PubMed Health⌕ Search

Biomedical subjects

M L Freedman

Publications and source records attributed to M L Freedman.

At least 73 records · Page 4Linked to original sources
PubMed1979

Ethanol inhibition of rabbit reticulocyte haem synthesis at the level of delta-aminolaevulinic acid synthetase.

Ethanol inhibition of rabbit reticulocyte synthesis occurs as a result of a decrease in haem synthesis. The present study therefore was undertaken in order to localize the inhibitory site of ethanol on the haem biosynthetic pathway. Ethanol (0.05--0.15 M) inhibition of reticulocyte protein synthesis was prevented by simultaneous incubation with 0.025--1 mM delta-aminolaevulinic acid (ALA). Ethanol inhibited both 14C-glycine and 14C-ALA incorporation into haem. However, the extent of haem formation with 14C-ALA as substrate in the presence of ethanol was still equal to that when 14C-glycine was used. These data suggest that ethanol inhibits the haem synthetic pathway at several loci, but that the decrease in haem synthesis, responsible for the decrease in protein synthesis, is due to the inhibition at the rate-limiting enzyme, delta-aminolaevulinic acid synthetase (ALA-S). To confirm this, ALA-S activity was then directly measured in intact reticulocytes, and it was shown that ethanol indeed inhibited its activity. The inhibition of ALA-S was prevented by 10(-4) M dibutyryl cyclic AMP (db cAMP) or theophylline, agents which elevate intracellular cAMP and which have previously been shown to prevent and reverse ethanol inhibition of haem and protein synthesis. Thus, it appears that cAMP protects against ethanol toxicity by preventing inhibition of ALA-S.

5-Aminolevulinate Synthetase↗
PubMed1979

Erythrocyte parameters in the elderly: an argument against new geriatric normal values.

Erythrocyte parameters in 292 unselected geriatric patients were studied retrospectively. Statistically significant mean decreases compared to the laboratories' normal mean values were found in the red blood cell count (RBC), hemoglobin level, hematocrit, and mean corpuscular hemoglobin concentration; there also was a slight increase in mean corpuscular volume and mean corpuscular hemoglobin. However, when 17 patients with a hemoglobin level less than 10 gm/dl or a hematocrit reading less than 35 percent were excluded, all of the mean values for erythrocyte parameters fell within the normal range. In 71 percent of these 17 patients the etiology of the anemia was documented. Although one cannot definitely exclude a slight change in erythrocyte parameters with aging, it is concluded that the establishment of so-called new geriatric norms is premature, as the population studied included patients with various diseases. Good medical practice dictates continued evaluation and monitoring of patients in whom erythrocyte values are outside the established normal ranges.

Aged↗
PubMed1978

Heat inhibition of reticulocyte protein synthesis. Evidence for a mechanism independent of the hemin-controlled repressor.

Incubation of rabbit reticulocytes at 45 degrees C results in a prompt but reversible decrease in protein synthesis and a concomitant conversion of polyribosomes to smaller aggregates. These effects occur even in the presence of 100 micrometer hemin in the incubation medium. There is also inhibition of heme synthesis but this occurs at a later time than the effect on protein synthesis. The inhibtion of heme synthesis results from a decrease in activity of beta-aminolevulinic acid synthetase. This decrease of heme synthesis appears to be secondary to the inhibition of protein synthesis with resultant accumulation of intramitochondrial heme (which will decrease beta-aminolevulinic acid synthetase activity). An inhibitor of reticulocyte cell-free protein synthesis formed in the postribosomal supernatants of cells incubated at both 45 and 37 degrees C but not at 0 degrees C. No temporal or quantitative differences in the amount of this inhibitor from cells treated at either 37 or 45 degrees C was apparent. The inhibitor was not found in the fraction where the hemin-controlled repressor is isolated. It is concluded that heat inactivation of intact reticulocyte protein synthesis does not depend upon a decrease in heme synthesis, heme concentration or generation of the hemin-controlled repressor. Furthermore, it appears that the inhibitor formed in the post-ribosomal supernatant cannot be the sole cause of the heat inhibition of protein synthesis.

5-Aminolevulinate Synthetase↗
PubMed1978

Genetic alterations of Streptococcus mutans' virulence.

The use of mutants defective in caries-associated traits has enabled the genetic dissociation of agglutination from adhesion, the demonstration of serotype-specific contributions of IPS to virulence, the importance of glucanohydrolase to virulence to a greater degree than to plaque formation, and the apparent lack of importance of agglutination to virulence. We have also been able to demonstrate the ability of plaque formation-defective mutants and other variants both to infect and to emerge, yet not to cause disease. Additional mutants, currently under study in our laboratory include fructanase, invertase, and sucrose permease-defectives. Ultimately, the identification of key, probably surface-associated virulence factors will offer more potent and specific antigens for directed immune responses by the host.

Adhesiveness↗
PubMed1978

In vitro colonization of Streptococcus mutans on enamel.

An in vitro model consisting of enamel from extracted human molars, suspended from wires in inoculated culture tubes, was used to study the adhesion of bacteria to enamel. Under conditions in which there was no macroscopically visible plaque formation, electron micrographs showed no bacterial deposits on the enamel surface. In samples where Streptococcus mutans attached to enamel, an extracellular, pellicle-like material was associated with the bacteria adjacent to the enamel. This material appeared to bind to the enamel surface and to mediate bacterial attachment. Membrane-filtered (Millipore Corp.) saliva deposited a thin surface layer on the enamel, but there were no observable alterations of S. mutans attachment to enamel pretreated with saliva. It was noted that Bratthall serotype c and e strains of S. mutans, when grown in glucose-containing medium, attached, although less tenaciously, to enamel and nichrome wires. Chemical and gas chromatographic analyses of cell-associated materials formed by serotype c and e strains cultured in glucose-containing medium revealed low amounts of glucose-positive material and no polymer linkages characteristic of glucan; yet the same strains cultured in sucrose-containing medium had relatively high amounts of glucose-positive material, with polymer linkages-characteristic of glucan. Serotype a, b, and d strains could attach only in sucrose-containing media.

Dental Enamel↗
PubMed1978

Hypersplenic thrombocytopenia differentiated from increased peripheral destruction by platelet volume.

Platelet volume distribution was examined in 16 patients with hepatosplenomegaly and platelet counts of 45 000 to 90 000/mm; 12 patients with autoimmune thrombocytopenia and randomly matched platelet counts; and 20 normal subjects. Five platelet volume variables of increasing platelet size were defined from the averages of 20 normal curves. Patients with hypersplenism had decreased volume values of 78% to 87% (mean, 83%) of the average normal population (P less than 0.001). Patients with autoimmune thrombocytopenic purpura had values significantly greater than normal by 124% to 149% (mean, 134%) (P less than 0.001). Patients with autoimmune thrombocytopenic purpura, when compared with hypersplenic patients, had significantly greater platelet volume values ranging from 154% to 174% (mean, 161%), P less than 0.001. We concluded that patients with hepatosplenomegaly have smaller platelets in their peripheral blood and a platelet volume distribution that can be distinguished easily from patients with autoimmune thrombocytopenic purpura, despite comparable platelet counts.

Autoimmune Diseases↗
PubMed1977

Fine structure of the Panorex image.

The plane of focus free of blur margin, as well as the rate of increase of blur margin both medial and lateral to the plane of focus, has been described in the Panorex panoramic x-ray machine by means of a continuous phantom. The average of readings made at ninety-eight separate film positions yields a focaltrough thickness of 4.3 mm. if a blur margin of 10 per cent of the phantom's film image is accepted, the percentage blur increasing directly with trough thickness. The factor enlargement of objects at the plane of focus is uniform at 1.25 and increases along with percentage blur more rapidly for objects medial to the plane of focus than for objects lateral to it.

Models, Anatomic↗
PubMed1977

Benzene inhibition of in vitro rabbit reticulocyte haem synthesis at delta aminolaevulinic acid synthetase: reversal of benzene toxicity by pyridoxine.

Benzene (0.113 M) inhibited haem and protein synthesis in rabbit reticulocytes. This inhibition of haem synthesis was found when L-2-[14C]-glycine was used as the radioactive precursor. However, when 4-[14C]delta-aminolaevulinic acid (ALA) was used, there was no significant inhibition. Since ALA measures the haem synthetic pathway beyond the enzyme delta-aminolaevulinic acid synthetase (ALA synthetase), these results suggest that benzene inhibits haem synthesis at or before ALA synthetase. This was confirmed by demonstrating that 1 mM ALA both protected against and reversed the benzene inhibition of reticulocyte protein synthesis. In addition, 1 mM pyridoxine both protected against and reversed the benzene inhibition of reticulocyte protein synthesis. In addition, ImM pyridoxine both protected against and reversed the benzene inhibition of reticulocyte haem and protein synthesis. These results indicate that benzene (or a metabolite) either competes with pyridoxal phosphate at ALA synthetase or competes with pyridoxine for pyridoxal phosphokinase. These results are discussed in terms of their implications for the possible roles of ALA synthetase and the haemin-controlled repressor in benzene-induced aplastic anaemia.

5-Aminolevulinate Synthetase↗
PubMed1976

Ferritin and sideroblastic anaemias: inhibition of protein synthesis by protease contaminants in commercial preparations of ferritin.

Commercial preparations of ferritin inhibited reticulocyte-lysate cell-free protein synthesis and disaggregated polyribosomes to monoribosomes and ribosomal subunits. These effects were prevented by addition of reduced glutathione (GSH) to the incubation medium, but ferritin did not lower GSH concentration in the lysates. The more purified the ferritin preparation, the less inhibition of protein synthesis was observed. These data suggested that the effect was due to a contamination of the ferritin with proteolytic activity. In confirmation of this proposal we demonstrated that there was protease activity in both the 2X and 5X crystalized ferritin preparations, with 2.5 times greater activity in the 2X preparation. The proteolytic activity in ferritin was inhibited by incubation with the protease inhibitor tosyl lysine chloromethyl ketone (TLCK). When an amount of trypsin equivalent to the protease activity of the ferritin was added to the incubation mixture, similar effects on protein synthesis and the ribosome-polyribosome component were found. Both GSH and TLCK prevented these effects of trypsin. These data suggest that the previously reported effect of ferritin on reticulocyte cell-free protein synthesis was due to contamination of the ferritin by a protease. It appears that ferritin does not play a direct role in the pathogenesis of sideroblastic anaemias.

Anemia, Sideroblastic↗
PubMed1976

A rabbit reticulocyte model for the role of hemin-controlled repressor in hypochromic anemias.

Hemin allows maximal protein synthesis in intact rabbit reticulocytes and their cell-free lysate preparations by retarding the formation of a translational repressor (HCR) found in the postribosomal supernate. In order to evaluate the role of HCR in the pathogenesis of hypochromic anemias, HCR was isolated and partially purified from intact rabbit reticulocytes incubated in vitro with either 0.1 mM alpha,alpha-dipyridyl (an iron-chelating agent) or 0.1 M ethanol. Both of these agents inhibit reticulocyte protein synthesis. Hemin (50 muM) protects against the inhibition by both agents. A ferrous iron-transferrin mixture, however, protects only against alpha,alpha-dipyridyl. Both alpha,alpha-dipyridyl and ethanol inhibit heme synthesis before the time that protein synthesis is affected, while neither lowers either ATP or GSH levels. These results indicate that while both agents inhibit heme synthesis, alpha,alpha-dipyridyl does so by inducing iron deficiency while ethanol works at a non-iron-requiring step. When HCR was isolated from intact cells and assayed in the reticulocyte cell-free systems, plus and minus hemin, premature appearance of HCR was found in cells incubated in vitro with alpha,alpha-dipyridyl or ethanol. When hemin was present in the intact cell incubation, the appearance of HCR was retarded. The HCR from alpha,alpha-dipyridyl ethanol-treated cells was partially purified and eluted at the same location on a Sephadex G-200 column (molecular weight approximately 3 x 10(5)) as that from postribosomal supernates incubated minus hemin. In addition rabbits with phenylhydrazine-induced hemolytic anemia were given intravenous ethanol in vivo at a dose of 0.4 ml/kg. This concentration of alcohol resulted in an inhibition of the rate of heme synthesis and protein synthesis as well as an acceleration of HCR formation in reticulocytes. The HCR from these in vivo treated rabbits was isolated, partially purified, and assayed in an identical fashion as the in vitro experiments. These in vivo experiments further support the physiological and pathophysiological role of HCR in reticulocytes. On the basis of these results a model for a role of HCR in some of the hypochromic anemias is proposed. In iron deficiency or chronic disease (where iron is not available to the erythroblast for heme synthesis) HCR appears prematurely and inhibits protein synthesis. When heme synthesis is inhibited by ethanol but there is sufficient intracellular iron, HCR appears prematurely and inhibits protein synthesis, iron accumulates in the erythroblast, and the end result is sideroblastic anemia.

2,2'-Dipyridyl↗
PubMed1976

Hemin reversal of benzene-induced inhibition of reticulocyte protein synthesis.

Benzene (0.056-0.113 M) rapidly and reversibly inhibited protein synthesis in anucleate human sickle cell and rabbit reticulocytes. Hemin (50 muM) both prevented and reversed this effect of benzene. The inhibition in rabbit reticulocytes was accompanied by a conversion of polyribosomal disaggregation required ribosomal movement along mRNA and was also prevented and reversed by 50 muM hemin. Benzene was also shown to inhibit heme synthesis in rabbit reticulocytes while neither ATP nor GSH levels were altered. A translational repressor (HCR) of reticulocyte cell-free protein synthesis was isolated from intact cells incubated with benzene, while no significant amount of HCR was found in cells incubated with both benzene and hemin. These results indicated that benzene inhibits translation at the heme-dependent site of initiation. The clinical implications of these experiments remain to be elucidated.

Adenosine Triphosphate↗
PubMed1976

Benzene and lead inhibition of rabbit reticulocyte heme and protein synthesis: evidence for additive toxicity of these two components of commercial gasoline.

Both benzene and lead are known hematopoietic toxins. These compounds are components of commercial gasoline, and therefore pose an environmental risk. The present study demonstrates that benzene and lead individually inhibit both intact reticulocyte heme and protein synthesis in the presence or absence of iron-transferrin. When these two compounds are present in the same incubation, their effects are additive. These in vitro results suggest that further evaluation of the combined risks of benzene and lead is indicated. Furthermore, the data shows the potential for additive interaction by chemically unrelated environmental compounds.

Anemia↗