Insulinomimetic effect on glucose transport by epidermal growth factor when combined with a major histocompatibility complex class I-derived peptide

Peptides derived from the alpha 1-region of the murine H-2Dk molecule enhance glucose uptake in rat adipose cells above the maximum obtained with insulin stimulation alone (Stagsted, J., Reaven, G. M., Hansen, T., Goldstein, A., and Olsson, L. (1990) Cell 62, 297-307). We now describe that epidermal growth factor (EGF) in combination with the same peptides, Dk-(61-85) and Dk-(62-85), stimulates cellular glucose uptake 5-7 times over the basal level, i.e. to 30-50% of the maximal insulin effect. EGF alone increased glucose uptake by only approximately 50% above basal and the peptide alone by 100% above basal. Maximal effect of EGF and peptide was reached in 10-20 min with 30 microM peptide (EC50 10-15 microM) and 50 nM EGF (EC50 1-2 nM). The effect of EGF and peptide on glucose uptake was additive to that of insulin and peptide until the maximal level attained with insulin and peptide was reached. The combined effect of EGF plus peptide on glucose transport was associated with a recruitment of GLUT4 molecules to the plasma membrane. However, the phosphatidylinositol (PI) kinase which is activated by insulin was not activated by EGF plus peptide. Thus, the effect of EGF plus peptide on glucose uptake seems independent of the activity status of the insulin receptor. 125I-Labeled EGF bound specifically to rat adipose cells with an apparent affinity of approximately 2 nM and Bmax approximately 5 x 10(3). However, the major histocompatibility complex (MHC) peptides did not affect EGF-stimulated internalization of EGF receptor, in contrast to their effect on the insulin receptors. Transforming growth factor alpha had an effect similar to EGF on glucose uptake. Three other peptides derived from other parts of murine MHC class I had no effect on glucose uptake in combination with EGF. Thus, EGF in combination with certain MHC class I-derived peptides is insulinomimetic concerning glucose transport and this effect is independent of the insulin receptor activity.     

Trends and variations in neonatal length of in-hospital stay in Canada

The objectives of this study were to evaluate the possible mechanisms involved in prolongation of bleeding time in pre-eclamptic patients receiving a magnesium sulfate infusion to prevent convulsions. Eighteen pre-eclamptic patients near term or at term (4 cases 33 to 35 weeks; the remainder > 36 weeks) were studied. Fifteen of them received magnesium sulfate infusion; 3 did not and served as controls. Bleeding time (modified Ivy method with Surgicutt), platelet count, platelet aggregation pattern, as well as serum arachidonic acid metabolites [thromboxane B2 (TxB2) and 6-Keto-prostaglandin F1 alpha (6-Keto-PGF1 alpha)] werde done on admission to the labor floor (before magnesium infusion) and repeated at discontinuation of the infusion, 12-24 hours postpartum; the controls received the second test 24 hours postpartum. Thirteen of 15 patients receiving magnesium sulfate had an increase in bleeding time from an average of 6 minutes 31 seconds to 11 minutes 56 seconds, an 82% rise (p < 0.004). In 2 there was a decrease. Among the 3 controls the averages were 6 minutes 38 seconds and 6 minutes 3 seconds. The total magnesium given ranged from 52.5 to 145 grams. Platelet counts averaged 251,000/mm3 (range 145,000-519,000). Platelet aggregation pattern done in 11 patients and was normal and unchanged after magnesium in 10 of the patients with increased bleeding time and one control. TxB2 and 6-Keto-PGF1 alpha levels did not change significantly either after magnesium administration (688 and 135 pgm/ml, to 654 and 117) or in controls (695 and 230 pgm/ml, to 445 and 225). Likewise, the ratio of these 2 substances did not change in either group (6.3 to 6.6, and 4.2 to 2.2). There was no correlation between duration of infusion or total magnesium given and directions of small changes observed. This study confirms a prior preliminary observation that magnesium sulfate infusion, as currently used to prevent eclamptic convulsions, induces a significant prolongation of bleeding time. This effect is mediated neither by changes in platelets count or aggregation pattern, nor by changing the level or ratios of serum arachidonic acid metabolites (TxB2 and 6-Keto-PGF1 alpha). Further studies are needed to clarify the mechanism of this clinically important observation of increased bleeding following magnesium sulfate infusion.     

Reliability Potential Of Epoxy Based Encapsulants For Automotive Applications

Automotive under-the-hood electronics often have to withstand temperatures up to 175 °C in combination with harsh environment conditions. This paper gives an overview about the reliability potential of epoxy based encapsulants for automotive applications. Therefore the resistance of epoxy molding compound against typical automotive fluids at temperature of use is analyzed. Six epoxy molding compounds available on the market, showing high temperature automotive potential have been carefully selected to undergo a media resistance testing. For this purpose a dedicated mold tool has been designed and manufactured to prepare these encapsulants for material testing. Thermo-mechanical, mechanical and thermal properties had been determined in initial state directly after molding and after storage in aggressive fluids typical for automotive applications as e.g. gas oil, automatic transmission fluid (ATF), brake fluid or accumulator acid at the respective temperature of use.     

A versatile data acquisition system for time resolved X-ray scattering using gas proportional detectors with delay line readout

A data acquisition system for time resolved X-ray scattering experiments using linear, quadrant or area gas proportional detectors with delay line readout based on commercially available hardware (National Instruments) is described. The system can easily be configured for recording data from point detectors (e.g. photomultipliers and photodiodes) and/or ancillary data only. Applications involving measurements with two different types of time to digital converters illustrate the features and performances of the system.     

Fluoride catalysis at 175°C. of the reaction: Glycerol plus fatty acid

1. In uncatalyzed reactions the esterification of stearic, oleic, and linolenic acid proceeded at the same general rate though the speed was somewhat enhanced as the degree of unsaturation of the fatty acid increased. None of the reactions went to completion, after 15 hours being 72%, 83%, and 84%, respectively, completed. Only a small fraction of the distearate was converted to the triglyceride; a greater amount of the dioleate was esterified to the trioleate and about half the dilinolenate was changed to the trilinolenate.
2. Of 15 metallic fluorides studied, all afforded some catalysis, but only antimony trifluoride and zinc and cobalt fluorides were outstanding. Antimony trifluoride was the most active although in large amount it caused severe decomposition and charring of the products. Zinc fluoride had the best general properties, approaching antimony trifluoride yet causing practically no decomposition even in high concentration. Its activity was enhanced with an increase in amount, the best concentration apparently being 0.1 mole per 100 grams fatty acid. Antimony trifluoride was also better in larger amount within limits, but cobalt fluoride did not exhibit this property.
3. With zinc fluoride as catalyst the reactions of stearic, oleic and linolenic acids after 15 hours were 95%, 94%, and 92%, respectively, completed. At least three-fourths of the diglycerides were converted to triglycerides. From a study of reaction velocities, zinc fluoride caused the monoglyceride to form at a rate approximately four times that of the uncatalyzed reaction and the diglyceride at a six-fold rate. The triglyceride was formed at an almost undiminished rate whereas in the uncatalyzed reaction the formation of triglyceride was diminished rapidly.
4. Unsaturation and viscosity were not affected seriously in the presence of the fluorides except antimony trifluoride. Certain other fluorides caused severe darkening but without accompanying serious decomposition.

     

Terahertz characterisation of building materials

To obtain realistic models for propagation channels in future pico-cellular indoor terahertz communication systems it is necessary to know the reflective properties of building materials found in a typical office environment. The angular dependent reflection coefficients of different building materials were determined using terahertz time-domain spectroscopy in transmission geometry and Fresnel's equations. This approach is more efficient than a set of measurements in reflection geometry for different angles. Verification of the method with a set of such reflection measurements shows an excellent agreement.     

Mutational analysis of the three cysteines and active-site aspartic acid 103 of ketosteroid isomerase from Pseudomonas putida biotype B

In order to clarify the roles of three cysteines in ketosteroid isomerase (KSI) from Pseudomonas putida biotype B, each of the cysteine residues has been changed to a serine residue (C69S, C81S, and C97S) by site-directed mutagenesis. All cysteine mutations caused only a slight decrease in the k(cat) value, with no significant change of Km for the substrate. Even modification of the sulfhydryl group with 5,5'-dithiobis(2-nitrobenzoic acid) has almost no effect on enzyme activity. These results demonstrate that none of the cysteines in the KSI from P. putida is critical for catalytic activity, contrary to the previous identification of a cysteine in an active-site-directed photoinactivation study of KSI. Based on the three-dimensional structures of KSIs with and without dienolate intermediate analog equilenin, as determined by X-ray crystallography at high resolution, Asp-103 was found to be located within the range of the hydrogen bond to the equilenin. To assess the role of Asp-103 in catalysis, Asp-103 has been replaced with either asparagine (D103N) or alanine (D103A) by site-directed mutagenesis. For D103A mutant KSI there was a significant decrease in the k(cat) value: the k(cat) of the mutant was 85-fold lower than that of the wild-type enzyme; however, for the D103N mutant, which retained some hydrogen bonding capability, there was a minor decrease in the k(cat) value. These findings support the idea that aspartic acid 103 in the active site is an essential catalytic residue involved in catalysis by hydrogen bonding to the dienolate intermediate.