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T W Steele

Publications and source records attributed to T W Steele.

At least 19 recordsLinked to original sources
PubMed2001

Metabolism of S-nitrosoglutathione in intact mitochondria.

S-nitrosation of protein thiol groups by nitric oxide (NO*) is a widely recognized protein modification. Only few intracellular S-nitrosated proteins have been identified and it has been reported that S-nitrosation/denitrosation can serve as a regulatory process in signal-transduction pathways. Given the potential physiological importance of S-nitrosothiols, and considering that mitochondria are endowed with high levels of thiols and the biochemical requisites for synthesizing NO*, we examined the occurrence of S-nitrosoglutathione (GSNO) in intact, coupled rat liver mitochondria. These organelles contained 0.34 nmol of GSNO/mg of protein, detected by HPLC with UV-visible and electrochemical detections. This concentration was dynamically modulated by the availability of NO*; its decay was affected mainly by GSH and superoxide dismutase in a reaction that entailed the generation of GSSG. On the basis of the relatively long half-life of GSNO and the negligible recovery of NO* during its decay, roles for GSNO as a storage and transport molecule for NO* are discussed. Moreover, the formation of GSNO and its reaction with GSH can be considered to be partly responsible for the catabolism of NO* via a complex mechanism that might result in the formation of hydroxylamine, nitrite or nitrous oxide depending upon the availability of oxygen, superoxide dismutase and glutathione. Finally, the high concentrations of GSH in the cytosol and mitochondria might favour the formation of GSNO by reacting with NO* 'in excess', thereby avoiding damaging side reactions (such as peroxynitrite formation), and facilitate the inactivation of NO* by generating other nitrogen-related species without the chemical properties characteristic of NO*.

Animals↗
PubMed1998

Sequence analysis of the mip gene of the soilborne pathogen Legionella longbeachae.

To understand the basis of pathogenesis by Legionella longbeachae serogroup 1, the importance of the Mip protein in this species was examined. Amino-terminal analysis of the purified, cloned L. longbeachae serogroup 1 ATCC 33462 Mip protein confirmed that the cloned gene protein was expressed and processed in an Escherichia coli background. DNA sequence analysis of plasmid pIMVS27, containing the entire L. longbeachae serogroup 1 mip gene, revealed a high degree of homology to the mip gene of Legionella pneumophila serogroup 1, 76% homology at the DNA level and 87% identity at the amino acid level. Primer extension analysis determined that the start site of transcription was the same for both species, with some differences observed for the -10 and -35 promoter regions. Primers designed from the mip gene sequence obtained for L. longbeachae serogroup 1 ATCC 33462 were used to amplify the mip genes from L. longbeachae serogroup 2 ATCC 33484 and an Australian clinical isolate of L. longbeachae serogroup 1 A5H5. The mip gene from A5H5 was 100% identical to the type strain sequence. The serogroup 2 strain of L. longbeachae differed by 2 base pairs in third-codon positions. Allelic exchange mutagenesis was used to generate an isogenic mip mutant in ATCC 33462 and strain A5H5. The ATCC mip mutant was unable to infect a strain of Acanthamoebae sp. both in liquid and in a potting mix coculture system, while the A5H5 mip mutant behaved in a manner siilar to that of L. pneumophila serogroup 1, i.e., it displayed a reduced capacity to infect and multiply within Acanthamoebae. To determine if this mutation resulted in reduced virulence in the guinea pig animal model, the A5H5 mip mutant and its parent strain were assessed for their abilities to establish an infection after aerosol exposure. Unlike the virulent parent strain, the mutant strain did not kill any animals under two different dose regimes. The data indicate that the Mip protein plays an important role in the intracellular life cycle of L. longbeachae serogroup 1 species and is required for full virulence.

Amino Acid Sequence↗
PubMed1996

Infection of Tetrahymena pyriformis by Legionella longbeachae and other Legionella species found in potting mixes.

All Legionella longbeachae strains, both serogroups of L. bozemanii, and three strains of L. anisa reproducibly infected washed Tetrahymena pyriformis at 30 degrees C. L. pneumophila serogroup 1 strains infected T. pyriformis less reproducibly than did L. longbeachae. Low-level concentrations of nutrients in cocultures inhibited infection. Four L. micdadei strains and L. anisa ATCC 35292 failed to infect T. pyriformis.

Animals↗
PubMed1994

Occurrence and distribution of Legionella species in composted plant materials.

Legionellae were found in many samples of composted plant matter obtained from home gardeners and from facilities which undertook bulk composting. The predominant species isolated from these composts was Legionella pneumophila, the strains of which belonged to serogroups other than serogroup 1. Other Legionella species were present in many samples. Legionella longbeachae serogroup 1, which is implicated in human infections in South Australia, was present in samples obtained from two of six facilities composting large volumes of material and from 3 of 30 gardeners. Many of the species or strains isolated from composts have not been implicated as causative agents of legionellosis in South Austrailia, but some cause infection in healthy and immunosuppressed persons.

Agriculture↗
PubMed1990

Isolation of Legionella longbeachae serogroup 1 from potting mixes.

Following a statewide outbreak of legionellosis due to Legionella longbeachae serogroup 1 in South Australia in 1988 and 1989, studies were performed to find a source of the organism. A number of water and soil samples with and without acid decontamination were examined for L. longbeachae by using a selective medium containing vancomycin, aztreonam, and pimafucin. There were no isolations of L. longbeachae from water samples. Organisms resembling L. longbeachae were isolated from a number of samples of potting mixes and from soil surrounding plants in pots collected from the homes of four patients. The organisms were found to persist for 7 months in two potting mixes stored at room temperature. Legionellae were isolated with difficulty from potting mixes which were allowed to dry out. Identification of isolates as L. longbeachae serogroup 1 was confirmed by quantitative DNA hybridization and serological tests. Restriction-fragment-length-polymorphism studies showed minor differences between patient and environmental isolates but differentiated these readily from L. longbeachae serogroup 2 and other antigenically related legionellae. The isolation of L. longbeachae from some potting mixes and the prolonged survival of the organisms in this medium suggest that soil rather than water is the natural habitat of this species and may be the source of human infections.

Blotting, Southern↗
PubMed1990

Distribution of Legionella longbeachae serogroup 1 and other legionellae in potting soils in Australia.

Legionella longbeachae serogroup 1 and other Legionella spp. were isolated from 73% of 45 potting soils made in Australia by 13 manufacturers but were not detected in 19 potting soils made in Greece, Switzerland, and the United Kingdom examined between March 1989 and May 1990. Several Legionella species were isolated from a small number of samples of uncomposted pine sawdusts, but it is not known whether sawdust was the source of some of the legionellae found in potting soils. Legionella spp. persisted for periods ranging from 3 to 10 months in a potting soil held at temperatures between -20 and 35 degrees C. Isolates of L. longbeachae serogroup 1 from soil did not grow at 43 degrees C, a temperature which was also lethal for this species in soil. Most Legionella spp. isolated from potting and natural soils belonged to one distinct group according to analysis of ubiquinones and were serologically related to several known species in this group. A small number of potting soils contained L. pneumophila and L. micdadei.

Australia↗
PubMed1990

Genetic relatedness of Legionella longbeachae isolates from human and environmental sources in Australia.

The genetic relatedness of Legionella longbeachae isolated in Australia since 1987 was investigated by restriction fragment length polymorphism (RFLP) analysis and allozyme electrophoresis. Three radiolabeled probes were used in Southern hybridizations for the RFLP studies. They were Escherichia coli 16S and 23S rRNA and cloned fragments of L. longbeachae selected empirically from genomal banks in lambda and a cosmid. The legionellae included in the study comprised 11 Legionella longbeachae serogroup 1 organisms isolated from humans, 28 L. longbeachae serogroup 1 isolates from environmental sources, and 3 L. longbeachae serogroup 2 environmental isolates. These were compared with the American Type Culture Collection reference strains of both serogroups and some other related Legionella species. Results of allozyme and RFLP analysis showed that all the isolates from humans and all but three of the environmental L. longbeachae serogroup 1 isolates were closely related. They were also closely related to L. longbeachae serogroup 1 ATCC 33462. There was wider variation among the three L. longbeachae serogroup 2 environmental isolates. One of these was closely related to L. longbeachae serogroup 2 ATCC 33484. RFLP studies with the rRNA probe provided the most discrimination among isolates but did not distinguish between the two serogroups.

Australia↗
PubMed1989

Legionnaires' disease in South Australia, 1979-1988.

Laboratory investigations supported the diagnosis of legionella pneumonia in 108 patients in South Australia over the past 10 years. Legionella pneumophila was responsible for 91 infections: the serogroup-1 strain caused 81 of these. L. pneumophila serogroup 2 was the only other strain of L. pneumophila that was isolated from patients; it caused infection in eight patients. In two patients, the serological results did not distinguish between infection with L. pneumophila serogroup 1 and serogroup 2. Legionella longbeachae serogroup 1 accounted for the remaining 17 infections. Serological tests were used to make the diagnoses in 77 cases. Legionella spp. were isolated from 24 patients and were identified in the respiratory-tract secretions of a further seven cases by direct immunofluorescence microscopy. L. longbeachae serogroup 1 first was isolated from a patient with pneumonia in South Australia in May, 1987. Since then it has been isolated from specimens from six other patients. No evidence exists for a common-source outbreak of L. longbeachae but an outbreak of Legionnaires' disease that was caused by L. pneumophila serogroup 1 occurred in South Australia in 1986.

Agglutination Tests↗
PubMed1988

Plasmid profile analysis of a salmonellosis outbreak and identification of a restriction and modification system.

After an outbreak of salmonellosis in humans caused by Salmonella typhimurium bacteriophage type 135, 62 isolates from human, animal, and water sources were retained for further analysis. Most of the isolates (92%) could be placed in one of five plasmid pattern groups, with a majority containing a common 60-kilobase plasmid and a smaller 3.8-kilobase-pair plasmid. This small plasmid, pIMVS1, was labeled with [32P]phosphate and used as a probe in subsequent colony and Southern hybridization studies. We concluded that pIMVS1 from isolates obtained from humans was genetically different from plasmids of a similar size found in isolates from chickens. Studies to characterize pIMVS1 were undertaken to determine if it codes for known virulence factors. It did not appear to be associated with the formation of attachment pili or major outer membrane proteins. By using transposon mutagenesis techniques, Tn3(Apr) was inserted into pIMVS1, and the existence of a restriction and modification system was deduced.

Animals↗
PubMed1988

Maturation of the rat small intestine at weaning: changes in epithelial cell kinetics, bacterial flora, and mucosal immune activity.

The relationship between maturation of the small intestine and change in mucosal immune activity was examined in the DA rat during the weaning period from 12 to 30 days. Two stages of jejunal maturation were observed: an initial stage of morphological development and crypt proliferation (days 12 to 22), followed by a period of stabilisation (days 24 to 30). By day 22 of the initial phase, villi increased principally in width but not in length, crypt length increased, and crypt cell production rate increased from 0.5 (day 12) to 11.1 (day 22) cells/crypt/hour. Various measures of mucosal immune activity showed a biphasic response. Up to days 20 to 22, the weight of the mesenteric lymph node increased seven-fold (p less than 0.0001), counts of jejunal eosinophils and goblet cells increased 3- (p less than 0.0001) and 19-fold (p less than 0.0001) respectively, and mean serum rat mucosal mast cell protease II, released from mucosal mast cells, increased from 24 (day 12) to 313 (day 22) ng/ml (p less than 0.0001). After day 22, mesenteric lymph node weight stabilised, eosinophil count stabilised and goblet cells decreased, serum rat mucosal mast cell protease II decreased three-fold (p less than 0.0001), and mean jejunal count of intraepithelial lymphocytes increased from 26 (day 22) to 54 (day 24) cells per mm of muscularis mucosae (p less than 0.0001), before stabilising. These results demonstrated a close association between maturation of the small intestine and change in activity of the mucosal immune system.

Animals↗
PubMed1985

DNA relatedness and biochemical features of Campylobacter spp. isolated in central and South Australia.

Investigations of the etiology of diarrhea in patients in South Australia and the Northern Territory showed that Campylobacter spp. other than Campylobacter jejuni and C. coli were common in children. Campylobacters which were hippurate positive, nitrate negative, and susceptible to cephalothin and polymyxins were shown to be closely related to C. jejuni by DNA studies. Thermotolerant catalase-negative campylobacters were also isolated. These were H2S negative and biochemically resembled the catalase-negative or weak strains found in dogs in Sweden. DNA studies showed these campylobacters to be distinct from C. sputorum subsp. sputorum and to form a homogeneous group distinct from the enteropathogenic catalase-positive campylobacters. Preliminary studies suggest that these campylobacters are related to the Swedish catalase-negative or weak strains.

Adult↗
PubMed1985

Acute enteric sepsis: bacteriology and antibiotic cover.

Most cases of enteric sepsis are caused by both aerobic and anaerobic organisms which form the normal flora of the mouth and lower gastrointestinal tract. This flora is extremely variable and subject to change due to disease and antimicrobial treatment. Bacteriological investigation of patients with severe enteric sepsis is important and should be undertaken before antibiotic treatment is commenced. The choice of antibiotics depends on the nature of the infection and its location. Initially they should be given in maximum dosage. If polymicrobial infection is suspected both aerobes and anaerobes should be covered to prevent bacteraemic shock and abscess formation. If abscesses have formed or the patient fails to respond to appropriate antibiotics, surgical exploration and drainage remain the treatment of choice. Antibiotics often fail to eradicate organisms from established abscesses and are responsible for some serious complications.

Abdomen↗
PubMed1984

The use of membrane filters applied directly to the surface of agar plates for the isolation of Campylobacter jejuni from feces.

Cellulose triacetate membrane filters applied directly to the surface of non-selective blood agar plates were found to be as effective as the use of antibiotic media in isolating Campylobacter jejuni from patients with diarrhea. This method was used in parallel with selective media in the examination of 1000 specimens of feces. Campylobacters were isolated from 56 specimens using all methods. The membrane filter method detected 50 (89%), 45 of which were C. jejuni, and selective media 45 strains of C. jejuni (80%). Membrane filters used in this way can result in the detection of most cases of campylobacter enteric infection and can be used by small laboratories with limited access to selective media. They may also facilitate the isolation of antibiotic sensitive campylobacters.

Agar↗
PubMed1980

Legionnaires' disease in South Australia. Prevalence and diagnosis.

Legionella pneumophila was successfully isolated from sputum, and from respiratory secretions obtained by tracheal aspiration, of two patients with Legionnaires' disease by means of guinea pigs and charcoal yeast extract agar. Direct culture of lung tissue from one of these patients gave a pure growth of L. pneumophila. In both cases, legionellas were isolated from specimens which were collected several days after treatment with erythromycin began. Direct fluorescent antibody tests on these specimens gave positive results. This test can result in the rapid diagnosis of legionellosis in carefully selected patients. Serological diagnosis by demonstrating a greater than fourfold rise in antibody level, particularly that of IgM antibody, is the method of choice for making the diagnosis of legionellosis in patients who do not have life-threatening infections or in whom a definitive bacteriological diagnosis cannot be made. Serological studies suggest that infection with L. pneumophila is endemic in South Australia.

Adult↗