CYTOGENETIC STUDIES IN THE GENUS TRIBOLIUM (POACEAE: ARUNDINEAE)

The genus Tribolium Desv. consists of nine species, i.e., T. utriculosum (Nees) Renv., T. ciliare (Stapf) Renv., T. echinatum (Thunb.) Desv., T. hispidum (Thunb.) Desv., T. acutiflorum (Nees) Renv., T. obliterum sensu Davidse, T. glomeratum sensu Davidse, T. uniolae (L.f.) Renv., and T. brachystachyum (Nees) Renv. The genus has a basic chromosome number of 6, and from diploid to hexaploid specimens have been examined. Precocious segregation of metaphase I bivalents were observed in four species. Multivalent formation results in unequal chromosome segregation during anaphase I, and several cells with an 11–13 chromosome distribution have been observed. The presence of univalents and anaphase I bridges in all T. brachystachyum specimens suggests a hybrid origin for the species. B-chromosomes were present in specimens from four species. The B-chromosomes are similar to the euchromosomes with the exception that they do not participate in meiosis. The B-chromosomes have a possible isochromosome origin. The cytogenetic evidence presented supports the combination of Plagiochloa and Lasiochloa into Tribolium and indicates that the genus is closely related to Urochlaena, whereas it is not closely related to Prionanthium.     

Vigilant Behaviour: Predictability or Randomness? Spectral Analysis of Series of Scan Durations and their Relationship with Inter-scan Intervals

While feeding, many animals frequently look up and visually scan their environment. Using spectral analysis of continuous series of scan durations from a purple sandpiper Calidris maritima and Barbary doves Streptopelia risoria, we show that there are sequential non-random patterns and significant periodicities in all the examined series such that the birds cycled regularly between short and long scans. The cycles are comparable to those for the continuous series of inter-scan intervals of the same behavioural sequences. We suggest a re-examination of the functional costs and benefits of instantaneous randomness versus sequential predictability in alternating between feeding and scanning and a revision of the models of the way animals alternate between these behaviour patterns.     

The Influence of Alarm Substance on Feeding in Zebra Danio Fish (Brachydanio rerio)

The effects of alarm substance on feeding behaviour of zebra danio fish (Brachydanio rerio) were tested by offering them high and low densities of enclosed waterfleas (Daphnia magna). Normally the fish attacked high densities of prey, but when exposed to alarm substance they preferred lower and presumably less confusing prey densities — also lowering their feeding rate.     

Total chemical synthesis and NMR characterization of the glycopeptide tx5a, a heavily post-translationally modified conotoxin, reveals that the glycan structure is alpha-D-Gal-(1-->3)-alpha-D-GalNAc.

The 13-amino acid glycopeptide tx5a (Gla-Cys-Cys-Gla-Asp-Gly-Trp*-Cys-Cys-Thr*-Ala-Ala-Hyp-OH, where Trp* = 6-bromotryptophan and Thr* = Gal-GalNAc-threonine), isolated from Conus textile, causes hyperactivity and spasticity when injected intracerebral ventricularly into mice. It contains nine post-translationally modified residues: four cysteine residues, two gamma-carboxyglutamic acid residues, and one residue each of 6-bromotryptophan, 4-trans-hydroxyproline and glycosylated threonine. The chemical nature of each of these has been determined with the exception of the glycan linkage pattern on threonine and the stereochemistry of the 6-bromotryptophan residue. Previous investigations have demonstrated that tx5a contains a disaccharide composed of N-acetylgalactosamine (GalNAc) and galactose (Gal), but the interresidue linkage was not characterized. We hypothesized that tx5a contained the T-antigen, beta-D-Gal-(1-->3)-alpha-D-GalNAc, one of the most common O-linked glycan structures, identified previously in another Conus glycopeptide, contalukin-G. We therefore utilized the peracetylated form of this glycan attached to Fmoc-threonine in an attempted synthesis. While the result-ing synthetic peptide (Gla-Cys-Cys-Gla-Asp-Gly-Trp*-Cys-Cys-Thr*-Ala-Ala-Hyp-OH, where Trp* =6-bromotryptophan and Thr* = beta-D-Gal-(1-->3)-alpha-D-GalNAc-threonine) and the native peptide had almost identical mass spectra, a comparison of their RP-HPLC chromatograms suggested that the two forms were not identical. Two-dimensional 1H homonuclear and 13C-1H heteronuclear NMR spectroscopy of native tx5a isolated from Conus textile was then used to determine that the glycan present on tx5a indeed is not the aforementioned T-antigen, but rather alpha-D-Gal-(1-->3)-alpha-D-GalNAc.     

Soil 13C–15N dynamics in an N2-fixing clover system under long-term exposure to elevated atmospheric CO2

Reduced soil N availability under elevated CO₂ may limit the plant's capacity to increase photosynthesis and thus the potential for increased soil C input. Plant productivity and soil C input should be less constrained by available soil N in an N₂‐fixing system. We studied the effects of Trifolium repens (an N₂‐fixing legume) and Lolium perenne on soil N and C sequestration in response to 9 years of elevated CO₂ under FACE conditions. ¹⁵N‐labeled fertilizer was applied at a rate of 140 and 560 kg N ha^?1 yr^?1 and the CO₂ concentration was increased to 60 Pa pCO₂ using ¹³C‐depleted CO₂. The total soil C content was unaffected by elevated CO₂, species and rate of ¹⁵N fertilization. However, under elevated CO₂, the total amount of newly sequestered soil C was significantly higher under T. repens than under L. perenne. The fraction of fertilizer‐N (f_N) of the total soil N pool was significantly lower under T. repens than under L. perenne. The rate of N fertilization, but not elevated CO₂, had a significant effect on f_N values of the total soil N pool. The fractions of newly sequestered C (f_C) differed strongly among intra‐aggregate soil organic matter fractions, but were unaffected by plant species and the rate of N fertilization. Under elevated CO₂, the ratio of fertilizer‐N per unit of new C decreased under T. repens compared with L. perenne. The L. perenne system sequestered more ¹⁵N fertilizer than T. repens: 179 vs. 101 kg N ha^?1 for the low rate of N fertilization and 393 vs. 319 kg N ha^?1 for the high N‐fertilization rate. As the loss of fertilizer‐¹⁵N contributed to the ¹⁵N‐isotope dilution under T. repens, the input of fixed N into the soil could not be estimated. Although N₂ fixation was an important source of N in the T. repens system, there was no significant increase in total soil C compared with a non‐N₂‐fixing L. perenne system. This suggests that N₂ fixation and the availability of N are not the main factors controlling soil C sequestration in a T. repens system.     

Systolic Blood Pressure is Related to Catecholamine Sensitivity in Subcutaneous Abdominal Fat Cells

A relationship between abdominal obesity and hypertension is well established. In search for an early-onset defect in adipocyte function linking these two conditions, we compared catecholamine sensitivity in subcutaneous abdominal fat cells with 24-hour systolic, mean arterial and diastolic blood pressure in 16 healthy, normotensive subjects. Clear inter-individual variations in the adipocyte lipolytic adrenoceptor sensitivity (pD₂) for noradrenaline were observed in dose-response experiments (i.e., about 4 log units). An inverse and independent correlation was found between the 24-hour systolic blood pressure and pD₂ for noradrenaline (r = ?0.67, p < 0.01). The mean arterial blood pressure was also negatively correlated to peripheral noradrenaline sensitivity (r = ?0.58, p < 0.05). However, no significant relationship between the 24-hour diastolic blood pressure and pD₂ for noradrenaline was demonstrated. In conclusion we suggest that catecholamine resistance in subcutaneous fat cells may be associated with autonomic dysfunction and impaired blood pressure regulation. This finding is supported by the fact that both noradrenaline sensitivity and 24-hour systolic blood pressure also are correlated to the individual orthostatic heart rate responses, reflecting the sympathetic nervous system tone (r=0.61, p=0.01 and r= ?0.53, p=0.03, respectively). The relationship between noradrenaline sensitivity and systolic blood pressure may be of importance in the early development of hypertension in man.     

Characterization of the activity of fatty-acid epoxide hydrolase in seeds of castor bean (Ricinus communis L.)

Epoxide hydrolase (EC 3.3.2.3) activity was measured with [1-¹⁴C]cis-9,10-epoxystearic acid as the substrate. Homogenates were prepared from the endosperm tissue of germinating seeds of castor bean (Ricinus communis L. zanzibariensis). The activity of fatty-acid epoxide hydrolase was characterized with respect to dependence on time, amount of protein, pH and temperature. Analyses of enzyme distribution in endosperm, cotyledons, root and hypocotyl showed the highest total activity in the endosperm, less in the cotyledons and low activity in the root and hypocotyl. The specific activity was similar for cotyledons and endosperm. Analysis of the temporal expression of the enzyme in the endosperm during germination revealed high activity already in the imbibed seed. Activity was maximal between days four to six and then decreased at the end of one week. Subcellular fractionation of endosperm revealed a dual distribution of activity between the glyoxysomal and the cytosolic fractions.     

In vitro influence of zinc and magnesium on the deformability of red blood cells artificially hardened by heating

Trace elements have been shown to improve red blood cell (RBC) deformability: zinc in sickle cell disease and magnesium in an in vitro model of chemically rigidified erythrocytes. In this study, we investigated the effect and the influence of incubation time of zinc or magnesium on an in vitro model of rigidified RBCs by heating. Erythrocyte rigidity was determined by viscosimetry at high shear rate by a falling ball viscosimeter MT 90. In the first part of the study, six normal volunteers participated. Viscosimetry was performed on native blood before and after heating the sample for 10 min at 50°C. Therefore, increasing concentrations of zinc gluconate (final concentration: 0.5–4 g/L) or isotonic NaCl as control medium were added to the sample. Heating induced a twofold increase in all indices of RBC rigidity (p<0.05). At all these concentrations of zinc, a highly significant, dose-related fluidifying effect was observed (40–70%): this effect was immediately obtained and did not change over 60 min. Even at the highest concentration, recovery was not complete. In the second part of the study, we studied magnesium’s effects on blood. In a first protocol, whole blood was rigidified by heating at 56°C for 10 min, and the correcting effect of 5 min of incubation at 37°C of RBCs in 150 mmol/L NaCl, MgSO₄, magnesium acetate, and magnesium gluconate was investigated. In a second protocol, the same incubation with NaCl and magnesium salts was made on blood that had not been previously heated. In a third protocol, the correcting effect of magnesium gluconate on heated red blood cells was tested at four concentrations (75, 150, 225, and 300 mmol/L) over 1 h, for evaluating the effects of both concentration and time. Erythrocyte rigidity by heating is corrected by the three salts employed in protocol 1 (compared to sodium). In protocol 2, the deformability of normal (nonheated) red cells is not modified by magnesium. In protocol 3, no marked modification over 1 h is observed. The correcting effect is not complete for 75 mmol/L Mg, but remains the same at the three other concentrations. This study shows that zinc and magnesium at supraphysiological concentration are able to reverse RBC’s rigidification induced by heating, but that magnesium does not modify the flexibility of normal RBCs. This article suggests that zinc and magnesium may be studied in vivo as potential pharmacologic tools for improving hemorheologic disturbances.     

Effects of oral zinc gluconate on glucose effectiveness and insulin sensitivity in humans

Zinc improves both insulin secretion and insulin sensitivity, and exerts insulin-like effects. We investigated its acute effects on the parameters of glucose assimilation determined with the minimal model technique from frequent sampling intravenous glucose tolerance test (FSIVGTT) in seven healthy volunteers. FSIVGTTs (0.5 g/kg of glucose, followed by 2 U insulin iv injection at 19 min) were performed after the subjects had taken 20 mg zinc gluconate twice (the evening before and 30 min before the beginning of the test) or placebo pills (simple blind randomized protocol). Glucose assimilation was analyzed by calculating Kg (slope of the exponential decrease in glycemia), glucose effectiveness Sg (i.e., ability of glucose itself to increase its own disposal independent of insulin response), and SI (insulin sensitivity, i.e. the effect of increases in insulinemia on glucose disposal). The two latter parameters were calculated by fitting the experimental data with the two equations of Bergman’s “minimal model”. Zinc increased Kg (p<0.05) and Sg (p<0.05), whereas SI and insulin first-phase secretion did not significantly increase. This study suggests that zinc improves glucose assimilation, as evidenced by the increase in Kg, and that this improvement results mainly from an increase in glucose effectiveness (insulin-like effect), rather than an action on insulin response or insulin sensitivity.