The role of the arginine-B22 residue in insulin action.

We describe the modification of the side chain of the arginine-B22 residue of insulin by the N8N9-(1, 2-dihydroxycyclohex-1,2-ylene) group and by the adipoyl group. These are the first insulin derivatives described that contain a modified arginine residue in an otherwise unaltered molecule. When tested for their ability to lower blood sugar concentration, both modified insulins showed a specific activity indistinguishable from that of insulin. In view of the fact that the substituent groups involved are very bulky and in one case of opposite charge to that of the side chain, the retention of biological activity casts doubt on the idea, previously generally accepted, that the arginine-B22 residue is essential to the activity of the hormone.     

Myocardial Revascularization in Patients 70 Years of Age and Older

Myocardial revascularization has been carried out by us in 67 patients 70 years of age or older. Advanced coronary artery disease was found at angiography in more than two thirds of the patients. The postoperative morbidity and mortality compare very favorably with those in younger patients. The early and late mortality in the 67 patients was 4.5 percent and 6.0 percent, respectively. Fifty-seven survivors have been followed an average of 21 months; for most patients there has been a pronounced improvement in clinical classification. Properly selected, patients of advanced age can undergo successful revascularization surgical procedures. The adequacy of function of the left ventricle, proper timing of the surgical operation and an aggressive yet realistic approach seem to be major determinants for a good result.     

Synthesis and characterization of 5-azido-UDP-glucuronic acid. A new photoaffinity probe for UDP-glucuronic acid-utilizing proteins.

A new active site-directed photoaffinity analogue, [beta-32P]5-azido-UDP-glucuronic acid (UDP-GlcA), was enzymatically synthesized from [beta-32P]5-N3UDP-Glc using UDP-glucose dehydrogenase. The product was characterized by its mobility on ion exchange and two thin-layer chromatographic systems, by its UV absorbance at 288 nm, and the loss of this absorbance after UV irradiation of the compound. Photoincorporation of [beta-32P]5-N3UDP-GlcA into bovine liver UDP-Glc dehydrogenase (EC 1.1.1.22) was saturable with an apparent Kd of 12.5 microM, and was inhibited by the known active-site effectors UDP-GlcA, UDP-Glc, and UDP-xylose. When human liver microsomes with known UDP-glucuronosyltransferase (EC 2.4.1.17) activities were photolabeled with [beta-32P]5-N3UDP-GlcA, major photolabeled bands of 35-37 and 50-54 kDa were detected. When rat liver microsomes from phenobarbital-injected rats were photolabeled with [beta-32P]5-N3UDP-GlcA, there was a marked increase in photoincorporation of a 51-kDa protein as compared with control animals. Evidence is presented which suggests that the photolabeled 51-54-kDa proteins in the liver microsomes from both tissues are UDP-glucuronosyltransferase and that [beta-32P]5-N3UDP-GlcA represents a new alternative approach in the study of UDP-glucuronosyltransferase and other UDP-GlcA-utilizing enzymes.     

Composition and seasonal fluctuations of the ichthyoplankton community in a shallow tidal channel of Cadiz Bay (S.W. Spain)

The composition and seasonal variation of the ichthyoplankton community in a shallow tidal channel in Cadiz Bay was studied for 5 years (1984 1988), taking fortnightly samples with a conical tide-strained net (0.5 mm mesh). Thirty-nine species belonging to 21 families of teleosts were caught. Four species ( Pomutoschistus microps Kroyer, Atherina hoyeri Risso, Liza rumuda Risso and Lizu a lira Ia Risso) totalled more than 90% of the annual ichthyoplankton abundance. Three species, assumed to be permanent residents, dominated the number offish (837%), and were represented by all their developing stages; 26 species (66.7%) were temporary residents, contributing 17.2% to the total abundance and were represented only by early developing stages; the remaining 10 species were classified as rare. Monthly numbers of species and individuals showed seasonal variation with a main annual peak in spring and in late spring-early summer, respectively. The diversity (H') and evenness (S) indices of this community were low (0.13 to 1.83 and 0.03 to 0.71. respectively), with higher values in winter and late summer, and both were significantly correlated with the log-transformed density of temporary residents (r = 0.43 and 0.41, respectively; P<0.0l) and permanent residents (r=−0.75 and −0.79, respectively; P<0.01). Multivariate classification analysis of samples revealed that similar cyclical changes were taking place in the structure of this ichthyoplankton community every year; three periods could be differentiated: (1) a cool period of low fish density but relatively high diversity; (2) moderate temperature and salinity with high numbers of species and density but low diversity and (3) a warm period of moderate diversity and fish density.     

Left handed alpha-helix formation by a bacterial peptide

The alpha-helix is a common element of secondary structure in proteins and peptides. In eukaryotic organisms, which exclusively incorporate L-amino acids into such molecules, stereochemical interactions make such alpha-helices, invariably right-handed. Pseudomonas tolaasii Paine is the causal organism of the economically significant brown blotch disease of the cultivated mushroom Agaricus bisporus (Lange) Imbach. P. Tolaasii proceduces an extracellular lipodepsipeptide toxin, tolaasin, which causes the brown pitted lesions on the mushroom cap. Circular dichroism studies on tolaasin in a membrane-like environment indicate the presence of a left-handed alpha-helix, probably formed by a sequence of 7 D-amino acids in the peptide. P. tolaasii represents the first reported example of an organism which has evolved the ability to biosynthesize a left-handed alpha-helix.     

Proliferation- and cell cycle-dependent differences in expression of the 170 kilodalton and 180 kilodalton forms of topoisomerase II in NIH-3T3 cells.

The cellular content of 170kD and 180kD topoisomerase II was studied as a function of the proliferation state and cell cycle position in NIH-3T3 cells. When the cells were synchronized by serum starvation and then stimulated to enter the cell cycle by addition of fresh growth medium, the amount of 170kD topoisomerase II present was undetectable until the cells reached late S phase, peaked in G2-M phase cells, and decreased as the cells completed mitosis. The amount of 180kD topoisomerase II was constant once the cells entered the cell cycle. When exponentially growing cells were induced to enter G0 by serum starvation, the amount of 170kD topoisomerase II decreased in parallel with the loss of cells from the S and G2-M phases of the cell cycle and was undetectable once all of the cells reached G0. In contrast, the 180kD enzyme was still present after all of the cells had entered G0. The tightness of association of the two enzymes with chromatin was measured by determining the concentration of salt required to extract them from isolated nuclei. The 180kD enzyme required a higher concentration of NaCl for extraction than did the 170kD enzyme. The different patterns of expression of the two forms of topoisomerase II suggest that they perform different functions in cells.     

Trace metals in the benthic habitat of the Maarsseveen Lakes system,The Netherlands

The cadmium, zinc, lead and copper concentrations in benthic invertebrates and sediment were determined during two consecutive winters in the Maarsseveen Lakes system. A sequential extraction procedure was applied to estimate the bioavailability of the trace metals in the sediment. Based on the trace metal analyses of organisms and sediment, it is concluded that the Maarsseveen Lakes system has background levels of cadmium, zinc, lead and copper. As the majority of metals was present in geochemically more stable sediment phases, the sequential extractions provided limited additional information on trace metal bioavailability.     

Application of 5-azido-UDP-glucose and 5-azido-UDP-glucuronic acid photoaffinity probes for the determination of the active site orientation of microsomal UDP-glucosyltransferases and UDP-glucuronosyltransferases.

A new approach to determining the active site orientation of microsomal glycosyltransferases is presented which utilizes the photoaffinity analogs [32P]5-Azido-UDP-glucose ([32P]5N3UDP-Glc) and [32P]5-Azido-UDP-glucuronic acid ([32P]5N3UDP-GlcA). It was previously shown that both photoprobes could be used to photolabel UDP-glucose:dolichol phosphate glucosyltransferase (Glc-P-Dol synthase), as well as the family of UDP-glucuronosyltransferases in rat liver microsomes. The effects of detergents, proteases, and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) on the photolabeling of these enzymes were examined in intact rat liver microsomes. Photolabeling of Glc-P-Dol synthase by either photoprobe was the same in intact or disrupted vesicles, was susceptible to trypsin digestion, and was inhibited by the nonpenetrating inhibitor DIDS. Photolabeling of the UDP-glucuronosyltransferases by [32P]5N3UDP-GlcA was stimulated 1.3-fold in disrupted vesicles as compared to intact vesicles, whereas photolabeling of these enzymes by [32P]5N3UDP-Glc showed a 14-fold increase when vesicles were disrupted. Photolabeled UDP-glucuronosyltransferases were only susceptible to trypsin digestion in disrupted vesicles, and this was further verified by Western blot analyses. The results indicate a cytoplasmic orientation for access of UDP-sugars to Glc-P-Dol synthase and a lumenal orientation of most UDP-glucuronosyltransferases.     

Algal pigment diversity in coastal sediments from Kerguelen (sub-Antarctic Islands) reflecting local dominance of green algae,euglenoids and diatoms

Summary High Performance Liquid Chromatography analysis of algal pigments from inter- and subtidal (deep and shallow) sediments from the Kerguelen Islands showed clear differences in the pigment composition at the different stations. High concentrations of chlorophyll c and fucoxanthin were present at all locations, indicating significant diatom densities, chlorophyll b was detected at all sites. At one station the other green algal pigments were also present; here green algae contributed more to chlorophyll a concentrations than diatoms, as estimated by using pigment ratios and microscopic observations. At another location chlorophyll b was associated with a high concentration of diadinoxanthin, indicating an abundance of euglenoids. This indicates that chemotaxonomy can be powerful tool in microphytobenthos studies since enumeration of living cells are difficult as many algae are attached to sediment particles (epipsammic algae). Ways of diagenesis, carotenoid degradation and the role of grazing are briefly mentioned. Phaeophorbide a-like pigments were the most significant chlorophyll a degradation products, with concentrations up to 110 g · g^–1 dry weight sediment, i.e. 10 times the chlorophyll a concentration. Some taxonomic estimations, based on pigments ratios, and their limits, are discussed.