AKT2, a member of the protein kinase B family, is activated by growth factors, v-Ha-ras, and v-src through phosphatidylinositol 3-kinase in human ovarian epithelial cancer cells

Three members have been identified in the protein kinase B (PKB) family, i.e., Akt/PKB alpha, AKT2/PKB beta, and AKT3/PKB gamma. Previous studies have demonstrated that only AKT2 is predominantly involved in human malignancies and has oncogenic activity. However, the mechanism of transforming activity of AKT2 is still not well understood. Here, we demonstrate the activation of AKT2 with several growth factors, including epidermal growth factor, insulin-like growth factor 1, insulin-like growth factor II, basic fibroblast growth factor, platelet-derived growth factor, and insulin, in human ovarian epithelial cancer cells. The kinase activity and the phosphorylation of AKT2 were induced by the growth factors and blocked by the phosphatidylinositol (PI) 3-kinase inhibitor, wortmannin, and dominant-negative Ras (N17Ras). Moreover, the activated Ras and v-Src, two proteins that transduce growth factor-generated signals, also activated AKT2, and this activation was not significantly enhanced by growth factor stimulation but was abrogated by wortmannin. These results indicate that AKT2 is a downstream target of PI 3-kinase and that Ras and Src function upstream of PI 3-kinase and mediate the activation of AKT2 by growth factors. The findings also provide further evidence that AKT2, in cooperation with Ras and Src, is important in the development of some human malignancies.     

Effects of surface electrostimulation on the structure and metabolic properties in monkey skeletal muscle

Adaptative changes in skeletal muscle following surface electrical stimulation (SES) were investigated in rhesus monkeys. SES was performed on the triceps brachialis muscle (TB) according to an intermittent pattern. The procedure was carried out for 3 wk, using a current with a medium frequency of 60 Hz normally observed in fast motor axons. The histochemical assays performed on biopsies taken from proximal and distal parts of the TB muscle, before and after the SES program, showed that the distribution of fibers typed by ATPase was unaffected. On the other hand, SES led to an overall increase in the mean fiber cross-sectional area (FCSA); P < 0.01 (+13.7%, NS, in proximal portion, vs +31%, P < 0.01 in distal portion). This increase in size occurred in all fiber types. SES was shown to induce an overall increase in capillary to fiber ratio (C/F; +11.06%, NS, in proximal portion, vs +25.93%, P < 0.05 in distal portion). The number of capillaries surrounding fiber Type II (CAFII) was significantly increased by SES (P < 0.05): +3.21%, NS, in proximal portion, versus +21.47%, P < 0.05 in distal portion. Moreover, the number of capillaries surrounding fiber Type I (CAFI) was statistically unaffected by SES. These results suggest that a stimulation of capillary growth may occur following SES-training. Citrate synthase activity was significantly increased after SES. This enhancement in oxidative potential was shown to occur in all fiber types (NADH-diaphorase staining).(ABSTRACT TRUNCATED AT 250 WORDS)     

FhuF, an iron-regulated protein of Escherichia coli with a new type of [2Fe-2S] center

We previously used fhuF as a sensitive reporter gene of the iron status of Escherichia coli. In this report, the fhuF gene was identified as open reading frame f262b at 99.2 min on the genome sequence map of E. coli K-12. The FhuF protein was labeled with a His-tag and then purified to electrophoretic homogeneity. Based on sulfur determinations and M?ssbauer and EPR spectroscopy, FhuF was identified as a [2Fe-2S] protein. The g values (gx = 1.886, gy = 1.961, gz = 1.994) and some of the M?ssbauer parameters of FhuF obtained [oxidized protein as isolated: delta EQ,4.2K = 0.474 mm s-1; Fe3+ (reduced protein): delta EQ = 0.978 mm s-1] are not typical of common [2Fe-2S] proteins and indicate that FhuF has unusual structural properties. The primary sequence of FhuF does not show any sequence similarities to known [2Fe-2S] proteins. By site-directed mutagenesis, each of the six cysteines of FhuF was replaced by serine. EPR of the six reduced mutant proteins revealed that the terminal cysteine residues 244, 245, 256, and 259 form the [2Fe-2S]Cys4 cluster. Mutants having the Cys-to-Ser replacement at positions 244, 245, 256, or 259 did not complement a fhuF mutant. The motif Cys-Cys-Xaa10-Cys-Xaa2-Cys in FhuF differs considerably from the motif Cys-Xaa2-Cys-Xaa9-15-Cys-Xaa2-Cys found in other [2Fe-2S] proteins. The unusual Cys-Cys terminal group of the cluster may explain the atypical EPR and M?ssbauer spectroscopic properties of the FhuF protein; possibly the tetrahedral symmetry at the ferric ion site is distorted. The phenotype of fhuF mutants and the structural features of the FhuF protein suggest that FhuF is involved in the reduction of ferric iron in cytoplasmic ferrioxamine B.     

Stability of Lycopene Emulsions in Food Systems

ABSTRACT The chemical stability of lycopene and the physical stability of lycopene emulsions diluted in 3 different food systems (skimmed milk, orange juice, and water as control) were studied. In these investigations, 3 different emulsifiers were used. It was found that lycopene stability strongly depends on the food system. In orange juice, lycopene is particularly stable. The emulsifiers used have only little influence on the stability of lycopene. Emulsions with α-tocopherol as an additional antioxidant showed a good lycopene stability in all food systems. Coalescence of oil droplets was not observed in any of the food systems investigated. Keywords: lycopene, stability, food systems     

Muscle unloading induces slow to fast transitions in myofibrillar but not mitochondrial properties. Relevance to skeletal muscle abnormalities in heart failure

Muscle deconditioning is a common observation in patients with congestive heart failure (CHF), chronic obstructive pulmonary disease, neuromuscular diseases or prolonged bed rest. To gain further insight into metabolic and mechanical properties of deconditioned slow-twitch (soleus) or fast-twitch (EDL) skeletal muscles, we induced experimental muscle deconditioning by hindlimb suspension (HS) in rats for 3 weeks. Cardiac muscle was also studied. Besides profound muscle atrophy, increased proportion of fast type II fibers as well as fast myosin isoenzymes, we found decreased calcium sensitivity of Triton X-100 skinned fiber bundles of soleus muscle directed towards the fast muscle phenotype. Glycolytic enzymes such as hexokinase and pyruvate kinase were increased, and the LDH isoenzyme pattern was clearly shifted from an oxidative to an anaerobic profile. Creatine kinase (CK) and myokinase activities were increased in HS soleus towards EDL values. Moreover, the M-CK mRNA level was greatly increased in soleus, with no change in EDL. However, oxygen consumption rate assessed in situ in saponin skinned fibers (12.5 +/- 0.8 in C and 15.1 +/- 0.9 micromol O2/min/g dw in HS soleus compared to 7.3 +/- 1.3 micromol O2/min/g dw in control EDL), as well as mitochondrial CK (mi-CK) and citrate synthase activities, were preserved in HS soleus. Following deconditioning no change in Km for ADP of mitochondrial respiration, either in the absence (511 +/- 92 in C and 511 +/- 111 microM in HS soleus compared to 9 +/- 4 microM in control EDL) or presence of creatine (88 +/- 10 in C and 95 +/- 16 microM in HS soleus compared to 32 +/- 9 microM in control EDL), was found. The results show that muscle deconditioning induces a biochemical and functional slow to fast phenotype transition in myofibrillar and cytosolic compartments of postural muscle, but not in the mitochondrial compartment, suggesting that these compartments are differently regulated under conditions of decreased activity.     

Extraction of sp18 family membrane proteins on posterior head of bull sperm and the effect of anti-sp18 IgG on sperm motility and murine in vitro fertilization

BACKGROUND AND PURPOSE: We evaluated sucralfate, well-known in the treatment of gastric ulcers, in relation to its possible reduction of radiation-induced acute complications in the treatment of head and neck cancers. MATERIALS AND METHODS: One hundred two patients were randomized in a double-blind placebo-controlled prospective setting. All patients were treated to a minimum dose of 55 Gy in 5 weeks. Oral intake of sucralfate was started at the beginning of radiotherapy and continued during the whole treatment at a dose of 1 g six times a day. All patients were scored according to a scoring system developed in our department. Weight was checked once a week. RESULTS: Comparing the time course of the mean scores for subjective intolerance, mucositis, dysphagia, dermatitis and nausea, no statistically significant differences between the two treatment arms (sucralfate, n = 38; placebo, n = 45) were observed. The mean weight loss in the sucralfate arm was 1.6 +/- 3.4 kg while it was 1.3 +/- 2.0 kg in the placebo arm. Apart from gastrointestinal upset, the administration of sucralfate did not cause any side-effects. CONCLUSION: This trial produced no clinical evidence indicating that the oral intake of sucralfate reduces the acute radiation-induced side-effects. Therefore, we do not recommend the prophylactic use of sucralfate in patients with head and neck cancer treated by radiotherapy.     

Increase in energy cost of running at the end of a triathlon

The purpose of the present study was to verify the increase in energy cost of running at the end of a triathlon. A group 11 trained male subjects performed a triathlon (15-km swimming, 40-km cycling, 10-km running). At least 1 week later the subjects ran 10-km as a control at the same pace as the triathlon. Oxygen uptake (VO2), ventilation (VE) and heart rate (HR) were measured during both 10-km runs with a portable telemetry system. Blood samples were taken prior to the start of the triathlon and control run, after swimming, cycling, triathlon run and control run. Compared to the control values the results demonstrated that triathlon running elicited a significantly higher (P < 0.005) mean VO2 [51.2 (SEM 0.4) vs 47.8 (SEM 0.4) ml.min-1.kg-1] VE [86 (SEM 4.2) vs 74 (SEM 5.3) l.min-1], and HR [162 (SEM 2) vs 156 (SEM 1.9) beats.min-1)]. The triathlon run induced a greater loss in body mass than the control run [2 (SEM 0.2) vs 0.6 (SEM 0.2) kg], and a greater decrease in plasma volume [14.4% (SEM 1.5) vs 6.7% (SEM 0.9)]. The lactate concentrations observed at the end of both 10-km runs did not differ [2.9 (SEM 0.2) vs 2.5 (SEM 0.2) m.mol.l-1]. Plasma free fatty acids concentrations were higher (P < 0.01) after the triathlon than after the control run [1.53 (SEM 0.2) to 0.51 (SEM 0.07) mmol.l-1]. Plasma creatine kinase concentrations rose under both conditions from 58 (SEM 12) to 112 (SEM 14) UI.l-1 after the triathlon, and from 61 (SEM 7) to 80 (SEM 6) UI.l-1 after the control run. This outdoor study of running economy at the end of an Olympic distance triathlon demonstrated a decrease in running efficiency.     

Variability in energy cost of running at the end of a triathlon and a marathon

The aim of this study was to investigate the increase in energy cost of running occurring at the end of a triathlon and a marathon event and to link them to the metabolic and hormonal changes, as well as to variations in stride length. Seven subjects took part in 3 experimental situations: a 2 h 15 min triathlon (30 min swimming, 60 min cycling and 45 min running), a 2 h 15 min marathon (MR) were the last 45 min were run at the same speed as the triathlon run (TR), and a 45 min isolated run (IR) done at triathlon speed. The results show that energy cost during MR was higher than during TR (p < 0.01) (+ 8.9%). Similar observations were made for pulmonary ventilation (+ 7.9%) and heart rate (+ 6.3%). Moreover, the values were significantly greater than the values obtained during the IR. TR and MR lead to greater weight loss (p < 0.01) (2.4 +/- 0.3 kg) than IR (1 +/- 0.2 kg). The triathlon and the marathon produced a large decrease in plasma volume (respectively 19.6 +/- 1.4% and 12.9 +/- 1.1%) compared to IR (2 +/- 0.4%). Plasma renin activity was higher for the triathlon and the marathon than for the IR (p < 0.01). MR produces a significantly greater increase in plasma free fatty acids (F.F.A.) than TR (p < 0.05) and IR (p < 0.01). In addition, the F.F.A. at the end of TR were significantly higher than IR (p < 0.05). At the end of the trial the mean stride lengths for TR and IR were greater (+ 15%) (p < 0.01) than for MR. This study, carried out with subjects running overground, confirms the decrease in running efficiency previously shown at the end of a laboratory triathlon, and demonstrates that this decrease is lower than that occurring during a marathon.     

Analysis of developing enamel of the rat. II. Electrophoretic and amino acid studies

Amino-acid analyses showed that proline, glutamic acid and leucine were the most common amino acids in immature or developing enamel and in each of its fractions (i.e., in the 1st and 14th water-extractable fractions of a sequential series of extractions, EDTA-water soluble and water-insoluble fractions. The immature enamel and its 1st and 14th water-extractable fractions were similar in their proportions of the basic amino acids (lysine, histidine and arginine), the beta-hydroxylated aliphatic amino acids (valine, leucine, and isoleucine). On the other hand, the immature enamel differed from the water-extractable fractions chiefly in its relative content of aspartic acid, glutamic acid, alanine, proline, glycine, tyrosine and methionine. Also the 1st water-extractable fraction differed from the 14th in its amino-acid profile. tthe EDTA-water-soluble fraction most closely resembled the 14th water-extractable fraction except for its proportion of arginine and alanine residues. Although with polyacrylamide gel electrophoresis the EDTA-water-soluble and the water-extractable fractions exhibited companion bands (at least 7 peaks were evident at pH 9.3) they differed decidely as to which band was the most prominent. The water-insoluble fractions compared with any of the soluble fractions or with immature enamel showed a higher percent of serine, threonine, glycine, aspartic acid, alanine, valine, lysine, and arginine but relatively less glutamic acid, proline, methionine and histidine. Neither hydroxyproline nor hydroxylsine were detected in any of the samples.