Lysosomal enzyme activities in polymorphonuclear leukocytes,macrophages, serum,and spleen of conventional,germ-free,and antigen-free minnesota miniature swine

The activities of lysozyme, myeloperoxidase, and elastase were lower in PMNs and AMs from GF and AF Minnesota miniature piglets than in the leukocytes from their CONV counterparts. In the spleen and serum of gnotobiotic piglets only the levels of lysozyme were slightly reduced. Substantially depressed activities of these LEs were found also in PMNs from precolostral piglets in comparison with PMNs from their CONV mother. The bisassociation of GF piglets with Enterococcus liquefaciens and Escherichia coli caused an increase of LE activities in their AMs, spleens, and sera. Fewer LEs were released after phygocytic stimulation with zymosan from PMNs of GF, AF, and precolostral piglets than from PMNs of CONV animals of the same age. These data suggest that the antigenic-microbial stimulation is important for the development of normal lysosomal enzyme activities in PMNs and AMs from gnotobiotic animals.     

Poly-3-hydroxybutyratc production and changes of bacterial community in the soil

Changes in the number of bacteria and relative distribution of strains producing poly-3-hydroxybutyrate (PHB) in soil were investigated. Samples of chernozem soil were cul tivated with glucose in the presence of a mineral nitrogen source (diammonium hydrogen phos phate) or in its absence, either in a batch or a heterocontinuous cultivation system. In both cultivation systems the addition of glucose resulted in a roughly ten-fold increase of bacteria concentration and an increase in the proportion of strains able to produce PHB granules. When the nitrogen source was added simultaneously with glucose, the concentration of bacteria increased by two orders of magnitude in both cultivation systems. In the batch system changes in the concentration of strains capable of PHB production were very small under these con ditions whereas in the heterocontinuous system their number decreased by almost 50 %. The survival of bacteria in the soil suspension after 57-d starvation was associated with PHB production which differed, depending on the previous treatment of the soil samples. The concentration of bacteria decreased least pronouncedly in the control with water and most significantly during cultivation with glucose and a nitrogen source, where the initial PHB content was very low in spite of high numbers of bacteria.     

Antigenic properties of T cell antigen-specific receptors isolated from the surface of rabbit and mouse spleen and lymph node cells

The presence ofa allotypic determinants was tested in fractions obtained by gel filtration of antigen-specific receptors isolated by immunoadsorption from lymphoid cells of antigen-stimulateda3-3 rabbits. This technique, as well as the inhibition of the reaction of isolated receptors with anti-T cell receptor antisera (anti R) by anti-a3 antibodies failed to demonstrate the presence of a allotypie determinants. The inhibitory effect of antigen-specific receptors isolated from the lymphoid cells of stimulated A/J mice on the cytotoxic effect of anti-Ia antibodies on mouse spleen cells in the presence of rabbit complement was tested. All preparations inhibited the cytotoxic reaction with the average effectivity of 60%. In order to confirm the presence of Ia determinants on the rabbit and mouse T cell receptor molecules it was shown that the reactions of three anti-R antisera with 12 different receptor preparations were inhibited by anti-la antibodies. SDS-PAGE analyses of¹²⁵I-labelled mouse specific receptors and the precipitate obtained by anti-R antisera showed that T cell receptors were present in fractions with molar mass 100 and 85 kg/mol. The molar mass of the former fraction after reduction and alkylation was 45 kg/mol.     

Bacteriocin production inAgrobacterium tumefaciens

Agrobacterium tumefaciens C58 forms “plaques” during layer cultivation. The “plaques” were shown not to be caused by the presence of a temperate bacteriophage or by random contamination. The “plaques” and their central microcolonies were used to repeatedly isolate cultures producing an antibiotic substance against the original strainA. tumefaciens C58, other nopaline strains, some octopine strains ofA. tumefaciens and some strains of the related genusRhizobium. The substance is thus a bacteriocin; in analogy to agrocins 84 and D286 it was named agrocin C58. The agrocin is not inactivated by trypsin. Its production by strain C58 was found only on cultivation on solid but not liquid media. The producing isolate ofA. tumefaciens C58 (strain C58i2) contains neither plasmid pTiC58 nor the plasmid analogous to pAgK84 which controls the production of agrocin 84 inA. radiobacter K84.     

Microgeographic and Temporal Genetic Differentiation in Natural Populations of DROSOPHILA SUBOBSCURA

Evidence of microgeographic and temporal genetic differentiation in natural populations of Drosophila subobscura is presented. The alcohol dehydrogenease locus was used as a genetic marker. Behavioral differences among the sexes and genotypes may explain these observations, although the molecular basis remains obscure.     

Evolution and Extinction of Transposable Elements in Mendelian Populations

A model of the evolution of a transposable element family in a Mendelian host population is proposed that incorporates heritable phenotypic mutations in the elements. The temporal behavior of the numbers of mutant and wild-type elements is studied, and the expected extinction time of the transposable element family is examined. Our results indicate that, if the mutant can be transposed equally well in the presence of the wild type, then it can be expected to be found in preponderance, whereas elements, such as retroviruses, where the transposing genome and its phenotypic expression are coupled, may be characterized by a low mutant frequency.     

The effect of lake restoration measures on the physical,chemical and phytoplankton variables of Lake Bled

Monthly changes of physical, chemical and biological variables due a combination of artificial inflow of clean water, removal of hypolimnetic water, and diversion of sewage were studied in Lake Bled from December 1980 to December 1982.During the winter period 1981/82 the species composition of the phytoplankton changed. New species replaced those observed in previous years. We conclude that the combined effect of these three lake restoration measures was responsible for the sudden disappearance ofOscillatoria rubescens D.C. A marked decrease in some nutrients and an increase in temperature and oxygen concentration also occurred.     

The effect of glucose on the metabolism and excretion of cortisol in man

The 24 hour urinary free cortisol and cortisone excretion after an oral 100 g glucose load was measured in 60 males (aged 22-56) divided into three groups. G-I consisted of 10 healthy men, G-II of 37 surgical patients and G-III comprised 23 patients with atherosclerotic peripheral vascular disease. The followed subjects responded to the glucose ingestion accordingly to their cortisol excretion. Subjects with an urinary cortisol excretion up to 200 micrograms/24 h responded to the glucose load with an increase of excretion in free cortisol and cortisone. Subjects with the excretion of cortisol above 200 micrograms/24 h responded unambiguously with a decrease in their excretion. We suggest that these changes in both directions can be explained by the available amount of NADPH in the liver. In patients with atherosclerotic peripheral vascular disease, in whom disturbances in lipid and carbohydrate metabolism can be proposed, the response of free corticoids, namely the respond of cortisone, are unequal.     

Methylation of type II and type I collagen genes in differentiated and dedifferentiated chondrocytes

The methyl-sensitive restriction endonucleases HpaII and HhaI as well as the methyl-insensitive enzyme MspI were used to examine the methylation status of the pro-alpha 1(II) collagen gene of cartilage. Five different cell types with varying abilities to express type II collagen were studied. Chick embryo chondrocytes express type II collagen, while 5-bromodeoxyuridine-treated chondrocytes, retinoic acid-treated chondrocytes, chick embryo fibroblasts, and erythrocytes do not synthesize type II collagen. Both cDNA and genomic probes for the pro-alpha 1(II) collagen gene were used, covering the complete 3' end of the gene and its flanking sequences. The pro-alpha 1(II) collagen DNA was undermethylated in chondrocytes, compared to either fibroblasts or erythrocytes. However, the methylation of the 5-bromodeoxyuridine-treated and retinoic acid-treated chondrocytes was identical to that of control chondrocytes. The methylation pattern of two regions of the gene of the pro-alpha 2(I) collagen chain was identical in all cell types tested, whether or not the gene was expressed. Our results indicate that genes for these collagen chains differ in their methylation pattern. The type II collagen gene shows reduced methylation in expressing cartilage, but does not acquire an increase in methylation in "dedifferentiated" chondrocytes. The changes in DNA methylation that occur during cell differentiation do not appear to be sufficient to explain gene activation and deactivation.     

Electrostatic effect of trypsin binding on the hydrogen exchange rate of bovine pancreatic trypsin inhibitor beta-sheet NH's

The changes of H-D exchange rates upon protein-protein interactions are generally interpreted as a result of the changes of the dynamic properties of the proteins. The effect of trypsin binding on the H-D exchange kinetics of some trypsin inhibitor amide H's was reported (Simon et al., 1984). In this paper the electrostatic potential originating from the trypsin molecule is calculated at the positions of the studied amide H's in the trypsin-trypsin inhibitor complex. We conclude that the observed decrease of the exchange rates is mainly due to the electrostatic field of the trypsin molecule.