Calcium activity and post-ischemic suppression of protein synthesis

Increase in intracellular calcium concentration is a prominent feature of ischemia and has been considered a major factor in the initiation of ischemic pathology, which involves inhibition of protein synthesis. A reduction of calcium ion activity during and immediately after in vitro ischemia did not prevent inhibition of protein synthesis in hippocampae slices. When slices were overloaded with calcium by NMDA receptor activation or by the calcium ionophore A23187, no significant inhibition of protein synthesis was observed. We conclude that calcium overload plays only a limited role in ischemic inhibition of protein synthesis.     

Incorporation d'acides aminés radioactifs dans les membranes des globules lipidiques du lait humain

Summary Human milk fat globule membranes (MFGM) can incorporate radioactive¹⁴C amino acids in a hot trichloracetic acid-insoluble material. Aspecific adsorption and bacterial contamination are unlikely. The products of protein synthesis were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate or by action of proteolytic enzymes. Various inhibitors of protein synthesis were assayed. Fragments of rough endoplasmic reticulum or mitochondria could be involved in this incorporation.     

Protein synthesis inhibition induced by dimethylnitrosamine and diethylnitrosamine on isolated rat hepatocytes.

Time- and dose-dependent protein synthesis inhibition takes place following exposure to high doses of dimethylnitrosamine (DMN) or diethylnitrosamine (DENA) in isolated rat hepatocytes. The ability of DENA to depress protein synthesis is 5-fold higher than that of DMN. Cells inhibited by 60 min exposure to DMN or DENA, and then incubated in a nitrosamine-free medium, regain their initial rate of protein synthesis. This recovery is faster and more complete for DENA-treated cells.     

Protein synthesis inhibition induced by dimethylnitrosamine and diethylnitrosamine on isolated rat hepatocytes

Summary Time- and dose-dependent protein synthesis inhibition takes place following exposure to high doses of dimethylnitrosamine (DMN) or diethylnitrosamine (DENA) in isolated rat hepatocytes. The ability of DENA to depress protein synthesis is 5-fold higher than that of DMN. Cells inhibited by 60 min exposure to DMN or DENA, and then incubated in a nitrosamine-free medium, regain their initial rate of protein synthesis. This recovery is faster and more complete for DENA-treated cells.     

Effect of nonprotein thiols on protein synthesis in isolated rat hepatocytes

The ability of nonprotein thiols to modulate rates of protein synthesis was investigated in isolated rat hepatocytes. Addition of cysteine stimulates protein labelling by [¹⁴C] Leucine. Glutahione depletion, induced by in vivod administration of L-buthionine sulfoximine and diethylmaleate, did not alter the effect of cysteine, although it decreased the rate of protein synthesis by 32%. The effect of cysteine on protein synthesis does not seem to be related to a perturbatin of the redox state of the NAD⁺/NADH system or to changes in the rate of gluconeogenic pathway. The following observations indicate that cysteine may stimulate protein syntheis by increasing intracellular levels of aspartate: 1. Amino-oxyacetate, an inhibitor of pyridoxyal-dependent enzymes, inhibits protein labelling and decreases aspartate cellular content, whereas most amino acids accumulate or remain unchanged; 2. Cysteine, in the absence or in the presence of amino-ocycetate, stimulates protein labelling and induces aspartate accumulation, although mot amino acids diminish or remain unchanged.     

Zum Proliferations-Stoffwechsel der Rattenleber nach Teilhepatektomie. Autoradiographische Untersuchungen mit3H-Cytidin,-Phenylalanin und -Thymidin

Summary During the first wave of parenchymal liver regeneration in adult rats after partial hepatectomy, the cellular synthesis and migration of RNA and the metabolism of protein were studied by autoradiography following an injection of³H-cytidine or³H-l-phenylalanine and double injections of 1 of these precursors +³H-thymidine. The following results were obtained: the synthesis and migration of RNA and the metabolism of protein are enhanced under these conditions of proliferation. In spite of this, the relation of metabolic activity in nucleolus, karyoplasm and cytoplasm remains constant. By double injection techniques it is proved that no differences exist in migration of RNA into the cytoplasm and cytoplasmic protein synthesis between cells with or without DNA synthesizing nuclei.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

The effect of temperature and protein synthesis on the renaturation of firefly luciferase in intact H9c2 cells

A mild increase in temperature that does not exert an effect on tolerance development or synthesis of heat shock proteins (Hsps) in control cells can stimulate these processes when applied to cells that have previously been heat shocked. To study the underlying mechanism of this effect, H9c2 cells were stably transfected with the gene encoding firefly luciferase (Luc). Heat-shock-induced inactivation of Luc and its subsequent reactivation is frequently used as a model for cellular protein denaturation and renaturation. Luc reactivation was determined following a damaging heat shock (43 or 44 degrees C for 30 min) in cells that were subsequently exposed to either control temperatures (37 degrees C) or various mild hyperthermic conditions (from 38.5 to 41.5 degrees C for 1 h). To prevent changes in Luc activity consequent to new synthesis of Luc, Luc reactivation was monitored in the presence of cycloheximide, an inhibitor of protein synthesis. The results showed that reactivation of Luc was inhibited when heat-treated cells were post-treated under mild hyperthermic conditions. The observed increase in Hsp synthesis under mild hyperthermic post-heat shock conditions therefore appears to be the result of an increase in the period during which denatured proteins are present. In addition, we studied Luc reactivation in the absence of protein synthesis inhibitors. This condition led to much higher Luc activity. By estimating half-life times of Luc, the contribution of new Luc synthesis in this recovery could be determined, and only partially explained the observed increase in Luc reactivation after heat shock. Thus the synthesis of other proteins must be important for the renaturation of heat-damaged proteins.     

Eiweissvermehrung in ein- und zweikernigen systemen vonAcetabularia

Summary (1) The rate of protein synthesis was found to be different inAcetabularia crenulata andAcetabularia mediterranea the higher cytoplasmic protein synthesis inA. crenulata depending upon the diameter of the stalk.(2) In systems containing one or two nuclei, there was no difference in the rate of cytoplasmic synthesis of proteins. This corresponds to the diminution of size and efficiency of the nuclei in binucleated systems.(3) In interspecific grafts, the rate of cytoplasmic protein synthesis corresponds nearly to the rate of protein synthesis ofAcetabularia crenulata. Corresponding to morphogenetic processes, thecren-action is prevalent.     

Interphase differential sensitivity to protein synthesis inhibition

Summary The effect of protein synthesis inhibition during interphase ofAllium cepa L. root meristem cells was studied. Anisomycin and cycloheximide were used in intermittent treatments during interphase of a synchronous cell subpopulation labelled as binucleate by caffeine, and the delays in reaching prophase were recorded. High sensitivity to protein synthesis inhibition was detected in G₁ and in the S/G₂ boundary, while protein synthesis in most of the S and G₂ phase was not required for the normal timing of next prophase.     

Possibility of using131I-albumin as a marker for the estimations of microbial protein synthesis rates in the rumen

Summary Total microbial protein synthesis rates in the rumen of buffaloes were estimated by isotope dilution technique, using¹³¹I-albumin treated with tannic acid as a marker. The animals were fed groundnut cake treated with formaldehyde to meet 50% of their digestible crude protein (DCP) requirement and 2.5% urea molasses mixture was given to meet the remaining requirement of DCP. Wheat straw was fed as the basal roughage. The total average microbial protein synthesis was 58.14 g/day.     

Der Einfluss von Halothan und Chloroform auf die Feinstruktur und Proteinsynthese der Rattenleber

Summary Examinations by electron microscopic and autoradiographic techniques of whether chloroform and halothane alter protein synthesis in rat liver, demonstrate, that chloroform causes an early destruction of the granular endoplasmatic reticulum accompanied by a marked decrease of the protein synthesis in the centre of the acini. After anaesthesia with halothane, the granular endoplasmatic reticulum and the protein synthesis proved to remain unaffected.     

Memory formation and the regulation of gene expression

On a cellular level, formation of memory is based on a selective change in synaptic efficacy that is both fast and, in case of important information, long-lasting. Rapidity of cellular changes is achieved by modifying preexisting synaptic molecules (receptors, ion channels), which instantaneously alters the efficacy of synaptic transmission. Endurance, that is the formation of long-term memory (LTM), is based on transient and perhaps also long-lasting changes in protein synthesis. A number of different methods exist to interfere with the synthesis of specific proteins or proteins in general. Other methods, in turn, help to identify proteins whose synthesis is changed following learning. These mostly molecular methods are briefly described in the present review. Their successful application in a variety of memory paradigms in invertebrates and vertebrates is illustrated. The data support the importance of selective changes in gene expression for LTM. Proteins newly synthesized during memory consolidation are likely to contribute to restructuring processes at the synapse, altering the efficiency of transmission beyond the scope of STM. Increased or, less often, decreased synthesis of proteins appears during specific time windows following learning. Recent evidence supports older data suggesting that two or even more waves of protein synthesis exist during the consolidation period. It is expected that the new molecular methods will help to identify and characterize molecules whose expression changes during LTM formation even in complex vertebrate learning paradigms.     

Effect of epidermal growth factor on growth and maturation of fetal and neonatal rat small intestine in organ culture

Small intestinal explants from pre- and post-natal rats were incubated in an organ culture system in the absence and presence of epidermal growth factor (EGF). The rate of synthesis of small intestinal DNA and protein as well as the activity of lactase and alkaline phosphatase increased rapidly between 17 and 20-day gestational age, whereafter they declined. The maximal incorporation of 3H-thymidine and 14C-alanine into DNA and protein, respectively, was significantly stimulated by EGF (100 ng/ml). EGF had no effect on the activity of either lactase or alkaline phosphatase in the small intestinal explants.     

Protein synthesis in vivo in muscles of the growing lamb]

The relationship between incorporation of intravenously injected 14C lysine and specific radio-activity of precursor was used to estimate protein synthesis in muscle of growing lambs. The rate of protein synthesis per unit of muscle weight in Supraspinatus and Extensor digitorum longus decreased strongly from one week of age to puberty (10 weeks); afterwards it decreased in supraspinatus and increased slightly in Extensor digitorum longus. The rate of protein synthesis increase in muscle protein weight was constant during the whole experiment (1 week-16 weeks). In preruminant Lambs )1 week-5 weeks) the rate of protein synthesis per unit of muscle weight decreased; however, due to the increase in muscle weight, the rate of protein synthesis in whole muscle remained relatively constant. In order Lambs the rate of protein synthesis in whole muscle decreased. The turnover time of protein increased with age. These results give some explanation on muscular development of Lambs.     

Analyse électrophorétique des protéines caroténogènes deNeurospora crassa

Summary In an effort to determine the number of proteins responsible for the synthesis of the neutral carotenoids inN. crassa, the soluble protein extracts from light-induced and dark-grown cultures of a neurosporaxanthinless strain were compared electrophoretically. The differences relevant to the carotenogenic proteins were identified by reference to an albino polarity mutant. A single, electrophoretically determinable, protein was found to be specific to carotenogenesis. This implication of a single (presumably an enzyme aggregate) protein controlling carotenoid synthesis is discussed and correlated to the existing genetic, complementation and biochemical evidence.     

Yeast as a sensor of factors affecting the accuracy of protein synthesis

The cell monitors and maintains the fidelity of translation during the three stages of protein synthesis: initiation, elongation and termination. Errors can arise by multiple mechanisms, such as altered start site selection, reading frame shifts, misincorporation or nonsense codon suppression. All of these events produce incorrect protein products. Translational accuracy is affected by both cis- and trans-acting elements that insure the proper peptide is synthesized by the protein synthetic machinery. Many cellular components are involved in the accuracy of translation, including RNAs (transfer RNAs, messenger RNAs and ribosomal RNAs) and proteins (ribosomal proteins and translation factors). The yeast Saccharomyces cerevisiae has proven an ideal system to study translational fidelity by integrating genetic approaches with biochemical analysis. This review focuses on the ways studies in yeast have contributed to our understanding of the roles translation factors and the ribosome play in assuring the accuracy of protein synthesis.Received 27 November 2002; received after revision 16 April 2003; accepted 25 April 2003     

Effect of acute administration of triiodothyronine in chicken. Liver glycogen depletion and amino acid incorporation to proteins

Summary Single injections of thyroid hormone (T₃) produce liver glycogen depletion in chickens. This effect cannot be suppressed by protein synthesis inhibitors and is previous to the hormone-induced increase in protein synthesis.     

Comparison of protein synthesis profiles in chronic lymphocytic leukaemia cells and B-lymphocytes from peripheral blood,cord blood and tonsil

2D-gel electrophoresis was used to investigate protein synthesis in leukaemic cells from a series of 15 chronic lymphocytic leukaemia (CLL) patients, and in non-malignant B-cell populations from different sources. The protein synthesis profiles of CD5+ B-cells from umbilical cord blood and from tonsil were determined, and the levels of expression of their proteins were observed to be similar to the CLL cells. The CD5-cells from cord blood resembled peripheral blood B-lymphocytes, and the protein synthesis profile of CD5-cells from tonsils was very complex. One protein was also identified which consistently appeared to be synthesised at a low level in CD5+ B-cells from tonsil but which was always more prominant in CLL cells and other non-malignant B-lymphocytes. On the basis of these data it is possible that the closest non-malignant counterpart to CLL is the CD5+ B-lymphocyte from cord blood.     

Contrasting effects of RNA and protein synthesis blocking on natural and lectin-dependent cell-mediated cytotoxicity against adherent HEp-2 cells

In this study, earlier observations concerning the independence of both natural (NCMC) and lectin-dependent cell-mediated cytotoxicity (LDCC) from DNA synthesis have been confirmed. In addition, blocking of RNA synthesis by actinomycin D and of protein synthesis, reversibly by puromycin (PM) and irreversibly by emetine (EM) had different effects on NCMC and LDCC against 3H-thymidine-prelabeled HEp-2 target cells. Similarly to the Con A-induced proliferation of lymphocytes, LDCC activity was also inhibited by blocking of RNA and protein synthesis. NCMC to HEp-2 target cells was not affected by blocking of RNA synthesis, while both PM and EM strongly enhanced NCMC activity.