Plant fructans stabilize phosphatidylcholine liposomes during freeze-drying.

Fructans have been implicated as protective agents in the drought and freezing tolerance of many plant species. A direct proof of their ability to stabilize biological structures under stress conditions, however, is still lacking. Here we show that inulins (linear fructose polymers) isolated from chicory roots and dahlia tubers stabilize egg phosphatidylcholine large unilamellar vesicles during freeze-drying, while another polysaccharide, hydroxyethyl starch, was completely ineffective. Liposome stability was assessed after rehydration by measuring retention of the soluble fluorescent dye carboxyfluorescein and bilayer fusion. Inulin was an especially effective stabilizer in combination with glucose. Analysis by HPLC showed that the commercial inulin preparations used in our study contained no low molecular mass sugars that could be responsible for the observed stabilizing effect of the fructans. Fourier transform infrared spectroscopy showed a reduction of the gel to liquid-crystalline phase transition temperature of dry egg PtdCho by more than 20 degrees C in the presence of inulin. A direct interaction of inulin with the phospholipid in the dry state was also indicated by dramatic differences in the phosphate asymmetric stretch region of the infrared spectrum between samples with and without the polysaccharide.     

Active hexose transport across cultured human Caco-2 cells: characterisation and influence of culture conditions

Human Caco-2 cells (passage 80 to 100) were seeded onto collagen-coated Millipore filter assemblies and these were maintained in culture either (a) floated on the surface of the medium or (b) submerged within the body of the medium. Structural and functional assessments were made over a 30-day period. After seeding, all cells assumed a flattened, squamous configuration and rapidly became confluent. Cells submerged within the medium formed polarised monolayers with well developed junctional complexes, abundant apical microvilli and increasing levels of alkaline phosphatase activity. Cells grown floated on the surface of the medium formed complex multilayers in which polarisation was confined to the surface layer. Junctional complexes and apical microvilli were similar to those seen in submerged monolayers but alkaline phosphatase activities were higher. Transepithelial electrical resistance increased rapidly from day 1, as the layers became confluent. Electrical resistance was higher and short-circuit current and potential differences were lower across monolayers than across multilayers. After 10 days in culture, the addition of D-glucose to the apical bathing solution, of all cell layers, caused a rapid rise in short-circuit current and potential difference. These changes were sodium-dependent and phlorizin-sensitive. Galactose and 3-O-methylglucose induced similar changes and the affinity constants for these hexoses ranked in the order reported for rat jejunum (Km glucose 2.44 +/- 0.52 mM; Km galactose 8.05 +/- 1.33 mM; Km 3-O-methylglucose 22.0 +/- 5.2 mM). Culture conditions had a marked effect on hexose maximum transport rates (glucose Vmax: submerged 2.94 +/- 0.20 microA/cm2; floated 9.94 +/- 0.82 microA/cm2, P less than 0.05) but affinity constants were unchanged. Apical to basolateral mannitol fluxes, used as an index of paracellular permeability, decreased from day 1 to day 5 and then remained steady. Fluxes across monolayers and multilayers were not significantly different. We conclude that sodium-dependent hexose transport occurs in cultured Caco-2 cell layers grown on permeable supports. Culture conditions, however, have a marked effect on both cell layer structure and function, and should be an important factor when considering Caco-2 cells as an in vitro model of enterocyte function.     

VOCAL BEHAVIOUR OF GREYWING FRANCOLIN FRANCOLINUS AFRICANUS CAN BE USED TO ESTIMATE POPULATION DENSITY

Little, R.M. & Crowe, T.M. 1992. Vocal behaviour of Greywing Francolin Francolinus africanus can be used to estimate population density. Ostrich 63:98-109.

Four common calls of the Greywing Francolin Francolinus africanus are described acoustically, and their functions discussed. Data from 166 crepuscular call count surveys (2 472 counts) and 540 diurnal call counts were analysed to investigate the temporal and meteorological effects on calling activity, and to assess the use of call counts as an index of between year and between area variation in population density. Calling remained at high levels from August to April during the breeding season. Calling was most frequent at sunrise and sunset. Calling frequency and the number of calling coveys were significantly higher at sunrise than at sunset. Calling was concentrated in the 30 minute periods straddling sunrise and sunset, and peaked during the 15-minute periods before sunrise and before sunset. Calling activity was negatively correlated with wind speed and positively correlated with relative humidity. Calling was spuriously negatively correlated with seasonal variation in Greywing population density and strongly positively correlated with between year and between area variations in population density. We therefore suggest that call counts collected during March-April could be used to index annual change in the population density in a particular area from year to year, as well as within-season variation among areas.     

Flux Analysis of Glucose Metabolism in Rhizopus oryzae for the Purpose of Increasing Lactate Yields

A flux analysis of glucose metabolism in the filamentous fungus Rhizopus oryzae was achieved using a specific radioactivity curve-matching program, TFLUX. Glycolytic and tricarboxylic acid cycle intermediates labeled through the addition of extracellular [U-14C]glucose were isolated and purified for specific radioactivity determinations. This information, together with pool sizes and the rates of glucose utilization and end product production, provided input for flux maps of the metabolic network under two different experimental conditions. Based upon the flux analysis of this system, a mutant of R. oryzae with higher lactate and lower ethanol yields than the parent was sought for and found.     

The effect of estradiol benzoate on synchrony of estrus and fertility in cattle after removal of a progesterone-releasing intravaginal device.

The aim was to determine the effect of estradiol benzoate (EDB) given after removal of a progesterone-releasing intravaginal device (PRID) at either emergence or dominance of a follicle wave, on the interval to estrus, variation in its onset and pregnancy rate in heifers. Heifers (n=186) were assigned randomly to four treatments in a 2 x 2 factorial design; emergence or dominance of a follicle wave at PRID removal, with or without 0.5 mg EDB 24 h after PRID removal. Ovarian ultrasonography was performed to confirm follicular status; data from heifers of undeterminable follicular status were excluded (n=36). Mean size of the largest follicle of the new wave at PRID removal was smaller (P < 0.01) in heifers given EDB at emergence (6.3 +/- 0.09 mm) compared with those given it at dominance (10.9 +/- 0.30 mm). The onset of estrus was earlier (P < 0.01) in heifers given EDB at dominance (median 42 h, range 13 h) compared with those not given EDB at dominance (median 43 h, range 42 h). The median interval to estrus was decreased (P < 0.01) in heifers given EDB at emergence (median 48 h, range 73 h) compared with those not given EDB at emergence (median 66 h, range 45 h). Variation in onset of estrus was reduced (P < 0.05) in heifers given EDB compared with those not given EDB. The pregnancy rate was not affected when EDB was given at dominance, however, it was decreased (P < 0.05) when given at emergence (23 of 40 vs 26 of 32, respectively). To determine the effect of EDB on follicular dynamics in heifers treated with EDB at emergence, heifers (n=37) were assigned to two treatments: at emergence with or without EDB and their ovaries were examined daily using ultrasonography. Follicular dynamics were not different (P > 0.05) in EDB-heifers compared with untreated controls. Mean serum estradiol was greater (P < 0.01) in EDB-treated heifers compared with controls. In conclusion, 0.5 mg EDB given 24 h after PRID removal to heifers decreased the interval to estrous onset at emergence or dominance, decreased variation in onset of estrus and decreased pregnancy rates when given at emergence of a follicle wave.