Antimicrob Agents Chemother, 1996 Oct, 40(10), 2252 - 7 ParC subunit of DNA topoisomerase IV of Streptococcus pneumoniae is a primary target of fluoroquinolones and cooperates with DNA gyrase A subunit in forming resistance phenotype; Munoz R et al.; The genes encoding the ParC and ParE subunits of topoisomerase IV of Streptococcus pneumoniae, together with the region encoding amino acids 46 to 172 (residue numbers are as in Escherichia coli) of the pneumococcal GyrA subunit, were partially characterized . The gyrA gene maps to a physical location distant from the gyrB and parC loci on the chromosome, whereas parC is closely linked to parE . Ciprofloxacin-resistant (Cpr) clinical isolates of S . pneumoniae had mutations affecting amino acid residues of the quinolone resistance-determining region of ParC (low-level Cpr) or in both quinolone resistance-determining regions of ParC and GyrA (high-level Cpr) . Mutations were found in residue positions equivalent to the serine at position 83 and the aspartic acid at position 87 of the E . coli GyrA subunit . Transformation experiments suggest that ParC is the primary target of ciprofloxacin . Mutation in parC appears to be a prerequisite before mutations in gyrA can influence resistance levels. Am J Obstet Gynecol, 1996 Oct, 175(4 Pt 1), 1036 - 42 Group B Streptococcus and preterm premature rupture of membranes: a randomized, double-blind clinical trial of antepartum ampicillin; Grable IA et al.; OBJECTIVE: Our purpose was to determine whether ampicillin prolongs the latency period after preterm premature rupture of membranes in patients colonized with group B Streptococcus . STUDY DESIGN: Sixty patients presenting at < or = 35 weeks' gestation with preterm premature rupture of membranes were included in the study . Cervical, vaginal, and perianal cultures for group B premature rupture were obtained . The participants then were randomized to receive either ampicillin or placebo intravenously for 24 hours and then orally until hospital discharge or delivery . All patients were treated without the use of tocolytic drugs . The chi(2) test, Fisher exact test, Student t test, and Wilcoxon signed-rank test were used for statistical analysis when appropriate . RESULTS: Fifteen patients had cultures positive for group B Streptococcus . Patients with cultures positive for group B Streptococcus who received ampicillin (n = 8) were more likely not to have been delivered of their infants 48 hours after preterm premature rupture of membranes than patients who received placebo (n = 7), a statistically significant difference (100% vs 43%; p = 0.01; relative risk 2.3; 95% confidence interval 1.2 to 4.5) . Seven days after preterm premature rupture of membranes, however, there was no significant difference in percentage of patients with cultures positive for group B Streptococcus who remained undelivered (63% vs 29%; p = 0.19; relative risk, 2.2; 95% confidence interval 0.7 to 7.1) . Among patients with cultures negative for group B Streptococcus, there was a trend for patients who received ampicillin to remain undelivered 48 hours after preterm premature rupture of membranes compared with those who received placebo, but the difference was not statistically significant (87% vs 64%; p = 0.07; relative risk, 1.4; 95% confidence interval 1.0 to 1.9) . There also was no difference in percentage of patients with cultures negative for group B Streptococcus who remained undelivered 7 days after preterm premature rupture of membranes 39% vs 27%; p = 0.40; relative risk, 1.4; 95% confidence interval 0.61 to 3.3) . There were no differences between the treatment and placebo arms of the group B Streptococcus positive and negative cohorts in incidence of cesarean section, chorioamnionitis, postpartum endometritis, or neonatal infectious morbidity . CONCLUSION: Use of antibiotics increases the percentage of patients with cultures positive for group B Streptococcus who remain undelivered 48 hours after preterm premature rupture of membranes . Antibiotic therapy may provide a window of opportunity for maternal treatment with corticosteroids to decrease the risk for neonatal morbidity among these preterm gestations. Microbiology, 1996 Oct, 142 ( Pt 10), 2747 - 57 Multiply antibiotic-resistant Streptococcus pneumoniae recovered from Spanish hospitals (1988-1994): novel major clones of serotypes 14, 19F and 15F; Coffey TJ et al.; We analysed a collection of 95 multiply antibiotic-resistant pneumococci, recovered since 1988 from 14 Spanish hospitals, that have MICs > or = 0.25 microgram benzylpenicillin ml-1 . The majority of the isolates were of serogroups 14, 23, 6, 19 and 15, which are currently the serogroups mainly associated with multiresistance in Spain . All of the serogroup 23 isolates were members of the major Spanish serotype 23F multiresistant clone . Similarly, most of the serogroup 6 isolates were members of the major multiresistant serotype 6B clone, or variants of this clone . Eighteen of the 24 isolates of serogroup 19 were members of a highly penicillin-resistant clone that appears to be a serotype 19F variant of the major Spanish serotype 23F multiresistant clone . Eighteen of the 25 isolates of serotype 14 were members of a previously uncharacterized highly penicillin-resistant clone . Thirteen of the 16 isolates of serogroup 15 were members of a single previously unreported clone of serotype 15F that had moderate levels of resistance to penicillin . Approximately 65% of the multiresistant pneumococci that are currently circulating in Spain were members of the three new clones of serotype 14, 15F and 19F that we describe here, or the previously described serotype 6B and 23F clones . The other 35% of isolates were minor variants of the major clones, unrelated minor clones, and unique isolates, many of which appeared to have arisen by horizontal gene transfer events. J Clin Microbiol, 1996 Oct, 34(10), 2609 - 12 Surrogate disks for predicting cefotaxime and ceftriaxone susceptibilities of Streptococcus pneumoniae; Barry AL et al.; Cefotaxime- and ceftriaxone-resistant Streptococcus pneumoniae is now appearing in some medical centers, but 30-micrograms cefotaxime or 30-micrograms ceftriaxone disks are not reliable for detecting such strains . Studies were undertaken to select another cephalosporin disk that might be used as a screening test that could be used in conjunction with a 1-micrograms oxacillin disk . A 30-micrograms cefuroxime disk is proposed: strains with zones > or = 28 mm in diameter are predictably susceptible to cefotaxime and ceftriaxone, and those with smaller zones should be further studied to confirm resistance to either drug . A 30-micrograms ceftizoxime disk may also be used as a screening test with zones > or = 26 mm indicating susceptibility, but cefuroxime disks are preferred. J Pediatr, 1996 Oct, 129(4), 602 - 4 Complications associated with severe invasive streptococcal syndrome; Montgomery VL et al.; Group A beta-hemolytic streptococcal sepsis may cause life-threatening disease . We describe a child with severe invasive streptococcal syndrome in whom severe respiratory failure and pulmonary pneumatoceles required extracorporeal life support . Physicians should be aware of the full spectrum of pathologic changes and life-threatening complications caused by group A beta-streptococcus. J Pediatr, 1996 Oct, 129(4), 529 - 36 Children hospitalized for varicella: a prevaccine review; Peterson CL et al.; OBJECTIVES: To describe varicella complications in healthy and previously ill children hospitalized for varicella and to explore trends in group A beta-hemolytic streptococcus complications of varicella . METHODS: A retrospective record review of children hospitalized for varicella between January 1, 1990, and March 31, 1994, was conducted in nine large acute care hospitals in Los Angeles County, California . RESULTS: We identified 574 children hospitalized for varicella in study hospitals during the 4.25-year study period (estimated risk of hospitalization, approximately 1 in 550 cases of varicella); 53% of the children were healthy before the onset of varicella and 47% were previously ill with underlying cancers or other chronic illnesses . Children were hospitalized for treatment of complications (n = 427, 74%) or for prophylactic antiviral therapy or observation (n = 147, 26%) . Systems involved in complications included skin/soft tissue (45%), neurologic (18%), respiratory (14%), gastrointestinal (10%), and hematologic, renal, or hepatic (8% or less) . The mean age of children with skin/soft tissue infections was 2.7 years (range < 1 to 16 years) compared with 4.7 years (< 1 to 18 years) for other complications . Children with skin/soft tissue and neurologic complications were more often previously healthy (p < 0.05), whereas those with respiratory complications were more often previously ill (p < 0.001) . Hospitalizations for skin/soft tissue infections increased during the study period . The proportion of complications as a result of group A beta-hemolytic streptococcus infection increased from 4.7% before 1993 to 12.2% for the remainder of the study period (p = 0.02) . CONCLUSIONS: Prior health status was predictive of the type of complications experienced by children with varicella requiring hospitalization . Our data suggest a recent increase in skin/soft tissue complications of varicella requiring hospitalization and an increase in the proportion of complications related to group A beta-hemolytic streptococcus . Wide-scale vaccine use should reverse this trend and reduce the overall impact of varicella on both healthy and previously ill children. Am Fam Physician, 1996 Oct, 54(5), 1634 - 6 Use of the rapid streptococcus test in extrapharyngeal sites; Bourgeois SD et al.; The rapid "strep" test is established as a valuable tool for the management of pharyngitis . Because of the recent increase in reports of serious, rapidly progressive streptococcal soft tissue infections, the test can also be useful in determining early management of skin infections . Patients with positive results can be treated immediately with specific therapy, usually with an excellent outcome . As an illustration, use of this technique in 15 patients is described. Obstet Gynecol, 1996 Oct, 88(4 Pt 1), 577 - 82 Sexual behavior and vaginal colonization by group B streptococcus among minority women; Newton ER et al.; OBJECTIVE: To test the hypothesis that sexual behaviors predict colonization of the vagina by group B streptococcus among minority women . METHODS: We conducted a prospective, descriptive study of 192 consecutive African-American (37%) and Hispanic women (63%) . Each woman underwent a detailed interview concerning sexual behavior . Separate specimens were taken from the endocervix, upper vagina, lower vagina, and anorectum and placed in selective broth media for isolation of group B streptococcus . Significant behavioral predictors of vaginal group B streptococcus colonization and heavy (3-4+) colonization were identified using stepwise logistic regression . RESULTS: The incidence of vaginal colonization was 39% and heavy colonization was 35% . Nineteen percent reported anal intercourse, 46% reported sex at least two times per week, and 21% reported more than one partner in the previous 30 days . The significant predictors of vaginal group B streptococcal infection were: African-American ethnicity, adjusted odds ratio (OR) 6.1 (95% confidence interval {CI} 2.5-15.1); presence of rectal group B streptococcus, adjusted OR 100.6 (95% CI 26.7-379.3); nulliparous, adjusted OR 3.6 (95% CI 1.4-9.5); and nonpregnant status, adjusted OR 3.9 (95% CI 1.3-12.2) . The significant predictors of heavy colonization were: more than one partner in the last 30 days, adjusted OR 2.6 (95% CI 1.2-5.6); and African-American ethnicity, adjusted OR 2.3 (95% CI 1.2-4.5) . Anal intercourse was associated with a reduced likelihood of vaginal group B streptococcal infection, adjusted OR 0.34 (95% CI 0.12-0.91) . CONCLUSION: Sexual behavior, especially anal intercourse, does not predict vaginal colonization by group B streptococcus . African-American women are more likely to have vaginal and heavy group B streptococcus colonization . Heavy vaginal colonization is associated with multiple partners in African-American women. FEMS Microbiol Lett, 1996 Oct 1, 143(2-3), 279 - 84 Investigation of a choline phosphate synthesis pathway in Streptococcus pneumoniae: evidence for choline phosphate cytidylyltransferase activity; Whiting GC et al.; In this study we have demonstrated the activity of a choline phosphate cytidylyltransferase in cell free extracts of Streptococcus pneumoniae . Southern blot analysis of restricted S . pneumoniae genomic DNA probed with the gene coding for the choline phosphate cytidylyltransferase of Saccharomyces cerevisiae demonstrated that there is homology between the S . cerevisiae cct gene and genomic DNA of S . pneumoniae . We believe that this enzyme is involved in the biosynthesis of the choline containing cell wall antigens, teichoic acid and lipoteichoic acid, catalysing the activation of choline phosphate to CDP-choline which is then incorporated into the polysaccharide moiety. J Bacteriol, 1996 Oct, 178(20), 6087 - 90 Competence for genetic transformation in encapsulated strains of Streptococcus pneumoniae: two allelic variants of the peptide pheromone; Pozzi G et al.; The nucleotide sequence of comC, the gene encoding the 17-residue competence-stimulating peptide (CSP) of Streptococcus pneumoniae (L . S . Havarstein, G . Coomaraswamy, and D . A . Morrison, Proc . Natl . Acad . Sci . USA 92:11140-11144, 1995) was determined with 42 encapsulated strains of different serotypes . A new allele, comC2, was found in 13 strains, including the type 3 Avery strain, A66, while all others carried a gene (now termed comC1) identical to that originally described for strain Rx1 . The predicted mature product of comC2 is also a heptadecapeptide but differs from that of comC1 at eight residues . Both CSP-1 and CSP-2 synthetic peptides were used to induce competence in the 42 strains; 48% of the strains became competent after the addition of the synthetic peptide, whereas none were transformable without the added peptides. J Bacteriol, 1996 Oct, 178(19), 5831 - 5 Natural genetic transformation in Streptococcus gordonii: comX imparts spontaneous competence on strain wicky; Lunsford RD et al.; Streptococcus gordonii Wicky becomes competent only after stimulation with conditioned medium from strain Challis as a source of competence factor (CF) . A 3.2-kbp genomic fragment from Challis was found to impart spontaneous competence on Wicky by a complementation assay . Wicky clones containing the fragment secreted a heat-sensitive activity that induced competence in Wicky and in a comA insertion mutant of Challis . Activity was localized to a putative open reading frame, comX, with the potential to encode a 52-amino-acid peptide . comX had no similarity to known sequences, and a comX::ermAM insertion mutant of Challis transformed normally and secreted CF . These data suggest that a CF-independent pathway for competence induction exists in S . gordonii. J Bacteriol, 1996 Oct, 178(19), 5826 - 30 Rgg is a positive transcriptional regulator of the Streptococcus gordonii gtfG gene; Sulavik MC et al.; The Streptococcus gordonii (Challis) glucosyltransferase-encoding determinant gtfG is regulated by the product of the adjacent gene rgg . Results of analyses described here showed that in both S . gordonii and Escherichia coli Rgg is a positive transcriptional regulator of glucosyltransferase expression . In addition, the transcriptional start sites of both gtfG and rgg were determined. Curr Microbiol, 1996 Oct, 33(4), 216 - 9 Catabolite regulation in a diauxic strain and a nondiauxic strain of Streptococcus bovis; Kearns DB et al.; Streptococcus bovis JB1 utilized glucose preferentially to lactose and grew diauxically, but S . bovis 581AXY2 grew nondiauxically and used glucose preferentially only when the glucose concentration was very high (greater than 5 mM) . As little as 0.1 mM glucose completely inhibited the lactose transport of JB1 . The lactose transport system of 581AXY2 was at least tenfold less sensitive to glucose, and 1 mM glucose caused only a 50% inhibition of lactose transport . Both strains had phosphotransferase systems (PTSs) for glucose and lactose . The glucose PTSs were constitutive, but little lactose PTS activity was detected unless lactose was the energy source for growth . JB1 had approximately threefold more glucose PTS activity than 581AXY2 (1600 versus 600 nmol glucose (mg protein)-1(min)-1 . The glucose PTS of JB1 showed normal Michaelis Menten kinetics, and the affinity constant (Ks) was 0.12 mM . The glucose PTS of 581AXY2 was atypical, and the plot of velocity versus velocity/substrate was biphasic . The low capacity system had a Ks of 0.20 mM, but the Ks of the high capacity system was greater than 6 mM . On the basis of these results, diauxic growth is dependent on the affinity of glucose enzyme II and the velocity of glucose transport. J Immunol, 1996 Oct 1, 157(7), 3021 - 9 A highly variable region in members of the streptococcal M protein family binds the human complement regulator C4BP; Johnsson E et al.; Strains of Streptococcus pyogenes express one or more molecules that are members of the M protein family, a group of surface proteins implicated in virulence . A characteristic property of the molecules in this family is the presence of a highly variable N-terminal region, whose function is unknown . Here we show that human C4b-binding protein (C4BP), a regulatory component of the complement system, binds to the highly variable region of many members of the M protein family . Chimeric molecules, in which the N-terminal regions of four different C4BP-binding proteins were combined with the C-terminal part of the non-binding M5 protein, had intact C4BP-binding ability, as judged by binding assays and Scatchard analysis with highly purified molecules . Moreover, work with the C4BP-binding Arp4 protein showed that an N-terminal 52-residue fragment retained binding ability, and that a 21-residue synthetic peptide derived from the variable region completely inhibited the binding of C4BP . Computer-assisted analysis of the four C4BP-binding regions studied here (45-66 amino acid residues) indicated that they lack residue identities that could explain their ability to bind the same ligand, but differ from the nonbinding M5 protein in their lower propensity to form a coiled-coil . Thus, the variable C4BP-binding regions have an extraordinary capacity for sequence variation, while retaining the ability to bind C4BP . These data indicate that an important function of the variable region in members of the M protein family is to bind a host protein that down-regulates the complement system. Vet Rec, 1996 Sep 28, 139(13), 308 - 13 Respiratory disease in thoroughbred horses in training: the relationships between disease and viruses, bacteria and environment; Burrell MH et al.; A longitudinal study of respiratory disease in racehorses was carried out to assess its relative associations with different infectious agents and to examine any role that the environmental conditions might play . The relationships between coughing, nasal discharge, pyrexia and lower respiratory tract disease were also examined to provide information for improving clinical diagnosis, particularly of disease of the lower respiratory tract . Lower airway disease was closely associated with infection with Streptococcus zooepidemicus . It was also found that equine herpesvirus seroconversions and S pneumoniae infections were independently associated with the development of nasal discharge . Coughing was a specific, but insensitive measure of lower respiratory tract disease (specificity 84 per cent, sensitivity 38 per cent) . However, horses that coughed were very likely to have had lower airway disease for more than one month . Horses housed on straw in loose boxes were twice as likely to suffer from lower airway disease as those kept on shredded paper in American barns . The study was not large enough to assess the significance of rarer infections but it did improve the definition of the problem of respiratory disease in racehorses and revealed some of the trends in the associations between viruses, bacteria and the environment in respiratory disease. Carbohydr Res, 1996 Sep 23, 291, 21 - 41 Efficient, convergent syntheses of oligosaccharide allyl glycosides corresponding to the Streptococcus group A cell-wall polysaccharide; Auzanneau FI et al.; Convergent syntheses of di-, tri, tetra-, penta-, and hexa-saccharide allyl glycosides corresponding to the beta-hemolytic Streptococcus Group A cell-wall polysaccharide are described . The strategy relies on the preparation of related di- and tri-saccharide building blocks: beta-D-Glc pNAc-(1-3)-alpha-L-Rhap and alpha-L-Rhap-(1-2)-{(beta-D-Glc p NAc-(1-3)}-alpha-L-Rhap, which could be used either as glycosyl donors or acceptors in subsequent glycosylation reactions . The protecting groups were chosen to allow the selective removal of the allyl aglycon to access the intermediate glycosyl donors but also to allow their own removal without affecting the allyl group . The allyl group was intended for use in conjugation of the oligosaccharides to soluble protein carriers or solid supports for the preparation of antigens and immunoadsorbents, respectively. J Biol Chem, 1996 Sep 20, 271(38), 23395 - 9 Functional cloning of the cDNA for a human hyaluronan synthase; Shyjan AM et al.; Hyaluronan is a constituent of the extracellular matrix of connective tissue and is actively synthesized during wound healing and tissue repair to provide a framework for ingrowth of blood vessels and fibroblasts . Changes in the serum concentration of hyaluronan are associated with inflammatory and degenerative arthropathies such as rheumatoid arthritis . In addition, hyaluronan has been implicated as an important substrate for migration of adhesion of leukocytes during inflammation . A human hyaluronan synthase (HuHAS1) cDNA was isolated by a functional expression cloning approach . Transfection of CHO cells conferred hyaluronidase-sensitive adhesiveness of a mucosal T cell line via the lymphocyte hyaluronan receptor, CD44, as well as increased hyaluronan levels in the cultures of transfected cells . The HuHAS1 amino acid sequence shows considerable homology to the hasA gene product of Streptococcus pyogenes, a glycosaminoglycan synthetase from Xenopus laevis (DG42), and is the human homolog of a recently described murine hyaluronan synthase. J Biol Chem, 1996 Sep 20, 271(38), 22945 - 8 Molecular identification of a putative human hyaluronan synthase; Watanabe K et al.; To identify the putative mammalian hyaluronan synthase, we cloned a human cDNA that is related to the Streptococcus hyaluronan synthase (HasA) and the Xenopus developmental protein DG42 which has been shown to have chitin synthase activity . The cDNA, for which we propose the name Has2, encodes a novel protein with a predicted molecular mass of 63.6 kDa . Has2 shows 55% amino acid identity with Xenopus DG42 and 52% identity with the mouse HAS protein, another putative hyaluronan synthase recently reported by Itano and Kimata (Itano, N., and Kimata, K . (1996) J . Biol . Chem . 271, 9875-9878) . The deduced primary structure revealed the presence of several hydrophobic stretches which can form multiple transmembrane domains . It also demonstrated the complete conservation of amino acid residues that are known to be critical for N-acetylglucosaminyltransferase activity of yeast chitin synthase . When the Has2 cDNA was transfected into human 293 and Chinese hamster ovary cells, the production of hyaluronan in the transfected cells increased up to 34- and 9-fold, respectively . Strong expression of Has2 mRNA was observed in exponentially proliferating human IMR-90 fibroblasts but not in growth-arrested IMR-90 cells . These results suggest that the Has2 protein is a crucial component of the human hyaluronan synthase system. FEMS Microbiol Lett, 1996 Sep 15, 143(1), 69 - 76 Cloning and characterization of chsD, a chitin synthase-like gene of Aspergillus fumigatus; Mellado E et al.; A chitin synthase-like gene (chsD) was isolated from an Aspergillus fumigatus genomic DNA library . Comparisons with the predicted amino acid sequence from chsD reveals low but significant similarity to chitin synthases, to other N-acetylglucosaminyltransferases (NodC from Rhizopus spp., HasA from Streptococcus spp . and DG42 from vertebrates . A chsD- mutant strain constructed by gene disruption has a 20% reduction in total mycelial chitin content; however, no differences between the wild-type strain and the chsD- strain were found with respect to morphology, chitin synthase activity or virulence in a neutropenic murine model of aspergillosis . The results show that the chsD product has an important but inessential role in the synthesis of chitin in A . fumigatus. J Immunol, 1996 Sep 15, 157(6), 2479 - 87 Species-dependent post-translational modification and position 2 allelism: effects on streptococcal superantigen SSA structure and V beta specificity; Stevens KR et al.; Epidemiologic and molecular population genetic analyses support a role for superantigens (SAg) in the pathogenesis of severe staphylococcal and streptococcal infections . To investigate how variations in SAg structure influence immunomodulatory activity, we examined the biochemical and functional properties of two allelic variants of streptococcal SAg SSA that differ at position 2 . Mass spectrometry revealed both recombinant (Escherichia coli) and native (Streptococcus pyogenes) SSA allelic variants to have significantly larger molecular masses than predicted by primary sequence alone and provided evidence that the proteins were modified by the addition of biochemical moieties, a phenomenon that has not been described for related SAg . Furthermore, the molecular masses of native and recombinant SSA were not the same; SSA was differentially post-translationally modified by the two bacterial genera . The substitution of E . coli-dependent processing for that of S . pyogenes altered both protease digestion and V beta specificity, suggesting that recombinant SAg from E . coli may not accurately represent the native toxin . In addition, the observation that SSA allelic variants differed in V beta specificity supports a role for position 2 in SSA-TCR interactions . That SSA position 2 contributes to V beta specificity could not have been predicted from functional or crystallographic studies of other SAg and suggests that SSA may adopt unique interactions with TCR and/or MHC class II molecules . Determining the structural basis for these differences should offer additional clues to the manner in which SAg exert their effects on the immune system during infection and may allow the designing of SAg mutants with specific quantitative and qualitative immunomodulatory properties. J Biol Chem, 1996 Sep 13, 271(37), 22414 - 21 The ColE1 unidirectional origin acts as a polar replication fork pausing site; Viguera E et al.; Co-orientation of replication origins is the most common organization found in nature for multimeric plasmids . Streptococcus pyogenes broad-host-range plasmid pSM19035 and Escherichia coli pPI21 are among the exceptions . pPI21, which is a derivative of pSM19035 and pBR322, has two long inverted repeats, each one containing a potentially active ColE1 unidirectional origin . Analysis of pPI21 replication intermediates (RIs) by two-dimensional agarose gel electrophoresis and electron microscopy revealed the accumulation of a specific RI containing a single internal bubble . The data obtained demonstrated that initiation of DNA replication occurred at a single origin in pPI21 . Progression of the replicating fork initiated at either of the two potential origins was transiently stalled at the other inversely oriented silent ColE1 origin of the plasmid . The accumulated RIs, containing an internal bubble, occurred as a series of stereoisomers with different numbers of knots in their replicated portion . These observations provide one of the first functional explanations for the disadvantage of head-to-head plasmid multimers with respect to head-to-tail ones. Arch Intern Med, 1996 Sep 9, 156(16), 1851 - 6 Roentgenographic findings of pneumonia caused by Chlamydia pneumoniae . A comparison with streptococcus pneumonia; Kauppinen MT et al.; BACKGROUND: Pneumonia caused by Chlamydia pneumoniae or Streptococcus pneumoniae cannot be reliably differentiated by clinical signs or symptoms . OBJECTIVE: To find differences in the roentgenographic patterns of community-acquired pneumonia caused by C pneumoniae, S pneumoniae, or both in hospitalized patients during a C pneumoniae epidemic in Finland . METHODS: The patients were divided into 3 groups: 24 patients with serologic evidence of C pneumoniae only; 8 patients with combined C pneumoniae and S pneumoniae infection; and 13 patients with infection caused by S pneumoniae only . The chest roentgenograms obtained on admission to the hospital, during the hospital stay, and at follow-up visits were reevaluated by one of us (S.L.) who was unaware of the causative organism . In the final study groups, other causes of community-acquired pneumonia were excluded by a large pattern of microbiological methods . RESULTS: Bronchopneumonia was observed in 21 (88%) of the group with C pneumoniae and 10 (77%) of the group with S pneumoniae (P = .67) . Lobar or sublobar (air space) pneumonia was seen in 7 (29%) of the patients with C pneumoniae compared with 7 (54%) with pneumonia caused by S pneumoniae . In the combined group, bronchopneumonia was seen as frequently as in the group with C pneumoniae, and air-space involvement was seen as frequently as in the group with S pneumoniae . The pneumonic shadowing was usually unilateral and in the lower lobes in all groups . Of the patients in the C pneumoniae group, 17% had residual abnormalities at follow-up visits . CONCLUSIONS: Roentgenographic changes cannot be used to differentiate pneumonia caused by C pneumoniae from that caused by S pneumoniae . Thus, initial antibiotic treatment should be directed at the pathogens that commonly cause community-acquired pneumonia. Vet Rec, 1996 Sep 7, 139(10), 225 - 8 Protection of experimentally infected pigs by suilysin, the thiol-activated haemolysin of Streptococcus suis; Jacobs AA et al.; Three groups of three pigs were vaccinated either with vaccine VAC-SLY, containing purified suilysin derived from Streptococcus suis strain P1/7 (serotype 2), or with vaccine VAC-SCF, containing most of the other extracellular antigens produced by strain P1/7 (but essentially free from suilysin), or with a placebo vaccine . The pigs were vaccinated twice at four weeks and six weeks of age and were challenged intravenously with S suis strain P1/7 at eight weeks of age . On the day of challenge, only the VAC-SLY vaccinated pigs showed an increase in haemolysin neutralisation titre . After challenge the placebo vaccinated pigs developed severe clinical signs characterised by lameness involving several joints, a depressed appearance, high temperatures and/or neurological signs . The VAC-SCF vaccinated pigs showed the same clinical signs but less severely . The VAC-SLY vaccinated pigs were the least affected and showed only mild signs which subsided more quickly than those of the other groups . A post mortem investigation and histology of brain tissue samples confirmed the clinical findings; fibrinous arthritis was less severe and less frequently observed in the VAC-SLY vaccinated pigs than in the VAC-SCF or placebo vaccinated pigs, and none of the VAC-SLY vaccinated pigs had meningitis whereas two of the VAC-SCF and two of the placebo vaccinated pigs did so . All the samples of brain, lung and tarsus taken from the VAC-SLY vaccinated pigs were sterile whereas S suis was reisolated from most of these tissues from the other groups. Vojnosanit Pregl, 1996 Sep-Oct, 53(5), 383 - 6 {Penicillin resistance in strains of Streptococcus pneumoniae}; Tomanovic B et al.; The values of minimal inhibitory concentration (MIC) of penicillin were determined using the agar-dilution method for 114 strains of Streptococcus pneumoniae isolated during 1994 in the patients treated in hospital and outpatient clinics . Diminished susceptibility to penicillin was determined in 50 (43.8%) strains . High level of penicillin resistance was observed in 12 (10.5%) strains . Streptococcus pneumoniae cannot any more be regarded as the agent with presumed susceptibility to penicillin, but every isolated strain must be tested. Mikrobiologiia, 1996 Sep-Oct, 65(5), 656 - 62 {Biology of lysogenic strains of Streptococcus bovis and virulent mutants of their temperate phages}; Tarakanov BV; Three lysogenic streptococcal strains and four virulent mutants of temperate phages were studied . The bacterial strains proved to be close to the type strain of Streptococcus bovis in their morphological, cultural, physiological, and biochemical properties . Investigation into the interaction of the lysogenic cultures with virulent mutants of temperate phages showed that the culture age optimal for infecting was 3-5 h; intense lysis began in the second hour after infection and was virtually completed by the end of the third hour, i.e., the procedure of obtaining phages in HMT medium took 7-8 h . Phage titers in phage lysates varied from 3.93 x 10(10) to 11.25 x 10(10) active phage particles per ml; residual amounts of viable bacteria varied from 0.7 x 10(6) to 34.0 x 10(6) cells per ml . In liquid HMT medium, phage production by lysogenic cultures was not limited by 0.3% glucose or maltose, 0.5% peptone, or casein hydrolysate . Descriptions of virulent mutants VM 6/6, VM 32/6, VM 28/28, and VM 54/54 of temperate phages of Str . bovis are presented. Zh Mikrobiol Epidemiol Immunobiol, 1996 Sep-Oct, (5), 7 - 10 {The phage sensitivity and lysogeny of cultures of Streptococcus pyogenes group A isolated in different streptococcal infections}; Asoskova TK et al.; The collection of moderate phages of S . pyogenes, group A, had been created earlier . As shown in this work, group A streptococcal cultures isolated from patients with rheumatism, glomerulonephritis and tonsillitis exhibited different sensitivity to the phages of this collection: the cultures were lyzed by phages of groups II and III in rheumatism, group III in tonsillitis and group I in glomerulonephritis . The study revealed that lysogeny was widely spread among S . pyogenes strains isolated from patients with different diseases under study . Most frequently occurred among cultures isolated from tonsillitis patients . In this disease only phage-resistant streptococcal cultures proved to be lysogenic . Lysogeny was found among both phage-sensitive and phage-resistant cultures in rheumatism and especially in glomerulonephritis. Pediatr Med Chir, 1996 Sep-Oct, 18(5), 433 - 50 {Colonization by group B hemolytic streptococcus in pregnancy . Note of prevention and therapy of the materno-neonatal infection . Casuistics}; Della Morte MA et al.; As several international studies show, the knowledge of the wide clinical spectrum of perinatal group B streptococcal infection, particularly of the early and of the late-onset neonatal diseases in GBS carrier mothers, is basically important for medical diagnosis . Risk factors analysis further determines both the diagnosis and the maternal intrapartum chemoprophylaxis . The considerable rate of neonatal disease without risk factors and its possible serious and fatal consequences bring to tendentially non selective prevention approaches that must consider the local background . At Merate Hospital, in a 3 years time, vaginal and rectal specimens for GBS cultures were obtained from 1766 pregnant women either at the 32nd or at the 36th week of gestation and regularly at the labor . 376 women (21.29 percent) resulted GBS carriers; the maternal-fetal contamination rate was 15.42 percent (58/376) i.e . 32.6 per 1000 live births (58/1769) . Intrapartum chemoprophylaxis was carried out with i.v . erytromycin, i.v . or i.m . cephalosporins, i.v . ampicillin and per os amoxicillin (which gave the most interesting results) . In infants born to mothers who received an antibiotic therapy at labor as compared with those who received no treatment, GBS neonatal colonization was present in 31 of 286 (10.8 percent) versus 27 of 90 (30 percent; P < 0.001); heavy colonization was observed in 10 of 286 (3.4 percent) versus 15 of 90 (16.6 percent; P < 0.001) and early-onset neonatal disease (both symptomatic and asymptomatic) occurred in none of 286 versus 4 of 90 (4.44 percent; P = 0.0031) . Perinatal risk factors (no-screened mothers, labor at 36th week of gestation, prolonged membrane rupture) were present only in 1 of 4 GBS infected infants (25 percent) . Intrapartum therapy both in carriers and in no-screened women significantly reduced GBS neonatal colonization, particularly the heavy one and, consequently, the early-onset neonatal group B streptococcal disease. Pathologe, 1996 Sep, 17(5), 391 - 5 {Polypoid ulcerating endocarditis aortalis caused by Streptococcus viridans with perforation of the right atrium . Pathology and clinical aspects of 2 autopsy cases}; Poremba C et al.; We report two cases of patients (one 65 and one 43 years of age, respectively) who died of Streptococcus-viridans induced endocarditis of the aortic valve with perforation into the right atrium . Whereas perforation in Staphylococcus-induced endocarditis is a common complication, it occurs rarely in Streptococcus-induced endocarditis . Because of its uncharacteristic symptoms, the endocarditis was clinically unknown in both cases and was recognized to be the cause of death only at autopsy . To reduce the large number of complications in patients suffering from endocarditis, it is necessary to confirm the diagnosis as soon as possible if endocarditis might be suspected. Minerva Pediatr, 1996 Sep, 48(9), 397 - 400 {Hepatic abscesses caused by Streptococcus intermedius}; Alessandri C et al.; Pyogenic abscess of the liver is uncommon child's pathology . The authors briefly describe a clinical picture characterized by beginning of an hepatic abscess dues to a germ that is not usually pathogen for men . It is often a mouth saprophyte. Acta Otorrinolaringol Esp, 1996 Sep-Oct, 47(5), 401 - 3 {Acute sinusitis, bacteremia and meningitis caused by group F beta-hemolytic Streptococcus}; Perez C et al.; A 32-year old man without immunodepression developed sinusitis followed by acute pyogenic meningitis and beta-hemolytic Streptococcus group F bacteremia . Amplicillin treatment produced a favorable outcome with sequela of righ neurosensorial deafness . Beta-hemolytic Streptococcus group F meningitis, sinusitis and bacteremia is a rare association. Rev Esp Enferm Dig, 1996 Sep, 88(9), 605 - 8 {Bacteremia and endocarditis caused by Streptococcus bovis in patients with alcoholic hepatopathy without evidence of colonic pathology}; Castroagudin JF et al.; The association of Streptococcus bovis bacteremia and endocarditis with colonic pathology, mainly neoplastic, is well known . Its relationship with liver disease without evidence of gastrointestinal disease has been rarely described . To analyze the association between S . bovis infection and liver disease, positive blood cultures for this microorganism in hospitalized patients in the Internal Medicine and Gastroenterology Departments from December 1993 until October 1995, have been reviewed . Three cases of S . bovis infection (one bacteremia, two endocarditis) were found . Alcoholic liver disease was diagnosed in all three patients, with associated hepatitis C virus in one of them . Colonic pathology was excluded by colonoscopy and/or barium enema . Other gastrointestinal disorders were excluded by means of gastroscopy, barium gastrointestinal study and abdominal ultrasonography . Antibiotic therapy was based in betalactamics, with associated aminoglycoside in two cases . One patient needed aortic and mitral valve replacement and another one needed orthotopic liver transplantation . No new gastrointestinal pathology emerged in the follow-up (5-23 months) . Cases of S . bovis bacteremia and endocarditis should be screened not also for colonic pathology, but also for liver disease, particularly in alcoholics. West Indian Med J, 1996 Sep, 45(3), 95 - 6 Penicillin-resistant Streptococcus pneumoniae in a Jamaican patient with homozygous sickle-cell disease; Wierenga KJ et al.; Penicillin prophylaxis against infection by Streptococcus pneumoniae is now routine in young children with homozygous sickle-cell (SS) disease and the emergence of penicillin-resistant strains is a serious clinical concern . The first death associated with such resistance in a Jamaican child with SS disease is reported. Diagn Microbiol Infect Dis, 1996 Sep, 26(1), 23 - 7 Tentative interpretive criteria for testing the susceptibility of Streptococcus pneumoniae to eight fluoroquinolones; Fuchs PC et al.; Broth microdilution and disk diffusion methods were used to test ciprofloxacin, clinafloxacin, fleroxacin, grepafloxacin, ofloxacin, PD131628, sparfloxacin, and trovafloxacin against Streptococcus pneumoniae isolates . With each fluroquinolone tested, there was a striking unimodal population . With the exception of fleroxacin, over 96% of pneumococci tested were inhibited by each drug's modal MIC +/-1 twofold concentration . Over 92% of pneumococci produced inhibitory zone diameters equal to the mode +/-4 mm for each drug . For most fluoroquinolones, the interpretive criteria approved or proposed for nonfastidious organisms appear to be appropriate for pneumococci . For clinafloxacin and trovafloxacin, however, the proposed MIC breakpoints for nonfastidious organisms are four-to eightfold greater than the highest MIC encountered with pneumococci . For future surveillance efforts, low concentrations must be evaluated to detect any subtle shift in pneumococcal populations that might be undetected by testing only breakpoint concentrations. Rev Prat, 1996 Sep 1, 46(13), 1593 - 8 {Erysipelas and impetigo}; Cribier B; Erysipela is a dermal or hypodermal infection of the skin, which predominantly involves the leg and is associated with high fever . Erysipela is most often caused by Streptococcus pyogenes . Venous insufficiency or lymphoedema are important local factors for the development of this infection which spreads from intertrigo, local wound or leg ulcer . Treatment is essentially based on parenteral penicillin G . Impetigo is a superficial infection of the skin due to Staphylococcus aureus or to Streptococcus pyogenes, and is frequent in children . Classical impetigo is made of yellow-brown crusts located around the mouth and nose, whereas bullous impetigo involves frequently the trunk and limbs . Secondary impetigo occurring in pediculosis or scabiosis is frequent . It is a contagious disease which is more frequent in patients with poor hygiene . It can be treated by general antibiotics, mainly macrolides, penicillin M or cephalosporins. Eur J Clin Microbiol Infect Dis, 1996 Sep, 15(9), 718 - 24 Azithromycin versus penicillin V in the treatment of paediatric patients with acute streptococcal pharyngitis/tonsillitis . Paediatric Azithromycin Study Group; O'Doherty B; The efficacy and safety of azithromycin and penicillin V in the treatment of acute streptococcal pharyngitis/tonsillitis in paediatric patients were compared in a double-blind, double-dummy prospective study . A total of 489 children (age range, 2-13 years) were randomized to receive treatment with penicillin V (125-250 mg 4 x daily for 10 days) or azithromycin in an oral suspension (10 or 20 mg/kg 1 x daily for 3 days) . Only patients with baseline cultures positive for Streptococcus pyogenes and complete clinical and microbiological assessments at the end of the therapy and follow-up one month later were included in the efficacy analysis . A satisfactory clinical response (cure or improvement) was recorded in 99% of the 10 mg/kg azithromycin group, 100% of the 20 mg/kg azithromycin group, and 97% of the penicillin V group at the end of therapy (day 12-14) . At the follow-up evaluation (day 28-30), relapse rates in patients cured or improved at the end of therapy were 6%, 5%, and 2%, respectively . Bacteriological eradication rates at the end of therapy were 98% in both azithromycin groups and 92% in patients who received penicillin V (p = 0.011); pathogen recurrence was recorded at follow-up in 4% of the 20 mg/kg azithromycin group and in 6% of both the 10 mg/kg azithromycin and penicillin V groups . Treatment-related adverse events, the majority of mild to moderate severity, occurred in 13% of patients in the 20 mg/kg azithromycin group, 9% in the 10 mg/kg azithromycin group, and 5% in the penicillin V group . Azithromycin in a dosage of 10 or 20 mg/kg/day one daily for three days was as safe and effective as penicillin V administered four times daily in the treatment of paediatric patients with acute pharyngitis/tonsillitis. J Nurse Midwifery, 1996 Sep-Oct, 41(5), 355 - 63 Group B streptococcus in the perinatal period . A review; Clay LS; Prior to the 1960s, little was known about morbidity and mortality in humans due to group B streptococcus (GBS) . Since that time, a thorough understanding of the organism's potential has been developed . Of pregnant women, 20-30% are prenatal carriers of GBS and 40-75% of these mothers will transmit GBS to their neonates . However, the actual incidence of GBS disease in infants is one to three cases per 1,000 live births . Several risk factors for transmission and development of early-onset GBS disease have been identified . The understanding of late-onset disease is less clear . Many strategies have been investigated to find an optimal protocol to reduce significantly the incidence of GBS disease in a cost-effective manner . The Centers for Disease Control and Prevention have recently made recommendations for prevention strategies against this disease. J Dermatol, 1996 Sep, 23(9), 628 - 30 Successful treatment of molluscum contagiosum in the immunosuppressed adult with topical injection of streptococcal preparation OK-432; Inui S et al.; A 37-year-old Japanese woman received an anticancer drug for 5 years following resection of mammary cancer and then developed widespread mollusca contagiosa, which we considered to be caused by immunosuppression induced by the chemotherapy . Because OK-432 (penicillin-treated and heat-treated lyophilized powder by a substrain of Streptococcus pyogenes A) was expected to be effective for immunosuppression, we tried its topical injection . The skin lesions disappeared almost completely within three months . OK-432 therapy is considered hopeful for treating viral skin diseases, even in immunosuppressed patients. J Perinatol, 1996 Sep-Oct, 16(5), 346 - 51 Effect of intravenous immunoglobulin G on the deposition of immunoglobulin G and C3 onto type III group B streptococcus and Escherichia coli K1; Lassiter HA et al.; Seventeen neonates with suspected or proven sepsis received either a 750 mg/kg dose of intravenous immunoglobulin G (IVIG) or placebo . Compared with values in adult serum, the preinfusion serum concentrations of immunoglobulin G (IgG) and complement component C3 were diminished; the concentrations were unaffected by the administration of placebo to nine infants . Fifteen minutes after infusion in the eight IVIG recipients, the serum concentration of IgG increased from 3.66 mg/ml to 16.58 mg/ml but the C3 concentration of 540 micrograms/ml was unaffected . Similarly, a radioimmunoassay revealed that during incubation of bacteria with sera from the neonates, the quantities of IgG and C3 bound to type III group B streptococcus and Escherichia coli O7:K1: NM were low and were unaffected by the infusion of placebo . During incubation of bacteria with the postinfusion sera from the IVIG recipients, the amount of IgG, but not C3, deposited onto the bacteria increased to a level equivalent to that observed in adult serum . Therefore IVIG enhanced the capacity of sera from ill neonates to deposit IgG but not C3 onto bacteria . We speculate that in neonates with sepsis, a diminished capacity to deposit C3 onto bacteria may possibly limit the therapeutic efficacy of IVIG. Immunol Invest, 1996 Sep-Nov, 25(5-6), 387 - 96 Nasal-associated lymphoid tissue is an inductive site for rat tear IgA antibody responses; Carr RM et al.; The role of nasal-associated lymphoid tissue (NALT) as a mucosal inductive site for tear IgA antibody responses was investigated in the rat model . Fluorescent microspheres were shown to access and be taken up by NALT after intranasal of ocular-topical administration, although fewer microspheres were found in the latter case . Tear IgA anti-DNP antibody responses to dinitrophenylated Streptococcus pneumoniae were 6 micrograms/ml at day 7, 10 micrograms/ml at day 10, and were still detectable on day 21 (5 micrograms/ml) following ocular or gastrointestinal immunization . Intranasal immunization induced tear IgA responses which were 1.7-fold higher at day 7 (10 micrograms/ml), peaked by day 10 (14 micrograms/ml) and were still 1.6-fold higher (8 micrograms/ml) at day 21 than responses of ocular or gastrointestinal groups . These findings suggest that intranasal immunization may be more effective than ocular or gastrointestinal administration in eliciting tear IgA antibody responses and, taken together with the microsphere data, indicate that NALT can serve as an inductive site for ocular mucosal IgA responses. Vet Microbiol, 1996 Sep, 52(1-2), 113 - 25 Detection of antibodies against Streptococcus suis capsular type 2 using a purified capsular polysaccharide antigen-based indirect ELISA; del Campo Sepulveda EM et al.; In the present study a purified capsular polysaccharide antigen-based indirect ELISA (CPS-ELISA) to detect antibodies against Streptococcus suis capsular type 2 was developed and compared with a whole cell antigen-based ELISA (WCA-ELISA) . The WCA-ELISA presented a very low specificity when rabbit antisera to other capsular types were tested . Most of these cross-reactions were due to common proteins . The standardized CPS-ELISA gave satisfactory results using a concentration of 0.1 micrograms/well; most cross-reactions decreased significantly, with some exceptions, such as those shared by capsular types 1/2, 12 and 17 . These cross-reactions were mainly due to common epitopes present in the capsule, as shown by immunoblotting . In a second experiment, the CPS-ELISA was used to detect antibodies in experimentally infected piglets . Despite the fact that capsular type 2 S . suis could be reisolated from all infected animals during and/or after the trial, antibody titers against a second infection . Sera from piglets experimentally infected were completely protected against a second infection . Sera from piglets experimentally infected with S . suis capsular types 1/2 or 12 presented cross-reactions at low dilutions, confirming data previously obtained with rabbit sera . Finally, sera of animals from herds with clinical signs associated with S . suis capsular type 2 did not present titers significantly different from those of disease free herds . From our results we concluded that the CPS-ELISA developed in this study can not be used as a diagnostic tool to identify infected animals. J Dairy Sci, 1996 Sep, 79(9), 1683 - 8 Efficacy of teat dips containing a hypochlorous acid germicide against experimental challenge with Staphylococcus aureus and Streptococcus agalactiae; Boddie RL et al.; Two teat dip formulations containing sodium dichloroisocyanurate, which released hypochlorous acid (2800 ppm) as the active ingredient, were tested for efficacy against new Staphylococcus aureus and Streptococcus agalactiae IMI using an experimental challenge model . Product 1 reduced the number of new Staph . aureus IMI by 73.6% and reduced the number of new Strep . agalactiae IMI by 65.1% . Product 2 reduced the number of new Staph . aureus IMI by 69.0% and reduced the number of new Strep . agalactiae IMI by 63.5% . No adverse effects on teat skin condition were observed over the course of the studies. J Antimicrob Chemother, 1996 Sep, 38(3), 507 - 21 Treatment of endocarditis with teicoplanin: a retrospective analysis of 104 cases; Wilson AP et al.; Infective endocarditis is an uncommon disease but retains a high mortality . Glycopeptides are used for patients with resistant pathogens, those allergic to penicillins or for those outside the hospital . The once daily administration of teicoplanin and its low toxicity suggest that it would be suitable for use in the long courses required for endocarditis . However, the dosage and combinations to be used require further study . A retrospective review has been made of 104 episodes of endocarditis treated with teicoplanin in 101 patients seen over 7 years . Most patients had been referred to major London hospitals following failure of medical treatment . After three loading doses of 400 mg, teicoplanin was given at a dose of 400 mg/day in combination with other antibiotics such as gentamicin . Follow up was for one year . The most common pathogens were Streptococcus sanguis (15 cases), Staphylococcus aureus (13 cases) and Staphylococcus epidermidis (10 cases) . Of 80 patients febrile at the start of treatment with teicoplanin, 63 (79%) lost their fever within a median of 2 days (1-35 days) . Cure without surgery was effected in 50 (48%) and 75% of patients survived . Other antibiotics, usually gentamicin or rifampicin, were used in 92 (90%) of patients . Two strains of Streptococcus spp . were said to be resistant but there was no relationship between MIC of teicoplanin and outcome . Pathogens with a high MBC tended to be more likely to resist treatment . Adverse effects resulted in the withdrawal of teicoplanin in 20 cases (19%) but most events were mild and renal deterioration occurred in only five patients . Teicoplanin was effective in the treatment of endocarditis and appeared to be safe given the severity of disease in the patients treated. Drug Metab Dispos, 1996 Sep, 24(9), 1028 - 31 Hemodynamic effects of N-acetylamrinone in a porcine model of group B streptococcal sepsis; Allen EM et al.; High plasma concentrations of N-acetylamrinone, a primary metabolite of amrinone, are measured in some children during prolonged amrinone infusion . The purpose of this investigation was to determine if N-acetylamrinone has direct hemodynamic effects independent of amrinone . Twenty neonatal piglets received an infusion of 6 x 10(9) colony-forming units/kg of group B Streptococcus to induce sepsis . Subsequently, they were divided into 1 of 3 groups and received a 1-hr infusion of either normal saline (N = 4); 8 mg/kg amrinone, followed by 20 micrograms/kg/min (N = 9); or 8 mg/kg N-acetylamrinone, followed by 20 micrograms/kg/min (N = 7) . Hemodynamic measurements and arterial/venous blood-gas determinations were obtained every 30 min during the study . Systemic vascular resistance and pulmonary vascular resistance were calculated . One milliliter of blood was obtained every 30 min during drug administration to determine plasma amrinone and N-acetylamrinone concentrations . The mean amrinone plasma concentrations measured at 30 and 60 min during the infusion time in the group receiving amrinone were 8.8 +/- 1.1 and 6.9 +/- 0.7 micrograms/ml, respectively . These animals experienced a significant decrease in mean pulmonary artery pressure and pulmonary vascular resistance, compared with saline controls after a 30-min infusion of amrinone . The mean N-acetylamrinone plasma concentrations measured at 30 and 60 min during the N-acetylamrinone infusion were 7.3 +/- 0.8 and 5.7 +/- 0.6 micrograms/ml, respectively . There was no difference between any hemodynamic parameter measured in these animals, compared with saline controls at any time during the infusion . We conclude that amrinone, but not N-acetylamrinone, causes pulmonary vasodilation in a porcine model of sepsis and that the parent drug is the sole active component in amrinone. Zentralbl Veterinarmed B, 1996 Sep, 43(7), 385 - 92 Adherence of Streptococcus uberis to bovine mammary epithelial cells and to extracellular matrix proteins; Almeida RA et al.; Adherence of an encapsulated (UT 101) and a non-encapsulated (UT 102) strain of Streptococcus uberis to a bovine mammary epithelial cell line (MAC-T) and to extracellular matrix proteins (ECMP) including fibronectin, collagen and laminin was investigated . S . uberis was co-cultured at 4 degrees C with MAC-T cell monolayers . Both strains of S . uberis adhered to MAC-T cells . However, the non-encapsulated strain of S . uberis adhered better to MAC-T cells than the encapsulated strain . Preincubation of MAC-T cells with lipoteichoic acid (LTA) and/or treatment of S . uberis with antibodies directed against the carboxyl-terminal half of type 24 M protein reduced adherence of both strains of S . uberis to MAC-T cells . Adherence to ECMP was measured by incubating bis-carboxyethyl-carboxyfluorescein acetomethyl ester (BCECF-AM) labelled S . uberis in 96-well plates coated with fibronectin, collagen or laminin . Both strains adhered to ECMP, however, the encapsulated strain adhered better to ECMP than the non-encapsulated strain . Results of this investigation demonstrated that both strains of S . uberis evaluated were capable of adhering to bovine mammary epithelial cells and to ECMP . Adherence of S . uberis to mammary epithelium may be an extremely important mechanism in the establishment and progression of bovine intramammary infections. FEMS Immunol Med Microbiol, 1996 Sep, 15(2-3), 81 - 91 Isolation of a new superantigen with potent mitogenic activity to murine T cells from Streptococcus pyogenes; Nemoto E et al.; A mitogenic substance on murine lymphocytes was detected in the culture supernate of Streptococcus pyogenes type 12 strain . This substance had a molecular weight of 28,000 and pI 9.2, and was designated as S . pyogenes mitogen (SPM) . The proliferative response of C3H/HeN spleen cells began at 1 ng ml-1 and reached a maximal response at 100 ng ml-1 of SPM for 4 days culture . Anti-Thy 1.2 mAb and complement-treated spleen cells abrogated the proliferative response to any dose of SPM . Although the anti-major histocompatibility complex class 1 mAbs had no blocking effect on proliferation by SPM, this proliferation was substantially inhibited by the addition of either anti-I-A or anti-I-E mAb, and complete inhibition was produced by the addition of both mAbs . Fixed antigen-presenting cells still induced T cell proliferation by SPM . A significant expansion of T cells bearing V beta 13 T-cell receptor was observed up to 73% among the Thy 1.2+ cells in cultures stimulated with SPM, indicating expansion in a V beta-specific manner . Immunoblotting of IEF-separated proteins showed that anti-streptococcal pyrogenic exotoxin (SPE) C reacted with a protein of pI 6.9 and anti-SPEB did not show any reactivity . SPEA was reported to expand V beta 8.1 and 8.2 bearing murine T cells, and SPM did not . SPM also exhibited potent mitogenic activity on human T cells and V beta 21+ T cells were selectively expanded . These results lead to the conclusion that SPM was neither SPEA, B nor C, but a new protein belonging to a group of streptococcal superantigens with activity on not only human but also murine lymphocytes. Artif Cells Blood Substit Immobil Biotechnol, 1996 Sep, 24(5), 553 - 66 Kinetic properties of glucosyltransferase adsorbed onto saliva-coated hydroxyapatite; Steinberg D et al.; Results from previous studies have shown that several properties of glucosyltransferase (GTF) adsorbed onto saliva-coated hydroxyapatite beads differ from those of GTF in solution . For example: thermostability, pH-activity dependency, sensitivity to inhibitors . The aim of this study was to compare the kinetics of the adsorbed GTF with its kinetic properties in solution . Hydroxyapatite beads were coated with human parotid saliva (sHA) . Following washes, cell-free GTF enzyme from Streptococcus sobrinus 6715 (S . sobrinus 6715) or Streptococcus mutans GS-5 (S . mutans GS-5) was adsorbed onto sHA . The GTF-coated sHA were then incubated with radiolabeled sucrose for intervals of 5-360 minutes and the amount of glucans synthesized in situ by the adsorbed GTF was determined and compared with that produced in solution . The adsorbed GTF (from S . sobrinus 6715) exhibited a sharp increase in glucan production within the first 5 minutes of incubation while surface-bound GTF of S . mutans GS-5 displayed an initial burst of activity within the first 15 minutes of incubation . During the next 6 hours (duration of experiment) the amount of glucan on the beads did not increase with either enzyme . In contrast, the kinetic profile of the two GTFs in solution demonstrated a linear increase in the amount of glucans formed, with no initial burst effect . The results indicate that the rapid formation of glucans by GTF adsorbed onto sHA could have implications for colonization by oral microorganisms on tooth surfaces . The accelerated synthesis of glucan on tooth surfaces may affect the microbiology of the dental plaque, and might also influence the movement of substances, such as acids and antiplaque agents, across the acquired pellicle and dental plaque. Antimicrob Agents Chemother, 1996 Sep, 40(9), 2190 - 3 Canadian national survey of prevalence of antimicrobial resistance among clinical isolates of Streptococcus pneumoniae . Canadian Bacterial Surveillance Network; Simor AE et al.; The antimicrobial susceptibilities of 1,089 clinical isolates of Streptococcus pneumoniae obtained from 39 laboratories across Canada between October 1994 and August 1995 were determined . A total of 91 isolates (8.4%) demonstrated intermediate resistance (MIC, 0.1 to 1.0 microgram/ml) and 36 (3.3%) had high-level resistance (MIC, > or = 2.0 micrograms/ml) to penicillin . Penicillin-resistant strains were more likely to have been recovered from normally sterile sites (P = 0.005) and to be cross-resistant to several beta-lactam and non-beta-lactam antimicrobial agents (P < 0.05) . These results indicate that there has been a recent significant increase in the prevalence of antibiotic-resistant S . pneumoniae in Canada. Antimicrob Agents Chemother, 1996 Sep, 40(9), 2147 - 51 Synergy between amoxicillin and gentamicin in combination against a highly penicillin-resistant and -tolerant strain of Streptococcus pneumoniae in a mouse pneumonia model; Darras-Joly C et al.; In vivo synergy with beta-lactam antibiotics and aminoglycosides has been studied only with penicillin-susceptible Streptococcus pneumoniae strains . We evaluated the interaction between amoxicillin (AMX) and gentamicin (GEN) on the basis of in vitro checkerboard and time-kill curves and of findings in a mouse model of acute bacteremic pneumonia due to a highly penicillin-resistant and -tolerant S . pneumoniae strain of serotype 19 (penicillin and AMX MICs of 4 micrograms/ml; gentamicin MIC of 16 micrograms/ml) . Checkerboard results at 18 h of incubation showed indifference . With regard to AMX alone, in vitro time-kill curves demonstrated synergy between AMX (1 microgram/ml) and GEN (16 micrograms/ml) at 5 and 8 h of incubation and for AMX (16 micrograms/ml) in combination with GEN (16 micrograms/ml) at 3, 5, and 8 h of incubation . In leukopenic mice, pulmonary killing curves after a single drug injection demonstrated that AMX (100 mg/kg of body weight) with GEN (16 mg/kg) was more effective than AMX alone (P = 10(-4) . With repeated-dose treatment, a synergy was apparent at 8 h after four injections with AMX (100 mg/kg) in combination with GEN (8 or 16 mg/kg) (P < or = 0.05) . The cumulative survival rate with AMX (100 mg/kg) every 8 h, combined with GEN (4 or 8 mg/kg) every 8, 12, or 24 h, was better than with AMX alone . Combined use of AMX and GEN may be a valuable therapeutic alternative for pneumococcal pneumonia due to highly penicillin-resistant S . pneumoniae strains. Antimicrob Agents Chemother, 1996 Sep, 40(9), 2062 - 66 In vivo activity and pharmacodynamics of amoxicillin in combination with fosfomycin in fibrin clots infected with highly penicillin-resistant Streptococcus pneumoniae; Chavanet P et al.; Using a clinical pneumococcal strain for which MICs were 4, 2, and 32 mg/liter for penicillin, amoxicillin, and fosfomycin, respectively, we studied the efficacies of these antibiotics alone and their combinations in the treatment of prolonged (48-h) experimental fibrin clot infection in rabbits . Treatments were as follows: amoxicillin IV at 20 mg/kg of body weight in one dose (Amo20), 50 mg/kg in one dose (Amo50), or two doses 6 h apart (Amo20 x 2 and Amo50 x 2); fosfomycin IV at a fixed dose of 50 mg/kg in one dose (Fos50) or two divided doses 6 h apart (Fos50 x 2); or the combinations of amoxicillin and fosfomycin with the same schedules . Maximum concentrations in clots were 2.03 +/- 1.02 and 2.13 +/- 0.33 mg/liter for Amo20 regimens, 3.7 +/- 1.9 and 4 +/- 1.3 mg/liter for Amo50 regimens, and 24 +/- 7 and 40 +/- 8 mg/liter for fosfomycin regimens, respectively . The mean half-lives of elimination from clots were between 2 and 3 h for amoxicillin regimens and between 5 and 7 h for fosfomycin . We observed the highest bacterial reductions (log10 CFU/gram) for Amo50 in two divided doses with or without fosfomycin . A significantly higher bacterial reduction than that with each monotherapy was observed when Amo20 was combined with fosfomycin in either one dose or two doses 6 h apart (0.16 +/- 0.8 and 1.64 +/- 1.6 log10 CFU/g for Amo20 in one and two doses, respectively, and 0.93 +/- 0.81 and 0.61 +/- 0.56 log10 CFU/g for fosfomycin in one and two doses, respectively, versus 3.46 +/- 1.26 and 3.16 +/- 1.31 log10 CFU/g for Amo20 plus fosfomycin in one and two doses, respectively {P < 0.001}) . A time-dependent effect was observed with amoxicillin regimens . The time of regrowth was significantly delayed when amoxicillin was combined with fosfomycin . By using a multivariate analysis, we demonstrated that the most important parameter correlated to efficacy of the combination amoxicillin-fosfomycin was the length of the period during which the concentration of amoxicillin remained above the MIC . We demonstrated that the in vivo efficacy of the combination of amoxicillin and fosfomycin gave higher antibacterial effect than each monotherapy. Antimicrob Agents Chemother, 1996 Sep, 40(9), 1977 - 82 Efficacy of single-dose ceftriaxone in experimental otitis media induced by penicillin- and cephalosporin-resistant Streptococcus pneumoniae; Barry B et al.; We used a gerbil model of otitis media to assess the efficacy of single-dose ceftriaxone against three Streptococcus pneumoniae strains highly resistant to penicillin (MICs, 4 to 8 micrograms/ml) and with various susceptibilities to ceftriaxone (MICs, 0.5, 4, and 8 micrograms/ml) . Middle ear infection was induced by bilateral transbullar challenge with 10(7) bacteria per ear . Antibiotic treatment was administered subcutaneously at 2 h postinfection . Infection status was checked 2 days later by counting the bacteria in middle ear and cerebrospinal fluid samples . With the cefriaxone-susceptible strain (MIC, 0.5 microgram/ml), we tested doses of 5 to 100 mg/kg of body weight . With a dose of 50 mg/kg, treatment outcome was equivalent to that with amoxicillin, which was used as a reference (25 mg/kg, two injections); no bacteria were recovered from 82% of the middle ear samples, and the rate of cerebrospinal fluid culture positivity was significantly reduced to 6%, relative to 59% for the untreated controls . Similar efficacy was obtained with a dose of 100 mg/kg against the two ceftriaxone-resistant strains . Pharmacokinetic study indicates that the values of the parameters in plasma after the administration of a dose of 100 mg/kg (peak level of total drug, 268 +/- 33 micrograms/ml; elimination half-life, 0.8 h; area under concentration-time curve, 488 micrograms.h.ml-1) were still suboptimal compared with the values of the parameters measured in pediatric patients after intravenous or intramuscular administration of a dose of 50 mg/kg . Our results indicate the efficacy of ceftriaxone against experimental cephalosporin-resistant pneumococcal otitis and provide a basis for the clinical use of single-dose ceftriaxone against pneumococcal otitis media. Pediatr Infect Dis J, 1996 Sep, 15(9), 791 - 5 Evaluation of the efficacy, safety and toleration of azithromycin vs . penicillin V in the treatment of acute streptococcal pharyngitis in children: results of a multicenter, open comparative study . The Swiss Tonsillopharyngitis Study Group; Schaad UB et al.; BACKGROUND: For many years alternatives to penicillin have been studied for the management of pediatric group A beta-hemolytic Streptococcus (GABHS) pharyngitis . As a result of its pharmacokinetic profile azithromycin is unique among these alternative antimicrobials in allowing once daily dosing and shorter duration of treatment . However, the optimum dose (e.g . 10 or 12 mg/kg/day) and duration (e.g . 3 or 5 days) of azithromycin therapy have not been defined yet . METHODS: An open, comparative multicenter study was conducted in 343 children with clinical symptoms of GABHS pharyngitis and a positive culture to evaluate the efficacy and safety of azithromycin (10 mg/kg) once daily for 3 days compared with penicillin V three times daily for 10 days . RESULTS: Among the evaluable patients bacteriologic eradication documented at follow-up visits was inferior with azithromycin when compared with penicillin V therapy: at Days 9 to 20 (mean, 12 days), negative cultures in 65% (99 of 152 patients) vs . 82% (128 of 126 patients) (P < 0.001); and at Days 17 to 57 (mean, 25 days), in 55% vs . 80% (P < 0.001) . Overall clinical success (cure or improvement) was achieved in 93% (149 of 160 patients) of azithromycin-treated and in 89% (143 of 160 patients) of penicillin-treated patients (P > 0.50) . There was no correlation between bacteriologic response and clinical outcome, as assessed shortly after completion of therapy or during 6-month follow-up . Both treatments were well-tolerated . CONCLUSIONS: In the present study on GABHS pharyngitis in children, a once daily (10-mg/kg), 3-day oral regimen of azithromycin was as clinically effective and as safe as traditional penicillin but appeared inferior in eliminating GABHS from the throat. Clin Diagn Lab Immunol, 1996 Sep, 3(5), 517 - 22 Clonal diversity of Streptococcus mitis biovar 1 isolates from the oral cavity of human neonates; Fitzsimmons S et al.; The clonal diversity of 101 isolates of the pioneer bacterium Streptococcus mitis biovar 1 obtained from the oral cavities of 40 human neonates 1 to 3 days, 2 weeks, and 1 month postpartum was examined by using rRNA gene restriction patterns . There was a high degree of genetic diversity, with the 101 isolates comprising 93 unique PvuII ribotypes . There were eight identical pairs of ribotype patterns, and seven of the eight pairs were obtained from individual neonates . Only one identical pair comprised isolates obtained from different neonates . In all but two cases, isolates with matching ribotypes were obtained at one visit . Two pairs of isolates with matching ribotype patterns were obtained from neonates on successive visits . The ribotype patterns of the isolates were examined by cluster analysis . The isolates forming each cluster were very similar, yet each cluster was well separated from its neighbors . When several isolates were obtained from individual neonates at a particular visit, in some instances they were contained in a single cluster, whereas in other cases each isolate was contained in a separate cluster . Isolates obtained from individual neonates on successive visits tended to be contained in different clusters . This high degree of diversity, which has been observed in other mucosal commensal bacteria, may serve as a mechanism for avoiding immune elimination of these bacteria. J Perinat Neonatal Nurs, 1996 Sep, 10(2), 1 - 16 Perinatal group B streptococcal infections: the nurse's role in identification and prophylaxis; Mahlmeister L; Group B Streptococcus (GBS) is a major cause of perinatal infectious morbidity and mortality in the United States . An estimated 50,000 women and 7,600 neonates experience GBS disease, and as many as 310 infants die each year . In 1992, the American Academy of Pediatrics published recommendations for identification and treatment of pregnant women colonized with GBS . In 1996, the Centers for Disease Control and Prevention and the American College of Obstetricians and Gynecologists developed more comprehensive guidelines . The article describes the central role of perinatal nurses in the identification of pregnant women colonized with GBS, patient education about GBS disease, and successful implementation of intrapartum GBS prophylaxis. Isr J Med Sci, 1996 Sep, 32(9), 771 - 4 Pneumococcal psoas abscess: report of a case and review of the world literature; Giladi M et al.; A 39-year-old man was admitted to the hospital with fever and right hip pain . CT scan of the pelvis showed a psoas muscle abscess . Pus, which was surgically evacuated, grew Streptococcus pneumoniae . The patient fully recovered after 30 days of intravenous antibiotics . Review of the literature revealed only nine similar cases of primary pneumococcal psoas abscess . Three cases were complicated with pneumococcal meningitis . In four cases there was a history of trauma to the involved area . Pneumococcal psoas abscess is a rare entity that carries a good prognosis provided early surgical drainage is performed and parenteral antibiotic therapy is administered to the patient. Can J Microbiol, 1996 Sep, 42(9), 927 - 33 Effects of a strain of Saccharomyces cerevisiae (Levucell SC1), a microbial additive for ruminants, on lactate metabolism in vitro; Chaucheyras F et al.; The effect of Levucell SC, a strain of Saccharomyces cerevisiae marked as a feed additive for ruminants, was investigated in vitro on lactate metabolism by the ruminal bacteria Streptococcus bovis and Megasphaera elsdenii . The coculture between 10(7) live cells x mL(-1) of SC and a Streptococcus bovis strain in the presence of glucose reduced lactate production by the bacterial strain . Live yeast cells were able to compete with Streptococcus bovis for glucose utilization in strictly anaerobic conditions, so less glucose was available for the bacterium . SC also stimulated L-lactate utilization by a strain of M . elsdenii . The effect depended on the concentration of yeast cells added . Bacterial growth and fermentation end-product concentrations were also increased in the presence of SC . Some amino acids and vitamins, but not dicarboxylic acids, stimulated the bacterial specific activity of L-lactate uptake . SC was able to provide amino acids to M . elsdenii . In a coculture of Streptococcus bovis and M . elsdenii on glucose, the reduction of lactate concentration was improved by SC, the same trend being observed when maltose or soluble starch were used as carbon and energy source . These results indicate that SC can be a very useful tool to reduce lactate accumulation in vitro during fermentation of soluble sugars. Br J Haematol, 1996 Sep, 94(4), 729 - 39 Evidence for the involvement of complement proteins in platelet aggregation by Streptococcus sanguis NCTC 7863; Ford I et al.; We investigated the mechanisms of platelet aggregation by the type strain of Streptococcus sanguis (NCTC 7863) . This species is one of the major aetiological agents of infective endocarditis . S . sanguis NCTC 7863 caused aggregation of normal human platelets in vitro following a lag period that varied between donors (7-19 min) . Platelet aggregation was dependent on one or more plasma constituents and all the necessary factors gradually became bound to the bacterial surface during the lag period . The length of the lag period was determined by the plasma of the donor and not by a feature of their platelets . Platelet aggregation by S . sanguis NCTC 7863 could be inhibited by heating plasma at 56 degrees C, by treating plasma with cobra venom factor, or by incubating with soluble Complement Receptor 1, all of which inhibit or deplete complement . Complement activation required Mg2+, but not Ca2+ ions and the the cleavage fragment, Ba, of factor B was produced, indicating that the alternative pathway was operative . Zymosan- and S . sanguis-induced aggregation showed similarities, including the same variability in lag times among donors, and absorption of plasma with zymosan prevented the plasma from supporting platelet aggregation by S . sanguis, C3, C9 and vitronectin were found to bind to S . sanguis NCTC 7863, but the latter two were present at very low levels on a non-aggregating strain of S . sanguis, SK96 . The rate of assembly of the C5b-9 complex on the NCTC 7863 bacterial surface correlated with the lag time . These data suggest a role for the complement pathway in platelet aggregation by the type strain of S . sanguis. Ann Allergy Asthma Immunol, 1996 Sep, 77(3), 167 - 73; quiz 173-5 Emerging resistance to antibiotics: impact on respiratory infections in the outpatient setting; Green M et al.; OBJECTIVE: To review the major mechanisms of antibiotic resistance and provide an overview of currently available oral antibiotics . This article discusses the impact of antibiotic resistance on respiratory infections (eg, otitis media, sinusitis, and pneumonia) encountered in the outpatient setting and appropriate management strategies for these infections are proposed . DATA SOURCES: A Medline search was performed to identify recent references from the English language pertinent to this topic . Proposed management schemes for infections due to antibiotic-resistant bacteria were derived from current literature as well as the opinion of the authors . RESULTS: Currently available oral antibiotics including penicillins, cephalosporins, macrolides, trimethroprim/sulfamethoxazole, and clindamycin continue to provide effective therapeutic options despite the increasing importance of antibiotic resistance in bacteria causing respiratory tract infections in the outpatient setting . The effectiveness of a given agent may no longer be assured, leading to the need for development of updated management strategies for patients experiencing treatment failure with antibiotics . Diagnostic strategies (ie, tympanocentesis) and novel treatment algorithms (eg, use of amoxicillin in combination with amoxicillin/clavulanate) for patients experiencing clinical failures potentially due to antibiotic-resistant bacteria are proposed . CONCLUSIONS: The increasing prevalence of antibiotic resistant bacteria (especially penicillin-resistant Streptococcus pneumoniae) is leading to new approaches to the management of common respiratory infections in the outpatient setting . Additional experience and careful clinical trials are necessary to identify optimal management strategies for these infections. J Med Microbiol, 1996 Sep, 45(3), 179 - 85 The effect of oral commensal bacteria on candidal adhesion to human buccal epithelial cells in vitro; Nair RG et al.; The effect of Streptococcus sanguis, S . salivarius, Escherichia coli and Porphyromonas gingivalis on the adhesion of Candida albicans and C . krusei to human buccal epithelial cells (BEC) was investigated with a modified membrane filter system . The filters (12 microns diameter pores) acted as a support for the BEC which were pre-exposed to known concentrations of bacterial suspensions (for 45 min-1 h), and then re-incubated with standardised concentrations of yeast suspensions for various periods . The BEC with adherent yeasts were then transferred on to a glass slide, gram-stained and counted by light microscopy . Three of the four bacterial species significantly suppressed adhesion of C . albicans to BEC; S . sanguis had no effect . Both S . sanguis and S . salivarius suppressed adhesion of C . krusei to BEC pre-exposed to three different bacterial concentrations, although variable results were obtained with P . gingivalis and E . coli . Significant differences in the relative adhesion of C . albicans and C . krusei to BEC were also recorded . These results indicate that the adhesion of yeasts to BEC is modulated both by the composition and the quantity of the pre-existing bacterial flora on the BEC. J Bacteriol, 1996 Sep, 178(18), 5546 - 9 Physical and genetic chromosomal map of an M type 1 strain of Streptococcus pyogenes; Suvorov AN et al.; A physical map of the chromosome of an M type 1 strain of Streptococcus pyogenes was constructed following digestion with three different restriction enzymes, SmaI, SfiI, and SgrAI, and separation and analysis of fragments by pulsed-field gel electrophoresis . The genome size of this strain was estimated to be 1,920 kb . By employing Southern hybridization and PCR analysis, 36 genes were located on the map. J Bacteriol, 1996 Sep, 178(18), 5402 - 9 The non-penicillin-binding module of the tripartite penicillin-binding protein 3 of Escherichia coli is required for folding and/or stability of the penicillin-binding module and the membrane-anchoring module confers cell septation activity on the folded structure; Goffin C et al.; The ftsI-encoded multimodular class B penicillin-binding protein 3 (PBP3) is a key element of the cell septation machinery of Escherichia coli . Altered ftsI genes were overexpressed, and the gene products were analyzed with respect to the level of production, stability, penicillin affinity, and cell septation activity . In contrast to the serine beta-lactamases and low-molecular-mass PBPs which are autonomous folding entities, the S-259-to-V-577 penicillin-binding module of M-1-to-V-577 PBP3 lacks the amino acid sequence information for correct folding . The missing piece of information is provided by the associated G-57-to-E-258 non-penicillin-binding module which functions as a noncleaved, pseudointramolecular chaperone . Key elements of the folding information reside within the motif 1-containing R-60-to-W-110 polypeptide segment and within G-188-to-D-197 motif 3 of the n-PB module . The intermodule interaction is discussed in the light of the known three-dimensional structure (at 3.5-A {0.35-nm} resolution) of the analogous class B PBP2x of Streptococcus pneumoniae (S . Pares, N . Mouz, Y . Petillot, R . Hakenbeck, and O . Dideberg, Nature Struct . Biol . 3:284-289, 1996) . Correct folding and adoption of a stable penicillin-binding conformation are necessary but not sufficient to confer cell septation activity to PBP3 in exponentially growing cells . The in vivo activity of PBP3 also depends on the M-1-to-E-56 amino-terminal module which encompasses the cytosol, the membrane, and the periplasm and which functions as a noncleaved pseudo-signal peptide. J Pediatr, 1996 Sep, 129(3), 396 - 402 An outbreak of M serotype 1 group A Streptococcus in a neonatal intensive care unit; Campbell JR et al.; OBJECTIVE: To describe the investigation and control of an outbreak of M serotype 1, Streptococcus pyogenes (group A Streptococcus, GAS) infections in a neonatal intensive care unit (NICU) . STUDY DESIGN: The study was conducted in an NICU in a large urban university-affiliated hospital . Retrospective review was performed of all infants and health care workers in the NICU, especially those either colonized or infected with GAS during the outbreak and the prospective surveillance period (July through September 1994) . Prospective epidemiologic investigation, including cultures of throat, umbilicus, and anorectum (infants), or throat and anus (NICU personnel), identified a possible common source of the disease in case infants . Antimicrobial susceptibility testing and serotyping of all GAS strains were performed; M serotype 1 isolates were examined by DNA analysis with restriction fragment length polymorphism . The M-1 GAS isolates were tested for streptococcal pyrogenic exotoxin (SPE) A and SPE B production . A retrospective chart review and analysis of infants with GAS infection or colonization was conducted . RESULTS: During a 1-week period, two very low birth weight infants more than 3 weeks of age had GAS septicemia and focal infection . Two additional very low birth weight infants with asymptomatic throat colonization were identified during the first week of surveillance . Benzathine penicillin G was administered to all NICU infants, but failed to eradicate throat colonization in the four case subjects . Seven days after completing parenteral antibiotic therapy, the index patient had a recurrence of GAS septicemia that was fatal . Eradication of throat colonization in the remaining three infants was achieved with a 10-day course of intravenous clindamycin therapy . Among 103 NICU personnel, five (4.9%) had asymptomatic GAS colonization with strains that were uniformly susceptible to penicillin . Each colonized adult was successfully treated with oral clindamycin therapy . Serotyping revealed that five isolates of GAS from four infants and one NICU respiratory therapist were M-1 isolates; DNA analysis confirmed that these were the same strain . The five M-1 isolates produced both SPE A and SPE B . CONCLUSIONS: The previously documented increase in prevalence of M-1 strains of GAS in the United States is likely to be associated with their introduction into closed populations including NICUs . Control of such outbreaks may be achieved by isolation, cohorting of case subjects and possible carriers, and successful eradication of colonization in case subjects and carriers . Although GAS organisms are uniformly susceptible to penicillin G, eradication may require agents other than penicillin. J Infect Dis, 1996 Sep, 174(3), 520 - 8 Mechanisms of the spread of penicillin resistance in Streptococcus pneumoniae strains causing meningitis in children in France; Doit C et al.; The molecular epidemiology of penicillin-resistant Streptococcus pneumoniae strains causing meningitis in children was studied in France . Typing procedures included analysis of total DNA polymorphism by random amplification of polymorphic DNA, restriction fragment length polymorphism (RFLP) analysis of the ribosomal RNA gene regions, and pulsed-field gel electrophoresis . Penicillin-binding protein (PBP) genes pbp2b and pbp2x were studied by RFLP analysis and DNA sequencing in selected cases . Statistical analysis of the data by factorial analysis of correspondence established that the emergence of penicillin-resistant pneumococci in this pathology is the result of the spread of two highly resistant closely related clusters and a cluster of serotype 23 strains with an intermediate level of resistance, the spread of genes confering high resistance to penicillin between the two highly resistant clusters, and complex genetic events involving the pbp genes in a heterogeneous population of strains leading to an intermediate level of resistance. J Infect Dis, 1996 Sep, 174(3), 513 - 9 Association of penicillin-resistant pneumococci with residence in a pediatric chronic care facility; Mannheimer SB et al.; Risk factors for the acquisition of penicillin-resistant pneumococci (PRP) were analyzed at a university hospital in New York City . Patients with PRP and control patients with penicillin-sensitive pneumococcal infections were compared . In 1994, 24 (21%) of 113 patients with Streptococcus pneumoniae infections had PRP; 13 PRP isolates were from children and 11 from adults . Only white race (P < .05) and residence in a pediatric chronic care facility (P < .05) were significantly associated with penicillin resistance . An investigation at one chronic care facility revealed that 33% of children (17/52) had PRP colonization . Fourteen of the 17 PRP isolates were also resistant to ceftriaxone . Prior antibiotic use and specifically beta-lactam use were associated with penicillin resistance . All typeable PRP isolates were multidrug-resistant serotype 23 . Pediatric residents in chronic care facilities may be an important reservoir of PRP and may serve as a source of PRP transmission when they are transferred to acute care hospitals. J Infect Dis, 1996 Sep, 174(3), 507 - 12 Ethanol ingestion reduces antipneumococcal activity of rat pulmonary surfactant; Rubins JB et al.; Because chronic ethanol ingestion decreases pulmonary clearance of Streptococcus pneumoniae in rats, and extracellular antipneumococcal factors in rat surfactant are important in the early clearance of pneumococci from the rat alveolus, the effects of ethanol ingestion on surfactant bactericidal activity were investigated . Normal surfactant from chow-fed rats showed potent anti-pneumococcal activity, even against bacteria growing in nutrient-rich media under favorable conditions . In contrast, surfactant from ethanol-fed rats and from calorie-restricted control-fed rats had significantly reduced antipneumococcal activity compared with surfactant from chow-fed rats . The reductions in surfactant bactericidal activity produced by ethanol ingestion or caloric restriction did not appear to be mediated through changes in either the total amount or the distribution of fatty acids, the antipneumococcal factors in normal surfactant . Rather, ethanol ingestion, and to a lesser extent caloric restriction, produced a surfactant inhibitor of free fatty acids that was partially characterized as a hydrophobic protein. J Infect Dis, 1996 Sep, 174(3), 500 - 6 High-resolution genotyping elucidates the epidemiology of group A streptococcus outbreaks; Stanley J et al.; Streptococcus pyogenes strains were genotyped by a combination of molecular methods for high- resolution epidemiologic studies of disease outbreaks . Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the emm gene is reported . Alone or in conjunction with other molecular techniques (16S ribotyping, pulsed-field gel electrophoresis, and detection of exotoxin genes), PCR-RFLP could differentiate outbreak-related strains from contemporaneous background strains of the same M serotype . Three outbreaks were studied: pharyngitis in a boarding school (serotype M5), cross-infection in a hospital burn unit (serotype M76), and severe invasive disease in two elderly care homes (serotype R28) . It was possible, for example, to identify within serotype R28 a clone with particular potential for invasive disease . In all cases, the four molecular methods yielded complementary results that were hierarchically related . Strains could be assigned to the outbreak or the background in a precise, reproducible, and rapid manner. Infect Immun, 1996 Sep, 64(9), 3772 - 7 Uptake of Streptococcus pneumoniae by respiratory epithelial cells; Talbot UM et al.; Although Streptococcus pneumoniae is the leading cause of community-acquired pneumonia in humans, the mechanism whereby the organism penetrates lung tissue is poorly understood . In the present study we have examined the capacity of pneumococci to penetrate A549 cells, a human lung alveolar carcinoma (type II pneumocyte) cell line . Not all clinical S . pneumoniae isolates initially tested were capable of penetration of the cells, as judged by resistance to extracellular antibiotics . The presence of a polysaccharide capsule also significantly reduced the capacity to both adhere to and penetrate A549 cells . Electron micrographs showed the presence of pneumococci enclosed within vacuoles of intact A549 cells, but bacteria were also seen free in the cytoplasm of damaged cells . Ongoing bacterial DNA, RNA, or protein synthesis was not essential for uptake of pneumococci by A549 cells, and uptake was not diminished by pretreatment of the pneumococci with trypsin . However, inhibition of A549 microfilament assembly with cytochalasin D abolished the phenomenon. Infect Immun, 1996 Sep, 64(9), 3659 - 65 The galactosyl-(alpha 1-4)-galactose-binding adhesin of Streptococcus suis: occurrence in strains of different hemagglutination activities and induction of opsonic antibodies; Tikkanen K et al.; The occurrence of the galactose-(alpha 1-4)-galactose-specific adhesin in Streptococcus suis, a pig and human pathogen causing sepsis, meningitis, and other serious infections, was studied . Poly- and monoclonal anti-bodies to the purified adhesin, as well as pigeon ovomucoid, a specific probe for the adhesin activity, detected one single protein band in extracts of S . suis . The adhesin was detected in all 23 strains studied, representing pathogenic serotypes (1, 2, 4, 5, 7, 8, and nontypeable) and including several weakly hemagglutinating or nonhemagglutinating strains and phase variants . The amount of adhesin detected was not correlated with the hemagglutination activity of the intact bacteria . Extraction of cells showing no binding of pigeon ovomucoid by ultrasonic treatment resulted in extracts with pigeon ovomucoid binding activity, suggesting that the adhesin was not accessible to the probe on the intact cells . Analysis of the amount of capsular polysaccharide revealed an inverse relationship between the hemagglutination activity and expression of capsular polysaccharide, thus suggesting a factor influencing adhesin accessibility . The purified adhesin was highly immunogenic and induced in preliminary experiments bactericidal activity in mice . Thus, the adhesin, with its specific binding mechanism to host cells and a proposed pathogenic role, is widely expressed among strains of different serotypes and therefore appears to represent a novel promising candidate for the development of a vaccine against S . suis. Infect Immun, 1996 Sep, 64(9), 3652 - 8 Characterization of a P1-deficient strain of Streptococcus mutans that expresses the SpaA protein of Streptococcus sobrinus; Kuykindoll RJ et al.; The Streptococcus sobrinus SpaA protein and the Streptococcus mutans P1 protein share 66% sequence homology at the amino acid level . To determine if the SpaA protein can be expressed in S . mutans and functionally replace the P1 protein, the spaA gene of S . sobrinus 6715 was isolated from plasmid pX1303 and inserted into the Escherichia coli-Streptococcus shuttle vector pVA838 . The resulting plasmid pX1600 was transformed into the P1-deficient strain S . mutans 834 that has defects in saliva-mediated aggregation and in the ability to adhere to saliva-coated hydroxyapatite surfaces . Western blot (immunoblot) analysis of cellular protein fractions of S . mutans 834 (pX1600) detected in mutanolysin-solubilized cell walls a major protein of 210 kDa with an electrophoretic mobility similar to that of S . sobrinus SpaA protein and a minor 210-kDa protein and a major 64-kDa protein in the extracellular protein fraction . Analysis of virulence traits showed that expression of SpaA protein by S . mutans 834(pX1600) cells had restored the ability of the S . mutans 834 cells to aggregate in the presence of saliva or salivary agglutinin but not to adhere to saliva-coated hydroxyapatite . This cell aggregation was inhibited specifically by antisera to S . sobrinus SpaA protein . These results indicate that SpaA plays a role in the virulence of S . sobrinus by specifically interacting with fluid-phase salivary agglutinin to mediate cell aggregation. Infect Immun, 1996 Sep, 64(9), 3518 - 23 Experimental vaccination against group B streptococcus, an encapsulated bacterium, with highly purified preparations of cell surface proteins Rib and alpha; Larsson C et al.; Encapsulated bacteria cause some of the most common diseases in humans . Although the polysaccharide capsules of these pathogens have attracted the most attention with regard to vaccine development, recent evidence suggests that bacterial surface proteins may also be used to confer protective immunity . We have analyzed this possibility in group B streptococcus (GBS), an encapsulated bacterium that is the major cause of invasive bacterial disease in the neonatal period . Previous work has shown that the majority of GBS strains causing invasive infections express the Rib protein, and that most strains lacking Rib express a protein designated alpha . Here we report that active immunization with highly purified preparations of Rib or alpha protected mice against lethal infection with strains expressing the corresponding protein . Vaccination with the Rib protein protected against two strains of capsular type III and two strains of type II, and vaccination with the alpha protein protected against one strain of type II and one strain of type Ib . The mice vaccinated with Rib or alpha showed a good immunoglobulin G response to the immunogen . These data suggest that a vaccine against GBS disease may be based on cell surface proteins and support the notion that proteins may be used for immunization against encapsulated bacteria. Vet Rec, 1996 Aug 31, 139(9), 204 - 7 Incidence of clinical mastitis in a random sample of dairy herds in the southern Netherlands; Miltenburg JD et al.; The incidence of clinical mastitis and distribution of pathogens in dairy cows was estimated in 171 randomly selected dairy herds in the southern Netherlands . A total of 1103 quarter cases were reported . The average annual incidence rate was 12.7 quarter cases per 100 cows per year . The most frequent isolates from clinical cases were Escherichia coli (16.9 per cent), Staphylococcus aureus (14.4 per cent), Streptococcus uberis (11.9 per cent) and Streptococcus dysgalactiae (8.9 per cent) . Most cases were reported in early lactation: 25.4 per cent in the first month of lactation for all cows, and 39.1 per cent in the first month for first lactation cows . The rear quarters had a significantly higher incidence rate than the front quarters . Cows with an E coli infection showed more general clinical signs than cows infected with S aureus, S uberis and S dysgalactiae . A significantly higher incidence was observed in herds with a low (< 150,000 cells/ml) bulk milk somatic cell count than in herds with a count above 250,000 cells/ml. Mol Gen Genet, 1996 Aug 27, 252(1-2), 207 - 11 Cloning of heterologous genes specifying detrimental proteins on pUC-derived plasmids in Escherichia coli; Muller J et al.; A system is described that enables the cloning of genes specifying detrimental proteins in Escherichia coli . The system is based on pUC plasmids and was developed for the expression of the Bacillus subtilis csaA gene, which is lethal when expressed at high levels . Suppressor strains that tolerate the presence of plasmids for high-level expression of csaA were isolated, which contained small cryptic deletion variants of the parental plasmid in high copy numbers . The cryptic plasmids consisted mainly of the pUC replication functions and lacked the csaA region and selectable markers . The co-resident, incompatible, cryptic plasmids enabled the maintenance of the csaA plasmids by reducing their copy number 20-fold, which resulted in a concomitant 3- to 7-fold reduction in the expression of plasmid-encoded genes . Strains carrying these cryptic endogenous plasmids proved to be useful for the construction of pUC-based recombinant plasmids carrying other genes, such as the skc gene of Streptococcus equisimilis, which cannot be cloned in high copy numbers in E . coli . Several strategies to reduce production levels of heterologous proteins specified by plasmids are compared. MMWR Morb Mortal Wkly Rep, 1996 Aug 2, 45(30), 650 - 3 Invasive infection with Streptococcus iniae--Ontario, 1995-1996; Identification of a family of streptococcal surface proteins with extremely repetitive structure; Department of Medical Microbiology, Lund University, Solvegatan 23, S-223 62 Lund, SwedenThe group B Streptococcus (GBS) causes the majority of life-threatening bacterial infections in newborn children . Most GBS strains isolated from such infections express a surface protein, designated Rib, that confers protective immunity and therefore is of interest for analysis of pathogenetic mechanisms . Sequence analysis demonstrated that Rib has an exceptionally long signal peptide (55 amino acid residues) and 12 repeats (79 amino acid residues each) that account for >80% of the sequence of the mature protein . The repeats are identical even at the DNA level, indicating that an efficient mechanism operates to maintain a highly repetitive structure in Rib . The structure of Rib is similar to that of alpha, a previously characterized surface protein that is common among GBS strains lacking Rib . However, highly purified preparations of Rib and alpha did not cross-react immunologically, although the two proteins show extensive amino acid residue identity (47% in the repeat region) . When analyzed in Western blots, Rib and alpha give rise to a regularly spaced ladder pattern, apparently due to hydrolysis of acid-labile Asp-Pro bonds in the repeats . We conclude that Rib and alpha are members of a novel family of streptococcal surface proteins with unusual repetitive structure. Bull Tokyo Dent Coll, 1996 Aug, 37(3), 119 - 28 Methicillin-resistant Staphylococcus aureus and Candida albicans on denture surfaces; Tawara Y et al.; Infectious diseases caused by methicillin-resistant Staphylococcus aureus, MRSA, and Candida albicans are often serious in compromised hosts . We enumerated MRSA and C . albicans on denture surfaces and in saliva samples from 29 adults . Staphylococcus species, MRSA, and methicillin-resistant Staphylococcus epidermidis, MRSE, were detected on 17, 3, and 1 of the 29 denture surfaces, respectively . C . albicans were detected on 22 denture surfaces . All saliva samples from patients whose dentures carried Staphylococcus species and C . albicans were also found to contain both microorganisms . Adherence of isolated 3H labeled cells of MRSA and C . albicans to resin beads and saliva-coated resin beads was examined . Cells of both microorganisms adhered in significantly higher numbers to saliva-coated resin beads than to resin beads . The hydrophobicity of the MRSA isolated from denture surfaces varied from strain to strain; that of C . albicans strains was moderately high . The zeta potentials of MRSA isolates and of C . albicans isolates determined in KCI buffer were significantly low . The potential of the resin beads decreased after treatment with saliva . Two out of 5 MRSA strains were found to be inhibited in growth by oral Streptococcus, Actinomyces, and gram-negative bacterial strains, suggesting that some oral bacterial species play a role in inhibiting the colonization of Staphylococcus species . No isolates of C . albicans were inhibited in their growth by any of the oral bacteria tested . Isolates of MRSA and C . albicans coaggregated with Porphyromonas gingivalis and Fusobacterium nucleatum strains . Using denture cleaners every night for 2 weeks did not reduce numbers of Staphylococcus species or C . albicans organisms in saliva. Int Dent J, 1996 Aug, 46(4), 350 - 6 The effect of areca nut on salivary and selected oral microorganisms; de Miranda CM et al.; The aim of this study was to investigate the effect on the growth of salivary and selected oral microorganisms of areca nut, aqueous extracts of the nut, its major alkaloid arecoline and the components tannic acid and catechin of its tannin fraction . The antibacterial properties of the above were tested on Streptococcus mutans, Streptococcus salivarius, Candida albicans and Fusobacterium nucleatum and, as a control, Staphylococcus aureus . This was followed by investigating its effect on salivary organisms cultured from the saliva after chewing boiled areca nut . Extracts inhibited the growth of the selected organisms in a concentration dependent manner, baked and boiled nuts being significantly more potent than raw nut . Growth of C . albicans was the least affected by the nut extracts . Tannic acid was strongly antibacterial but not catechin or arecoline . No antibacterial effect could be demonstrated on salivary organisms after chewing the nut for 5 minutes but exposure of saliva to the cud for 1 hour caused a significant depression of bacterial growth . It is concluded that the hydrolysable tannins in the tannin fraction, which include tannic acid, are responsible for the antibacterial properties of the nut and that prolonged intraoral exposure to the nut can suppress bacteria in the mouth. Oral Microbiol Immunol, 1996 Aug, 11(4), 254 - 8 Effect of antibodies on chemiluminescence and on killing of Streptococcus sobrinus by polymorphonuclear leukocytes; van Raamsdonk M et al.; The effect of a polyclonal antiserum and OMVU10, a monoclonal antibody reactive with Antigen B of Streptococcus sobrinus, on the interaction of polymorphonuclear leukocytes with S . sobrinus was studied, using chemiluminescence and bacterial killing assays . Increased stimulation of neutrophils as measured in the chemiluminescence assays was established when S . sobrinus was preincubated with polyclonal antiserum or when polyclonal antiserum was added to the reaction mixture . Higher counts were measured in comparison to preimmune serum . After 90 min, 52% of S . sobrinus preincubated with polyclonal antiserum was killed . Killing was also increased when polyclonal antiserum was added to the reaction mixture in comparison to the controls . No killing was found when bacteria were preincubated with OMVU10 or when OMVU10 was added to the reaction mixture in comparison to Clone 24, a control antibody. Enferm Infecc Microbiol Clin, 1996 Aug-Sep, 14(7), 422 - 5 {Activity of erythromycin and clarithromicin against isolates of Streptococcus pneumoniae with decreased sensitivity to penicillin obtained from healty carriers . Spanish Collaborative Group}; Garcia de Lomas J et al.; BACKGROUND: Empirical treatment of pneumococcal upper and lower respiratory infections must be chosen on the basis of the susceptibility patterns of nasopharyngeal colonizing strains isolated from healthy carriers . METHODS: The susceptibility to erythromycin and clarithromycin was investigated by a conventional microdilution method among 103 pneumococci isolates recovered from healthy children (n = 63) and adults (n = 40) exhibiting decreased susceptibility to penicillin (MIC > or = 0.12 mg/l) . RESULTS: 63% of penicillin -resistant pediatric isolates were susceptible to erythromycin and 78.9% were susceptible to clarithromycin . Among isolates with diminished susceptibility to penicillin , 77.7% were susceptible to erythromycin and 86.3% to clarithromycin . 90% of adult isolates were susceptible to erythromycin and to clarithromycin . Overall, clarithromycin exhibited a better activity than erythromycin . CONCLUSIONS: Clarithromycin , and to a lesser extent erythromycin , are good alternatives to penicillin in the empirical treatment of respiratory tract infections caused by pneumococci with diminished susceptibility to penicillin . Neurologia, 1996 Aug-Sep, 11(7), 268 - 71 {Bilateral epidural psoas abscess simulating bilateral proximal diabetic plexopathy}; Roquer J et al.; We describe a diabetic woman with an epidural abscess and associated bilateral psoas abscesses which simulated the clinical course of diabetic bilateral proximal plexopathy . We emphasize three aspects: 1) the rarity of associated epidural abscess and bilateral psoas abscesses, 2) the rarity of the germ isolated (Streptococcus bovis), and 3) the diagnostic difficulties when both processes are associated, particularly in patients with an underlying disease that may simulate other common, clinical pictures. An Esp Pediatr, 1996 Aug, 45(2), 153 - 6 {Neonatal infections by Streptococcus agalactiae}; Juncosa Morros T et al.; OBJECTIVES: The objectives of the study were to evaluate: 1) The incidence and characteristics of neonatal infections by S . agalactiae in the Hospital Sant Joan de Deu of Barcelona, 2) the efficiency of a microbiological control during the third quarter of pregnancy in order to detect colonization by this microorganism; and 3) the efficiency of intrapartum prophylaxis . MATERIAL AND METHODS: Neonatal infections that took place between May 1991 and December 1994 have been studied . Children were born from women controlled in our hospital (7.772 pregnant women) or from women who delivered in other health centers . RESULTS: Nineteen newborn children with an invasive infection and four asymptomatic bacteriemias were diagnosed and treated during the above mentioned period of time . Early forms of the illness were sepsis (eight cases), meningitis (four cases) and arthritis (one case), whereas late forms were comprised of four cases of meningitis and two of arthritis . Three of the neonates died (mortality rate of 15.7%) and two of them developed neurologic sequelae . CONCLUSIONS: The microbiological control during pregnancy in order to detect S . agalactiae carriers, as well as intrapartum antibiotic prophylaxis contributes to reduce the number of this sort of infections . Negative results of cultures carried out during the third quarter of pregnancy do not exclude a carrier state at delivery, therefore, so posterior controls are necessary until the end of pregnancy. Tohoku J Exp Med, 1996 Aug, 179(4), 291 - 4 Use of fura-2/AM to measure intracellular free calcium in Selenomonas ruminantium; Nakamura I et al.; This paper describes a procedure for loading the acetoxymethyl ester of fura-2 (fura-2/AM), and the subsequent measurement of the concentration of intracellular free Ca2+ ({Ca2+}i) in Selenomonas ruminantium (S . ruminantium) using this technique . To ascertain the optimal loading conditions, the effect was examined on the loading of fura-2/AM of ethylenediamine-tetraacetic acid (EDTA), lysozyme, pluronic F127 alone, or the simultaneous application of EDTA and pluronic F127 . Individual administration of either EDTA, lysozyme or pluronic F127 did not produce an optimal loading of fura-2/ AM . The co-application of pluronic F127 and bovine serum albumin after treatment with EDTA increased the ratio of the fluorescence due to excitation at 340 nm to that at 380 nm (R340/380) markedly, with a high signal intensity for intracellular fura-2, indi