deficient alcoholics suggest that ethanol interferes with the recovery of folate status and the hematopoietic response to folate. Previous animal studies have suggested diverse effects of ethanol on intestinal absorption, hepatic metabolism, and urinary excretion of folate. In order to examine the effects of ethanol on folate distribution during folate deficiency, tissue incorporation of a tracer dose of folate was studied in rats chronically fed ethanol-containing and/or folate-deficient diets. Rats fed these diets were also used to study the effect of chronic ethanol consumption on the dietary reversal of folate deficiency by changing the diets (adding folate or replacing ethanol) from 12 to 16 weeks. After 16 weeks, tissue folate depletion was severe in rats fed folate-deficient diets. Plasma and whole body retention of the tracer dose of folate was decreased in folate-deficient rats consuming ethanol. In folate-deficient rats, ethanol consumption increased the incorporation of folate by the kidney and brain, but had no effect in other tissues (liver, lung, spleen, intestine, testis). In ethanol-fed folate-deficient rats that continued to consume ethanol, but with added folate in their diets, urine, plasma, liver, and kidney folate levels returned to control levels in 4 weeks. In the rats that stopped ethanol, but continued low folate diet consumption, no recovery of tissue folate levels was seen in 4 weeks. These results suggest that chronic ethanol consumption can exacerbate folate requirements by inhibiting body retention of small doses of folate. However, these effects are minor because ethanol consumption does not block recovery from folate deficiency when rats are fed sufficient amounts of folate.     

Roentgenographic Bone Changes in Chronic Iron Deficiency Anemia: A Study in Twelve Patients

Radiologic bone changes of the skull and long and short bones similar tothose of thalassemia are described in 12 patients with chronic iron deficiencyanemia. Certain differences between the roentgen findings of the present studyand those of the cases reported by other investigators are discussed.

Submitted on August 17, 1965 Accepted on September 15, 1965     

Relationship Between Gestational Iron Deficiency and Iron Deficiency in the Newborn; Erythrocytes

Objective: To investigate the iron status of women at term and its impact in their neonates. Study design: Serum ferritin (SF) was measured in 201 women at term and in their newborns. The relationship between maternal and infant SF and hemoglobin (Hb) and the effects of partial prenatal care and iron supplementation were analyzed. Results: 86% of the women had iron deficiency (ID) at term and 46% were anemic. 13 (7.5%) of the babies born to the 172 iron deficient mothers were also iron deficient, but none of the babies born to the 29 mothers with SF levels > 12 &mgr;g/L. The mean cord ferritin level (103.6 +/- 75 &mgr;g/L) and Hb (164 +/- 20 g/L) were significantly higher in babies born to mothers who had SF > 12 &mgr;g/L, compared to babies born to iron deficient mothers (73.5 +/- 49 &mgr;g/L and 156 +/- 16 g/L). The respective values in iron deficient babies were 7.1 +/- 3.5 &mgr;g/L and 157 +/- 9.0 g/L, compared to 82.8 +/- 52.8 &mgr;g/L and 162 +/- 17.5 g/L in the iron sufficient babies. Conclusion: ID was diagnosed in 86% of women at term and in 7.5% of their neonates. A placental iron threshold, limiting iron acquiscition by the fetuses of women with severe ID, is suggested.     

Effect of Granulocyte Colony-Stimulating-Factor Administration on Tissue Regeneration Due to Thioacetamide-Induced Liver Injury in Rats

It has been shown recently that granulocytecolony-stimulating factor (G-CSF) accelerates andenhances the hepatocyte proliferative capacity ofpartially hepatectomized rats. In the present study, weinvestigated the effect of G-CSF administration in a ratmodel of liver injury and regeneration, induced bythioacetamide (TAA) injection. TAA (300 mg/kg bodyweight) was injected intraperitoneally in male Wistarrats, and this was followed by administration ofeither saline (group A) or G-CSF at a dose of 150g/kg body weight (group B), 24 hr later. The animalswere killed at different time points after TAA treatment and the rate of tritiated thymidineincorporation into hepatic DNA, the activity of theenzyme thymidine kinase (EC 2.7.1.21) in the liver, andthe assessment of the mitotic index of hepatocytes, wereemployed to estimate liver regeneration. Theadministration of TAA caused severe hepatic injury,recognized histopathologically and by the raisedactivities of the serum hepatic enzymes aspartate andalanine aminotransferases. The hepatic injury, which peaked 36 hr afterTAA injection, was followed by a regenerative process ofhepatocytes presenting peaks at time points of 48 and 60hr (group A). The administration of G-CSF 24 hr after the injection of TAA (group B) causeda statistically significantly increase in the hepatocyteproliferation indices examined (P < 0.001), comparedto those found in group A at the same time points. It was concluded that G-CSFadministration enhanced the hepatocyte proliferativecapacity in this model of liver injury induced by TAAadministration.     

Utilization of Iron Dextran in Recurrent Iron Deficiency Anaemia

Previously published studies have documented a reduction in the rate at which iron stores laid down by iron dextran therapy can be utilized for haemoglobin synthesis after the acute demands of haemorrhage and phlebotomy. In order to determine if a defect in the mobilization of these stores exists in the face of a chronic stimulus to red cell production, 93 patients who had previously received a total dose infusion of iron dextran were examined for a recurrence of iron deficiency anaemia, and in those in whom anaemia had recurred, iron stores were assessed by marrow aspiration. Twenty of the 93 patients were found to have recurrent iron deficiency anaemia, and marrow aspiration in all failed to demonstrate stainable iron stores. Although the rate at which iron dextran can be mobilized from storage sites is reduced, the present study demonstrates that ultimately these stores are fully utilizable.     

Increased Formation of Early-Labeled Bilirubin in Rats with Iron Deficiency Anemia: Evidence for Ineffective Erythropoiesis

The production of early-labeled bilirubin and erythrocyte hemoglobin hemewas measured in rats with iron deficiency anemia, using glycine-2-¹⁴C asprecursor. The erythropoietic component of the early pigment fraction wassignificantly augmented and the formation of labeled hemoglobin depressed inthe anemic animals, findings characteristic of ineffective erythropoiesis. Bycontrast, the hepatic component of early-labeled bilirubin was substantiallyenlarged during the acute response to iron therapy. These experiments illustrate that overproduction of bilirubin may originate from both erythropoieticand hepatic sources of the early-labeled fraction of bile pigment, as well asfrom hemolysis of circulating red blood cells.

Submitted on November 14, 1968 Accepted on January 21, 1969     

Coexistence of Megaloblastic Anemia and Iron Deficiency Anemia in a Young Woman with Chronic Lymphocytic Thyroiditis

Pernicious anemia is a megaloblastic anemia caused by vitamin B12 deficiency, and is the end-stage of autoimmune gastritis that typically affects persons older than 60 years. It is the most common cause of vitamin B12 deficiency. Pernicious anemia can also be diagnosed concurrently with other autoimmune diseases. We report the occurrence of megaloblastic anemia in a 22-year-old woman with chronic autoimmune thyroiditis for 10.5 years. Recently, she presented with microcytic anemia, and iron deficiency anemia was diagnosed initially. After administration of ferrous sulfate, macrocytic anemia was revealed and vitamin B12 deficiency was detected. Pernicious anemia was highly suspected because of the endoscopic finding of atrophic gastritis, and high titer of antigastric parietal cell antibody, as well as elevated serum gastrin level. After intramuscular injections of hydroxycobalamine 100 microg daily for 10 days, and monthly later, her blood counts returned to normal.     

The Effect of Injected Iron on the Absorption of Iron in Iron Deficiency

S ummary . The effects of injected iron on the absorption of iron by iron deficient animals have been studied. When iron deficiency is partly or fully corrected by injection of iron dextran, prolonged and marked depression of iron absorption follows despite the persistence of iron deficiency. Although total absorption increases with decreasing size of parenteral replacement dose, the depression of absorption per unit of iron injected increases.     

Globin Chain Biosynthesis in Iron Deficiency

Globin chain synthesis was studied in seven severely iron-deficient patients before and after treatment with iron. There was no appreciable difference between the individual pre- and post-treatment α/β specific activity ratios and the mean α/β ratio for each group was 1·00±SD 0·04. In a further six untreated iron-deficient patients the mean α/β ratio was 1·00±SD 0·04. There was therefore no evidence that iron deficiency caused a reduction in α/β ratio.
Three patients with β thalassaemia trait and coexistent iron deficiency had lower α/β ratios before treatment than after treatment with iron. It appeared that iron deficiency had caused reduced α chain synthesis in this group.
Preliminary experiments have shown that the α/β specific activity ratio of purified haemoglobin A is decreased in iron deficiency, indicating an increase in the size of the free α chain pool. It is suggested that iron deficiency may interfere with the proteolytic mechanism normally responsible for the destruction of excess α chains. In combined iron deficiency and β thalassaemia trait, the resulting increase in free α chains might act by negative feedback to inhibit further α chain synthesis (Blum et al , 1970) thereby reducing the pre-treatment α/β ratio.     

Plasma and Erythrocyte Folate in Iron Deficiency and Folate Deficiency

Erythrocyte and plasma folate levels were studied before treatment in 20patients with iron deficiency anemia and in 23 patients with megaloblasticanemia due to folate deficiency. Fourteen of the cases of iron deficiency anemiawere also studied after treatment with oral iron alone. Fifty-seven normalpersons were used as controls.

The mean erythrocyte folate (ng./ml. packed cells) was significantly increased in iron deficiency anemia and significantly depressed in folate deficiency anemia. After treatment with oral iron alone, the mean erythrocytefolate level fell to normal in the iron deficiency anemia group. The mean corpuscular folate (ng. x 108^-8) was also significantly raised in iron deficiency:in eight of 10 cases this fell after treatment, but the overall fall was not significant. The plasma folate rose in iron deficiency anemia after oral irontreatment.

Submitted on August 8, 1969 Accepted on November 17, 1969     

Myoglobin Depletion in Childhood Iron Deficiency

S ummary . Serum concentrations of myoglobin are significantly lower in iron-deficient than in non-iron-deficient children. In the iron-deficient subjects there is a correlation between haemoglobin concentration and serum myoglobin suggesting that the synthesis of both haem compounds is affected in parallel. No abnormality of serum myoglobin concentration was found in adults with iron deficiency anaemia.     

Histological Study of Bone Marrow Regeneration following Chemotherapy for Acute Myeloid Leukaemia and Chronic Granulocytic Leukaemia in Blast Transformation

S ummary Haemopoietic regeneration following chemotherapy was studied on serial bone-marrow trephine specimens from 10 patients with acute myeloid leukaemia (AML), and seven with chronic granulocytic leukaemia (CGL) in blast transformation. In AML following the stage of treatment-induced hypoplasia during which the marrow was extremely hypocellular, oedematous, and contained widely dilated sinuses, areas of large, uniform, unilocular fat cells, designated 'structured fat' developed from multilocular precursor fat cells. Early foci of haemopoietic regeneration were present almost exclusively in areas of structured fat: only scattered small erythropoietic foci were seen in the earliest specimens taken 1 week after the completion of treatment; small granulopoietic foci were first seen 2 weeks later, and the first megakaryocytes were not seen until the third week. Transient regeneration was observed in two patients who subsequently failed to enter remission and died.
The pattern of haemopoietic regeneration in CGL in blast transformation treated by intensive therapy and autografting with cryopreserved buffy-coat cells differed from that seen in AML. Bone marrow regeneration occurred earlier than in AML and appeared to be independent of the presence of structured fat. Regeneration in all three cell lines appeared simultaneously within 2 weeks of autografting. Regeneration tended to be focal, each aggregate consisting almost exclusively of erythroid, granulocytic or megakaryocytic cells.
The proximity of regenerating haemopoietic foci to structured fat in AML suggests that, in some circumstances, haemopoietic stem cells require the presence of fat cells in their immediate environment before they proliferate.     

Studies in Iron Metabolism. IV. Iron Absorption in Experimental Iron Deficiency

1) Whole body counting by means of a large phosphor well scintillationcounter has been used to measure the absorption of Fe⁵⁹-tagged inorganic iron,and shown to compare favourably with other methods.

2) There is a delay in the fecal elimination of the unabsorbed portion ofthe dose of Fe⁵⁹ by iron-deficient rats on iron-deficient diet. The cause of thisdelay is unknown but it may be associated with the marked cecal enlargementwhich exists in these animals.

3) It is confirmed that iron deficiency is associated with striking enhancement of absorption of ferrous and ferric inorganic iron.

4) When a series of doses of ferrous iron of increasing size from 5 to 1,000µg. was given, there was a progressive increase in absorption for each increasein dose in both iron-supplemented and iron-deficient rats. The relationshipbetween amount of iron given and amount absorbed suggests that two processesmay be involved: 1) simple diffusion, and 2) a carrier mechanism.

5) The effect on iron absorption of a sudden change in iron intake hasbeen investigated. Switch from a low to high iron diet reduces absorption,and from a high to a low iron diet increases absorption, too rapidily for hemoglobin level or body iron stores alone to be the most important governingfactors and this finding emphasizes the importance of local changes in theintestine.

Submitted on April 23, 1962 Accepted on June 25, 1962