Two-dimensional impedance studies of BSA buoyant density separated human erythrocytes

Combined DC (Coulter Volume) and radio frequency impedance studies were performed on human erythrocytes which had been separated by buoyant density in linear, neutral, isotonic bovine serum albumin gradients. The individual buoyant density fractions showed no reproducible shift in volume with buoyant density but did show a shift with opacity, radio frequency impedance divided by dc impedance. This new electronic parameter of opacity can be related to cell age, since both it and cell age are directly related to buoyant density. This increase in opacity with buoyant density is correlated with a change in shape.     

Differences in reactivity of confluent and nonconfluent cultures of human endothelial cells toward thrombin-stimulated platelets or heparinized salt solution

Summary This study examined whether nonconfluent endothelial cell cultures reacted differently than confluent ones toward thrombin-stimulated platelets or a heparinized salt solution. The adherence to the endothelial cell cultures of⁵¹Cr-labeled human platelets stimulated at different thrombin concentrations was studied. There was significantly higher adherence of stimulated platelets to nonconfluent cultures compared with confluent ones. This was confirmed by scanning electron microscopy, which also revealed a tendency for the platelets to adhere at the cell periphery. Electron microscopy also showed that thrombin-stimulated platelets induced endothelial cell contraction. Part of the peripheral endothelial cell surface toward the bottom of the culture dish was inverted, facing the lumen of the dish. This phenomenon was particularly seen in nonconfluent cultures. When⁵¹Cr-labeled endothelial cultures were incubated with a mildly injurious fluid as heparinized sodium acetate and 20% serum, at 20° C for 30 min, the nonconfluent cultures showed significantly more cell detachment and release of⁵¹Cr than the confluent ones. We conclude that under the conditions of the present experiments there are differences in the reactivity of confluent and nonconfluent endothelial cell cultures. These differences probably reflect biological dissimilarities. In experiments where properties of cultured endothelium are studied, care should be taken that the degree of confluency is standardized.     

Resource partitioning of roach Rutilus rutilus and bleak Alburnus alburnus in two eutrophic lakes in SE Norway

In this paper I investigate the resource partitioning of roach Rutilus rutilus and bleak Alburnus alburnus in two eutrophic lakes in SE Norway. Zooplankton was the preferred food item for both species during summer, whereas blue-green algae dominated the diet in September/October. There was no diet overlap in May, with the roach feeding mainly on detritus and the bleak feeding on surface insects. During summer the diet overlap increased, reaching high values in autumn. Habitat overlap was close to unity in spring and early summer. Coinciding with increasing diet overlap during summer and autumn the overlap in habitat decreased.     

Genetic Linkage in the Horse. II. Distribution of Male Recombination Estimates and the Influence of Age, Breed and Sex on Recombination Frequency

In the present study an extensive amount of data, comprising more than 30,000 offspring in total, was analyzed to evaluate the influence of age and sex on the recombination frequency in the K-PGD segment of the equine linkage group (LG) I and the influence of age, breed and sex on recombination in the Al-Es segment of LG II. A highly significant sex difference is reported for both segments. Male and female recombination values in the K-PGD segment were estimated at 25.8 ± 0.8 and 33.3 ± 2.5%, respectively. Similarly, recombination was less frequent in the male (36.6 ± 0.7%) than in the female (46.6 ± 1.2%) in the Al-Es segment. Comparison of data from two Swedish horse breeds revealed no significant breed differences in either sex for recombination in the Al-Es segment. No evidence of an age effect was found in any segment or sex. The distribution of individual male recombination estimates was also investigated, and a significant heterogeneity among stallions was revealed in the K-PGD segment. The results are discussed in relation to previous studies on factors affecting recombination in mammals.     

Boar sperm surface glycoproteins: Isolation,localization, and temporal expression during spermatogenesis

Boar sperm glycoprotein fractions were isolated by Lens culinaris hemagglutinin affinity chromatography of detergent-solubilized ejaculated spermatozoa, followed by preparative sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. In order to develop methods for further investigations of the sperm proteins, we proceeded with two of the isolated glycoproteins. Antibodies were raised in female rabbits against each of the two sperm glycoproteins. By a combination of immunosorbent chromatography, using the antibodies obtained, and preparative SDS polyacrylamide gel electrophoresis, highly purified sperm proteins were isolated. The sperm proteins were immobilized on Sepharose gel columns and specific immunoglobulin Fab fragments were enriched by affinity chromatography. The specificity of the Fab fragments was ascertained by immunoprecipitation analysis. The Fab fragments were used in indirect immunofluorescence analysis to localize the corresponding antigens on the surface of boar spermatozoa. Both antigens were exclusively confined to the postacrosomal region. Immunohistochemical staining of boar testis sections revealed that both antigens are expressed from the spermatid stage. This technique also revealed that one of the antigens congregated at the Golgi complex-acrosome region during spermatogenesis.     

Similarities of adenosine uptake systems in astrocytes and neurons in primary cultures

Uptake of extracellular adenosine was studied in primary cultures of astrocytes or neurons. Both cell types showed a high affinity uptake. TheK _m values were not significantly different (6.5±3.75 M in astrocytes and 6.1±1.86 M in neurons), but the intensity of the uptake was higher in astrocytes than in neurons (V _max values of 0.16±0.030 and 0.105±0.010 nmol×min^–1×mg^–1 protein, respectively). The temperature sensitivity was similar in the two cell types. Adenosine uptake inhibitors and benzodiazepines inhibited the adenosine uptake systems in both astrocytes and neurons with IC₅₀ values in the high nanomolar or the micromolar range and the rank order of potency was similar in the two cell types. In both cell types the (–) isomers of two sets of benzodiazepine stereoisomers were more potent than the (+) isomers. Dixon analysis showed that dipyridamole, papaverine, hexobendine and chlordiazepoxide inhibited the adenosine uptake competitively and clonazepam noncompetitively in both cell types.     

The role of secreted acid hydrolases in the utilization of complex nutrients byTetrahymena

In an attempt to extend our knowledge of the biology of feeding of the ciliateTetrahymena thermophila, this organism was grown axenically on complex organic material. The nutrient substrate was based on autoclaved wheat grains and adjusted to either pH 5.5 or 7.5. In wild type cultures the cells grew and multiplied only under acidic conditions. In cultures of a mutant cell line blocked in the secretion of acid hydrolases the cells did not grow at either pH value. Thus released acid hydrolases may play a key role in the utilization of complex nutrients in combination with uptake of small organic molecules. Mechanisms in the feeding biology ofTetrahymena thermophila andParamecium tetraurelia are compared.     

The importance of macrophyte bed size for cladoceran composition and horizontal migration in a shallow lake

Cladoceran composition and diel horizontal migration were studiedin 2, 10 and 25 m diameter macrophyte exclosures establishedin the littoral zone of shallow Lake Stigsholm, Denmark. Theexclosures were protected from waterfowl grazing, but open tofish. The macrophyte community cornprized Potwnogeton pectinatus,Potamogeton pusillus and Callitriche hemaphroditica. Cladoceranswere sampled randomly every third hour inside and outside themacrophyte exclosures during a 24 h period. In the 2m exclosure,the pelagic species Ceriodaphnia spp. and Bosmina spp. dominatedduring the day, mean density being as high as 3430 indiv. l^–1During the night, density decreased to 10–20% of the daytimedensity thus indicating diel horizontal migration. In the 10and 25 m exciosures, the daytime mean density of Ceriodaphniaspp. was 865 and 202 indiv. l^–1, respectively, and didnot decrease at night. In contrast to the pelagic species, thedensity of macrophyte-associated species tended to be higherin the 10 and 25 m exclosure than in the 2 m exclosure. In thedaytime, Eurycercus lamellatus density in the 2, 10 and 25 rnmacrophyte exclosures was 7, 28 and 16 indiv. l^–1 respectively,while that of Simocephalus vetulus was 11, 171 and 92 mdiv.l^–1, respectively. There was no thy-night difference inthe density of macrophyte-associated species. We conclude thatcladoceran community composition varies with macrophyte bedsize, and that the edge zone between the bed and open wateris an important daytime refuge for potentially migrating pelagiccladocerans.