Changes In Rate Of Synthesis Research Articles

Fibrinogen and albumin synthesis rates in major upper abdominal surgery.

Plasma fibrinogen and albumin concentrations initially decrease after abdominal surgery. On postoperative days 3–5 fibrinogen concentration returns to the preoperative level or even higher, while albumin stays low. It is not known if these altered plasma concentrations reflect changes in synthesis rate, utilization, or both. In particular a low albumin plasma concentration has often been attributed to a low synthesis rate, which is not always the case. The objective of this study was to determine fibrinogen and albumin quantitative synthesis rates in patients undergoing major upper abdominal surgery with and without intact liver size. Patients undergoing liver or pancreatic resection (n = 9+6) were studied preoperatively, on postoperative days 1 and 3–5. De novo synthesis of fibrinogen and albumin was determined; in addition, several biomarkers indicative of fibrinogen utilization were monitored. After hemihepatectomy, fibrinogen synthesis was 2-3-fold higher on postoperative day 1 than preoperatively. On postoperative days 3–5 the synthesis level was still higher than preoperatively. Following major liver resections albumin synthesis was not altered postoperatively compared to preoperative values. After pancreatic resection, on postoperative day 1 fibrinogen synthesis was 5-6-fold higher than preoperatively and albumin synthesis 1.5-fold higher. On postoperative days 3–5, synthesis levels returned to preoperative levels. Despite decreases in plasma concentrations, de novo synthesis of fibrinogen was markedly stimulated on postoperative day 1 after both hemihepatectomies and pancreatectomies, while de novo albumin synthesis remained grossly unchanged. The less pronounced changes seen following hepatectomies were possibly related to the loss of liver tissue.

The total mRNA concentration buffering system in yeast is global rather than gene-specific.

Gene expression in eukaryotes does not follow a linear process from transcription to translation and mRNA degradation. Instead it follows a circular process in which cytoplasmic mRNA decay crosstalks with nuclear transcription. In many instances, this crosstalk contributes to buffer mRNA at a roughly constant concentration. Whether the mRNA buffering concept operates on the total mRNA concentration or at the gene-specific level, and if the mechanism to do so is a global or a specific one, remain unknown. Here we assessed changes in mRNA concentrations and their synthesis rates along the transcriptome of aneuploid strains of the yeast Saccharomyces cerevisiae. We also assessed mRNA concentrations and their synthesis rates in nonsense-mediated decay (NMD) targets in euploid strains. We found that the altered synthesis rates in the genes from the aneuploid chromosome and the changes in their mRNA stabilities were not counterbalanced. In addition, the stability of NMD targets was not specifically compensated by the changes in synthesis rate. We conclude that there is no genetic compensation of NMD mRNA targets in yeast, and total mRNA buffering uses mostly a global system rather than a gene-specific one.

Effects of Protein in Combination with Carbohydrate Supplements on Acute or Repeat Endurance Exercise Performance: A Systematic Review

Protein supplements are consumed frequently by athletes and recreationally active adults for various reasons, including improved exercise performance and recovery after exercise. Yet, far too often, the decision to purchase and consume protein supplements is based on marketing claims rather than available evidence-based research. The purpose of this review was to provide a systematic and comprehensive analysis of the literature that tested the hypothesis that protein supplements, when combined with carbohydrate, directly enhance endurance performance by sparing muscle glycogen during exercise and increasing the rate of glycogen restoration during recovery. The analysis was used to create evidence statements based on an accepted strength of recommendation taxonomy. English language articles were searched with PubMed and Google Scholar using protein and supplements together with performance, exercise, competition, and muscle, alone or in combination as keywords. Additional articles were retrieved from reference lists found in these papers. Inclusion criteria specified recruiting healthy active adults less than 50 years of age and evaluating the effects of protein supplements in combination with carbohydrate on endurance performance metrics such as time-to-exhaustion, time-trial, or total power output during sprint intervals. The literature search identified 28 articles, of which 26 incorporated test metrics that permitted exclusive categorization into one of the following sections: ingestion during an acute bout of exercise (n = 11) and ingestion during and after exercise to affect subsequent endurance performance (n = 15). The remaining two articles contained performance metrics that spanned both categories. All papers were read in detail and searched for experimental design confounders such as energy content of the supplements, dietary control, use of trained or untrained participants, number of subjects recruited, direct measures of muscle glycogen utilization and restoration, and the sensitivity of the test metrics to explain the discrepant findings. Our evidence statements assert that when carbohydrate supplementation was delivered at optimal rates during or after exercise, protein supplements provided no further ergogenic effect, regardless of the performance metric used. In addition, the limited data available suggested recovery of muscle glycogen stores together with subsequent rate of utilization during exercise is not related to the potential ergogenic effect of protein supplements. Many studies lacked ability to measure direct effects of protein supplementation on muscle metabolism through determination of muscle glycogen, kinetic assessments of protein turnover, or changes in key signaling proteins, and therefore could not substantiate changes in rates of synthesis or degradation of protein. As a result, the interpretation of their data was often biased and inconclusive since they lacked ability to test the proposed underlying mechanism of action. When carbohydrate is delivered at optimal rates during or after endurance exercise, protein supplements appear to have no direct endurance performance enhancing effect.

Erythrocyte glutathione concentration and production during hyperinsulinemia, hyperglycemia, and endotoxemia in healthy humans

In diabetes mellitus and sepsis, low erythrocyte glutathione (GSH) concentrations are found. Whether this is caused by lowered GSH production has not been clarified. To obtain insight in the relationship between erythrocyte GSH concentrations and GSH production, GSH kinetics were measured in healthy male volunteers during 4 different clamps (low-dose or medium-dose insulin [100 or 400 pmol/L] and euglycemia or hyperglycemia [5 or 12 mmol/L]) in a control setting (n = 6; all 4 clamps in the same subject) or after systemic administration of lipopolysaccharide (to mimic sepsis) (4 groups of n = 6; each clamp in a different subject). Hyperinsulinemia decreased erythrocyte GSH concentration (P = .042), but did not affect fractional synthetic rate (FSR) of GSH. Hyperglycemia did not affect erythrocyte GSH concentration, but decreased FSR of GSH (P = .025). Lipopolysaccharide decreased erythrocyte GSH concentration (P < .001), but increased FSR of erythrocyte GSH (P = .035). Depending on the metabolic circumstances, we found either stable GSH concentrations with lower production rates or decreased levels with either no change or an increase in production rate. Based upon these data, it seems inappropriate to infer conclusions about changes in synthesis rate of GSH from changes in its concentration.

Rapid Dark Repression of 5-Aminolevulinic Acid Synthesis in Green Barley Leaves

In photosynthetic organisms chlorophyll and heme biosynthesis is tightly regulated at various levels in response to environmental adaptation and plant development. The formation of 5-aminolevulinic acid (ALA) is the key regulatory step and provides adequate amounts of the common precursor molecule for the Mg and Fe branches of tetrapyrrole biosynthesis. Pathway control prevents accumulation of metabolic intermediates and avoids photo-oxidative damage. In angiosperms reduction of protochlorophyllide (Pchlide) to chlorophyllide is catalyzed by the light-dependent NADPH:Pchlide oxidoreductase (POR). Although a correlation between down-regulated ALA synthesis and accumulation of Pchlide in the dark was proposed a long time ago, the time-resolved mutual dependency has never been analyzed. Taking advantage of the high metabolic activity of young barley (Hordeum vulgare L.) seedlings, in planta ALA synthesis could be determined with high time-resolution. ALA formation declined immediately after transition from light to dark and correlated with an immediate accumulation of POR-bound Pchlide within the first 60 min in darkness. The flu homologous barley mutant tigrina d(12) uncouples ALA synthesis from dark-suppression and continued to form ALA in darkness without a significant change in synthesis rate in this time interval. Similarly, inhibition of protoporphyrinogen IX oxidase by acifluorfen resulted in a delayed accumulation of Pchlide during the entire dark period and a weak repression of ALA synthesis in darkness. Moreover, it is demonstrated that dark repression of ALA formation relies rather on rapid post-translational regulation in response to accumulating Pchlide than on changes in nuclear gene expression.

Effects of Infliximab and Parenteral Nutrition on Albumin and Fibrinogen Synthesis Rates in Pediatric Crohn Disease

Tumor necrosis factor-alpha (TNF-alpha) may play a significant role in growth disturbance in pediatric Crohn disease. The aim of this study was to determine the effects of anti-TNF-alpha therapy on albumin and fibrinogen synthesis during both fasting and parenteral nutrition infusion in pediatric patients with active Crohn disease. Children with active Crohn disease scheduled for their initial dose of infliximab underwent assessment immediately before and 2 weeks following infliximab infusion. Using the stable isotope [d5] phenylalanine, rates of fractional and absolute albumin and fibrinogen synthesis were calculated. Measurements were made in both the fasting and parenterally fed states. Fifteen children (mean age 14.9 +/- 0.3) completed the study. The mean serum albumin changed from 3.59 +/- 0.08 to 3.66 +/- 0.04 g/dL, and the mean fibrinogen level decreased from 230 +/- 17 to 187 +/- 8 mg/dL (P < 0.05) following infliximab therapy. During fasting, there were no changes in albumin and fibrinogen synthesis rates following infliximab. During parenteral nutrition infusion, the fractional albumin synthesis rate changed from 11.8% to 15.1%/day (P = 0.06), and the absolute albumin synthesis rate increased from 192 to 248 mg x kg(-1) x day(-1) (P < 0.05), whereas no changes in fibrinogen synthesis rates were observed. Synthesis rates of albumin and fibrinogen were increased during parenteral nutrition infusion compared with the fasting state. Following infliximab therapy, during parenteral nutrition infusion, albumin synthesis increased significantly. Conversely, serum fibrinogen levels decreased following infliximab therapy in the absence of significant change in synthesis rates.

Cold tolerance and changes in rates of synthesis of individual proteins of Clostridium perfringens as a result of cold pre-treatment

Clostridium perfringens is a widely distributed foodborne pathogen. Its ability to survive cold encounters could contribute to its persistence in foods and the potential to cause disease. In this work five cold-shock proteins (101, 82, 70, 45 and 10 kDa) were induced by cold-shocking C. perfringens FD-1041 from 43 °C to 28 °C, as revealed by labeling with L-[35S]methionine and cysteine followed by gel electrophoresis. Cold shock also increased the cold tolerance of the cells at least fifteen fold. The acquired tolerance was maintained for 2 h after the cold treatment. This ability of C. perfringens could improve the survival in foods and present a significant hazard.

Cold tolerance and changes in rates of synthesis of individual proteins of Clostridium perfringens as a result of cold pre-treatment

Clostridium perfringens is a widely distributed foodborne pathogen. Its ability to survive cold encounters could contribute to its persistence in foods and the potential to cause disease. In this work five cold-shock proteins (101, 82, 70, 45 and 10 kDa) were induced by cold-shocking C. perfringens FD-1041 from 43 °C to 28 °C, as revealed by labeling with L-[ 35S]methionine and cysteine followed by gel electrophoresis. Cold shock also increased the cold tolerance of the cells at least fifteen fold. The acquired tolerance was maintained for 2 h after the cold treatment. This ability of C. perfringens could improve the survival in foods and present a significant hazard. _

The effect of rate of weight loss on erythrocyte glutathione concentration and synthesis in healthy obese men.

Obesity is commonly associated with a high incidence and prevalence of dyslipidaemia, cardiovascular disease and Type II diabetes. Interestingly, studies have also reported decreased antioxidant levels in obese subjects. This may constitute an independent risk factor in the pathogenesis of coronary artery disease as obese subjects would have a decreased capacity to prevent the oxidative modification of low-density lipoproteins, which is a mechanism suggested as central to the development of atherogenesis. As part of a study to investigate responses to weight loss, we have assessed the effects on GSH status of a decrease in body mass of 5%, either after 6 days of complete starvation or 11 days of a very low calorie diet (2.55 MJ/day). There were significant differences between the two groups in the synthesis rate of erythrocyte GSH in response to weight loss. Both the fractional and the erythrocyte synthesis rate of GSH decreased significantly (P<0.01) in the starvation group by 22% and 16% respectively. In contrast, no change in synthesis rates was observed in the very low calorie diet group (P>0.05). Total erythrocyte concentration of GSH was unaffected by the weight loss within both groups. These results suggest that erythrocyte GSH synthesis is depressed in response to a very rapid weight loss induced by fasting. An acute reduction in GSH synthesis in response to a rapid weight loss may constitute a risk factor during periods of increased GSH demands.

Erratum

AbstractElhadj Selim, Mousa Shaker A., Forsten‐Williams Kimberly. Chronic pulsatile shear stress impacts synthesis of proteoglycans by endothelial cells: Effect on platelet aggregation and coagulation. J Cell Biochem 86 (2) 239‐250, 2002. An unnecessary correction was added to the last sentence of the last paragraph of the Discussion section of the above article. The correct paragraph is: We have found both structural and overall quantity/synthesis difference in proteoglycans isolated from cells cultured under low and high shear stress. These structural changes manifest in differences in activity with regard to inhibition of platelet aggregation and clot formation. We also found, however, a difference in synthesis rate of proteoglycans from cells cultured under the high and low shear stress. If we, therefore, looked at the level of activity on a per cell basis, the differences in effect on aggregation and clot formation disappear and the curves are essentially superimposable (data not shown). This suggests that either the effect of shear stress on the proteoglycan fine structure is being compensated for by a change in proteoglycan synthesis per cell or that the change in synthesis rate is being compensated for by a change in the fine structure of the material. Our data can not allow us to distinguish between the two, however, either way it suggests that the cells are capable of compensating for changes in response to shear stress so as to maintain a desired level of proteoglycan activity. We have only investigated this with regard to platelet activity and further studies would be needed to determine if this compensatory mechanism, potentially important for vascular homeostasis, is general with regard to the myriad of other proteoglycan activities. The publisher regrets this error.

Acute modulation of the extent of apoB mRNA editing and the relative rates of syntheses of apoB48 and apoB100 in cultured rat hepatocytes by osmotic and other stress stimuli.

The mRNA for apolipoprotein B (apoB) is edited by the enzyme APOBEC-1, which acts as part of a multiprotein complex or editosome. In cultured rat hepatocytes obtained from fed animals this results in the presence of edited and unedited apoB mRNA in a ratio of approximately 3:2 in the basal state. In this study we show that hyper-osmotic media, which induce cell shrinkage, resulted in an acute increase in the degree of editing of apoB mRNA (hypo-osmotic conditions had no effect). This increase was accompanied by a parallel and highly positively correlated change in the ratio of the rate of synthesis of apoB48 relative to that of apoB100. These changes occurred in the absence of any changes in the overall APOBEC-1 mRNA levels, indicating that the activation of editing occurred at a post-transcriptional level. Levels of total apoB mRNA were also unaffected by hyper-osmotic exposure of the cells indicating that changes in the relative rates of synthesis of apoB48 and apoB100 were due to post/translational events. Exposure of cells to anisomycin at concentrations (50 micrograms/ml) that inhibit protein synthesis or to the transcriptional inhibitor actinomycin D produced changes in the degree of apoB mRNA editing that were similar to those given by hyper-osmotic shock indicating that editing is able to respond acutely to transcriptional or translational inhibition. Anisomycin, at concentrations (50 ng/ml) that activate SAPK/JNK but do not inhibit protein synthesis, gave only a fraction of the effect of hyper-osmotic shock. SB203580, an inhibitor of p38 kinase, did not attenuate the effects of hyper-osmotic conditions on APOBEC-1 editing. These observations suggest that these MAPkinase pathways play a relatively minor part in the transduction of the osmotic stimulus to the editing mechanism. The hyper-osmotically-induced increase in apoB mRNA editing was also insensitive to PD98059 and wortmannin (inhibitors of MEK and PI3 kinase, respectively). These data provide evidence that apoB mRNA editing is capable of acute modulation independently of transcriptional or translational mechanisms and suggest that one or more components of the editosome may undergo post-translational activation.

Age effect on fibrinogen and albumin synthesis in humans.

A strong association has been reported between atherosclerotic diseases and fibrinogen levels, and a decreased whole body protein synthesis has also been reported with aging. We investigated the effect of age on fractional synthesis rates (FSR) of fibrinogen and albumin in 12 human subjects of young (20-30 yr), middle (45-60 yr), and old (65-79 yr) age by use of L-[1-13C]leucine and L-[15N]phenylalanine as tracers. An age-related decline in FSR of fibrinogen (P < 0.01) was observed with use of both tracers, with the maximal decrease (average 37% with alpha-[13C]ketoisocaproate as the precursor) occurring by middle age and with no further changes thereafter. In contrast, plasma concentrations of fibrinogen increased with age (P < 0.002). There was no age-related change in synthesis rate and concentrations of albumin. An age-related decline in fibrinogen FSR, but not FSR of albumin, indicates a differential effect of age on synthesis rate of these two liver proteins. This study also demonstrated that the increased circulating levels of fibrinogen represent a slower rate of disposal of fibrinogen rather than an increased production rate.

Dietary carbohydrate and postexercise synthesis of proglycogen and macroglycogen in human skeletal muscle.

This study examined the role of carbohydrate (CHO) ingestion on the resynthesis of two pools of glycogen, proglycogen (PG) and macroglycogen (MG), in human skeletal muscle. Nine males completed an exhaustive glycogen depletion exercise bout at 70% maximal O2 consumption on two occasions. Subsequent 48-h dietary interventions consisted of either high (HC, 75% of energy intake) or low (LC, 32% of energy intake) CHO diets. Muscle biopsies were taken at exhaustion (EXH) and 4, 24, and 48 h later. The total muscle glycogen (Gt) at EXH for the HC and LC conditions was not significantly different, and the MG represented approximately 12% of the Gt. From EXH to 4 h, there was an increase in the PG only for HC and no change in MG in either diet (P < 0.05). From 4 to 24 h, the concentration of PG increased in both conditions (P < 0.05). Between 24 and 48 h, in HC the majority of the increase in Gt was due to the MG pool (P < 0.05). The MG and PG concentrations for HC were significantly greater than for LC at 24 and 48 h (P < 0.05). At 48 h the MG represented 40% of the Gt for the HC diet and only 21% for the LC diet. There was no change in the net rates of synthesis of PG or MG over 48 h for LC (P < 0.05). The net rate of PG synthesis from 0 to 4 h for HC was 16 +/- 1.68 mmol glucosyl units . kg dry wt-1 . h-1, which was threefold greater than for LC (P < 0. 05). The net rate of PG synthesis decreased significantly from 4 to 24 h for HC, whereas the net rate of MG synthesis was not different over 48 h but was significantly greater than in LC (P < 0.05). The two pools are synthesized at very different rates; both are sensitive to CHO, and the supercompensation associated with HC is due to a greater synthesis in the MG pool.

EFFECT OF HIGH AND LOW CARBOHYDRATE DIET ON THE SYNTHESIS OF PROGLYCOGEN AND MACROGLYCOGEN IN HUMAN SKELETAL MUSCLE

1471 The present study examined the role of carbohydrate(CHO) on the resynthesis of the two pools of glycogen, proglycogen(PG) and macroglycogen(MG), in human skeletal muscle. Ten subjects completed an exhaustive glycogen depletion exercise bout at 70% VO2 max on two occasions separated by 2 weeks. The diet was controlled by providing weighed, pre-packaged food such that either 75%(HC) or 32%(LC) of the energy was CHO. Muscle biopsies were taken at exhaustion(EXH), 4, 24 and 48 hours. The total muscle glycogen(TotG) at EXH was 79±15.6 and 113±20.1mmol glucosyl units/kg dw for HC and LC respectively. At exhaustion the MG concentration represented about 12% in each group. From EXH-4h there was an increase in the PG only for HC, and no change in MG in either diet(p<0.05). From 4-24h the concentration of PG increased in both conditions(p<0.05). Between 24-48h in HC all of the increase in TotG was due to the MG fraction(p<0.05). The MG and PG concentrations for HC were significantly greater than in LC at 24 and 48h (p<0.05). At 48h the MG represented 40% of the TotG for the HC diet and only 21% for LC. There was no change in the net rates of synthesis of PG or MG over 48h in LC(p<0.05). The net rate of PG synthesis from 0-4h in the HC was 16±1.68mmol glucosyl units/kg dw/h which is 3-fold greater than in the LC(p<0.05). The rate of PG synthesis decreased significantly 4-24h while the rate of MG synthesis was not significantly different over 48h but was significantly greater than LC(p<0.05). It appears that the two pools are dynamic and sensitive to CHO, especially the PG pool. The glycogen supercompensation is first generated in PG and the final increase in TotG occurs exclusively in MG. These data suggest that the MG pool is an important reservoir for glycogen synthesis at later time points if the CHO availability is high.

Hypoxia-induced changes in radiation sensitivity in human melanoma cells: importance of oxygen-regulated proteins, adenylate energy charge and cell cycle distribution

Background and purpose: The effects of transient hypoxia on the radiation sensitivity of human tumour cells have so far been investigated only to a limited extent, and only up to 12 h after reoxygenation. We irradiated cells shortly after reoxygenation (<1 h) or at prolonged times after reoxygenation (24 h and 48 h) in order to examine possible relationships between changes in radiation sensitivity on the one hand and changes in rates of synthesis of oxygen-regulated proteins, changes in energy metabolism and changes in cell cycle distribution on the other. Materials and methods: Four human melanoma cell lines (A-07, D-12, R-18 and U-25) were included in the study. After hypoxia treatment (4 h or 16 h) and reoxygenation, cells were either irradiated as monolayers at a dose rate of 2.0 cGy/min or prepared for protein analysis, energy charge measurements or flow cytometric measurements of DNA. Results: U-25 was the only line that showed increased radiation sensitivity shortly after reoxygenation, possibly because of extensive energy depletion. A-07 was the only line that showed increased radiation sensitivity at prolonged times after reoxygenation, possibly because of hypoxia-induced changes in the cell cycle distribution. The rates of synthesis of oxygen-regulated proteins (GRP78, GRP94, HSP70 and HSP90) were transiently perturbed to a similar extent in all lines after hypoxia treatment. Conclusion: The radiation sensitivity of the human melanoma cell lines was changed only to a minor extent by transient exposure to hypoxia.

Transferrin Kinetics Are Altered in Children with Severe Protein-Energy Malnutrition , ,

This study was undertaken to determine the following: 1 ) the kinetic changes responsible for the depletion and repletion of plasma transferrin (Tr) concentration in children with protein-energy malnutrition (PEM); 2 ) the role of infection in mediating these changes; and 3 ) whether plasma Tr concentration is related to body protein status. We measured plasma Tr concentration, and fractional (FSR) and absolute (ASR) Tr synthesis rates with the use of a constant intragastric infusion of 2H3-leucine in 14 children with PEM, at 2 d postadmission (study 1), 8 d postadmission when infections were under control (study 2), and at recovery (study 3). In studies 1 and 2, the children synthesized less Tr and had lower Tr concentrations compared with values at recovery. When infections were controlled, plasma Tr concentration rose, but Tr synthesis was unchanged. There were only fair correlations (P < 0.05) between plasma Tr concentrations and indices of wasting. Concerning malnourished children, we reached the following conclusions: 1 ) changes in the Tr pool size are achieved mainly through changes in synthesis rate; 2 ) infections play a minor role in reducing the Tr pool through either changes in the rate of catabolism or loss from the intravascular space; and 3 ) Tr concentration is not a very good indicator of protein nutritional status.

Protein expression during exponential growth in 0.7 M NaCl medium of Saccharomyces cerevisiae

Saccharomyces cerevisiae exponentially growing in basic or 0.7 M NaCl medium were isotopically labelled with 35S-methionine, followed by protein separation and quantification by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) combined with computerised image analysis. The electrophoretic separation resolved about 650 proteins of which 13 displayed significant and at least 2-fold changes in rate of synthesis during saline growth. By sequencing of 2D-PAGE resolved proteins, one of the 8 induced spot, p42.9/5.5, was shown to correspond to the full length (containing the N-terminal extension) product of the GPD1 gene encoding the cytoplasmic glycerol 3-phosphate dehydrogenase. The expression of the TDH3 gene, glyceraldehyde 3-phosphate dehydrogenase, and the ENO2 gene, enolase, decreased during growth in NaCl medium, declines hypothesised to have an impact on the flux to glycerol.

Protein expression during exponential growth in 0.7 M NaCl medium of Saccharomyces cerevisiae.

Saccharomyces cerevisiae exponentially growing in basic or 0.7 M NaCl medium were isotopically labelled with 35S-methionine, followed by protein separation and quantification by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) combined with computerised image analysis. The electrophoretic separation resolved about 650 proteins of which 13 displayed significant and at least 2-fold changes in rate of synthesis during saline growth. By sequencing of 2D-PAGE resolved proteins, one of the 8 induced spot, p42.9/5.5, was shown to correspond to the full length (containing the N-terminal extension) product of the GPD1 gene encoding the cytoplasmic glycerol 3-phosphate dehydrogenase. The expression of the TDH3 gene, glyceraldehyde 3-phosphate dehydrogenase, and the ENO2 gene, enolase, decreased during growth in NaCl medium, declines hypothesised to have an impact on the flux to glycerol.

Global changes in protein synthesis during adaptation of the yeast Saccharomyces cerevisiae to 0.7 M NaCl.

Exponentially growing Saccharomyces cerevisiae was challenged to increased salinity by transfer to 0.7 M NaCl medium, and changes in protein synthesis were examined during the 1st h of adaptation by use of two-dimensional gel electrophoresis coupled to computerized quantification. An impressive number of proteins displayed changes in the relative rate of synthesis, with most differences from nonstressed cells being found at between 20 and 40 min. During this period, 18 proteins exhibited more than eightfold increases in their rates of synthesis and were classified as highly NaCl responsive. Only two proteins were repressed to the same level. Most of these highly NaCl-responsive proteins seemed to constitute gene products not earlier reported to respond to dehydration. Applying a selection criterion to subsequent samples of a twofold change in the relative rate of synthesis, 14 different regulatory patterns were discerned. Most identified glycolytic enzymes exhibited a delayed response, and their rates of synthesis did not change until the middle phase of adaptation, with only a minor decrease in the rate of production. A slight salt-stimulated response was observed for some members of the HSP70 gene family. Overall, the data presented indicate complex intracellular signalling as well as involvement of diverse regulatory mechanisms during the period of adaptation to NaCl.

Fermentation Kinetics and the Production of Volatiles During Alcoholic Fermentation

Relationships between the kinetics of CO2 evolution and the kinetics of fusel alcohol and ester synthesis in lager fermentations were examined to assess whether on-line CO2 evolution rate measurements might provide a tool for the control of flavor development. Changes in synthesis rates of volatiles were correlated with the point of maximum CO2 production rate, but the process parameters that most strongly affected the kinetics of fermentation did not greatly influence the kinetics of alcohol or ester expression. Differential effects of temperature profile on the expression of alcohols and esters were also observed. The relationship between fermentation kinetics and volatiles production appears to be indirect, and the potential value of kinetic data appears to be in predicting anomalies in alcohol and ester content, rather than in controlling their expression.