Increasing incidence of meningococcal disease in Spain associated with a new variant of serogroup C

Serogroup B has been the main cause of meningococcal disease in Spain since at least 1979, but in recent years an increase in the prevalence of infection due to serogroup C meningococci has been detected. In 1996, for the first time, most cases of meningococcal disease were caused by serogroup C strains. The sero/subtype of all serogroup C meningococci received from 1993 to June 1996 was determined, and the results showed that C2bP1.2,5, the most common phenotype in 1995 and 1996 (63% and 65%, respectively), represented only 4.8% of strains in 1993. The C2bP1.2,5 epidemic strains appear to be responsible for the high prevalence of serogroup C in Spain. One hundred fifty-one randomly selected serogroup C strains were analyzed by multilocus enzyme electrophoresis, ribotyping, and pulsedfield gel electrophoresis. Pulsed-field gel electrophoresis provided the most accurate information: more than 80% of the C2bP1.2,5 and C2bP1.2 isolates exhibited one of two very closely related profiles, while most of the C:2b:NST and C2bP1.5 strains had a pattern located at a genetic distance of 0.24 from those two profiles. The results show that C2bP1.2,5 strains represent a subclone or a genetic variant of the previously identified Spanish epidemic clone C2bnon-subtypable strains.     

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.     

An autoradiographic identification of Purkinje axon terminals in the cat

Summary The Purkinje axon boutons terminating in nuclei fastigii and interpositus in the cat have been identified after injection of³H leucine into the cerebellar cortex overlying the nuclei. The animals survived from 4–48 h after injection of the isotope. Semithin and ultrathin sections were coated and exposed for 3 and 14 weeks, respectively. The electron micrographs showed labelling over myelinated axons down to a diameter of 0.8 m, and over boutons.Fifty labelled boutons were used for identification of the shape of their synaptic vesicles. The statistical analysis including the test for skewness showed that 39 boutons (78%) fall in one group. Most of the synaptic vesicles in this group are elliptical (ration from 11.1 – 11.7). Slightly ovoid vesicles (ration up to 11.3) are frequent, but flattened vesicles (ration above 11.7) are relatively few in this group of boutons. 8% of the boutons have a rather homogeneous vesicle population (prevalence of round vesicles, ratio 11).Synaptic specializations of Gray's type II or of an intermediate type were found in the boutons belonging to the first group (78% of the boutons). Specializations of Gray's type I were found in the other bouton groups (8% of the boutons).     

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).     

Effect of adjuvant arthritis on the disposition of acebutolol enantiomers in rats

Disease states such as arthritis may interact with the kinetics of -blockers. Acebutolol (AC) is a chiral -blocker which is available as a racemate. The beneficial properties of AC, however, is attributed mainly to theS-(+)-enantiomer. The disposition of AC enantiomers and their active, chiral metabolites, diacetolol (DC) were examined after oral administration to healthy and adjuvant-induced arthritic (AA) female Sprague-Dawley rats. Arthritis was induced by tail base injection ofMycobacterium butyricum. Swelling of hind and forepaws were apparent in 10–16 days in AA but not controls. Control and AA rats were sacrificed at 0.5, 1, 2, 4, 6 and 8 h after a 25 mg/kg oral AC dose and blood was collected (n=6). Significant three to tenfold increases in the initial plasma concentrations (0.5–2h) of AC were observed in AA. Enantiomers were equally affected, thus ACSR ratio was not changed. Higher plasma concentrations of the metabolite were only significant at 2h. The ratio of DCAC, however, was unaffected by AA. The DCSR ratio was significantly decreased at 0.5 and 1 h in AA. The limited protein binding of AC (10%) was neither stereoselective nor affected by AA. Reduced intrinsic clearance in AA may be responsible for these observations.     

Secretin and VIP-stimulated adenylate cyclase from rat heart

Cardiac adenylate cyclase activity was normal in 3 weeks-old spontaneously hypertensive rats of the Wistar-Okamoto substrain. The hormone-sensitive adenylate cyclase activity was reduced in 10 weeks-old or older animals, and secretin- and VIP-activations were definitely more impaired (by 64% and 69%, respectively) than isoproterenol- and glucagonactivation (17% and 22%, respectively). By contrast, the fluoride- and p[NH]ppG-stimulations of the enzyme were unaffected. These alterations in the adenylate cyclase system coupled to secretin and VIP appeared specific to the heart as the isolated pancreatic acinar cells from spontaneously hypertensive animals responded normally to secretin, as a liver particulate fraction responded normally to secretin and VIP, and both brain synaptic membranes and a particulate fraction of anterior pituitary to VIP.Abbreviations VIP vasoactive intestinal peptide - cyclic AMP cyclic adenosine 35-monophosphate - p[NH]ppG guanosine 5-(, -imido)triphosphate - EGTA ethylene-glycol-bis-(2-amino-ether)-N,N,N,N-tetraacetic acid - IBMX 3-isobutyl-1-methylxanthine     

Ein neuer Salmonella-Serotyp: (6), 7∶1, v∶z6 (S. gelsenkirchen)

Zusammenfassung Beschreibung eines neuenSalmonella-Typs mit der Seroformel (6), 71, vz₆ (S. gelsenkirchen).     

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.     

Establishment of a cell line fromculex bitaeniorhynchus

Summary A cell line from embryonic tissue ofCulex bitaeniorhynchus Giles was established using Mitsuhashi and Maramorosch's medium, containing 20% fetal bovine serum (FBS). In the primary culture a monolayer was obtained in 17 to 20 d. During initial subcultures the split ratio was 12 and a monolayer was formed in 7 to 10 d. In subsequent passages, the split ratio was gradually increased to 160, with about 4 to 5 × 10⁵ cells as inoculum in 4 ml of medium. From the 10th passage level, the concentration of FBS in the medium was reduced to 15%.     

Virulence factors and phenotypical traits of verotoxigenic strains of Escherichia coli isolated from human patients in Germany

Fecal isolates of Escherichia coli which were collected from human patients in different parts of Germany between 1985 and 1992 were examined for production of verotoxins (VT). Among 2165 isolates 54 (2.5%) verotoxigenic E. coli (VTEC) were found. The 54 VTEC belonged to 13 different serotypes, 46 (85.2%) of these were enterohemorrhagic E. coli (EHEC) types as O157H7, O157H-, O145H-, O111[H8] and O26[H11]. Of the 54 VTEC 50 (92.6%) hybridized with one or both of the DNA probes specific for VT1 and VT2. The 4 VTEC strains which were negative for VT1 and VT2 differed from all other VTEC by many phenotypical trains such as serotype, production of -hemolysin and absence of EHEC-plasmid and attaching and effacing (eae)-specific DNA sequences. In contrast, VTEC which were positive for VT1, VT2 or both were frequently positive for eae sequences (92.0%), EHEC-plasmids (90.0%) and for production of enterohemolysin (88.0%). With enterohemolysin as an epidemiological marker more VTEC strains (81.5%) could be identified than with others such as the absence of -glucuronidase activity (61.1 %) or non-fermentation of sorbitol (48.1%). Case reports were available for 42 of the 54 VTEC strains. The clinical presentation of 42 cases with VTEC ranged from uncomplicated diarrhea to severe diseases as hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). However, bloody diarrhea, HC and HUS were more associated with the O157 group than with other VTEC groups.     

Genetic determinants ofPseudomonas aeruginosa colonization in cystic fibrosis patients in Canada

The present study was aimed at analyzing whether the rate of colonization and the age at colonization withPseudomonas aeruginosa was genetically determined in cystic fibrosis (CF) patients. These two variables were calculated among 127 CF patients whose genotypes were known and who were monitored at the Clinique de Fibrose Kystique in Saguenay Lac-Saint-Jean. No statistically significant differences were found in the rate or the age at colonization when the patients were grouped by genotype; however, this result could be due to the small number of patients in each genotype group. The rate of colonization was significantly lower among CF patients carrying the A455E mutation, a mild allele with respect to exocrine pancreatic function, than among those carrying either the F508 or the 621+1G->T mutation, both of which are severe alleles. The results confirm previous reports that the rate of colonization withPseudomonas aeruginosa is, at least in part, genetically determined.     

Eosinophil migration in response to three molecular species of platelet activating factor

Multiple molecular species of the eosinophil chemoattractant platelet activating factor (PAF) are produced as a result of inflammatory processes. We therefore compared the ability of three naturally occurring PAF species (C160, C180, and C181), which only varied at carbon 1, to induce eosinophil chemotaxis through naked 3-m pore polycarbonate filters. Timecourse experiments indicated that all species of PAF tested induced significant and equivalent eosinophil migration at 1 h which peaked at 2 h. Overall, the rank order of chemotactic potency for the PAF species was relatively equivalent. The specific PAF antagonist WEB 2086 inhibited eosinophil migration induced by all three PAF species equally. We conclude that the degree of PAF-induced eosinophil migration is not dependent upon the molecular species of PAF.accepted by G. W. CarterThis work was supported in part by a Veterans Administration Merit Review Award.     

Bone metastasizing renal tumour of childhood

Summary The histopathological spectrum of a large series of a recently described tumour entity is presented. Seven diagnostic features which may be encountered are described and their frequency discussed. The most striking clinical feature was the marked male preponderance (MF=7.61). It is suggested that an appreciation of the full histopathological spectrum is necessary to ensure adequate diagnosis.Consultant Pathologist and Honorary LecturerLecturer in Pathology     

Decreased tissue immunoregulatory index in rheumatoid synovitis,determined by in-situ T-cell phenotyping with monoclonal antibodies

Conclusion The phenotypic tissue immunoregulatory index in rheumatoid synovitis is locally different and varies significantly between given areas. Ectopic lymphoid follicles show normal to high T4T8 ratios, whereas within the sublining perivascular and diffuse infiltrate T4T8 ratios are decreased. Thus the frequency reported immunoregulatory indices of eluted cells can't be representative for an immunoregulatory tissue disequilibrium in rheumatoid arthritis. An elevated T4T8 ratio in eluted cells may correlate with a follicular B-cell hyperplasia in rheumatoid synovitis (low local activity), a low T4T8 ratio may be related to rheumatoid synovitis with diffuse sublining T-cell infiltrates (high local activity). Our results are in principal accordance with the findings of Kurosaka and Ziff [15] and Meijer et al. [13]. They also found different T-cell distributions in synovial tissue depending on the histopathological localization.For these topographical reasons, immunoregulatory indices determined in eluted cells from tissue samples cannot be representative for the immunopathological reactions at the synovial membrane tissue level.The decrease in the phenotypic tissue immunoregulatory index in the rheumatoid synovium from deeper to superficial areas may be explained as a compensatory influx of peripheral, functionally defect T8-cytotoxic/suppressor cells (see Fig. 2).     

Sequence analysis of the cystic fibrosis gene in patients with disseminated bronchiectatic lung disease

Summary The diagnosis of classical cystic fibrosis (CF) is easily made by clinical assessment alone, but may be missed or delayed in cases with an atypical clinical course. In a recent major study the age at diagnosis varied between 2 months and 47 years. For diagnostic purposes we have investigated the cystic fibrosis transmembrane regulator (CFTR) gene in 10 adult patients (age 18 to 45 years) with chronic obstructive pulmonary disease since childhood or adolescence and bronchiectases disseminated through both lungs. Only one subject (a 29-year-old male) had exocrine pancreatic insufficiency (PI); all others were pancreatic-sufficient (PS). The first nucleotide (ATP)-binding fold of the CFTR was analyzed by direct sequencing of polymerase chain reaction (PCR)-amplified genomic DNA in these cases. Two patients with different phenotypes (one PI, one PS) were found to be homozygous for the common F508 mutation of the CFTR gene, which proved the diagnosis of cystic fibrosis in their cases and allowed genetic counselling. The PS patient had normal sweat tests and had not previously been recognized as having CF. Four other patients were heterozygous for F508, with no other mutation in exons 10 or 11 of the gene, and four patients had normal sequences of these exons. Because only about 70% of all CF chromosomes carry F508, the unexpectedly high frequency (4/8=50%) of heterozygosity for F508 among the non-F508/F508 patients with bronchiectases suggests that some of these might also have unrecognized CF with rare genotypes and mutations in any of the 22 exons not sequenced. About 90 different mutations have already been detected in coding regions of the CFTR gene and a very broad and so far not fully recognized spectrum of clinical phenotypes may be caused by mutations of this gene. Screening for the most frequent CFTR gene mutations, which is practicable using recent technology, may provide significant new diagnostic information in patients with CF-like pulmonary phenotypes, especially if they have normal or borderline sweat tests and no pancreatic insufficiency.Abbreviations PCR polymerase chain reaction - CF cystic fibrosis - CFTR cystic fibrosis transmembrane conductance regulator - COPD chronic obstructive pulmonary disease - PI pancreatic insufficiency - PS pancreatic sufficiency     

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.     

The influence of Poly I∶C on the course of infection in mice inoculated with West Nile Virus

Summary In mice infected intraperitoneally with a hundred per cent lethal dose of West Nile virus a significant reduction in mortality was found if treatment with the complex of synthetic polyriboinosinic and polycytidylic acids (Poly IC) was given four hours prior to or twenty hours after virus challenge.Treatment induced large amounts of circulating interferon a few hours after inoculation. Only a slight difference in maximum viraemia in the various groups was found, but viraemia developed later in the mice given Poly IC a few hours before virus injection. Infection of the brain developed later in the groups treated with Poly IC.Using various doses of West Nile virus almost the same mortality was found in the group given a lethal virus dose but treated with Poly IC and the group receiving sublethal virus dose and no Poly IC treatment. Maximum of viraemia was high in the former group, while in the latter group it was significantly lower. Therefore it is supposed that Poly IC in these experiments did not protect through an interferon mediated suppression of the viraemia but rather through an effect of the interferon exerted directly upon the target organ. A reduction of circulating HI antibodies was found in the group on which Poly IC had the most pronounced effect.     

Characterization ofEscherichia coli serotype O12∶K1∶H7 isolates from an immunocompetent carrier with a history of spontaneous abortion and septicemia

Escherichia coli isolates from an immunocompetent woman with a history of repeated amnion infections and spontaneous abortion were characterized.Escherichia coli were isolated from stool, blood and cervix swab samples taken over a 21-month period after the last abortion which followed septicemia during pregnancy. All samples except the last cervix swab contained isolates of serotype O12K1H7, which produced adhesins, P fimbriae, type I fimbriae and the iron-chelator aerobactin. Pulsed-field gel electrophoresis revealed identical Xbal restriction patterns of the O12K1H7 isolates, suggesting that one particularEscherichia coli strain was responsible for the severe extraintestinal infections during pregnancy. The strain was able to persist in the intestine of the woman despite antibiotic therapy.     

Analysis of γδ+ T cells in peripheral blood of children with perinatal human immunodeficiency virus (HIV) infection

The present study examined CD8 antigen expression and variable (V) gene segment usage by T cell receptor (TCR)-⁺ lymphocytes in peripheral blood of symptomatic children with perinatal HIV infection. The relative number of ⁺, CD8⁺ T cells in most of the infected children was higher than that in uninfected children from HIV⁺ or HIV^– mothers and correlated with the immunodeficiency status of the patients. Infected infants and children over 1 year old also showed an increased proportion of V1-J1⁺ T lymphocytes. CD8 expression on those cells was higher in infected than in uninfected infants and children. Sequence analysis of the gene rearrangement of the predominant V1 family in peripheral blood of three HIV⁺ donors revealed extensive junctional diversity. These results suggest that the V skewing in the majority of HIV⁺ children reflects peripheral expansion of V1-J1⁺ T lymphocytes early in life, which might be involved in the mechanisms of HIV-induced immunodeficiency.     

Mitoses in hypertrophied smooth muscle tissue of the rat posterior vena cava

Injection of colchicine into rats revealed mitotically dividing leiomyocytes in the hypertrophied muscle tissue of the posterior vena cava of rats. The frequency of dividing cells under these conditions did not exceed 11000–110,000. Many of these smooth muscle cells were in prophase.Laboratory of Experimental Pathomorphology and Ultrastructural Characteristics of Pathological Processes, Institute of General Pathology and Pathophysiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR, N. A. Kraevskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 4, pp. 485–487, April, 1976.