In vivo formation of omega-oxidized metabolites of leukotriene C4 in the rat

[3H8]Leukotriene C4 was administered to germfree rats and to conventional rats having a bile duct cannula. Several radioactive metabolites were isolated. Two polar biliary metabolites from conventional rats were identified as N-acetyl-omega-carboxy-leukotriene E4 and N-acetyl-omega-hydroxy-leukotriene E4. A polar fecal metabolite from germfree rats was found to be N-acetyl-omega-carboxy-leukotriene E4. Chemical identities were established using UV spectroscopy and cochromatographies with authentic compounds in several HPLC systems. The fecal metabolite was further characterized by reductive desulfurization followed by gas-liquid-radiochromatography. The yield of the two biliary metabolites was 5% of the administered tritium after three hours and the yield of fecal N-acetyl-omega-carboxy-leukotriene E4 was 3.5% after three days.     

Infections with Francisella tularensis biovar palaearctica in hares (Lepus timidus, Lepus europaeus) from Sweden

The occurrence of tularemia was studied in 1,500 hares submitted to the National Veterinary Institute, Uppsala, Sweden for postmortem examination during 1973 through 1985. A total of 109 tularemia cases was recorded based on the fluorescent antibody (FA) test for Francisella tularensis and on the gross and microscopic pathology. Tularemia was diagnosed only in the varying hare (Lepus timidus) and not in the European brown hare (Lepus europaeus). The geographical distribution of the 109 cases indicates that tularemia has not spread in Sweden during the last 45 yr, with the exception of an endemic occurrence of the disease on the island of Stora Karls? in the Baltic sea. The disease was most frequent in the autumn and only a few cases were recorded during winter. Cases were not seen in the spring. The annual prevalence varied, with several cases in 1974 and 1981, but there were no cases in 1976 and 1980. The postmortem findings in hares dying of tularemia in the autumn were characterized by focal coagulative necrosis in liver, spleen and bone marrow, with high numbers of bacteria FA-positive for F. tularensis. In hares dying during winter months, the most characteristic findings were hemorrhagic enteritis and typhlitis, although necrotic lesions could occur in liver, spleen and bone marrow. Diseased hares on the island of Stora Karls? were demonstrated to be infected with ticks, while hares on the mainland of Sweden generally were fed upon by mosquitoes. Twenty-six of the 109 hares with tularemia were examined bacteriologically and F. tularensis biovar palaearctica was isolated from eight. The lung extract antibody test for F. tularensis was performed in 18 of the 109 hares. All were negative. In addition to the field study, an experimental study with F. tularensis biovar palaearctica was performed. Four varying hares and three European brown hares were inoculated. None of the hares died from tularemia, and generalized infection was not demonstrated.     

Experimental infection of five species of raptors and of hooded crows with Francisella tularensis biovar palaearctica

Sixteen raptors and three hooded crows were infected experimentally with Francisella tularensis biovar palaearctica. The birds were infected parenterally or per os. One goshawk, one sparrow hawk and one hooded crow died during the experimental period, and the remaining 16 birds were killed 14-77 days after the first infection. Francisella tularensis was not isolated from any bird. Antibody levels against F. tularensis measured in nine birds varied from 0 to 1:1,280. In one goshawk with a titer of 1:1,280, positive fluorescent antibody reactions against F. tularensis were seen in the liver and spleen. These results are similar to those found by other authors indicating that raptors and corvids are normally resistant to infections with F. tularensis.     

An altered pattern of cross-resistance in multidrug-resistant human cells results from spontaneous mutations in the mdr1 (P-glycoprotein) gene

Multidrug resistance in human cells results from increased expression of the mdr1 (P-glycoprotein) gene. Although the same gene is activated in cells selected with different drugs, multidrug-resistant cell lines can be preferentially resistant to their selecting agent. The mdr1 cDNA sequence from vinblastine-selected KB cells, which are uniformly resistant to different lipophilic drugs, was compared with the corresponding sequence from colchicine-selected KB cells preferentially resistant to colchicine. These sequences differ at three positions, resulting in a single amino acid change in P-glycoprotein. These differences result from mutations that occurred during colchicine selection. The appearance of these mutations coincides with the emergence of preferential resistance to colchicine. We have constructed biologically active mdr1 cDNA clones that express either wild-type or mutant P-glycoprotein. Multi-drug-resistant transfectants obtained with the mutant sequence were characterized by increased relative resistance to colchicine compared with transfectants obtained with wild-type sequence. mdr1 mutations are therefore responsible for preferential resistance to colchicine in multidrug-resistant KB cells.     

The low-frequency EEG component in the healthy subject and its changes as affected by acutely developing foci at the level of the diencephalon and brain stem structures

The data are given of the analysis of the low-frequency EEG component (LF EEG--0.2--2.0 Hz) of 34 healthy subjects and 36 patients examined in dynamics in the acute period after operations: ablation of a tumour localized at the level of the diencephalon and the level of the brainstem. The LF EEGs were analyzed by a special program: auto- and crosscorrelation functions and power spectra were estimated. In the norm LF EEGs were characterized by mosaics, various frequencies and periodicity in various cortical areas, they appeared with interhemispheric asymmetry, depended on the EEG type--dominance or lack of the alpha-rhythm. In patients in the post-operative period the LF EEGs were changed; in favourable terminations, at first the period of oscillations and the coefficient of correlation increased, then they tended to normalization. In unfavourable terminations, with gross pathological foci in these areas, the LF EEGs acquired a synchronized character, with a higher frequency than in the norm.     

The lateralization of hemispheric control over pain-induced vocalizations in rats

Study was carried out on 232 white rats, male and female, fixed in a hammock and subjected to painful stimulation of the tail. The first impulse of pain vocalization was analyzed. The rats were divided into three groups--with intact brain, with inactivated left or right hemisphere. The first impulses of rats pain vocalizations may be divided into short screams with 40-140 ms duration and long squeals of 140-420 ms duration. Unilateral cortex inactivation led to a reduction of the latency, increase of the peak amplitude and change of the per cent both of the pain screams and pain squeals. After elimination of the left hemisphere as compared with that of the right one, a shorter latency and a greater per cent of the pain screams was observed and a smaller per cent of pain squeals. The obtained data testify to the dominance of the left hemisphere in the control of pain vocalizations in rats.     

Monoclonal antibodies localize events in the folding, assembly, and intracellular transport of the influenza virus hemagglutinin glycoprotein

We used monoclonal antibodies that recognize monomeric and/or trimeric forms of the influenza virus hemagglutinin (HA) to study biosynthesis of this integral membrane protein in influenza virus-infected cells. We find the following: First, the globular head of the HA folds into its mature conformation in the endoplasmic reticulum prior to the assembly of HA monomers into trimers. Second, trimerization begins within 1 to 2 min following synthesis, with a half-time of approximately 5 min. Third, trimerization occurs only after the HA has been transported from the endoplasmic reticulum. Fourth, newly formed trimers are sensitive to acid-induced conformational alterations associated with viral fusion activity.     

Production and Purification of d-Aminoacylase from Alcaligenes denitrificans and Taxonomic Study of the Strain

A d-aminoacylase-producing microorganism, strain DA181, isolated from soil was identified as Alcaligenes denitrificans subsp. denitrificans. This strain produced about 29,300 units (micromoles of product formed per hour) of d-aminoacylase and 2,300 units of l-aminoacylase per gram of cells (wet weight) when cultivated in a medium containing 1% N-acetyl-dl-leucine as the carbon source. The d-aminoacylase was purified 345-fold. The specific activity of the purified enzyme was 108,600 units per mg of protein when N-acetyl-d-methionine was used as a substrate. The apparent molecular weight was 58,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. N-Acetyl-d-methionine was the favored substrate, followed by N-acetyl-d-phenylalanine. This enzyme had a high stereospecificity, and its hydrolysis of N-acetyl-l-amino acids was almost negligible.