Persistent changes of corticostriatal plasticity in dt(sz) mutant hamsters after age-dependent remission of dystonia.

Abnormal plasticity in the cortico-basal ganglia-thalamocortical loop has been suggested to represent a key factor in the pathophysiology of dystonia. In a model of primary paroxysmal dystonia, the dt(sz) mutant hamster, previous experiments have shown a strongly increased long-term potentiation (LTP) in comparison to non-dystonic control hamsters. These basal changes, i.e. in the absence of dystonia, were found in young animals at an age of 5 weeks, when the age-dependent dystonia in dt(sz) mutant reaches highest severity. In the present study we examined in corticostriatal slices (1) whether the increases in synaptic plasticity can be modulated by stressful stimuli which induce dystonic episodes in young mutant hamsters, and (2) whether increases of LTP persist after spontaneous remission of dystonia in animals older than 10 weeks. The present data show that in slices of young mutant hamsters the extent of LTP was not influenced by the presence of dystonia: In comparison to age-matched control hamsters, LTP was increased in mutant hamsters independent of preceding stressful stimulation. After remission of dystonia, i.e., in older dt(sz) mutant hamsters >10 weeks, only LTP could be elicited, while in preparations from age-matched control hamsters, either LTP or long-term depression developed, depending on previous behavioral challenge. We conclude that in mature brain, corticostriatal connections have the potential for changes in metaplasticity, while in dt(sz) mutant hamsters this metaplasticity is persistently infringed even though stress-inducible dystonic symptoms are lost.

Biosynthetic capacities of actinomycetes. 2. Juglomycin Z, a new naphthoquinone antibiotic from Streptomyces tendae.

A new juglomycin-type antibiotic was identified by a HPLC-diode array screening technique in the culture filtrate of Streptomyces tendae Tü 901/8c. Juglomycin Z (1) differs from all other known juglomycin compounds by an additional methyl group in position 3 of the naphthoquinone ring system. Juglomycin Z is antibiotically active against Gram-positive and Gram-negative bacteria and against yeasts.

Stabilization of the yeast desaturase system by low levels of oxygen.

The stability of particulate palmitoyl-CoA desaturase preparations from anaerobically grown yeast cells was increased by exposure to low levels of oxygen. The stabilizing effect of oxygen may be based upon the increased amounts of palmitoleic acid and ergosterol that become available to the cells. These results suggest the evolutionary appearance of this system at a time when atmospheric oxygen was at a low level.

Biochemical changes in rat liver after 18.5 days of spaceflight.

The effect of "weightlessness" on liver metabolism was examined using tissue from rats flown in earth orbit for 18.5 days aboard the Soviet Cosmos 936 biosatellite. Changes in the activities of certain carbohydrate and lipid enzymes were noted. Of the 28 hepatic enzyme activities assayed, two, palmitoyl-CoA desaturase and lactate dehydrogenase, increased, whereas five, glycogen phosphorylase, 6-phosphogluconate dehydrogenase, both acyltransferases which act on alpha-glycerolphosphate and diglycerides, and aconitate hydratase decreased. The remaining enzyme activities measured were unchanged. In addition, increased levels of liver glycogen and palmitoleate were noted which probably resulted from the lowered glycogen phosphorylase and increased palmitoyl-CoA desaturase activities, respectively, in those animals that experienced weightlessness. These changes caused by weightlessness were transient since all of the aforementioned alterations returned to normal values when measured in the livers of other rats which had flown in the biosatellite 25 days after recovery.

Pathogen factors and host factors in murine pulmonary blastomycosis.

Factors related to pathogenesis were studied in a murine model of pulmonary fungal infection with strains of Blastomyces dermatitidis. Strain 26199V was greater than or equal to 10(5)-fold more lethal in pulmonary challenge than strain GA-1. This relationship between the two strains also occurred after intraperitoneal challenge. Serial studies with lung cultures and histology of sacrificed animals after pulmonary challenge with similar doses, and challenge studies in mice of differing maturity, indicated GA-1 is impaired in initial replication in situ, although the two strains grow equally well in the presence of murine substrates in vitro. In vitro studies indicated the two strains were indistinguishable in most studies of biochemical and metabolic markers, and in studies with chemical and physical inhibitors. The exceptions were their morphologic appearance in the yeast phase, ability to hydrolyze esculin, and susceptibility to crystal violet dye and bile. The last finding and previous work suggested possible differences in lipid content; these were confirmed in studies which also included two other strains. Greater phospholipid and palmitic acid content were associated with greater virulence.

Spontaneous mutant of Blastomyces dermatitidis attenuated in virulence for mice.

A spontaneous mutation occurred in cultures of a virulent strain of Blastomyces dermatitidis during serial passage at 35-37 degrees C. The mutant strain was shown to be 10,000-fold less virulent for mice than the parent strain, and this was independent of the challenge dose, the age of the challenged host, and the route of challenge (although attenuation was greater for pulmonary than for peritoneal challenge). The mutant grows as well as the parent in fungal media or in the presence of murine substrates. Studies of the chronology of pulmonary infection indicate the mutation affects an early event (first week after challenge) in establishment of infection and interaction with host defenses. Scanning electron microscopy showed that the mutant lacks deep irregular surface ridges and grooves present in the parent. It had increased resistance to crystal violet in vitro, increased susceptibility to bile, and could hydrolyze esculin. The latter findings are of interest because they place the mutant in an intermediate position with respect to these properties between the virulent parent and a previously studied avirulent strain. Lipid analyses indicated a marked increase in fatty acids in the mutant. Studies of four B. dermatitidis strains have also associated increasing palmitic acid and phospholipid content with increasing virulence.

The effects of space flight on some rat liver enzymes regulating carbohydrate and lipid metabolism.

We have examined, in the livers of rats carried aboard the Cosmos 936 biosatellite, the activities of about 30 enzymes concerned with carbohydrate and lipid metabolism. In addition to the enzyme studies, the levels of glycogen and of the individual fatty acids in hepatic lipids were determined. Livers from flight and ground control rats at recovery (R0) and 25 days after recovery (R25) were used for these analyses. For all parameters measured, the most meaningful comparisons are those made between flight stationary (FS) and flight centrifuged (FC) animals at RQ. When these two groups of flight rats were compared at R0, statistically significant decreases in the activity levels of glycogen phosphorylase, alpha-glycerol phosphate acyl transferase, diglyceride acyl transferase, aconitase and 6-phosphogluconate dehydrogenase and an increase in the palmitoyl CoA desaturase were noted in the weightless group (FS). The significance of these findings was strengthened by the fact that all enzyme activities showing alterations at R0 returned to normal 25 days postflight. When liver glycogen and total fatty acids of the two sets of flight animals were determined, significant differences that could be attributed to reduced gravity were observed. The weightless group (FS) at R0 contained, on the average, more than twice the amount of glycogen than did the centrifuged controls (FC) and a remarkable shift in the ratio of palmitate to palmitoleate was noted. These metabolic alterations, both in enzyme levels and in hepatic constituents, appear to be characteristic of the weightless condition. Our data seem to justify the conclusion that centrifugation during flight is equivalent to terrestrial gravity.

Factors affecting the palmitoyl-coenzyme A desaturase of Saccharomyces cerevisiae.

The activity and stability of the palmitoyl-coenzyme A (CoA) desaturase complex of Saccharomyces cerevisiae was influenced by several factors. Cells, grown nonaerobically and then incubated with glucose, either in air or under N2, showed a marked increase in desaturase activity. Cycloheximide, added during such incubations, prevented the increase in activity, suggesting de novo synthesis. The stability of the desaturase from cells grown nonaerobically was affected by subsequent treatment of the cells; enzyme from freshly harvested cells, or from cells that were then shaken under nitrogen, readily lost activity upon washing or during density gradient analysis, whereas aerated cells, in the presence or absence of glucose, yielded stable enzyme preparations. The loss of activity in nonaerobic preparations could be reversed by adding soluble supernatant from these homogenates and could be prevented by growing the cells in the presence of palmitoleic acid and ergosterol, but not with several other lipids tested.

Membranes of Saccharomyces cerevisiae.

A crude small particle pellet, obtained from postmitochondrial supernatant fractions of Saccharomyces cerevisiae, contains about half the ergosterol and phospholipid of crude cell homogenates. Most of the phospholipid of this pellet is in a "heavy" fraction which, with the aid of electron microscopy, shows membranous elements in addition to discrete particles. The "heavy" fraction, upon treatment with deoxycholate, can be freed of membranes, or, with ribonuclease treatment, ribosomes can be removed, leaving relatively clean membranes. The "heavy" fraction resembles the microsomes of animal cells, but contains considerably less lipids, including phospholipids, thus suggesting a less well-developed intracellular membrane system.

Subcellular sites involved in lipid synthesis in Saccharomyces cerevisiae.

When the crude ribosomal fraction of Saccharomyces cerevisiae was separated into "light" and "heavy" fractions, fatty acid synthetase was concentrated in the former, whereas acetyl-Coenzyme A synthetase, fatty acid "desaturase," and squalene oxidocyclase were found in the latter. The "desaturase" sedimented with the ribosomal material and was not solubilized by low concentrations of sodium deoxycholate (DOC). The other two systems found in the "heavy" fraction sedimented with the membranes, but, upon solubilization of the membranes by DOC, these enzyme systems remained as particles.

Fatty acid synthetase of Saccharomyces cerevisiae.

A light particle fraction of Saccharomyces cerevisiae, obtained from the crude ribosomal material, and containing the fatty acid synthetase, consisted primarily of 27S and 47S components. This fraction has a protein-ribonucleic acid ratio of about 13. Electron micrographs showed particles ranging in diameter between 100 and 300 A in this material. By use of density gradient analysis, the fatty acid synthetase was found in the 47S component. This component contained particles which were predominantly 300 A in diameter and which were considerably flatter than ribosomes, and it consisted almost entirely of protein.