Requirement for activation of the serine-threonine kinase Akt (protein kinase B) in insulin stimulation of protein synthesis but not of glucose transport

A wide variety of biological activities including the major metabolic actions of insulin is regulated by phosphatidylinositol (PI) 3-kinase. However, the downstream effectors of the various signaling pathways that emanate from PI 3-kinase remain unclear. Akt (protein kinase B), a serine-threonine kinase with a pleckstrin homology domain, is thought to be one such downstream effector. A mutant Akt (Akt-AA) in which the phosphorylation sites (Thr308 and Ser473) targeted by growth factors are replaced by alanine has now been shown to lack protein kinase activity and, when overexpressed in CHO cells or 3T3-L1 adipocytes with the use of an adenovirus vector, to inhibit insulin-induced activation of endogenous Akt. Akt-AA thus acts in a dominant negative manner in intact cells. Insulin-stimulated protein synthesis, which is sensitive to wortmannin, a pharmacological inhibitor of PI 3-kinase, was abolished by overexpression of Akt-AA without an effect on amino acid transport into the cells, suggesting that Akt is required for insulin-stimulated protein synthesis. Insulin activation of p70 S6 kinase was inhibited by approximately 75% in CHO cells and approximately 30% in 3T3-L1 adipocytes, whereas insulin-induced activation of endogenous Akt was inhibited by 80 to 95%, by expression of Akt-AA. Thus, Akt activity appears to be required, at least in part, for insulin stimulation of p70 S6 kinase. However, insulin-stimulated glucose uptake in both CHO cells and 3T3-L1 adipocytes was not affected by overexpression of Akt-AA, suggesting that Akt is not required for this effect of insulin. These data indicate that Akt acts as a downstream effector in some, but not all, of the signaling pathways downstream of PI 3-kinase.     

Cardiac hemangioma of the right ventricle--report of a case

OBJECTIVE: The prevalence of obesity and thinness is unknown among Iranian high-school age girls. We determined the prevalence of overweight and underweight among Iranian high-school girls from Kerman (south-east province of Iran). DESIGN: A cross-sectional study of indicative cluster sample. MEASUREMENTS: Weight, height, body mass index (BMI), chest, waist, abdomen, hip and thigh circumference of 1000 Iranian high-school girls aged 14-21 y (mean (standard deviation, s.d.) 16.2 (1.3)) were measured. The criteria for very underweight, underweight, desirable weight, grade 1, 2 and 3 overweight used in the present study were: BMI in kg/m2 < 15, 15-19.9, 20-24.9, 25-29.9, 30-39.9 and > or = 40, respectively. RESULTS: 4.6% (95% confidence interval (CI): 3.4%-6.1%) were grade 1 overweight (BMI = 25.0-29.9), 0.7% (95% CI: 0.3%-1.4%) were grade 2 overweight (BMI = 30-39.9), and none were grade 3 overweight (BMI > or = 40), while 54.6% (95% CI: 51.5%-57.7%) were underweight (BMI = 15-19.9) and 1.6% (95% CI: 0.9%-2.6%) were very underweight (BMI < 15). The mean (s.d.) BMI was 19.8 (2.9) (95% CI: 19.6-20.0). The mean (s.d.) waist-to-hip ratio (WHR), abdomen-to-hip ratio and chest-to-hip ratio values were 0.8 (0.06) (95% CI: 0.8-0.81), 0.8 (0.07) (95% CI: 0.8-0.81) and 0.9 (0.07) (95% CI: 0.9-0.91), respectively, in this sample. CONCLUSION: The results suggest a low prevalence of overweight among Iranian young women.     

Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (1996)

The bacteria isolated from the patients with lower respiratory tract infections were collected by institutions located throughout Japan, since 1981. Ikemoto et al. have been investigating susceptibilities of these isolates to various antibacterial agents and antibiotics, and characteristics of the patients and isolates from them each year. Results obtained from these investigations are discussed. In 16 institutions around the entire Japan, 557 strains of presumably etiological bacteria were isolated mainly from the sputa of 449 patients with lower respiratory tract infections during the period from October 1996 to September 1997. MICs of various antibacterial agents and antibiotics were determined against 98 strains of Staphylococcus aureus, 93 strains of Streptococcus pneumoniae, 84 strains of Haemophilus influenzae, 84 strains of Pseudomonas aeruginosa (non-mucoid strains), 17 strains of Pseudomonas aeruginosa (mucoid strains), 31 strains of Moraxella subgenus Branhamella catarrhalis, 21 strains of Klebsiella pneumoniae etc., and the drug susceptibilities of these strains were assessed except for those strains that died during transportation. 1) S. aureus S. aureus strains for which MICs of oxacillin (MPIPC) were higher than 4 micrograms/ml (methicillin-resistant S. aureus) accounted for 67.3%. The frequency of the drug resistant bacteria increased comparing to the previous year's 52.7%. Arbekacin (ABK) and vancomycin (VCM) showed the highest activities against both S. aureus and MRSA with MIC80s of 1 microgram/ml. 2) S. pneumoniae Imipenem (IPM) and panipenem (PAPM) of carbapenems showed the most potent activities with MIC80s of 0.063 microgram/ml. Faropenem (FRPM) showed the next potent activity with MIC80 of 0.125 microgram/ml. The other drugs except erythromycin (EM), clindamycin (CLDM) and tetracycline (TC) were active against S. pneumoniae tested with MIC80s of 8 micrograms/ml or below. 3) H. influenzae The activities of all drugs were potent against H. influenzae tested with MIC80s of 4 micrograms/ml or below. Cefotiam (CTM), cefmenoxime (CMX), cefditoren (CDTR) and ofloxacin (OFLX) showed the most potent activities with MIC80s of 0.063 microgram/ml. 4) P. aeruginosa (mucoid strains) Tobramycin (TOB) showed the most potent activity against P. aeruginosa (mucoid strains) with MIC80 of 1 microgram/ml. Ceftazidime (CAZ), cefsulodin (CFS), IPM, gentamicin (GM), ABK and ciprofloxacin (CPFX) showed the next potent activities, with MIC80s of 2 micrograms/ml. The MIC80s of the other drugs ranged from 4 micrograms/ml to 16 micrograms/ml. 5) P. aeruginosa (non-mucoid strains) TOB and CPFX showed the most potent activities against P. aeruginosa (non-mucoid strains) with MIC80s of 1 microgram/ml. The MIC80s of piperacillin (PIPC) and cefoperazone (CPZ) were 16 micrograms/ml in 1995, and they were 64 micrograms/ml in 1996. 6) K. pneumoniae All drugs except ampicillin (ABPC) were active against K. pneumoniae. CMX, cefpirome (CPR), cefozopran (CZOP) and carumonam (CRMN) showed the most potent activities against K. pneumoniae with MIC80s of 0.125 microgram/ml. The MIC80s of the other drugs ranged from 0.25 microgram/ml to 2 micrograms/ml. 7) M.(B) catarrhalis Against M.(B.) catarrhalis, all drugs showed good activities with MICs of 4 micrograms/ml or below. IPM and minocycline (MINO) showed the most potent activities with MICs of 0.063 microgram/ml. Also, we investigated year to year changes in the characteristics of patients, their respiratory infectious diseases, and the etiology. Patients' backgrounds were examined for 557 isolates from 449 cases. The examination of age distribution indicated that the proportion of patients with ages over 60 years was 71.0% of all the patients showing a slight increase over that in 1994. Proportions of diagnosed diseases were as follows: Bacterial pneumonia and chronic bronchitis were the most frequent with 35.9% and 30.3% respectively. They were followed by bronchiectasis with a proportion of 10.     

Low-threshold and high temperature single-longitudinal-mode operation of 1.55 ?m-band DFB-DC-PBH LDs

1.55 ?m-band distributed-feedback laser diodes with double-channel planar buried heterostructure (DFB-DC-PBH LDs) have been developed. As well as low threshold current, 19 mA at room temperature, stable CW single-longitudinal-mode operation up to the high power level of 23 mW and the high temperature of 108°C has been obtained.     

Involvement of neutrophil elastase in crescentic glomerulonephritis

To elucidate the role of neutrophils in the tissue damage of crescentic glomerulonephritis (GN), we examined neutrophils infiltrated in renal tissues and the localization of neutrophil elastase (NE), as a neutrophil-derived tissue destructive mediator, using an immunohistochemical technique with antibodies specific for neutrophils and neutrophil elastase; the enzyme histochemical technique (chloroesterase staining) also was used to detect neutrophils. In normal controls, neutrophil infiltration was scarce, and NE was localized in neutrophil cytoplasm. Neutrophils were abundant in crescentic GN and infiltrated in the glomerulus and interstitium; the infiltrating neutrophils were often aggregated. NE was localized in the cytoplasm of neutrophils and also appeared extracellularly (in granular or diffuse patterns) in glomerular necrotizing lesions, crescents, ruptured portions of Bowman's capsules, and in periglomerular and perivascular sites of the interstitium. Moreover, urinary concentration of NE measured by enzyme-linked immunosorbent assay (ELISA) in crescentic GN patients was significantly higher than in normals (93.6 +/- 13.3 v 1.4 +/- 0.5 microg/g x Cr, respectively; P < .001). These data suggest that NE plays a significant role in renal tissue damage, especially in the formation of glomerular necrotizing and crescentic lesions and in periglomerular interstitial lesions of crescentic GN.     

Change in fine structure of nylon 6 gut yarn in twisting,annealing, and untwisting processes. I. Observation by WAXD,SAXD, and EM

To obtain information on the change in fine structure of nylon 6 taking place during practical false-twisting processes, the manner of change in the three elemental processes, i.e., twisting, annealing, and untwisting, was studied. For simplicity, nylon 6 gut yarn was used instead of multifilament yarn. Wide- and small-angle x-ray diffraction (WAXD and SAXD) together with electron microscopy (EM) were used here. The degree of molecular orientation in the crystalline region of the twisted yarn gradually decreases with increase of the twist number (TN) in the region of TN ? 100. The long spacing, determined by SAXD, of the twisted yarn increases with increases in TN. The increase in long spacing cannot be interpreted only by macroscopic strain or elongation of the yarn in the twisting process. This difference seems to arise from the contribution of the decrease of lateral size of lamellae to the average long spacing; therefore the increase in long spacing should be attributed to the elongation of the amorphous region, deduced from the crystallinity measured by WAXD and the long spacing diffraction. The angle between the streak line on the surface of twisted yarn observed by EM and the fiber axis agrees well with the twist angle of the yarn. The crystal lamellae are stacked normal to the streak line at the initial stage, i.e., at a low value of TN, but they begin to deviate from the normal direction with increase in TN, accompanied by their partial destruction. Based on SAXD and density measurements, the internal strain of yarn annealed after twisting is fully relaxed. When the yarn is untwisted after twisting and annealing, the crystal orientation recovers gradually to that of the untreated yarn. The chain axis within the lamellae in the center region of the yarn becomes nearly parallel to the fiber axis, but the chain axis in the outer region does not.     

Two-dimensional multiarray formation of hepatocyte spheroids on a microfabricated PEG-brush surface

A two-dimensional microarray of ten thousand (100 x 100) hepatocyte heterospheroids, underlaid with endothelial cells, was successfully constructed with 100 microm spacing in an active area of 20 x 20 mm on microfabricated glass substrates that were coated with poly(ethylene glycol) brushes. Cocultivation of hepatocytes with endothelial cells was essential to stabilize hepatocyte viability and liver-specific functions, allowing us to obtain hepatocyte spheroids with a diameter of 100 microm, functioning as a miniaturized liver to secret albumin for at least one month. The most important feature of this study is that these substrates are defined to provide an unprecedented control of substrate properties for modulating cell behavior, employing both surface engineering and synthetic polymer chemistry. The spheroid array constructed here is highly useful as a platform of tissue and cell-based biosensors and detects a wide variety of clinically, pharmacologically, and toxicologically active compounds through a cellular physiological response.