A proton nuclear magnetic resonance study of gamma-glutamyl-amino acid cyclotransferase in human erythrocytes

Spin-echo NMR spectroscopy was shown to be a reliable technique for the monitoring of the in situ cleavage of gamma-Glu-Ala by gamma-glutamyl-amino acid cyclotransferase in whole erythrocytes and hemolysates. Of particular importance was the difference in chemical shifts between peptide resonances and those of the constituent amino acids. Using lysates of varying dilution, it was shown that the specific activity of the enzyme was not concentration-dependent, thus suggesting a lack of cytosolic low-molecular-weight-effectors or enzyme dissociation. Furthermore, the initial velocities of the reaction as a function of substrate concentration obeyed Michaelis-Menten kinetics with a Km = 2.0 +/- 0.3 mmol/l and Vmax = 137 +/- 7 mmol/h/l of cell water in 1H2O medium. Similar analysis in 2H2O medium revealed a solvent kinetic isotope effect of 1.9 +/- 0.4 at low substrate concentrations. The implications of this observation for the mechanism of the reaction are discussed. Cleavage of the peptide by a suspension of intact erythrocytes was at a rate 300 times less than the corresponding lysate flux, thus indicating the rate limitation by transport in the coupled system.     

Paleoceanographic interpretations of coccoliths and oxygen-isotopes from the sediment surface of the southwest Indian Ocean

The distribution of forty-four coccolithophore species in one hundred deep-sea core-tops from the southwest Indian Ocean is described. Three coccolith assemblages have been recognised (Maputo, Agulhas Current and deep water) by the relative abundances of four ecologically significant coccolithophore species (Gephyrocapsa oceanica, Emiliania huxleyi, Calcidiscus leptoporus and Umbilicosphaera sibogae). Their biogeographical distribution appears to be related to water temperature, nutrient concentration and dissolution.The degree of preservation of coccoliths and foraminifera indicates that the carbonate lysocline lies somewhere between 3500 and 4000 m, resulting in the concentration of dissolution-resistant microfossils below this depth.Stable oxygen isotope ratios of the planktonic foraminiferal species Globigerinoides sacculifer range between −1.5 to −1.0‰ PDB (equal to 22.8–25.1°C) and occur in a narrow band on the sea floor beneath the “A” route of the Agulhas Current.These values are about 0.5 per mil PDB lighter than samples analyzed on either side of this band and can be explained by the Agulhas Current's elevated temperature at the ocean surface of 2–3°C. Thus an oxygen isotope imprint of the Agulhas Current exists beneath it on the sea floor.The Agulhas Current is probably the major factor influencing sedimentation, sediment-distribution patterns and geological features in the study area. At present it is voluminous and fast flowing, possibly eroding sediments up to 2500 m below the surface.The oxygen-isotope ratios and nannoplankton counts obtained in this study indicate, however, that the majority of samples are most probably recent or at least not older than 85,000 years. This implies that sediments are accumulating on the ocean floor and that the Agulhas Current does not have a pronounced erosional influence, at least in areas from which cores were retrieved for this study.     

Altitudinal changes in the incidence of crassulacean acid metabolism in vascular epiphytes and related life forms in Papua New Guinea

Summary The occurrence of Crassulacean acid metabolism (CAM), as judged from ¹³C values, was investigated in epiphytes and some related plant species at a series of sites covering the approximate altitudinal range of epiphytes in Papua New Guinea. Comprehensive collections were made at each site and the occurrence of water storage tissue and blade thickness was also determined. Some 26% of epiphytic orchids from a lowland rainforest (2–300 m.a.s.l) showed ¹³C values typical of obligate CAM and possessed leaves thicker than 1 mm. A second group of orchids, mostly with succulent leaves, possessed intermediate ¹³C values between -23 and -26% and accounted for 25% of the total species number. Some species of this group may exhibit weak CAM or be facultative CAM plants. The remainder of the lowland rainforest species appeared to be C₃ plants with ¹³C values between -28 and -35%. and generally possessed thin leaves. Obligate CAM species of orchids from a lower montane rainforest (1175 m.a.s.l) comprised 26% of the species total and mostly possessed thick leaves. The remainder of the species were generally thin-leaved with ¹³C values between -26 and -35%. largely indicative of C₃ photosynthesis. Orchids with intermediate ¹³C values were not found in the lower montane rainforest. Obligate CAM appeared to be lacking in highland epiphytes from an upper montane rainforest and subalpine rainforest (2600–3600 m.a.s.l). However the fern, Microsorium cromwellii had a ¹³C value of -21.28%. suggesting some measure of CAM activity. Other highland ferns and orchids showed more negative °¹³C values, up to-33%., typical of C₃ photosynthesis. The highland epiphytic orchids possessed a greater mean leaf thickness than their lowland C₃ counterparts due to the frequent occurrence of water storage tissue located on the adaxial side of the leaf. It is suggested that low daytime temperatures in the highland microhabitats is a major factor in explaining the absence of CAM. The increased frequency of water storage tissue in highland epiphytes may be an adaptation to periodic water stress events in the dry season and/or an adaptation to increased levels of UV light in the tropicalpine environment.     

Hydrolysis of dietary flavonoid glycosides by strains of intestinal Bacteroides from humans.

Rutin and quercitrin are hydrolysed to quercetin, and robinin is hydrolysed to kaempferol, by faecal flora from healthy subjects. The enzymes required for these hydrolyses, namely alpha-rhamnosidase and beta-galactosidase, were produced by some strains of Bacteroides distasonis; other strains, however, synthesized beta-glucosidase. The last-named enzyme was also elaborated by Bacteroides uniformis and Bacteroides ovatus. All the enzymes were produced constitutively. A cell-free extract of B. distasonis containing beta-glucosidase displayed an enzymic activity of 1 mumol/10 min per 10 mg of protein.     

Population genetics implications of the premutation hypothesis for the generation of the fragile X mental retardation gene

Summary The population genetics implications of the premutation hypothesis for the generation of the fragile X mental retardation gene are explored. With some broad assumptions, the consequences of the model are that (a) 50% of mothers of probands carry the premutation; (b) 6.5% of mothers of probands receive the premutation from their mothers, 18.9% from their fathers, and 24.6% as a new mutation; (c) the incidence of carriers for the full mutation equals the incidence of affected males, whereas the incidence of carriers for the premutation is 1.35 times the incidence of affected males; (d) assuming mutation rates are equal in eggs and sperm, the mutation rate from normal to premutation alleles is 1.67x10^-4; (e) the expected segregation ratio in sibs of probands is 0.44, which corresponds to observed values. In addition, predictions using the premutation hypothesis of the expected segregation ratio in sibs of mothers of probands fits well with the data of Vogel and coworkers.     

Effect of winter high temperatures on reproduction and circannual rhythms in hibernating ground squirrels

We tested whether prevention of hibernation in ground squirrels by midwinter exposure to high ambient temperatures influenced timing of the spring phase of reproductive maturation and the phase and period of subsequent circannual rhythms of reproduction and body mass. Exposing hibernating adult male Spermophilus lateralis to 30 degrees C for 6 weeks beginning December 4 advanced the timing of testicular recrudescence by 4-5 weeks, compared to controls left at 4 degrees C. Males exposed to 30 degrees C for 6 weeks beginning at the average time of spontaneous end of hibernation (January 15) reached reproductive maturation at a time intermediate to those of controls and of the December 4 experimental group. However, neither the date of the subsequent fall's body mass peak, the date of the next year's reproductive maturation, nor the periods of circannual rhythms of body mass and reproduction differed among groups. Premature interruption of hibernation appears to allow early expression of reproduction, but does not affect the underlying timing mechanism.