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Biomedical subjects

P Huovinen

Publications and source records attributed to P Huovinen.

At least 19 recordsLinked to original sources

Pneumonia in childhood: etiology and response to antimicrobial therapy.

A prospective eight-month study was carried out in 50 children admitted to hospital for radiologically confirmed community-acquired pneumonia. A potential causative agent of infection was identified in 44 (88%) cases. Using virus isolation, virus antigen detection and enzyme immunoassay serology, respiratory virus infection was diagnosed in 30 (60%) patients. Antibody assays for seven bacteria and antigen detection from serum and urine for Streptococcus pneumoniae produced evidence of bacterial infection in 31 (62%) cases. Streptococcus pneumoniae (38%), respiratory syncytial virus (30%) and Mycoplasma pneumoniae (20%) were the most common causative agents. A mixed infection was diagnosed in 25 (50%) episodes. Nine patients failed to respond to antibiotics within 24 h after onset of treatment. Three of them had a pure viral infection, three a mixed viral-bacterial infection, two a Mycoplasma pneumoniae infection mixed with other bacteria and one a pure Mycoplasma pneumoniae infection. All three Mycoplasma pneumoniae infections were initially treated with penicillin.

Adolescent

Synergism of trimethoprim and ciprofloxacin in vitro against clinical bacterial isolates.

For the first time, the effects of combinations of trimethoprim and a fluoroquinolone (ciprofloxacin) against gram-positive and gram-negative bacterial isolates were evaluated in vitro. Synergism was found in 31% (fractional inhibitory concentration, FIC) and 33% (fractional bactericidal concentration, FBC) of 121 clinical isolates of various bacterial strains, most often in Escherichia coli, staphylococci, and enterococci. Antagonism occurred in 1% (FIC) and 3% (FBC). The combination of trimethoprim and ciprofloxacin merits further evaluation for potential usefulness as a clinical regimen.

Ciprofloxacin

Epidemiology of plasmid-mediated beta-lactamases in enterobacteria Swedish neonatal wards and relation to antimicrobial therapy.

TEM-1, OXA-1, SHV-1, and related beta-lactamases in fecal isolates from 953 infants in 22 Swedish neonatal intensive care units were studied by DNA hybridization. TEM-1- and OXA-1-positive isolates were always Escherichia coli and represented 86 and 8%, respectively, of the ampicillin-resistant isolates of this species. SHV-1 was found in 16% of the Klebsiella sp. (mainly Klebsiella pneumoniae) isolates. TEM-1 and SHV-1 occurred in 14 and 16 units and in up to 64 and 26% of the neonates, respectively. On average, two to four different biochemical phenotypes per species per ward were positive for each beta-lactamase. All but 1 of the 33 E. coli phenotypes found to be TEM-1 positive were uniformly positive for the beta-lactamase gene, whereas some of the phenotypes found to be positive for OXA-1 (2 of 3) and SHV-1 (6 of 70) were occasionally negative for the respective genes. The occurrence of the three beta-lactamases studied tended to be associated with local ampicillin usage (correlation coefficient, 0.31 to 0.39; P greater than 0.05). Of the neonates receiving ampicillin, 30% carried TEM-1-positive E. coli, compared with 13% for cephalosporin-treated neonates and 15% for untreated neonates (P less than or equal to 0.001). The corresponding rates for SHV-1 in Klebsiella spp. were 18, 13, and 9% (P less than or equal to 0.01). Ampicillin is thus a significant risk factor for the maintenance of the most prevalent gram-negative plasmid-mediated beta-lactamases in hospitalized neonates.

Anti-Bacterial Agents

[Pharyngitis].

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Adenovirus Infections, Human

Problems in interpretation of piperacillin susceptibility of TEM-1 producing Escherichia coli in the disk diffusion test.

The susceptibility of 455 Escherichia coli blood culture isolate to piperacillin was tested with the disk diffusion test. The presence of different beta-lactamase genes in these strains was also studied using DNA hybridization. Of the TEM beta-lactamase producing isolates, 64% (61/95) were interpreted as intermediately susceptible to piperacillin. Because piperacillin is hydrolyzed by TEM-type beta-lactamases, we suggest that the intermediate susceptibility category should be reduced or omitted in testing piperacillin susceptibility of Escherichia coli isolates.

Drug Resistance, Microbial

Analysis of beta-lactamase production in ampicillin-resistant Escherichia coli isolated from blood cultures 1983-1989.

From 1983 to 1989, 520 Escherichia coli blood culture pathogens were isolated from two hospitals in Turku, Finland. Ampicillin resistance (MIC greater than or equal to 16 micrograms/ml) of these isolates increased from 33% in 1983 to 66% in 1987, but decreased to 38-49% in 1988-1989. Occurrence of TEM-1 beta-lactamase producing isolates increased only slightly from 14% in 1983 to 25% in 1989 among all Escherichia coli strains studied. Strains with ampicillin MIC values of 16 micrograms/ml and 32 micrograms/ml were mostly responsible for the increase in resistance. Among these isolates, TEM-1 or any other of the well known plasmid-mediated beta-lactamases were not found by hybridization or isoelectric focusing.

Ampicillin Resistance

Nosocomial Staphylococcus epidermidis septicaemia among very low birth weight neonates in an intensive care unit.

A cluster of cases of septicaemia caused by coagulase-negative staphylococci was observed among the infants at the Neonatal Intensive Care Unit (NICU) of the Karolinska Hospital in May 1987. The presence of a unique antibiogram among the blood culture isolates prompted an investigation to determine whether an epidemic strain existed or not, using antibiogram, biotyping and plasmid profiles as epidemiological markers. All 14 isolates with a unique antibiogram were investigated, and 22 isolates without the unique antibiogram served as controls. Of the 14 isolates, 11 were Staphylococcus epidermidis and had similar plasmid patterns. Of the 22 control isolates, 15 were S. epidermidis, and none had the special plasmid pattern nor any other recurring plasmid patterns. The use of plasmid profile analysis together with antibiograms thus identified a possible epidemic strain of S. epidermidis which may have been responsible for the upsurge of septicaemia cases at the NICU. The presence of an epidemic strain implies that hospital cross infection control could be important in preventing neonatal septicaemia caused by S. epidermidis.

Coagulase

Sequence of the PSE-1 beta-lactamase gene.

The nucleotide sequence of the PSE-1 beta-lactamase gene from Tn1403 indicates that it is contained in an integron and encodes a class A enzyme differing from PSE-4 and CARB-3 by single amino acid substitutions.

Amino Acid Sequence

Analysis of genetic localization of the type I trimethoprim resistance gene from Escherichia coli isolated in Finland.

Among a collection of clinical Escherichia coli isolates, the type I dihydrofolate reductase (DHFR) mediating trimethoprim resistance was generally observed to be chromosomally determined. Only a minority of isolates carried the type I DHFR gene simultaneously on a plasmid. The majority of E. coli isolates studied also hybridized with a probe specific for the transposition gene tnsC of transposon Tn7; and in most of these isolates, Tn7 was found to be inserted into a preferred site in the E. coli chromosome. A minority of isolates that harbored the type I DHFR gene in the chromosome lacked a complete Tn7. Some of these harbored the type I DHFR gene inserted in a structure similar to that containing the gene for streptomycin resistance in Tn21. In the other isolates that were negative for a complete Tn7, the sequences upstream of the type I DHFR gene were demonstrated to be homologous to those flanking the type I DHFR gene in Tn7. This could indicate that the antibiotic resistance region of Tn7 may occur independently of this transposon. In two isolates, no sequences resembling Tn7 or Tn21 were found adjacent to the type I DHFR gene.

Bacterial Proteins

Application of gas-liquid chromatographic analysis of cellular fatty acids for species identification and typing of coagulase-negative staphylococci.

Gas-liquid chromatography (GLC) of bacterial cellular fatty acids was used to analyze 264 isolates of coagulase-negative staphylococci, of which 178 were Staphylococcus epidermidis. The presence and amounts of individual fatty acids were determined to generate fatty acid profiles for each of the seven coagulase-negative species tested. The fatty acid profiles were then analyzed by computerized correlation and cluster analysis to calculate mean correlation values between isolates belonging to the same or different species, as well as to establish cluster analysis dendrograms. These data ultimately allowed the clustering of individual samples into species-specific clusters. Species identification by the GLC clustering was highly consistent with species identification by biochemical assays; the results were similar in 92.4% of the cases. The GLC profile correlation analysis was further used to analyze multiple blood isolates from 60 patients in order to determine the usefulness of this methodology in establishing identity, as well as differences, between consecutive patient isolates. The correlation between those multiple S. epidermidis isolates determined to be identical by standard techniques (such as the antibiogram, biotype, and plasmid profile) was significantly (P less than 0.001) higher than that between random isolates of the same species. The correlation coefficient was greater than 97 for 40 (97.6%) of the 41 patients with multiple identical blood isolates, compared with less than 95 in all 19 (100.0%) patients with multiple nonidentical isolates. The successful use of the computerized GLC analysis in this study demonstrated its appropriate application for species identification and typing of coagulase-negative staphylococci.

Bacterial Typing Techniques

Antibiotic susceptibility of coagulase-negative staphylococcal blood isolates with special reference to adherent, slime-producing Staphylococcus epidermidis strains.

The epidemiologic relationship between slime-producing coagulase-negative staphylococci and antibiotic resistance was evaluated by analyzing the in vitro susceptibility of 278 blood isolates to 10 different antimicrobial agents. The strain collection consisted of 60 coagulase-negative staphylococcal septicemia isolates, recovered from at least 2 successive blood cultures from 59 septicemia patients and 218 single blood isolates defined as insignificant. When the slime-producing Staphylococcus epidermidis isolates were compared with all non-slime-producing S. epidermidis isolates, the slime-producing isolates exhibited higher MIC50 and/or MIC90 values than the non-slime-producing isolates, and were also more commonly resistant against many of the antimicrobials tested. The association between slime production and resistance against tobramycin, gentamicin and rifampicin was statistically significant (p less than 0.05). A similar finding was observed among the S. epidermidis septicemia isolates when they were analyzed separately from the single (suspected blood contaminant) S. epidermidis isolates. Further characterization of the septicemia isolates revealed that the positive correlation between slime production and antibiotic resistance reflected, to some extent, multiresistance of 1 epidemic slime-producing S. epidermidis strain. No difference in resistance was observed between the slime-producing and non-slime-producing single S. epidermidis blood isolates.

Anti-Bacterial Agents

Outbreak of diarrhoea due to Escherichia coli O111:B4 in schoolchildren and adults: association of Vi antigen-like reactivity.

During six days in November, 1987, 611 pupils (age range 7-19 years) and 39 adults (23-57) at a school complex in southern Finland had diarrhoea due to Escherichia coli O111:B4. Diarrhoea developed in 137 other household members during the two weeks after the school outbreak. The source of the organism remains unknown. The outbreak strains, when incubated at 22 degrees C or exposed to ampicillin, lost the lipopolysaccharide O antigen and began to react with antisera against Salmonella typhi Vi antigen. The Vi antigen-like reactivity increased the adherence of the organisms to Hep-2 cells. These results indicate that E coli O111:B4, and possibly other enteropathogenic E coli strains, should be considered in the diagnosis of all diarrhoea cases and not only in infantile diarrhoea. Expression of Vi antigen in E coli may play a part in virulence by enhancing adherence to the intestinal epithelium.

Adolescent

Analysis of the relationship between bacterial adherence and extracellular production of mannose, galactose, glucose and ribose in Staphylococcus epidermidis and Staphylococcus hominis.

Gas-liquid chromatography-mass spectrometry was used to analyze the extracellular extracts of 108 coagulase-negative staphylococcal strains for the presence of mannose, galactose, glucose and ribose, in order to determine whether production of these four monosaccharides, regarded as potential staphylococcal slime components, was associated with the adherence capacity of the individuals strains. A total of 90 Staphylococcus epidermidis and 18 Staphylococcus hominis strains were studied. Using the quantitative spectrophotometric assay, 21 Staphylococcus epidermidis strains were classified as strongly adherent, 12 as moderately adherent, 11 as weakly adherent, and 46 as nonadherent. All 18 Staphylococcus hominis strains were nonadherent. Mannose, galactose, glucose and ribose were detected as the main monosaccharide components in the extracellular extracts of all strains examined. Moreover, the mean relative concentrations of these monosaccharides were essentially the same for the different adherence phenotypes within the species Staphylococcus epidermidis. These results showed that there was no causal connection between the adherence of coagulase-negative staphylococci and the extracellular production of any of the four monosaccharides analyzed.

Bacterial Adhesion

Immunochemical analysis of the extracellular slime substance of Staphylococcus epidermidis.

To analyze immunochemically the extracellular slime substance of Staphylococcus epidermidis, rabbits were repeatedly immunized with the crude slime extract isolated from an adherent, slime-producing clinical Staphylococcus epidermidis strain. Immunoelectron microscopy demonstrated that the target antigens of the resulting antibodies were located in the extracellular slime-like layer of bacterial cells. When these target antigens were characterized by immunoblotting, a variety of antigens were detected, including many with molecular masses higher than 80 kilodaltons and also a predominant one with a molecular mass of 30 kilodaltons. No characteristic differences were observed between the tube adherence test positive and negative Staphylococcus epidermidis isolates. Although there was variation in the number and intensity of high molecular mass antigens, such variations did not correlate with the tube adherence test. Of the 110 Staphylococcus epidermidis isolates studied, 106 (96%) expressed the 30-kilodalton antigen. This component was found in no other Staphylococcus spp. examined in the study. The bacterial component was not only easily detached from bacterial cells but also water-soluble, characteristics implicating a slime-like nature. Further studies are needed to definitively establish the origin and nature of the 30-kilodalton Staphylococcus epidermidis-specific component, and determine its potential benefit as a diagnostic tool.

Antibodies, Bacterial