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A study of erythropoiesis and iron metabolism in the rabbit in vivo. II. Dependence of the response on iron storage and transport.

Rabbits subjected to a daily bloodletting schedule of 10 ml blood per kg body weight increase four- or fivefold their erythropoietic production compared to normal non-bled animals. The maximum response they can reach under these conditions mobilizes more than 9 mg of iron per day per rabbit into hemoglobin. When fed ad libitum with their regular diet, they do not need any further iron supplement for full erythropoiesis. The experimental increment in iron body stores and/or serum iron levels does not enhance their erythropoietic response, demonstrating that iron is not rate limiting under the conditions studied. Furthermore, although serum iron levels are elevated onefold in the controls under chronic anemia with respect to non-bled animals, the concentration of serum transferrin is only slightly increased; hence, the iron saturation of this protein changes from a 50% to an 80% level. In the absence of an extra supplement of iron, rabbits subjected to chronic bloodletting show no signs of body iron depletion, as judged by their continuous macrocytic RBC production.

Anemia↗

Iron dextran treatment in peritoneal dialysis patients on erythropoietin.

STUDY OBJECTIVE: To evaluate maintenance parenteral iron dextran in chronic peritoneal dialysis (PD) patients receiving erythropoietin (rHuEPO). DESIGN: Parenteral iron dextran was investigated in PD patients with poor response to rHuEPO and/or side effects of oral iron. SETTING: Tertiary-referral university hospital PATIENTS AND METHODS: Seven ESRD patients (five males and two females) were studied. A test dose of 25 mg iron dextran was given before starting a maintenance dose. Iron dextran 100 mg was given intramuscular weekly or biweekly. Six patients received rHuEPO and one patient was on decadurabolin. RESULTS: Hematocrits increased significantly (p < 0.01) from 29 +/- 2% to 38 +/- 2% and serum ferritin increased from 267 +/- 104 to 660 +/- 104 ng/dl after iron dextran. Serum albumin increased from 3.1 +/- 0.3 to 3.6 +/- 0.2 g/dl (p < 0.05). No patient developed an anaphylactic reaction or delayed reaction. Mean duration of parenteral iron dextran treatment was 7 +/- 1 months. Mean dose of erythropoietin was reduced significantly (p < 0.05) from 119 +/- 20 units/kg/week to 87 +/- 20 units/kg/week before and during fifth month of iron dextran therapy. CONCLUSIONS: Weekly/biweekly maintenance intramuscular iron dextran injection was effective and safe iron supplemental therapy in PD patients with poor response or side effects to oral iron.

Adult↗

[An experimental study on MR lymphography with various iron colloid agents].

Magnetic resonance (MR) lymphography with iron colloid agents was evaluated in an animal model. The agents examined were ferric gluconate (FeG), saccharated ferric oxide (SFO), iron chondroitin sulfate (ICS) and cideferron (CiF), which were used as intravenous medication for iron deficiency anemia. The author performed time-dose-response and correlative histologic studies. MR images of the popliteal lymph nodes of rabbits were obtained at 1.5 T with a spin-echo sequence (TR = 300, TE = 30 msec) before and after subcutaneous injection of the agents to dorsal hind-feet. The images were evaluated by signal intensity (SI). Histologic specimens were evaluated for distribution and relative quantity of stained iron with a color image analyzer. The SI with FeG 4 mgFe increased by 50% at 5 to 60 minutes after injection but returned to the pre-contrast level 48 hours after. The SIs with SFO 8 mgFe, ICS 1 mgFe and CiF 1 mgFe decreased to the background level at 60 minutes and 48 hours after. The histologic study in combination with the images indicated that in case of FeG the particles in lymph increased SI while in the other agents those within macrophages decreased SI.

Animals↗

Hereditary hemorrhagic telangiectasia. A case with hepatocellular carcinoma and acquired hepatocerebral degeneration.

A patient with hereditary hemorrhagic telangiectasia (Osler-Weber-Rendu disease), treated with ethinyl estradiol, multiple blood transfusions, and iron-dextran, developed hepatocellular carcinoma and acquired hepatocerebral degeneration. In addition to the carcinoma, the liver contained extensive arteriovenous maliformations, telangiectasis, and changes of Osler atypical cirrhosis. The carcinoma possibly had its genesis in the presence of an ocongenic serum hepatitis virus, or the cirrhosis, or both.

Aged↗

[Investigation of impairment of neutrophil's phagocytosis and bactericidal function in rats with iron deficiency].

Studies were made to determine the neutrophil's phagocytosis and bactericidal function in three groups of rats (control, iron deficiency, and iron supplement). Results showed that there were significant differences in values of chemiluminescence (CL) among three groups. The values of peak CL and five minutes integrated CL were markedly decreased in neutrophils of iron-deficient rats, accounting for only 41% and 32% of the control's values respectively. These suggested that the activity of NADPH oxidase was decreased, and the function of respiratory burst of neutrophils was impaired. The activity of myeloperoxidase in the iron-deficient neutrophils was also significantly lower than that in the control cells. It constituted only 30% of the control's value, indicating that the bactericidal function of neutrophils was injured. One week after iron administration, the low values of the peak CL, the five minutes integrated CL and the activity of myeloperoxidase all went up apparently, but not reached the normal levels yet. The time the function of neutrophils in iron-deficient rats returned to normal may be related to the process of neutrophil maturation in bone marrow.

Anemia, Hypochromic↗

Ligand-specific isolation of endosomes and lysosomes using superparamagnetic colloidal iron dextran glycoconjugates and high gradient magnetic affinity chromatography.

We have developed a ligand-specific method for the visualization, isolation, and biochemical characterization of cell surface and intracellular membranes mediating endocytic transport. Iron dextran particles (FeDex) bearing either covalently conjugated galactosyl bovine serum albumin (GalBSA/FeDex) or asialofetuin (ASF/FeDex) are bound by the asialoglycoprotein receptor (ASGP-R) of HepG2 cells and transported to lysosomes with kinetics indistinguishable from those of free GalBSA or ASF. FeDex particles, which have a 3 to 5 nm electron-dense colloidal iron core, can be visualized by electron microscopy. Following incubation of GalBSA/FeDex with HepG2 cells at 37 degrees C, FeDex particles are seen at the cell surface, in endosomes, and in lysosomes. Surface membrane and intracellular organelles bearing a sufficient number of FeDex particles can be efficiently isolated from disrupted cells by high gradient magnetic affinity chromatography (HIMAC). Plasma membranes and endosomal/lysosomal membranes isolated by HIMAC are 35 to 40-fold enriched for GalBSA/FeDex or ASF/FeDex relative to the postnuclear supernatant. Alkaline phosphodiesterase I (APDE) and galactosyltransferase are each enriched 8-fold in the plasma membrane fraction prepared by HIMAC whereas neither beta-galactosidase nor glucose-6-phosphatase are detected in this fraction. The intracellular membrane fraction, containing both endosomes and lysosomes, is enriched for galactosyltransferase and beta-galactosidase but not for APDE or glucose-6-phosphatase. Use of FeDex conjugates in conjunction with HIMAC provides an effective method for ligand-specific isolation of membranes and correlation of morphological and biochemical characteristics.

Animals↗

Wheat germ agglutinin is selectively transported to multivesicular bodies.

Colloidal iron dextran particles bearing wheat germ agglutinin (WGA/FeDex) were bound by glycoconjugates expressed at the surface of HepG2 cells. Bound WGA/FeDex was internalized when cells were incubated at 37 degrees C and accumulated in intracellular structures which have the same buoyant density as the plasma membrane when examined on Percoll density gradients. The intracellular structures containing WGA/FeDex were identified as multivesicular bodies (MVB) by transmission electron microscopy. WGA/FeDex was not transported to lysosomes nor did it interfere with uptake and transport of GalBSA to lysosomes by the asialoglycoprotein receptor. WGA/FeDex was seen predominantly in non-coated invaginations at the cell surface, suggesting it may enter cells at a different site than GalBSA/FeDex. Highly enriched plasma membranes and MVBs containing superparamagnetic [125I]WGA/FeDex particles were prepared by high gradient magnetic affinity chromatography (HIMAC). Plasma membranes prepared by HIMAC were enriched 30-fold for [125I]WGA/FeDex, 15-fold for alkaline phosphodiesterase I, and 9-fold for galactosyltransferase relative to the crude post-nuclear homogenate and consisted entirely of plasmalemmal sheets. Intracellular structures containing WGA/FeDex were enriched 35-fold for [125I]WGA/FeDex, 10-fold for alkaline phosphodiesterase I, and 10-fold for galactosyltransferase but did not contain lysosomal beta-galactosidase. WGA/FeDex has a different ultimate destination in HepG2 cells than ligands internalized by the asialoglycoprotein receptor and can be used to obtain highly enriched plasma membranes and MVBs from cultured cells.

Animals↗

Thalamic afferents from the brain stem. An experimental study using retrograde single and double labelling with HRP and iron-dextran in the rat. I. Medial and lateral reticular formation.

The reticulo-thalamic projection of the rat was studied in single and double experiments using 51 injections of HRP and/or iron-dextran. The medial reticular formation (excluding the ncl. pedunculo-pontinus, subncl. compactus, see Part II) sends out projections to the ncll. intralaminares (IL), ncl. mediodorsalis (MD), ncl. ventralis medialis (VM), ncl. ventralis lateralis (VL), ncl. ventralis basalis (VB) and ncl. posterior (PO) of the thalamus. The mesencephalic RF (ncl. cuneiformis and ncl. subcuneiformis) projects to all the above mentioned nuclei. The rhombencephalic RF (ncl. pontis oralis, ncl. pontis caudalis, ncl. gigantocellularis et paragigantocellularis dorsalis et lateralis) projects to the posterior IL (ncl. parafascicularis and ncl. centromedianus) and to the MD, pars paralaminaris. The pontine RF (ncl. pontis oralis et caudalis) projects to the posterior IL and to the medial part of the ncl. parafascicularis. The RF of the pons and the upper oblongata projects to the VL, VB and PO. The lateral RF (ncl. parvocellularis) projects solely from the tiers around the ncl. n. VII. to the posterior IL in what is a scanty projection. We found no connections projecting from the medial or lateral RF to the midline nuclei, to the lateral nuclei or to the ncl. commissurae posterioris; the projection to the ncl. reticularis thalami was left unstudied. The reticulo-thalamic pathways exhibit marked laterality. The mesencephalic RF projects to the thalamus in what is pronounced ipsilateral predominance, the pontine RF in ipsilateral predominance, the upper oblongata RF equally ipsi- and contralaterally, and the lower oblongata RF with a predominance of the contralateral component. Collateralization (double labelled cells) was found projecting solely to the IL and MD regions. It was both ipsilateral and contralateral. The discoveries included the topographical organization of the reticulo-thalamic projection: a major antero-posterior in the rhombencephalon and a mediolateral in the projections from the mesencephalon and from the pons. The most substantial projection was found aiming at the posterior IL nuclei (ncl. parafascicularis-centrum medianum), a less substantial one to the ventral and posterior nuclei, and the scantiest to the anterior IL nuclei (ncl. paracentralis, ncl. centralis lateralis).

Animals↗

Investigation on the dosage/efficacy relationship of iron dextran in veal calves.

The efficacy of a single dose of 800 mg resp. 1600 mg iron in the form of an intramuscularly administrable iron (III)-dextran complex (Anaemex, CAS 9004-66-4) has been tested. On 3 groups of 13 calves each, 0 ml (comparing group), 4 ml resp. 8 ml iron dextran 20% have been applied. All calves received iron-containing food during the test period of 10 weeks. At the beginning of the therapy, 5 weeks and 8 weeks after application, the parameters: weight, hemoglobin, erythrocytes, hematocrit, mean corpuscular hemoglobin (MCH), MCH concentration, mean corpuscular volume, plasma protein, fibrinogen, leukocytes and serum iron were measured. After 10 weeks the dead weight has been determined and the spleen of some calves tested histologically. The study shows that, by the administration of 1600 mg iron as a depot injection, a better growth results with the same quality of veal. The red-coloring of the veal was not significantly different from that of the comparing group. The histological findings show especially that the iron depots of the spleen were empty in all three groups and thereby in this collective no connection exists between the color of the veal and the tested dosage of iron dextran 20%. It is considered meaningful and economic to renounce in future the iron-containing food and in its place to apply intramuscular a single dose of 1600-2400 mg iron per calf. The results are compared with a study on full term infants, which has shown that a intramuscular single dose of 150 mg of iron as iron dextran at birth affords a nutritional advantage in iron status for up to 15 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Iron and the liver: subcellular distribution of iron and decreased microsomal cytochrome P-450 in livers of iron-loaded rats.

To understand better the intracellular iron distribution and metabolic consequences of chronic hepatic iron overload, rats were given large doses of iron dextran or ferric citrate intraperitoneally. They accumulated large quantities of iron within Kupffer cells and hepatocytes. The relative subcellular iron distributions were similar in controls and iron-loaded rats, despite a ten- to 20-fold difference in hepatic iron concentration. Electron microscopy of whole liver and subcellular particulate fractions suggested that iron was present in highest concentration in lysosomes, which were rendered more labile by its presence. Nevertheless, quantitative iron determinations on all subcellular fractions, obtained by two preparative methods, showed that most of the iron was present in the "soluble" fraction. The amount of iron in the "microsomal" fraction varied, depending on the techniques used for preparation of this fraction. Cytochrome P-450 and total heme concentrations were decreased 40% to 50% in microsomes isolated from iron-loaded livers.

Animals↗

[Effect of various preparations of iron on the growth and haemoglobin levels of newborn piglets (author's transl)].

Four preparations of iron dextran, which differed markedly in price, were compared in an experimental study. 100 or 200 mg. of iron were injected intramuscularly on the first, second or third day of life of the piglets. Differences in weight or concentration of haemoglobin were not detectable during the first eight weeks of life, neither at a dosage of 100 mg. nor at a dosage of 200 mg. of iron.

Anemia↗

Morphologic and functional characteristics of bone marrow macrophages from imferon-treated mice.

The iron storage macrophage has been isolated from the marrow of Imferon-treated mice and studied in vitro by morphologic, histochemical, and functional tests and isotope labeling techniques. These macrophages on stained preparations are large, many times binucleate cells (up to 150 mu), and show Prussian blue reactivity. In Epon-embedded, stained thick sections they contain elongated narrow basophilic inclusions. These macrophages are actively phagocytic and pinocytic; histochemical studies show that these cells are rich in acid phosphatase, nonspecific esterase, and PAS diastase-resistant activity. Iron storage macrophages do not incorporate the 3H-thymidine. The electron microscopic appearance of this macrophage shows that the cell has ferritin free in the cytoplasm and several types of cytoplasmic granules: those with large quantities of electron-dense ferritin and/or hemosiderin (type A), elongated granules (type B) with moderately electron dense homogeneous matrix and some ferritin at the periphery, and granules with heterogeneous content (type C). The above findings demonstrate that the iron storage cell is a mature macrophage which contains hydrolases, ferritin, and a unique population of cytoplasmic granules which are lysosomal in nature. There is some evidence to suggest that the unusual lysosome (type B granule) occurs after macrophages have ingested erythrocytes.

Acid Phosphatase↗