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[Transcatheter exclusion of a giant aneurysm of the left pulmonary artery using covered stents].

An 11 year-old boy with an Alagille syndrome was referred for an urgent echocardiography because of new onset of febrile hemoptysis. Follow-up in our institution to that point had shown severe hypoplastic pulmonary arteries. The diagnosis of tricuspid endocarditis complicated by pulmonary embolisms and aneurysm of the left pulmonary artery was done. Despite effective antiobiotics, the evolution was marked by rapid progression in size of the left pulmonary aneurysm. Since the surgery was not an option because of high risk of death, this aneurysm was treated by a trans-catheter technique using covered stents (CP stents, Numed Inc). Its exclusion allowed to stop its progression in size and to avoid its rupture.

Aneurysm, Infected↗

Anterior mitral valve leaflet aneurysm due to infective endocarditis detected by cardiac magnetic resonance imaging.

We report a case of mitral valve aneurysm in a 30-year-old man presenting with fatigue, malaise, and fever 10 weeks after emergent aortic valve replacement for endocarditis. The transthoracic echocardiogram demonstrated perivalvular aortic regurgitation, but no abscess cavity was defined. Cardiac magnetic resonance imaging (MRI) revealed a mitral valve leaflet aneurysm. Both aortic and mitral valves were replaced with mechanical prostheses. Pathology of the excised mitral valve showed a focally hemorrhagic aneurysm of the anterior leaflet with myxoid degeneration and focal calcification. Early diagnosis and intervention are important to treat this rare, potentially fatal complication of aortic valve endocarditis. Cardiac MRI provided an accurate and useful preoperative diagnostic evaluation.

Abscess↗

Mitis salivarius-bacitracin 10% sacarose agar for oral streptococci and Streptococcus mutans counts.

The MSB Agar (mitis salivarius-bacitracin) 20% sacarose medium is frequently used for the isolation and count of total streptococci and Streptococcus mutans. Although it is considered a selective culture medium for this micro-organism, S. mutans recovery in this medium is much lower than in this Mitis Salivarius Agar (MSA). Because the number of S. mutans in saliva is used for estimating caries risk and activity from a microbiological stand point, the aim of this work was to find a modification of the MSB 20% sacarose medium so that it would offer not only selectivity in the isolation but also maximum recovery. This would detect people at risk more efficiency and would evaluate the preventive odontological treatments more accurately. The results show that: 1) the greatest recovery of total streptococci and S. mutans is obtained in the MSB 10% sacarose medium, 2) S. mutans must be incubated in aerobiosis and the total streptococci in a candle jar (10% CO2). MSB 10% sacarose medium is proposed as a choice medium for the microbiological estimation of cariogenic risk and activity, to detect infection levels and evaluate preventive odontological treatments.

Adolescent↗

Prophylactic teicoplanin during insertion of Hickman catheters.

With the extensive use of Hickman catheters in patients requiring cytotoxic chemotherapy there is now a resurgence of Gram-positive septicaemia among these patients during the neutropenic periods. We are currently running a prospective randomized study of prophylactic teicoplanin during insertion of Hickman catheters to determine if it reduces the incidence of Gram-positive septicaemia. A total of 44 patients have completed the study (23 in the teicoplanin group and 21 in the control group). The diagnoses were: acute myelogenous leukaemia (13), non-Hodgkin's lymphoma (16), Hodgkin's disease (11), chronic granulocytic leukaemia (3) and acute lymphoblastic leukaemia (1). The number of days between insertion of Hickman catheters and the development of neutropenia was: teicoplanin group: mean 14.7, median 11, range 0-53; control group: mean 11.8, median 10, range 0-37. In the treated patients, there were four episodes of line-associated Gram-positive septicaemia and a total of four organisms were isolated. In the control group, 10 organisms were isolated from a total of nine episodes of line-associated Gram-positive septicaemia. The organisms were: coagulase-negative staphylococci (8), Streptococcus B, (1), Strep. faecalis (1), Strep. mitis (1), alpha streptococcus (1), diphtheriods (1) and Staphylococcus aureus (1). All organisms were sensitive to teicoplanin. No adverse reaction was observed in any patient. Prophylactic teicoplanin during insertion of Hickman catheters may therefore reduce the incidence of line-associated Gram-positive septicaemia in neutropenic patients.

Anti-Bacterial Agents↗

Autolysis in strains of viridans streptococci.

Seven strains of viridans streptococci of the species Streptococcus sanguis, S. mutans and S. mitis were investigated for autolysis. The effect of pH, salt concentration and temperature on the autolytic process was studied in Na2HPO4/NaH2PO4 buffer. Whole cells and walls of all strains autolysed most rapidly at pH values above 7. Autolysis of whole cells of S. sanguis and one strain of S. mitis (ATCC15909) was maximal in 0-05 TO 0-2 M buffer, while the two S. mutans strains and S. mitis ATCC15912 showed maximal autolysis in 0-5 and 1-0 M buffers. Cultures harvested in the stationary phase of growth possessed only slightly decreased autolytic activity compared with those from the exponential phase. Whole cells autolysed more rapidly at 37 degrees C Than at 45 degrees C and 10 degrees C. Autolysis of isolated walls of three strains of S. mitis (ATCC903, ATCC15909 and ATCC15912) was maximal at pH 7-0 AND 7-5 and in 1-0 M buffers. Streptococcus mitis ATCC15909 also showed maximal lysis in 0-01 M and 0-5 M buffers. An endopeptidase action of the autolytic system of S. mitis ATCC15912 was indicated by the progressive release of soluble amino groups during autolysis of the walls. No release of reducing groups was observed. Several free amino acids were released during autolysis of these walls, alanine, lysine and glutamic acid being in greatest quanitity.

Amino Acids↗

Glucose-sucrose-potassium tellurite-bacitracin agar, an alternative to mitis salivarius-bacitracin agar for enumeration of Streptococcus mutans.

An agar medium for selective recovery and enumeration of Streptococcus mutans was developed as an alternative to mitis salivarius-bacitracin (MSB) agar. Combinations of dyes, antibiotics, and tellurite were added to a nonselective medium which, because of its sucrose content, allowed easy recognition of S. mutans colonies. Candle jar incubation for 2 days, by comparison with anaerobic incubation, reduced background flora but did not diminish S. mutans recoveries from clinical samples. Quantitative comparisons were made of the simultaneous recoveries of a number of authentic S. mutans serotype representatives and fresh clinical isolates, using various glucose-sucrose-potassium tellurite-bacitracin (GSTB) formulations and mitis salivarius, MSB, and blood agars. Mitis salivarius counts were not detectably different from blood counts, but counts on MSB were distinctly lower. A formulation of the new medium containing 5% glucose 5% sucrose, 0.001% potassium tellurite, 0.3 U of bacitracin per ml (hence GSTB), and 2% agar gave recoveries nearly equal to those on mitis salivarius agar and much greater than those on MSB. The medium yielded readily recognized S. mutans colonies and facilitated detection of intracellular polysaccharide formers upon flooding with I2 reagent. Freshly isolated serotype c, E, and f colonies could often be distinguished from serotype d and g colonies, a distinction made reliable by testing for intracellular polysaccharide. A study of 300 salivary samples revealed GSTB to give significantly higher recoveries than MSB. About 72% of all samples were substantially underestimated for S. mutans with MSB, and 6.7% of samples were falsely negative for S. mutans with MSB. Recovery of background flora on GSTB was as low or lower than on MSB, and both types of agar could be stored for at least 9 weeks without notable change of selectivity. Thus, GSTB agar appears to be simple and reliable to use and requires no anaerobic incubation. Caution is voiced about interpretation of data previously reported which evaluated S. mutans on MSB agar.

Agar↗

Allelic variation of polymorphic locus lytB, encoding a choline-binding protein, from streptococci of the mitis group.

The choline-binding protein LytB, an N-acetylglucosaminidase of Streptococcus pneumoniae, is the key enzyme for daughter cell separation and is believed to play a critical pathogenic role, facilitating bacterial spreading during infection. Because of these peculiarities LytB is a putative vaccine target. To determine the extent of LytB polymorphism, the lytB alleles from seven typical, clinical pneumococcal isolates of various serotypes and from 13 additional streptococci of the mitis group (12 atypical pneumococci and the Streptococcus mitis type strain) were sequenced. Sequence alignment showed that the main differences among alleles were differences in the number of repeats (range, 12 to 18) characteristic of choline-binding proteins. These differences were located in the region corresponding to repeats 11 to 17. Typical pneumococcal strains contained either 14, 16, or 18 repeats, whereas all of the atypical isolates except strains 1283 and 782 (which had 14 and 16 repeats, respectively) and the S. mitis type strain had only 12 repeats; atypical isolate 10546 turned out to be a DeltalytB mutant. We also found that there are two major types of alternating repeats in lytB, which encode 21 and 23 amino acids. Choline-binding proteins are linked to the choline-containing cell wall substrate through choline residues at the interface of two consecutive choline-binding repeats that create a choline-binding site. The observation that all strains contained an even number of repeats suggests that the duplication events that gave rise to the choline-binding repeats of LytB involved two repeats simultaneously, an observation that is in keeping with previous crystallographic data. Typical pneumococcal isolates usually grew as diplococci, indicating that an active LytB enzyme was present. In contrast, most atypical isolates formed long chains of cells that did not disperse after addition of purified LytB, suggesting that in these strains chains were produced through mechanisms unrelated to LytB.

Alleles↗

Clinical, phenotypic, and genotypic evidence for Streptococcus sinensis as the common ancestor of anginosus and mitis groups of streptococci.

In 2002, we reported the discovery of a novel species of viridans streptococcus, Streptococcus sinensis. Recently, we reported the isolation of two more strains of S. sinensis. Clinically, S. sinensis is a definite cause of infective endocarditis, a characteristic mainly pertaining to the mitis group of streptococci. Phenotypically, two of the three S. sinensis isolates were Lancefield group F, a characteristic of the anginosus group. However, none of the three strains possess the caramel smell typical of this group of streptococci. Biochemically, S. sinensis was identified in 56% of the time as members of the anginosus group, and in 33% of the time as members of the mitis group. These clinical and phenotypic properties should be governed by the presence/absence or expressivity of particular genes in the S. sinensis genome. Genotypically, phylogenetic analysis using 16S rRNA gene sequences showed that S. sinensis branched out as the first branch in the anginosus group, implying that it is the ancestor of the other members of this group. However, the bootstrap value for S. sinensis clustered with members of the anginosus group is only 47%, meaning that it is often not clustered with members of this group, but the mitis group. Furthermore, the differences in the 16S rRNA gene sequences between S. sinensis and Streptococcus intermedius (3.7%) and those between S. sinensis and Streptococcus gordonii (3.6%) are almost the same. All these indicated that it is very likely that S. sinensis is the common ancestor of the anginosus and mitis groups of streptococci. Complete genome sequencing of S. sinensis and comparative genomics studies on the S. sinensis genome and genomes of members in the anginosus and mitis groups should reveal clues to the underlying genotypic differences that govern the different phenotypic properties of the two groups of streptococci, such as why streptococci of the anginosus group are prone to cause abscess formation but not infective endocarditis as compared to other viridans streptococci.

Genotype↗

dnaJ and gyrB gene sequence relationship among species and strains of genus Streptococcus.

The dnaJ and gyrB nucleotide sequences were determined for members of the genus Streptococcus. The average similarity between the species tested was 76.4% (69.7-100%) for dnaJ and 75.9 (70.1-98.7%) for gyrB. These data indicated that the dnaJ and gyrB genes are more divergent and more discriminatory than the 16S rDNA gene. Furthermore, the variation in the dnaJ nucleotide sequences among the mitis group was greater than that of the gyrB nucleotide sequences, especially between Streptococcus pneumoniae and Streptococcus mitis. Subsequently, the high discrimination power of dnaJ within the mitis group was confirmed. Thus, we conclude that the dnaJ and gyrB genes are efficient alternative targets for the classification of the genus Streptococcus, and that dnaJ is suitable for phylogenetic analysis of closely related Streptococcus strains.

Bacterial Proteins↗

Species of alpha-hemolytic streptococci possessing a C-polysaccharide phosphorylcholine-containing antigen.

An enzyme-linked immunosorbent assay technique was used to detect and quantify C-polysaccharide-like antigen in strains of alpha-hemolytic streptococci classified into species by following the latest taxonomic recommendations. C-polysaccharide-like antigen is found only in Streptococcus oralis, S. mitis, and S. pneumoniae, which are genetically closely related.

Antigens, Bacterial↗

In vitro exchange of fluoroquinolone resistance determinants between Streptococcus pneumoniae and viridans streptococci and genomic organization of the parE-parC region in S. mitis.

Transfer of fluoroquinolone (FQ) resistance determinants between Streptococcus pneumoniae and viridans streptococci was explored by transformation in vitro. One-step FQ-resistant parC mutants were selected, and resistance could be transferred from DNA from S. oralis, S. mitis, S. sanguis, and S. constellatus to S. pneumoniae, with frequencies of 10(-3) to <10(-7) in correlation with the homologies of their quinolone resistance determining region sequences (95%, 91%, 85%, and 81%, respectively). Reciprocal transfers of mutated parC from DNA from S. pneumoniae to S. mitis and S. oralis were also observed. Simultaneous transfer of mutated parC and gyrA genes from S. mitis to S. pneumoniae yielded high-level-resistant pneumococcal transformants in one step at low frequencies. The parE-parC region of the type strain S. mitis 103335T had >90% homology with that of S. pneumoniae. The efficient interspecific transfer of quinolone resistance determinants in vitro leads us to anticipate their dissemination in the clinical setting.

Anti-Infective Agents↗

Adsorption of salivary proteins to the surface of oral streptococcal cells.

Oral tissues, especially tooth surfaces, are covered with a layer of salivary proteins. Oral bacterial cells that adsorb to salivary components accumulated on the tooth surface are, as a rule, covered with the same components, especially proteins. Thus, it is possible that the salivary proteins covering the bacterial cells are related to the adhesion of bacteria to oral tissues. The aim of this study was to clarify the mechanisms of adsorption of salivary proteins to the surface of Streptococcus sanguis, S. mitis and S. salivarius using an adsorption assay with salivary proteins labeled with tritiated formaldehyde. The results showed that salivary proteins adsorbed more to S. salivarius than to S. mitis, and least to S. sanguis. It was evident that hydrophobic bonding was involved in the adsorption of salivary proteins to the bacterial cells tested. The amount of salivary proteins adsorbed to S. mitis and S. salivarius was decreased by the presence of phosphate, that to S. sanguis was increased by the presence of a divalent cation such as Ca2+, and that to all bacteria tested was inhibited in different ways by the presence of sugars. The amount of salivary proteins adsorbed to S. sanguis and S. salivarius was reduced effectively by pretreatment of the cells with trypsin, chymotrypsin and papain. In the case of S. mitis, the amount of adsorbed salivary proteins was decreased by pretreatment of the cells with chymotrypsin only, and was increased by pretreatment with lipase. These results indicate that there are different mechanisms of adsorption of salivary protein to the cell surfaces of oral streptococci.

Acetylglucosamine↗

Alpha-hemolytic streptococcal bacteremia: a review of 203 episodes during 1980-1991.

This is a review of 203 episodes of alpha-hemolytic streptococcal bacteremia during a 12-year period (1980-91) in a large community teaching hospital in the USA. There were 87 male and 116 female patients, with ages ranging from newborn infants to a 96-year-old; 118 patients (58.1%) were 60 years or older. 31 patients had endocarditis, 93 had bacteremia, and 79 had polymicrobial bacteremia. Streptococcus sanguis, S. mitis, S. salivarius and S. intermedius accounted for 80% of the strains. Some strains were resistant to tetracycline, nafcillin, erythromycin or clindamycin. The overall mortality was 29.6% (60 of 203 patients). There was no significant difference in the mortality among patients with endocarditis, bacteremia, and polymicrobial bacteremia. Factors that adversely influenced the mortality rate were old age (60 years and older) and rapidly fatal or ultimately fatal underlying conditions.

Adolescent↗

Hydrogen peroxide excretion by oral streptococci and effect of lactoperoxidase-thiocyanate-hydrogen peroxide.

Approved type strains of Streptococcus sanguis, S. mitis, S. mutans, and S. salivarius were grown under aerobic and anaerobic conditions. The rate of hydrogen peroxide excretion, oxygen uptake, and acid production from glucose by washed-cell suspensions of these strains were studied, and the levels of enzymes in cell-free extracts which reduced oxygen, hydrogen peroxide, or hypothiocyanite (OSCN-) in the presence of NADH or NADPH were assayed. The effects of lactoperoxidase-thiocyanate-hydrogen peroxide on the rate of acid production and oxygen uptake by intact cells, the activity of glycolytic enzymes in cell-free extracts, and the levels of intracellular glycolytic intermediates were also studied. All strains consumed oxygen in the presence of glucose. S. sanguis, S. mitis, and anaerobically grown S. mutans excreted hydrogen peroxide. There was higher NADH oxidase and NADH peroxidase activity in aerobically grown cells than in anaerobically grown cells. NADPH oxidase activity was low in all species. Acid production, oxygen uptake, and, consequently, hydrogen peroxide excretion were inhibited in all the strains by lactoperoxidase-thiocyanate-hydrogen peroxide. S. sanguis and S. mitis had a higher capacity than S. mutans and S. salivarius to recover from this inhibition. Higher activity in the former strains of an NADH-OSCN oxidoreductase, which converted OSCN- into thiocyanate, explained this difference. The change in levels of intracellular glycolytic intermediates after inhibition of glycolysis by OSCN- and the actual activity of glycolytic enzymes in cell-free extracts in the presence of OSCN- indicated that the primary target of OSCN- in the glycolytic pathway was glyceraldehyde 3-phosphate dehydrogenase.

Depression, Chemical↗

Distribution of environmentally regulated genes of Streptococcus suis serotype 2 among S. suis serotypes and other organisms.

The occurrence of 36 environmentally regulated genes of Streptococcus suis strain 10 among all 35 S. suis serotypes was determined by using hybridization with the amplified genes as probes. In addition, the distribution of these genes among the virulence phenotypes of serotypes 1 and 2 was assessed. Hybridization was also performed with various other streptococcal species and nonstreptococcal bacterial species which may be present in pigs. Interestingly, probe ivs-25/iri-1, similar to agrA and sapR, hybridized only with S. suis serotype 1 and 2 strains with virulent phenotypes and is therefore suitable as a diagnostic parameter. Only one probe was specific for S. suis. This probe's sequence was identical to the epf gene, a putative virulence factor of S. suis. Probe ivs-31 was similar to a virulence factor of S. suis, namely, a gene encoding a fibronectin- and fibrinogen-binding protein. This probe hybridized only with oral streptococci. Nearly half of the probes (45%) hybridized with the oral streptococci (S. oralis, S. milleri, S. sanguis, S. gordonii, and S. mitis) and with Streptococcus pneumoniae. This indicates a close relationship between S. suis, the oral streptococci, and S. pneumoniae with respect to the selected environmentally regulated genes. One probe only hybridized with gram-negative species and therefore seems to be obtained by S. suis from a gram-negative organism by horizontal transfer.

Animals↗

Cell surface proteins of oral streptococci.

Whole cells of representative strains of oral streptococci (Streptococcus sanguis, S. mitis, and S. salivarius) were radiolabeled by the lactoperoxidase method of radioiodination. The labeled polypeptides obtained by extraction of whole cells with boiling sodium dodecyl sulfate were analyzed by polyacrylamide gel electrophoresis and autoradiography. Of the total radioactivity, ca. 70% was released by treating whole cells with trypsin, suggesting that the labeling was confined to proteins located on the cell surface. Most S. sanguis strains studied gave a characteristic banding pattern consisting of a high-molecular-weight (120,000 [120K] to 63K) group of six proteins. Three low-molecular-weight (12K, 16K, and 18K) proteins were also detected in many strains.

Bacterial Proteins↗

Rapid identification of Streptococcus and Enterococcus species using diffuse reflectance-absorbance Fourier transform infrared spectroscopy and artificial neural networks.

Diffuse reflectance-absorbance Fourier transform infrared spectroscopy (FT-IR) was used to analyse 19 hospital isolates which had been identified by conventional means to one Enterococcus faecalis, E. faecium, Streptococcus bovis, S. mitis, S. pneumoniae, or S. pyogenes. Principal components analysis of the FT-IR spectra showed that this 'unsupervised' learning method failed to form six separable clusters (one of each species) and thus could not be used to identify these bacteria base on their FT-IR spectra. By contrast, artificial neural networks (ANNs) could be trained by 'supervised' learning (using the back-propagation algorithm) with the principal components scores of derivatised spectra to recognise the strains from their FT-IR spectra. These results demonstrate that the combination of FT-IR and ANNs provides a rapid, novel and accurate bacterial identification technique.

Algorithms↗

Identification of clinically relevant viridans group streptococci to the species level by PCR.

A PCR assay that allows identification of clinically relevant viridans group streptococci (Streptococcus gordonii, S. mitis, S. mutans, S. oralis, S. salivarius, and S. sanguis) to the species level and identification of milleri group streptococci (S. anginosus, S. constellatus, and S. intermedius) to the group level was developed. This assay was based on specific amplification of internal fragments of genes encoding D-alanine:D-alanine ligases which are species specific and ubiquitous in prokaryotes possessing peptidoglycan. The specificity of this assay was tested on 9 reference strains and 91 characterized clinical isolates. This assay offers a specific and rapid alternative to phenotypic or DNA-DNA hybridization methods for identification of clinically relevant viridans group streptococci.

Alanine↗