Relationship between plasma PGE2 level and changes in hemoglobin proportions during bleeding-induced anemia in adult rats

Reversion of hemoglobin proportions toward newborn values is a characteristic change found in blood of acutely bled adult rats. In this study, adult Sprague-Dawley rats were bled over a period of time until they became anemic by hematologic parameters. We measured plasma prostaglandin E2 levels of anemic and control rats using double-antibody technique. A significant increase was recorded in bled plasma, and the value returned to unbled level when anemia was corrected. Acetylsalicylic acid intake during bleeding-induced anemia abolished the process of reverse switching of hemoglobin, as well as inhibited the increase in plasma PGE2 levels. Changes in hemoglobin proportions due to phlebotomy were also blocked when acetylsalicylic acid was replaced by indomethacin. These observations are of significance in understanding, at least in part, the mechanism of reverse hemoglobin switching in adult rats undergoing erythropoietic stress.     

Identification of rat and human hemoglobin acetilation sites after its interaction with acetylsalicylic acid

The possibility of interaction of 0.1 mg/mL acetylsalicylic acid with purified human and rat globin in vitro during 24 h at 37 degrees C was investigated. The rat globin can be modified with acetylsalicylic acid on aminoacid residues K-17, K-57, K-91, K-140 in alpha subunit as well as on K-18, K-77 in beta subunit. The human globin can be modified with acetylsalicylic acid on aminoacid residues K-17, K-41, K-57 and K-91 in alpha subunit as well as on K-18, K-96 and K- 133 in beta subunit. We identified of acetetylated lysines K-17 and K-57 in alpha subunit of human hemoglobin after incubation whole blood with 0.1 mg/mL acetylsalicylic acid during 3 h.     

Globin proteins of the normal and anemic duck

The red blood cells of normal adult ducks contain two main hemoglobins. The most abundant type, HbA, comprises approximately 80% of the total, with the remaining 20% being made up of HbD. An attempt was made to determine whether during hemolytic anemia a special alpha globin chain (alpha s) replaces the alpha chain of HbA found in normal animals. This special stress alpha globin, whose existence has been seriously questioned, was originally postulated to explain the sequence discrepancies obtained between alpha chains of normal and anemic chickens and ducks. Using gel electrophoresis, isoelectric focusing, and HPLC peptide mapping techniques no qualitative differences between the alpha A globins of normal and anemic animals were found. The nature of the beta globin chains present in adult ducks has also never been rigorously established. In this work, a variety of techniques, including HPLC, gel electrophoresis, and microcolumn amino acid analysis, were used to examine the beta chains from each hemoglobin. Using these methods, no differences were found between the beta globin chains of the two hemoglobins.     

The accumulation of basic nonhemoglobin proteins during the differentiation of primitive (embryonic) red cells of amphibia

The preferential accumulation of hemoglobin is a characteristic of the differentiation of definitive (adult) red blood cells. Since primitive (embryonic or larval) red blood cells have many properties which contrast with definitive red cells, the accumulation of red cell proteins was analyzed during the differentiation of primtive red cells to determine whether or not hemoglobin was the only protein which showed a substantial increase in amount. Primitive red cells of amphibia were used because the mature circulating cell retains large amounts (13%) of specific, well characterized, nonhemoglobin proteins (CP). Preparations of primitive red cells enriched in immature cells were obtained from the circulation of bullfrog tadpoles recovering from phenylhydrazine-induced anemia. The amount of CP, determined by electrophoresis, imunodiffusion, and ion-exchange chromatography, was compared for red cells from normal animals and anemic animals. Mature cells contained three to six times the amount of CP and three times the amount of hemoglobin found in the population of red cells enriched in immature cells. The accumulation of CP during maturation of primitive red cells indicates that differentiation of primitive red cells is less restrictive than differentiation of definitive red cells. Since the primitive red cell is less specialized than the definitive red cell, it is possible that primitive red cells have several roles in the developing animal, in contrast to the single role of synthesizing and maintaining hemoglobin in the adult animal.     

Studies on the erythropoietic effect of plasma from anemic toads both with and without testis.

Injection of plasma from experimentally induced anemic toad with intact testis increases erythropoiesis in starved toads evidenced by the increase of red blood cell, hemoglobin and hematocrit, whereas the plasma of castrated and phenylhydrazine-HCl treated anemic toad failed to do so. It can be suggested that the erythropoiesis stimulating factor (ESF) is produced from the testis of toad and the production of this factor (ESF) was found to increase during anemia in an attempt to correct the anemic condition.     

Patterns of hemoglobin production in the adult newt (Triturus cristatus) erythron following induction of anemia

• 1.1. Changes in the hemoglobins present in many vertebrates have been observed during development and during anemic episodes.
• 2.2. A change in the number of hemoglobins present and their relative amounts was observed when adult Triturus cristalus newts were made anemic by injection of acetylphenylhydrazine.
• 3.3. Hemoglobin IV, which is a minor hemoglobin in healthy adults, was found to be a major component during the subsequent erythropoietic response to hemolytic anemia.
• 4.4. No new hemoglobin not already present in the non-anemic state was detected during the response to induced anemia.

     

Sterol and bile acid metabolism during development. 1. Studies on the gallbladder and intestinal bile acids of newborn and fetal rabbit

Bile acid composition and content in the intestine and gallbladder of newborn and fetal rabbits were investigated. Unlike the circumstances in adult rabbits, the bile acids were conjugated with both taurine and glycine. The major bile acids of the fetus and newborn rabbit were cholic acid, chenodeoxycholic acid, and deoxycholic acid. This is different from the known bile acid composition of adult rabbits, in which deoxycholic acid is the major bile acid (> 80%). The proportion of chenodeoxycholic acid was higher in the fetal than in the newborn tissues. The total bile acid pool in the newborn was higher than in the fetus. In the fetus, large proportions of bile acids (60.9%) were associated with the gallbladder fraction, whereas in the newborn the bulk of the bile acids were found with the intestinal fraction (64.4%),     

The effects of acetylsalicylic acid on phosphoenolpyruvate carboxykinase activity and acinar heterotopy in livers from juvenile and adult rats

Male and female juvenile as well as adult rats were treated with acetylsalicylic acid in order to examine the effect of the drug on over-all activity and activity distribution of phosphoenolpyruvate carboxykinase (PEPCK) in the liver acinus. Upon administration of acetylsalicylic acid PEPCK activity increased in juvenile males and adult females, but was reduced in juvenile females and adult males. The periportal-perivenous activity gradient along the sinusoidal length, which is flatter in untreated juvenile rats compared to the livers of adult rats, was distinctly steepened by acetylsalicylic acid treatment. Acetylsalicylic acid did not affect the gradient in adult rats.     

Changes in the hemoglobin system of the coho salmon Oncorhynchus kisutch during smoltification and triiodothyronine and propylthiouracil treatment

Three electrophoretically separable forms of blood hemoglobin were found in fry of coho salmon (Oncorhynchus kisutch). Ten hemoglobin forms were found in juvenile and adult salmon. Treatment of yearling salmon with triiodothyronine (T3) at 12 ppm in the diet accelerated the increase in concentration of adult forms of hemoglobin, while treatment with T3 at 4 ppm had no effect. Dietary propylthiouracil (1.5 or 5 mg/g) reduced the increases in adult forms of hemoglobin during smoltification. These results implicate thyroid hormones in the expression of adult forms of hemoglobin during the parr to smolt transformation of juvenile salmon.     

Polyunsaturated fatty acids in serum lipids of reindeer during the close postnatal period

We examined serum fatty acid composition in reindeer during the close postnatal period (from <8 h to 3 weeks) by using maternal serum as a reference point and focusing on the proportions of polyunsaturated fatty acids (PUFAs) in serum lipids. A striking dissimilarity was found in the serum PUFAs between the neonatals and their mothers. In particular, the proportions of linoleic acid (18:2) and α-linolenic acid in serum cholesteryl esters and phospholipids of the newborn reindeer were significantly lower than those of the mothers. Furthermore, serum phospholipids of the newborns had lower arachidonic acid and docosapentaenoic acid but higher docosahexaenoic acid proportions than the maternal phospholipids. Although the proportions of the principal C18 PUFAs were low in reindeer milk, they increased sharply in serum cholesteryl esters and phospholipids of the calves during the first few days after birth. In particular, there were significant positive correlations in the proportions of 18:2 between serum and milk lipids. We conclude that the proportions of the serum C18 PUFAs are low in the newborn reindeer, but they are increased during the close perinatal period by a rate which suggests an efficient mechanism for selective retention of these fatty acids from milk lipids. Accepted: 10 August 1998     

Metabolism of [3H]Nipecotic Acid in the Rabbit Retina

Nipecotic acid has been demonstrated to block the gamma-aminobutyric acid transport systems. Previous studies have shown that the uptake system is the first transmitter-specific parameter to appear during the development of the rabbit retina. Use of these observations has been made to study the influence on the development of gamma-aminobutyric acid receptors of altering the uptake mechanism by treating newborn pups with nipecotic acid to block GABA transport. The present study of the in vivo metabolism of [3H]nipecotic acid in the CNS measured the changes in the levels of [3H]nipecotic acid in both adult and newborn rabbit retinas after injection of the label into the vitreal chamber. It was found that the effective half-life of [3H]nipecotic acid in the vitreous is about 5 h for adult tissue and 3 h for newborn. In contrast, all retinal fractions retained the label longer, the effective half-lives being about 60 h (adult) and 45 h (newborn). Further, no labeled metabolites of nipecotic acid were detected in either adult or newborn tissue. This study gives evidence that the degradation of nipecotic acid in nervous tissue is minimal and suggests that, although the rate of clearance is faster in neonates, the fate of nipecotic acid in vivo may be similar in both adult and newborn tissues.     

Effects of experimental anemia on blood ion and acid-base status of turtles during submergence in aerated water at 3 degrees C

The importance of blood hemoglobin to aquatic oxygen uptake by turtles (Chrysemys picta bellii) submerged in aerated water at 3 degrees C was tested by comparing the responses of anemic turtles (hematocrit approximately 6%) to turtles with normal hematocrits (hematocrit approximately 33%). All turtles were submerged for 42 days and blood samples were collected at 0, 7, 21, 32 and 42 days. Blood was analyzed for pH, PCO(2), PO(2), hematocrit, hemoglobin concentration ([Hb]) and plasma was analyzed for concentrations of lactate, glucose, Na(+), K(+), Ca(2+) and Mg(2+). Plasma [HCO(3)(-)] was calculated. [Hb] correlated closely with hematocrit levels. [Lactate] reached higher final values in anemic turtles (34.5+/-5.3 mmol l(-1)) than in normal turtles (14.5+/-4.6 mmol l(-1)) indicating a greater reliance of the anemic animals on anaerobic metabolism. Both groups compensated for acidosis by reduced PCO(2) and anemic turtles also had increased [Ca(2+)] and [Mg(2+)]. Blood pH fell significantly in the anemic turtles but not in the controls. Although the data indicate that the anemic turtles relied more on anaerobic metabolism than the controls, the effect was much less than expected on the basis of the reduced blood O(2) carrying capacity. Possible compensatory mechanisms utilized by the anemic turtles to minimize anaerobic metabolism are discussed.     

Mouse hemoglobin during gestation. Absence of fetal hemoglobin

A "fetal hemoglobin' has been reported to exist during mouse gestation, Investigations using CMC chromatography, starch gel electrophoresis or isoelectric focusing have shown a hemoglobin band from fetal tissues, and blood was obtained which was different from the adult hemoglobin and designated a "fetal hemoglobin'. In the current study isoelectric focusing was used to study the hemoglobins existing in the tissues and blood during fetal and neonatal development and the results suggest there is no "fetal hemoglobin' present during gestation. It appears that the hemoglobin designated as "fetal' in our laboratory was a methemoglobin formed by an incomplete reaction of KCN with the hemoglobin. The additional hemoglobin bands which were obtained from fetal liver or neonatal spleen tissues appeared to be a modified adult hemoglobin.     

Analysis of the recombination zone in hereditary persistence of fetal hemoglobin with fetal gene rearrangement of the G gamma-G gamma-A gamma type

An increased synthesis of fetal hemoglobin in adult life is a common feature of the genetically determined severe disorders like beta thalassemia and sickle cell anemia. A continued synthesis of fetal hemoglobin in adults is also characteristic of clinical or subclinical syndromes like respectively delta beta thalassemia or hereditary persistence of fetal hemoglobin (HPFH). These disorders are highly heterogeneous with respect to their molecular defects as well as to the composition of Hb F. We report here a novel case of hereditary persistence of fetal hemoglobin in heterozygous state discovered by chance, in a young perfectly healthy french man. The gamma chain of his fetal hemoglobin was almost entirely composed of G gamma chains. Molecular analysis of the DNA revealed the existence of triplicated gamma genes on one chromosome with the genotype arrangement of G gamma-G gamma-A gamma. A polymorphic Xmn I restriction site (at position -158 5' to the cap site) was present in 5' of both of these G gamma genes. The presence of this site in front of G gamma gene had previously been shown to be associated both with high G gamma phenotype constitutively and also with high fetal hemoglobin level only in case of anemic stress. In the absence of any anemic stress in this individual, the constitutive increase of both fetal hemoglobin and G gamma chains could be due to the presence of a chromosome with triplicated arrangement of gamma genes. The classical triplication (G gamma-A gamma-G gamma-A gamma) does not result in HPFH phenotype.(ABSTRACT TRUNCATED AT 250 WORDS)     

The life span of red blood cells in the amphibian larvae, Rana catesbeiana

Nearly one hundred amphibian tadpoles were made anemic by phenylhydrazine injection. During the recovery period radioactive thymidine was incorporated into the DNA of the nucleated amphibian red blood cells. Over the next 135 days blood was drawn and smears of circulating red blood cells were prepared. The blood smears from each tadpole were autoradiographed and percentage of labeled nuclei (PLN) determined. The average life span of tadpole red blood cells was calculated from the change in PLN, and is about 100 days. It is concluded that during the transition from tadpole to frog, the tadpole red blood cell life span must be drastically shortened to account for the hemoglobin transition observed during amphibian metamorphosis.     

Magnetic behavior of human erythrocytes at different hemoglobin states

The effect of a static magnetic field on human erythrocytes at different hemoglobin states (normal, oxidized and reduced hemoglobin) was investigated. Three different blood samples, normal, iron deficiency anemic and beta thalassemia minor, were studied. Measurements of the magnetization curves of the erythrocytes for all blood samples in all states showed diamagnetic behavior; however, oxidation was found to enhance this behavior. These measurements have also shown that the normal and iron deficiency samples in the reduced states exhibit a less diamagnetic response in comparison with the normal state. This result indicates that the reduction process gave rise to a paramagnetic component of the magnetization. Analysis of the measured paramagnetic behavior, using a Brillouin function, gave an effective magnetic moment of 8 muB per reduced hemoglobin molecule for both normal and anemic samples. This result shows that both anemic and normal blood have similar magnetic behavior and the only difference is the number of hemoglobin molecules per erythrocyte. For the beta thalassemia minor blood sample, magnetic measurements showed that both the normal and reduced states have almost the same diamagnetic behavior. However, this diamagnetic response is less than that for the normal state of the iron deficiency anemic sample. This result may indicate a low oxygen intake for the blood in the normal state for the beta thalassemia minor blood. All magnetic measurements were made using a vibrating sample magnetometer using field steps of 0.001 T from 1 T to -1 T.     

Anemia associated with anestrus in beef heifers inoculated with Anaplasmamarginale: Endocrinology and ovarian changes

Eleven cyclic Hereford heifers were synchronized to estrus with a commercial luteolytic agent. Eight heifers had been inoculated with $\text{Anaplasma}$$\text{marginale}$. Six of the eight inoculated heifers became anemic, i.e. packed cell volume, red blood cell counts and hemoglobin decreased. White blood cell counts were elevated during severe anemia. Anemic heifers did not show behavioral estrus or ovulate, and during this stage, serum concentrations of progesterone and estradiol-17β were significantly depressed. Mature beef heifers that have acute anaplasmosis suffer from inhibited ovarian function and anestrus.     

Prediction of carbon monoxide diffusing capacity of the lung in splenectomized sheep.

The steady state diffusing capacity of the lung for carbon monoxide (DLCO) was studied in 18 splenectomized adult ewes. Seven animals were anemic when studied. Weight (Wt) and, to a lesser extent, hemoglobin (Hb) level were the key predictive variables of DLCO. Sheep DLCO can be expected to range between 15 and 28 ml/min/mmHg in adult ewes which are not anemic. When DLCO measurements were repeated up to three times on the same day no significant decreases occurred. Thus, the data demonstrated no CO back-pressure caused by preceding DLCO determinations. This paper's importance is in defining a normal predictive range for this sensitive parameter of pulmonary function.     

Study of maturation of membrane transport function in red blood cells by X-ray microanalysis

Summary Red blood cells of certain species of animals, such as dogs and cats, contain low potassium and high sodium, whereas the erythropoietic stem cells giving rise to these cells are of high potassium type. This paper examines the sequence of membrane transport changes during erythropoiesis by analyzing the K, Na and Fe in single bone marrow cells, reticulocytes and mature red blood cells with X-ray microanalysis. The relationship between K/Na ratios and Fe/(K+Na) ratios were examined by X-ray microanalysis. The K/Na ratios give a measure of the membrane cation transport function. The Fe/(K+Na), which is analogous to hemoglobin concentration, gives an index of maturation stage. The relationships between K/Na and Fe/(K+Na) in the marrow cells of normal adult dog and those of a phenylhydrazine-injected dog with accelerated erythropoiesis show that the modification of cation composition occurs after the initiation of hemoglobin synthesis but before its completion. Similar relationships in the reticulocytes obtained from phenylhydrazine-injected dogs as well as from newborn dogs show a consistent decrease in K/Na with increased Hb, indicating a drastic change in cation composition during the maturation of the reticulocytes. Therefore the modification in membrane transport function must have occurred before or during the formation of reticulocytes.