Recognition by peptide mapping of three different structural groups of outer membrane protein YOP-1 of Yersinia enterocolitica and Yersinia pseudotuberculosis

The structural relation of YOP-1 of european and american Yersinia enterocolitica serotypes O3, O9, O5, 27, and O8 and O20, respectively, and Y. pseudotuberculosis serotypes I, II, and III was compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis and peptide mapping using Staphylococcus aureus protease V8. Apparent molecular weights of YOP-1 ranged from 206,000 (O3) to approx. 180,000 (O8). According to their respective peptide maps YOP-1 of the european and american Y. enterocolitica serotypes and Y. pseudotuberculosis serotypes could be assigned to three different groups. Evaluation of several isolates of Y. enterocolitica serotypes O3, O9, and O8 by peptide mapping indicated that YOP-1 is conserved within a serotype. However, one serotype O8 isolate differed from the consensus peptide pattern of the other serotype O8 and O20 isolates. The similarity of the peptide patterns of Yersinia serotypes which predominate in certain geographical locations, i. e., european and american Y. enterocolitica serotypes, suggest common evolution of YOP-1 of these serotypes independent of the evolution of the other serotypes.     

Selective lectin reactions of two stocks of Leishmania enriettii with differing pathogenicity

Five days old promastigote culture forms of two stocks of Leishmania enriettii pathogenic and non-infective for Cavia procellus, were tested with the lectins of Canavalia ensiformis, Ricinus communis-120, Soja hispida (Glycine maxima), Arachis hypogaea, Ulex europaeus, Ulex europaeus I, Ulex europaeus II, Laburnum alpinum, Lotus tetragonolobus, Euonymus europaeus and with a monoclonal antibody against blood group H. The pathogenic stock reacted with the anti-H lectin of Lotus tetragonolobus and the monoclonal anti-H and the monoclonal anti-H but not with anti-H of U. europaeus I and E. europaeus. The non-infective stock reacted with none of these anti-H specific agglutinins. They had strong agglutination reactions to C. ensiformis (500 g/ml, 1,000 g/ml, 10 mg/ml) R. communis-120 (110), S. hispida (1,000 g/ml) and A. hypogaea (500 g/ml, 1,000 g/ml, 2,000 g/ml). They showed moderate agglutinations to U. europaeus, L. alpinum and U. europaeus II. The non-infective stock showed only moderate reactions to C. ensiformis (10 mg/ml), R. communis-120 (110), A. hypogaea (2,000 g/ml) and S. hispida (1,000 g/ml). Neither N-acetylneuraminic acid nor neuraminidase was detected on the cell surface of both stocks. This result demonstrates clearly that both stocks of L. enriettii differ in their cell surface carbohydrates. The agglutination reactions with lectins of the non-infective stock of L. enriettii are very similar to L.t. major.Supported in the context of a Research Programme of the Commission of the European Communities and the Bernhard-Nocht-Institute for Nautical and Tropical Medicine, Department of Protozoology, Hamburg, TSD-005-D(B)The work was supported by the Ministry for Youth, Family Affairs and Health     

Effect of ivermectin on filariae ofMastomys natalensis

Summary The efficacy of ivermectin (Iv) was evaluated against four species of filariae,Litomosoides carinii, Acanthocheilonema viteae, Brugia pahangi andBrugia malayi inMastomys natalensis. Animals with patent infections, induced with L₃ larvae, by intravenous (iv) infusion of the respective microfilariae (Mf) (5×10⁴ Mf per animal) or by intraperitoneal (ip) route (2×10⁴ Mf per animal) were used in this study. A single dose of Iv (100 g·kg^–1) given subcutaneously (sc) toMastomys infected withL. carinii orA. viteae resulted in the disappearance of microfilaremia within 2 h of treatment. Iv treatment of sc-infected animals withBrugia spp. had no immediate effect on the circulating Mf 60 days posttreatment. In contrast, such treatment of animals infected with Mf by intravenous infusion completely eliminated the larvae of all four species from the circulation. Iv treatment had no significant effect on the Mf ofL. carinii, B. pahangi andB. malayi in animals infected by the ip route. However, the drug had dramatic effect in killing the Mf ofA. viteae in the peritoneal cavity. Sera from Iv-treated normal or fromL. carinii- orA. viteae-infectedMastomys were effective in clearing the circulating Mf of the species when administered to animals with the respective infections. Similar rapid clearance of Mf was seen when the sera were administered to animals infected iv with these larvae. Furthermore, adult females ofL. carinii andA. viteae recovered fromMastomys on different days after Iv treatment released smaller numbers of Mf in vitro. Thus, Iv effect was most pronounced on the Mf ofA. viteae and ofL. carinii and less so on the Mf ofBrugia. Iv kills the adults ofA. viteae and to a lesser extent those ofL. carinii and arrests the release of Mf from them. Iv had no effect on adults ofBrugia parasites.     

Biochemical characteristics,phage patterns,and O1 factor analysis ofEscherichia coli O1∶K1∶H7∶F11 and O1∶K1∶H−∶F9 strains isolated from patients with urinary tract infections

Biochemical characteristics, O1 antigen factors and phage patterns were examined in 35 urinary O1K1 isolates ofEscherichia coli different in H and F antigen. Fermentation of dulcitol, decarboxylation of ornithine, requirement for nicotinamide, and determination of O1 factor d allowed maximum differentiation. On the basis of these tests the strains could be divided into two major groups which are obviously of different clonal origin. Members of clone 1 represented by serotypes O1K1H7F11 (12 strains) and O1K1H^–F11 (5 strains) were characterized by positive biochemical reactions and absence of O1 antigen factor d. Negative biochemical tests and presence of O1 antigen factor d were shown by strains of clone 2 which were of serotypes O1K1H^–F9 (14 strains) and O1K1H^–F^– (3 strains). Phage patterns are less well correlated with clonal assignment. However, strains of clone 2 were not susceptible to K1-specific phage D and were non-typable with another set of 13 phages.     

Cloning and characterization of a repetitive sequence fromPneumocystis carinii

Four repetitive sequence clones measuring 10.9–23.4 kb in length were isolated from the genomic library ofPneumocystis carinii. Restriction enzymes mapping and cross-hybridization studies revealed that these clones are interrelated and that they derive from the common repeat unit, which is specific forP. carinii. Dot-blot analysis suggested that the copy number of the repeat sequence is about 100, assuming that the genome size is 1.5×10⁷ bp. Interestingly, the repetition unit extended over at least 23.4 kb and included long, 5.2-kb inverted repeats, for example, A-B-A-C, in which A is the inversion of A.     

SpecialEscherichia coli serotypes among enterotoxigenic strains from diarrhoea in adults and children

106 enterotoxigenicEscherichia coli strains from children and adults from many parts of the world were serotyped for O and H antigens. Some OH types,i.e. O6H16, O8H9, O15H11, O25H42, O78H11 and O78H12, were found repeatedly from different geographical locations. Some of these OH serotypes were only found rarely among more than 20000E. coli strains collected over many years from different locations and sources. It is suggested that these special OH serotypes represent clones which have been selected to the special conditions in the small intestine and selected to carry the plasmids necessary to provoke diarrhoea.     

Differences in antigenic properties of Fc-binding activity during infection with herpes simplex virus type 1

Summary The antigenic properties of the Fc receptor induced by herpes simplex virus type 1 (HSV-1) were studied with anti-HSV F(ab)₂ and pFc from infected rabbits.It appeared that the HSV-induced Fc-binding receptor had different antigenic characteristics at different times after infection. The Fc receptor present early in the infection (0.5 hours), during the adsorption period, most probably is the result of a fusion event between the virus envelope and the infected cell. We found that this Fc receptor reacted with anti-HSV F(ab)₂ and thus showed HSV-antigenic properties in such a way that binding of anti-HSV F(ab)₂ prevented the binding of pFc fragments.Later on in the infection (5 hours), the Fc-binding activity present on the surface of the infected cell is the result of newly synthesized and in the plasma membrane integrated polypeptides. The Fc-binding activity present on the cell surface of 5 hours infected cells could not be inhibited by anti-HSV F(ab)₂ and did not interfere with the binding of pFc to the Fc receptor.With 1 Figure     

An α-mating-type-specific mutation causing specific defect in sexual agglutinability in the yeast Saccharomyces cerevisiae

We have identified a recessive -mating-type-specific gene agl causing agglutinability defect without significant effects on other sexual activities. a cells carrying agl showed sexual agglutination with cells but cells carrying agl showed sexual agglutination with neither cells nor a cells. cells carrying agl produced pheromone and responded to a pheromone just like wild cells. cells carrying agl showed a little decreased but significant mating ability when tested on solid media or membrane filter. The agl mutant is different from any -specific ste mutants found so far in many sexual activities. The agl gene is recessive, and unlinked to the mating type locus. Biological significance of the mating type agglutinability is discussed based on the results obtained with the mutant.     

Lichtmikroskopische Untersuchungen über die Entwicklung vonTheileria annulata (Dschunkowsky und Luhs, 1904) inHyalomma anatolicum excavatum (Koch, 1844)

Summary Fully differentiated kinetes, average length 17.6m, appeared in the haemolymph of engorged nymphs usually 17 to 20 days after repletion. Kinetes were observed at first in the salivary glands on day 18 after repletion. The kinetes then transformed into fission bodies of about 10m in diameter, mainly in type III alveoli and less frequently in type II alveoli. The fission bodies grew up to a size of about 20m after several divisions of their nucleus. At this time the ticks moulted and no further development occurred until activation. Shortly before infestation the salivary glands began to proliferate, and rapid growth of the fission bodies was observed, especially in young ticks where development of infective particles (sporozoites) was concluded within two days. Development in feeding adult ticks apparently occurred in four major steps: (1) Division of primary fission bodies (sporonts) into numerous secondary fission bodies (primary sporoblasts), (2) division of secondary fission bodies into tertiary fission bodies (secondary sporoblasts), (3) production of particles (sporozoites) by tertiary fission bodies and release of particles into the saliva, and (4) degeneration of fission bodies and their host cell but further release of particles.The host cell was stimulated to giant growth, thus its diameter increased, on average, from 15 to 110 m. Heavy infections resulting from parasitaemias of >40% caused disease and mortality in the tick population. Development was much retarded by aging. In ticks starved for six months sporozoites did not develop before day five to seven of infestation. Sporozoites may not develop at all in six to nine month old female ticks during the infestation period. The significance of the described developmental stages ofT. annulata was discussed and a sexual generation postulated. The hypothetic development ofT. annulata in its tick vector was illustrated.

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Gefördert von der Deutschen Forschungsgemeinschaft (DFG)     

Studien zur Frage des Alexingehaltes in den Jahren 1939–1949

Zusammenfassung Die Höhe des Alexingehaltes im Blute wurde bei erwachsenen Personen in 4415 Untersuchungen während eines Zeitraumes von 11 Jahren verfolgt. In der Beobachtungszeit traten starke Schwankungen der Alexin-Titerhöhe auf (1939136,94±0,42; 1945113,98±0,54; 1949134,84±0,76), deren Ursache in einer Änderung der Umweltbedingungen zu suchen ist. Die hohen Werte des Variationskoeffizientenv im Jahre 1945 (v=73,23%) und des StreuungsexzessesQ ₁₂ im Jahre 1946 (Q ₁₂=1,87) deuten auf eine unterschiedliche Anpassungsfähigkeit der Versuchspersonen an die veränderten Umwelteinflüsse.     

Synergistic effects of tumour necrosis factor-α and interferon-γ on feline haemopoietic progenitors

Pathogenic mechanisms that underlie feline leukaemia virus subgroup-C (FeLV-C) induced erythroid aplasia are unknown. FeLV-C infection is associated with higher serum levels of interferon- (IFN-) and tumour necrosis factor- (TNF-), which may act synergistically to cause haemopoietic suppression. In the present studies, the synergistic effects of TNF- and IFN- on feline bone marrow progenitors in vitro were evaluated. Bone marrow mononuclear cells from specific-pathogen-free cats were exposed to TNF- (100 and 200 pg/ml) and IFN- (100 or 200 units/ml), alone or in combination, for 2 h before plating for clonal assays of colony forming units. Our results show that TNF- and IFN- in combination caused marked suppression of feline colony forming units-erythroid (CFU-E), burst forming units-erythroid (BFU-E), and colony forming units-fibroblasts (CFU-F), whereas colony forming units-granulocyte/macrophage (CFU-GM) were minimally affected. The same concentrations of TNF- and IFN- alone had minimal effects on CFU-E, BFU-E and CFU-F. These results suggest that TNF- and IFN- may play a significant role in regulating haemopoiesis in cats and may be involved in the pathogenesis of erythroid aplasia in cats infected with feline leukaemia virus.     

Induction of interferon and host resistance in vivo by double-stranded complexes of copolyribonucleotide of inosinic and guanylic acids with polyribocytidylic acid

Summary Several double-stranded complexes of copolyribonucleotide of inosinic and guanylic acids with polyribocytidylic acid (poly IGC) were found to possess interferon inducing activity stronger than poly ICin vivo, Their activity increased in parallel with increase in the ratio of guanine base to hypoxanthine base in these copolymers as far as double-strand formation was observed with polyribocytidylic acid. Many other combinations of copolyribonucleotide with homopolyribonucleotide were also investigated, and several of them were found to induce interferon. However, the interferon inducing effects of these combinations including complementary base-pairings of hypoxanthine and cytosine increased in parallel with the length of the base-pairings, thus approaching to that of poly IC. It is, therefore, supposed that the activity of poly IG C is somewhat different from poly IC and that those of other combinations owe to the essential structure of poly IC. Furthermore, kinetics of interferon induction, cross tolerance to reinduction, and antiviral effectsin vivo of poly IGC and poly IC were studied.     

The effectiveness of the control of pH in the extracellular fluid of the brain by the respiratory control system

Summary The effectiveness of the respiratory control system as a regulator of the pH in the extracellular fluid of the brain is defined by pH_{ECF op}/pH_{ECF cl} where pH_{ECF op} means the primary or open loop shift and pH_{ECF cl} the final or closed loop shift of brain extracellular fluid pH. The analysis of a steady state model described in a preceding paper (Loeschcke, 1973) allows, in the limits of the suppositions and simplifications, to calculate the effectiveness of the feedback regulator in the cases of increased metabolism, metabolic acidosis-alkalosis and inhalation of CO₂. The effectiveness is diminished if CO₂ production is increased, it drops in metabolic acidosis and rises in metabolic alkalosis and it drops steeply if CO₂ is inhaled. The effectiveness of this control system depends on the controlling action of the controlling element (the ventilation) rather than on varying sensitivity of the sensing element. The controlling effect is defined asC=–d P _CO2/d V _A orC=d pH_ECF/d V _A.Im Rahmen des Programms des Sonderforschungsbereiches 114 (Bionach) der Deutschen Forschungsgemeinschaft.     

Influence of muscle fibre type and pedal rate on the V̇O2-work rate slope during ramp exercise

We hypothesised that the ratio between the increase in oxygen uptake and the increase in work rate (O₂/WR) during ramp cycle exercise would be significantly related to the percentage type II muscle fibres at work rates above the gas exchange threshold (GET) where type II fibres are presumed to be active. We further hypothesised that ramp exercise at higher pedal rates, which would be expected to increase the proportional contribution of type II fibres to the total power delivered, would increase the O₂/WR slope at work rates above the GET. Fourteen healthy subjects [four female; mean (SD): age 25 (3) years, body mass 74.3 (15.1) kg] performed a ramp exercise test to exhaustion (25 W min^–1) at a pedal rate of 75 rev min^–1, and consented to a muscle biopsy of the vastus lateralis. Eleven of the subjects also performed two further ramp tests at pedal rates of 35 and 115 rev min^–1. The O₂/WR slope for exercise <GET (S ₁) was significantly correlated with O₂ peak in ml kg^–1 min^–1 (r=0.60; P<0.05), whereas the O₂/WR slope for exercise >GET (S ₂) was significantly correlated to percentage type II fibres (r=0.54; P=0.05). The ratio between the O₂/WR slopes for exercise above and below the GET (S ₂/S ₁) was significantly greater at the pedal rate of 115 rev min^–1 [1.22 (0.09)] compared to pedal rates of 35 rev min^–1 [0.96 (0.02)] and 75 rev min^–1 [1.09 (0.05), (P<0.05)]. The greater increase in S ₂ relative to S ₁ in subjects (1) with a high percentage type II fibres, and (2) at a high pedal rate, suggests that a greater recruitment of type II fibres contributes in some manner to the xs O₂ observed during ramp exercise.     

The influence of congenitalToxoplasma infection on the spontaneous running activity of mice

Home-cage running-wheel activity of mice congentally infected withToxoplasma was recorded over 24 days. Infected mice were consistently more active than uninfected controls over the entire testing period. This finding extends previous studies and indicates that such increased activity levels occur not only in novel but also in familiar environments, and suggests that congenital toxoplasmosis tends to render mice hyperactive. If such behavioural alterations occur in wild mice, it is likely that infected mouse intermediate hosts would be more susceptible to predation by cats, the definitive hosts ofToxoplasma.     

Is potassium co-transported by the cardiac Na-Ca exchange?

It has been suggested that the stoichiometry of the electrogenic Na-Ca exchange is 3Na1Ca. Recently, however, it was reported in rod outer segments that the stoichiometry of Na-Ca exchange is not 3Na1Ca but 4Na1Ca+1K. In cardiac cells, the reversal potential has always been measured in the absence of K or at a very low K concentration. We have, therefore, re-examined the reversal potential of the Na-Ca exchange current by whole-cell voltage clamp in single guinea-pig ventricular cells in the presence of K on both sides of the membrane. The Na-Ca exchange current reversed at potentials close to the calculated values for 3Na1Ca stoichiometry even in the presence of K. The magnitude of the Na-Ca exchange current did not change in 1 and 10 mM [K]₀. We therefore conclude that K is not co-transported by cardiac Na-Ca exchange and its stoichiometry is 3Na1Ca.     

Characterization of the cellular response of spleen cells in BALB/c mice inoculated with Mycoplasma pneumoniae or the P1 protein

BALB/c mice were intranasally infected or intraperitoneally inoculated with Mycoplasma pneumoniae whole cells or were immunized with the isolated adhesin (P1 protein). Spleen cells were isolated and tested in vitro for proliferation activity after stimulation with the P1 protein and sonicated M. pneumoniae whole antigen preparations. In frequency analysis experiments the P1 protein-specific proliferative response of spleen lymphocytes increased from 1/11494 in mice immunized once to 1/3246 in eightfold-inoculated mice, demonstrating that the P1 protein is a prominent immunogen of M. pneumoniae cells. Depletion experiments showed that T and B cells are activated in a 21 relation. Fluorescence-activated cells sorting analysis revealed a shift of the CD4/CD8 ratio from 21 in control mice up to 31 in M. pneumoniae-, and to 3.41 in P1 protein-immunized mice, as well as an increase in interleukin 2 receptor-bearing cells and macrophage cell populations. The results indicate that this animal model is appropriate to study host-M. pneumoniae interactions and vaccination schedules.     

Effects of sonication on physicochemical and biological properties of synthetic double-stranded polyribonucleotides as interferon inducer

Summary Two kinds of synthetic double-stranded polyribonucleotides, poly IGC¹ and poly I:C and their constituent single-stranded polymers were subjected to sonication. Sonication of both poly IGC and poly IC resulted in decreases in viscosity, molecular size and heterogeneity in the size distribution. In poly IGC, whose average sedimentation constant was larger than or around 11 S, these changes were accompanied with enhancements of interferon inducing activity in rabbits and mice and antiviral activity in mice, and moreover with a decrease in the systemic toxicity in mice. In poly IC, however, such an enhancement in the interferon inducing activity was observed only when its molecular size corresponded to that of poly IGC. Previous sonication of poly C of the relatively large molecular size (> 10 S) has also been shown effective, to a certain extent, in obtaining double-stranded RNA of smaller size distribution with increased interferon inducing activity and lowered toxicity. It has been shown that these changes induced by sonication were based on the breakage of phosphodiester bonds of both double and single-stranded polyribonucleotides. On the basis of the analyses of the correlations between molecular sizes and the biological activities, it has been suggested that, while toxicity decreases always when the molecular size becomes smaller, the optimal size of the double-stranded polyribonucleotide complexes for interferon production ranges roughly from 10 S (9.1×10⁵ daltons) to 5 S (1.2 ×10⁵ daltons).     

Trait anxiety,submaximal physical exercise and blood androgens

Summary This study evaluates the relationship between trait anxiety and both androgen and gonadotrophic hormone levels at rest and during severe physical exercise. Twelve volunteers were selected among 160 untrained male collegial students and classified as anxious (N=6) or non-anxious (N=6) subjects according to their scores on three trait-anxiety tests (STAI, IPAT, 16 PF). Serum ⁴-androgen (testosterone and ⁴-androstenedione), ⁴-androgen (DHEA and DHEA-SO₄) and gonadotrophin (LH and FSH) concentrations were measured by radioimmunoassay before, during and after 20 minutes of intensive bicycle exercise (80% of maximal heart rate). Results indicate significantly lower serum ⁴-androgens in anxious subjects before exercise. However, for each subject and irrespective of his anxiety level, all measured serum androgen concentrations increased significantly during exercise, although ⁴-androstene-dione remained lower in anxious subjects than in non-anxious ones. Serum LH concentrations (but not FSH) were signicantly higher in anxious subjects throughout the observation periods. However, exercise induced in each subject a significant decrease in the serum level of both gonadotrophic hormones. The results suggest that trait anxiety level may constitute an important factor that affects both pre-exercise and exercise serum androgen concentrations in untrained subjects.