Induction of microcin B17 formation in Escherichia coli ZK650 by limitation of oxygen and glucose is independent of glucose consumption rate

We examined the consumption of glucose from the media in which Escherichia coli ZK650 was grown. This organism, which produces the polypeptide antibiotic microcin B17 best under conditions of limiting supplies of glucose and air, was grown with a low level of glucose (0.5 mg/ml) as well as a high level (5.0 mg/ml) under both high and low aeration. Glucose consumption rates were virtually identical under both high and low aeration. Thus, glucose consumption rate is not a regulating factor in microcin B17 formation. Journal of Industrial Microbiology & Biotechnology (2001) 26, 341–344. Received 25 September 2000/ Accepted in revised form 16 April 2001     

Oxidative removal of lactate after strenuous exercise

Metabolic fate of lactate after strenuous exercise which lasted 2-3 min was investigated in rats and mice. 14C-labeled lactate or glucose was injected into the aorta of rats through an catheter. 14C-glucose was injected intraperitoneally into the mice after supramaximal exercise. The mice ran twice with a 4 hr interval to investigate muscle 14C-lactate metabolism which was produced from muscle 14C-glycogen. A great deal of blood and muscle 14C-lactate was expired as 14CO2 after the exercise. The results indicate that oxidative removal is the major fate of lactate metabolism after strenuous exercise and that blood glucose is the major substrate for muscle glycogen resynthesis. Light intensity exercise after strenuous exercise (active recovery) enhances oxidative removal of blood and muscle lactate. Gluconeogenesis from lactate to glycogen within the skeletal muscle is not a major pathway of muscle lactate metabolism, while high intensity training can activate this pathway.     

Biphasic changes in leukocytes induced by strenuous exercise

Seven healthy male volunteers participated in short- (STR, 1.7 km), middle- (MTR, 4.8 km) and long- (LTR, 10.5 km) term runs at a speed close to their maximum. A prompt mobilization of white cells, and lymphocytes in particular, appeared following the exercise. The initial increase in the number of lymphocytes was succeeded by a significant decrease [(P less than 0.03) lymphopenial], which on average was 32%-39% of the pre-exercise values in all groups. A close correlation was found between the initial increase in plasma cortisol concentration after exercise and the subsequent lymphopenia. A modest enhancement in the number of granulocytes immediately after the exercise was accompanied by a comprehensive increase in polymorphonuclear (PMN) elastase concentration accounting for 78.6%, SEM 16.3%, 140.7%, SEM 31.8% and 241.3%, SEM 48.1% in the STR, MTR and LTR groups. No correlation was found between granulocyte number and the plasma PMN elastase concentration. A delayed granulocytosis was noted in all subjects, reaching a peak between 2 and 4 h after the exercise. The magnitude of the granulocytosis varied among subjects and peak values of the number of circulating granulocytes were found to be 5.7 x 10(9) cells.l-1, SEM 0.5, 6.7 x 10(9) cells.l-1, SEM 0.6 and 8.8 x 10(9) cells.l-1, SEM 0.5 in STR, MTR and LTR respectively, whereas the mean baseline value was 3.6 x 10(9) cells.l-1, SEM 0.4. The neutrophilic granulocytosis was not accompanied by a corresponding enhancement in PMN elastase concentration. The plasma cortisol concentration reached a peak 30 min after exercise and declined below the control level in 4 h. Neither the initial increase, nor the subsequent decrease in plasma cortisol concentration were found to be essential for the magnitude of the delayed leukocytosis.     

Inflammatory response to strenuous muscular exercise in man

Based on the humoral and cellular changes occurring during strenuous muscular work in humans, the concept of inflammatory response to exercise (IRE) is developed. The main indices of IRE consist of signs of an acute phase response, leucocytosis and leucocyte activation, release of inflammatory mediators, tissue damage and cellular infiltrates, production of free radicals, activation of complement, and coagulation and fibrinolytic pathways. Depending on exercise intensity and duration, it seems likely that muscle and/or associated connective tissue damage, contact system activation due to shear stress on endothelium and endotoxaemia could be the triggering mechanisms of IRE. Although this phenomenon can be considered in most cases as a physiological process associated with tissue repair, exaggerated IRE could have physiopathological consequences. On the other hand, the influence of several factors such as age, sex, training, hormonal status, nutrition, anti-inflammatory drugs, and the extent to which IRE could be a potential risk for subjects undergoing intense physical training require further study.     

The effect of atrial natriuretic peptide on small intestinal contractility and transit.

Isometric tension in response to ANF (10(-10) to 10(-4) M) was recorded from longitudinally and circularly oriented rat jejunal smooth muscle strips. Conscious, fasted rats received an IV infusion of 1.25 nmol ANF/100 g body weight in 0.5 ml normal saline and controls received saline alone. Five minutes later 10 muCi Na2 51CrO4 in 0.5 ml saline was instilled through a jejunostomy. Fifteen minutes later animals were sacrificed, and the gut divided into 8 equal segments of small intestine, cecum and remaining colon. The radioactivity of each segment was measured and a geometric center of transit determined for each group. ANF induced relaxation of longitudinally oriented strips (Tm = -72.3 +/- 10.7 mN/g, ED50 7.3 +/- 3.6 x 10(-8) M), and contraction of circularly oriented strips (Tm = 35.0 +/- 5.0 mN/g, ED50 1.3 +/- 1.0 x 10(-8) M). This response was unaffected by 10(-6) M tetrodotoxin. The geometric mean center of transit was significantly (p less than 0.001) further aboral in ANF-treated compared to control animals (intestinal segment 4.2 +/- 0.2 vs. 3.2 +/- 0.2).     

Cardiovascular limitations of active recovery from strenuous exercise.

Recovery from muscle fatigue after exercise is known to have two beneficial effects: improved blood lactate elimination and a central nervous recuperation of the capacity for exercise. This study indicates circulatory mechanisms that might limit active recovery. Ten subjects were seated on a cycle ergometer and performed arm cranking exercise at an anaerobic intensity which was for each individual in three periods of 6 min, alternating with recovery intervals of 14 min. In two randomly assigned tests, recovery consisted either of passive sitting (control) or cycling at 80 W for 12 min. Both tests terminated with an identical final passive rest period of 25 min. In the cycling test arm cranking led to a heart rate increase which was further elevated with each repetition, while in the control test no differences were shown among the cranking periods. No corresponding difference was found for oxygen consumption. During the 25 min of final rest, the cycling test showed arterial hypotension and elevated heart rate both of which were absent in the control tests. Venous-occlusion-plethysmography revealed a postcranking forearm hyperaemia. In the cycling test hyperaemia was markedly reduced with the onset of cycling due to vasoconstriction; this effect was absent in the control test. A reduction in blood lactate occurred faster in the cycling test, mainly at the onset of cycling. Total plasma fluid loss combined with forearm fluid uptake was accentuated and prolonged by cycling recovery. Recovery exercise performed by muscles other than those that were fatigued could have led to arterial hypotension (shock-index about 1) through both plasma fluid loss and additional vasodilatation depending on the muscle mass involved.(ABSTRACT TRUNCATED AT 250 WORDS)     

Small intestinal transit in diabetics.

Small intestinal transit was assessed in diabetic patients and healthy controls by measuring the breath hydrogen appearance time after the ingestion of lactulose. Transit in diabetics with autonomic neuropathy was significantly slower than in diabetics without neuropathy and controls. Delayed transit is probably due to vagal denervation. These slower transit times would allow bacteria to proliferate, which might explain why some diabetics have diarrhoea. The test cannot be used in patients with bacteria in the small bowel because these may metabolise lactulose and release hydrogen prematurely.     

Changes in blood cell response following strenuous physical exercise.

The generation of tumour necrosis factor (TNF) and tissue factor activity in lipopolysaccharide (LPS) stimulated blood were studied in 25 healthy subjects before and after physical exercise of different intensities. Of the subjects a group of 9 were athletes who trained once to twice every day of the week, a second group of 8 exercised 3-7 times a week, and a third group of 8 exercised 4-5 times a month. The production of TNF in freshly drawn LPS stimulated blood in heparin, drawn from top athletes at rest was significantly lower than in the other subjects. The LPS induced concentrations of TNF-alpha of 2.73 (SEM 1.05) ng.ml-1 in the blood of the top athletes compared to 5.08 (SEM 0.7) ng.ml-1 and 7.6 (SEM 1.6) ng.ml-1, respectively, in the other two groups. The group that trained the least had the highest values. Immediately after exercise, the monocytes appeared to be less responsive to LPS stimulation, as a reduction of 47%-48% was observed in the top athletes and in the other group of well-trained individuals. The group that trained the least, which was also subjected to the least stressful exercise, had a 33% reduction in TNF production. Within 6 h the TNF concentration was back to pre-exercise values. Within 6 h the TNF concentration was back to pre-exercise values.(ABSTRACT TRUNCATED AT 250 WORDS)     

Autophagic response to strenuous exercise in mouse skeletal muscle fibers

Strenuous physical exercise induces necrosis of skeletal muscle fibers and increases lysosomal enzyme activities in surviving muscle fibers. This study examines the ultrastructural basis of the stimulation of the lysosomal system in mouse vastus medialis muscle during the appearance and repair of exercise-induced (9 h of running) injuries. Necrotic fibers appeared the day after exercise and an inflammatory response with the replacement of necrotic fibers by phagocytes was highest 2-3 days after exertion. Ultrastructural study of surviving muscle fibers revealed numerous autophagic vacuoles, residual bodies, and spheromembranous structures at the periphery of myofibers, especially in fibers adjacent to necrotic fibers. The autophagic response was most prominent between 2 and 7 days after exertion. Autophagic vacuoles with double or single limiting membranes contained mitochondria at various stages of degradation. Vacuolar and multilamellar structures were also observed in regenerating muscle fibers. The structure of injured skeletal muscle fibers returned to normal within 2 weeks. It is proposed that increased autophagic activity could be related to the breakdown of cellular constituents of surviving muscle fibers to provide structural elements for regenerating muscle fibers.     

Response of erythrocytic 2,3-diphosphoglycerate to strenuous exercise.

Since increases of erythrocytic 2,3-diphosphoglycerate (2,3-DPG) have been shown to enhance the release of oxygen from hemoglobin, experiments were designed to evaluate the response of 2,3-DPG to two different work-loads in 13 fasted human subjects. No signficant mean change in 2.3-DPG was found following 16 min of strenuous exercise on a bicycle ergometer, but when the subjects were subjected later to a greater workload for 20 min, there was a significant mean decrease in 2.3-DPG despite much individual variation. In addition, there was a significant positive correlation of 2.3-DPG reduction with increases in postexercise lactate, and a significant inverse correlation of oxygen consumption during exercise with postexercise lactate. The data suggest that the 2.3-DPG mechanism may not be compensating in exercise when the workload requires a preponderance of anaerobic metabolism promoting lactacidaemia.     

Effect of small intestinal glucose load on plasma ghrelin in healthy men

Postprandial ghrelin suppression arises from the interaction of meal contents with the small intestine and may relate to elevations in blood glucose and/or plasma insulin. We sought to determine whether the suppression of ghrelin by small intestinal glucose is dependent on the glucose load and can be accounted for by changes in blood glucose and/or plasma insulin. Blood glucose, plasma insulin, and plasma ghrelin levels were measured in 10 healthy males (aged 32+/-4 yr; body mass index: 25.1+/-0.4 kg/m2) during intraduodenal glucose infusions at 1 kcal/min (G1), 2 kcal/min (G2), and 4 kcal/min (G4), as well as intraduodenal hypertonic saline (control) for 120 min. There was a progressive decrease in ghrelin with all treatments, control at 45 min and between 90 and 120 min (P<0.05) and G1 (P<0.05), G2 (P<0.0001), and G4 (P<0.0001) between 30 and 120 min to reach a plateau at approximately 90 min. There was no difference in plasma ghrelin between G1, G2, or G4. Control suppressed ghrelin to a lesser extent than intraduodenal glucose (P<0.05). The suppression of ghrelin was not related to rises in blood glucose or plasma insulin. Suppression of ghrelin by intraduodenal glucose in healthy males is apparently independent of the glucose load and unrelated to blood glucose or insulin levels.     

Indomethacin modulates circulating cytokine responses to strenuous exercise in humans

Physical stress is associated with circulating cytokinemia. However the mechanisms of cytokine regulation during such stress are not clearly defined. Non-steroidal anti-inflammatory drugs (NSAIDs), including indomethacin, are widely used in countering the effects of excessive exercise, but their impact on circulating pro- and anti-inflammatory cytokine production in healthy humans also remains unclear. This study investigated the effect of five days of oral indomethacin treatment (75 mg per day) on the serum concentrations of IL-6, IL-10, IL-12, and TNF-alpha induced by exercising healthy volunteers. The results demonstrate that indomethacin does not alter resting serum cytokine concentrations. Increased circulating levels were noted, however, for all four cytokines with exercise, but with a different time-course. During and after strenuous physical exercise, indomethacin treatment blunted serum IL-6, and augmented TNF-alpha and IL-10. These findings may have important implications for both host defense and the injuries associated with excessively vigorous exercise.     

Excess postexercise oxygen consumption is unaffected by the resistance and aerobic exercise order in an exercise session

The main purpose of this study was to compare the magnitude and duration of excess postexercise oxygen consumption (EPOC) after 2 exercise sessions with different exercise mode orders, resistance followed by aerobic exercise (R-A); aerobic by resistance exercise (A-R). Seven young men (19.6 ± 1.4 years) randomly underwent the 2 sessions. Aerobic exercise was performed on a treadmill for 30 minutes (80-85% of reserve heart rate). Resistance exercise consisted of 3 sets of 10 repetition maximum on 5 exercises. Previous to the exercise sessions, V(O2), heart rate, V(CO2), and respiratory exchange rate (RER) were measured for 15 minutes and again during recovery from exercise for 60 minutes. The EPOC magnitude was not significantly different between R-A (5.17 ± 2.26 L) and A-R (5.23 ± 2.48 L). Throughout the recovery period (60 minutes), V(O2) and HR values were significantly higher than those observed in the pre-exercise period (p < 0.05) in both exercise sessions. In the first 10 minutes of recovery, V(CO2) and RER declined to pre-exercise levels. Moreover, V(CO2) and RER values in A-R were significantly lower than in R-A. In conclusion, the main result of this study suggests that exercise mode order does not affect the EPOC magnitude and duration. Therefore, it is not necessary for an individual to consider the EPOC when making the decision as to which exercise mode is better to start a training session.     

Adaptation of intestinal glucose transport in rats with diabetes mellitus occurs independent of hyperphagia.

Chronic diabetes enhances intestinal absorption of glucose and induces hyperphagia. We examined the enhanced intestinal absorption of glucose in ad libitum-fed rats with streptozocin-induced diabetes mellitus and compared these results with those obtained from pair-fed diabetic animals. Maximal transport capacity (Vmax) and carrier affinity (K0.5) were determined by measuring jejunal and ileal short circuit current (Isc) responses to varying concentrations of 3-O-methyl-D-glucopyranose and D-glucose. Pair-fed diabetic animals maintained the same body weight as animals fed ad libitum, although ad libitum-fed diabetic rats had an increased oral chow intake. Age-matched control rats maintained a constant jejunal and ileal Vmax and K0.5 throughout the study. Diabetic rats fed ad libitum demonstrated an enhanced Vmax and K0.5 in both jejunum and ileum. Pair feeding diabetic animals further enhanced jejunal Vmax while lowering jejunal K0.5 levels. In contrast, pair feeding diabetic animals delayed and blunted changes in ileal Vmax and prevented changes in ileal K0.5. In conclusion, signals other than those of hyperphagia regulate kinetic changes in glucose absorption during diabetes mellitus. Furthermore, these changes have differing effects on jejunum and ileum.     

Rat proximal small intestinal Golgi membranes: lipid composition and fluidity

The present studies were conducted to examine and characterize the lipid composition and physical state of the membrane lipids of rat proximal small intestinal Golgi membranes. Golgi membranes were purified from isolated enterocytes; lipids were extracted from these membranes and analyzed by thin-layer and gas-liquid chromatography. The 'static' and 'dynamic' components of fluidity of Golgi membranes and their liposomes were assessed by steady-state fluorescence polarization techniques utilizing r infinity and S values of 1,6-diphenyl-1,3,5-hexatriene and r values of DL-2-(9-anthroyl)- and DL-12-(9-anthroyl)stearic acid, respectively. Additional studies were also performed on these membranes, using benzyl and methyl alcohol, to examine the relationship between alterations in lipid fluidity and glycosphingolipid glycosyltransferase activities. The results of these studies demonstrated that: (1) the principal phospholipids and neutral lipids of intestinal Golgi membranes, respectively, were phosphatidylcholine, phosphatidylethanolamine and sphingomyelin, and unesterified cholesterol and fatty acids; (2) the major fatty acids of Golgi membranes were palmitic (16:0), stearic (18:0), linoleic (18:2), arachidonic (20:4) and oleic (18:1) acids; (3) fluorescence polarization studies using diphenylhexatriene detected a thermotropic transition at 24-26 degrees C in Golgi membranes and liposomes prepared from lipid extracts of these membranes; (4) benzyl alcohol (25 and 50 mM) but not methyl alcohol (50 mM) significantly increased the fluidity of these membranes; and (5) at these same concentrations, benzyl alcohol was also found to increase significantly the specific activity of UDP-galactosyllactosylceramide galactosyltransferase but not CMP-acetylneuraminic acid: lactosylceramide sialyltransferase. Methyl alcohol was not found to influence either enzyme's activity in these membranes.     

Biogenesis of Y RNA-derived small RNAs is independent of the microRNA pathway

Y RNAs are approximately 100 nucleotide long conserved cytoplasmic non-coding RNAs, which produce smaller RNA fragments during apoptosis. Here we show that these smaller RNA molecules are also produced in non-stressed cells and in a range of human cancerous and non-cancerous cell types. Recent reports have speculated that the cleavage products of Y RNAs enter the microRNA pathway. We tested this hypothesis and found that Y5 and Y3 RNA fragments are Dicer independent, they are in different complexes than microRNAs and that they are not co-immunoprecipitated with Ago2. Therefore we conclude that Y RNA fragments do not enter the microRNA pathway.