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Biomedical subjects

D J Brenner

Publications and source records attributed to D J Brenner.

At least 163 records · Page 9Linked to original sources

Legionella shakespearei sp. nov., isolated from cooling tower water.

A Legionella-like organism (strain 214T [T = type strain]) was isolated from a cooling tower in Stratford-upon-Avon, England. This strain required L-cysteine and contained cellular branched-chain fatty acids that are typical of the genus Legionella. Strain 214T produced pink colonies on buffered charcoal-yeast extract agar. Ubiquinone Q-12 was the major quinone. Strain 214T was serologically distinct from other legionellae as determined by a slide agglutination test. The results of DNA hybridization studies showed that strain 214T (= ATCC 49655T) is a member of a new Legionella species, Legionella shakespearei.

Base Composition↗

Recognition of Morganella subspecies, with proposal of Morganella morganii subsp. morganii subsp. nov. and Morganella morganii subsp. sibonii subsp. nov.

The genus name Morganella was established within the family Enterobacteriaceae in 1978. Morganella morganii is the only species described thus far within this genus, and the name M. morganii has been accepted by usage in the scientific community for strains previously known as Proteus morganii. M. morganii isolates differ in their abilities to ferment trehalose and exhibit variable lysine and ornithine decarboxylase patterns, emphasizing the phenotypic heterogeneity within this species. Previous genetic studies failed to reveal separate entities within the genus Morganella. We observed some trehalose-fermenting strains with different lysine and ornithine decarboxylase patterns. Two strains were lysine and ornithine positive, 3 were lysine positive and ornithine negative, and 29 were lysine negative and ornithine positive. These strains and 25 non-trehalose-fermenting strains with different lysine and ornithine decarboxylase patterns were investigated. DNA-DNA hybridization studies and phenotypic characterizations revealed that M. morganii can be separated into three DNA relatedness groups and seven biogroups. Strains from DNA relatedness group 1 were trehalose negative, and strains from DNA relatedness groups 2 and 3 were trehalose positive. One biogroup from DNA relatedness group 2 was phenotypically indistinguishable from DNA relatedness group 3. On the basis of these studies, we propose that M. morganii be subdivided into M. morganii subsp. morganii (type strain ATCC 25830) containing biogroups A, B, C, and D (DNA relatedness group 1) and M. morganii subsp. sibonii (type strain 8103-85; = ATCC 49948) containing biogroups E, F, and G (DNA relatedness groups 2 and 3).

Bacterial Typing Techniques↗

Characterization of Neisseria meningitidis serogroup C by multilocus enzyme electrophoresis and ribosomal DNA restriction profiles (ribotyping).

We compared multilocus enzyme electrophoresis (MEE) and ribosomal DNA fingerprinting (ribotyping) for subtyping 44 strains of Neisseria meningitidis serogroup C that were isolated in Los Angeles County, California, between December 1985 and July 1986. The isolates were divided into six enzyme types (ETs) by MEE, but 36 of the isolates were clustered in one ET, 3. The same isolates were divided into 17 ribotypes by use of restriction endonucleases ClaI, EcoRI, and XhoI. Twenty of the 36 ET 3 isolates were divided into 17 ribotypes by use of restriction endonucleases ClaI, EcoRI, and XhoI. Twenty of the 36 ET 3 isolates were grouped in a single ribotype, J. The rate of infection with ribotype J strains was higher in the southern part of the study area than in the northern part. Isolates from each of eight pairs (each isolate pair was cultured from the same patient from the same or different sites) were found identical by MEE, but ribotyping revealed a difference in one pair. In this study, ribotyping showed a greater discriminating capacity than MEE for subtyping N. meningitidis serogroup C, but the epidemiologic relevance of this increased sensitivity needs further assessment.

Adolescent↗

Rochalimaea henselae sp. nov., a cause of septicemia, bacillary angiomatosis, and parenchymal bacillary peliosis.

Nine strains of Rochalimaea spp. that were isolated from patients over a period of 4.5 years were characterized for their enzyme activities, cellular fatty acid compositions, and DNA interrelatedness among Rochalimaea spp., Bartonella bacilliformis, and Afipia felis (cat scratch disease bacillus). All except one isolate, which was Rochalimaea quintana, were determined to belong to a newly proposed species, Rochalimaea henselae sp. nov. After recovery from clinical material, colonies required 5 to 15 days of incubation to become apparent. Cells were small, gram-negative, curved bacilli and displayed twitching motility. Enzyme specificities for amino acid and carbohydrate substrates showed that R. henselae could be distinguished from Rochalimaea vinsonii by L-arginyl-L-arginine and L-lysyl-L-alanine peptidases, but not all strains could be distinguished from R. quintana on the basis of peptidases or carbohydrate utilization. R. henselae also closely resembled R. quintana in cellular fatty acid composition, with both consisting mainly of C18:1, C18:0, and C16:0 fatty acids. However, the strains of R. henselae all contained C18:0 in amounts averaging greater than or equal to 22%, in contrast to R. quintana, which contained this cellular fatty acid in amounts averaging 16 and 18%. DNA hybridization confirmed the identification of one clinical isolate as R. quintana and showed a close interrelatedness (92 to 100%) among the other strains. Under optimal conditions for DNA reassociation, R. henselae showed approximately 70% relatedness to R. quintana and approximately 60% relatedness to R. vinsonii. Relatedness with DNA from B. baciliformis was 43%. R. henselae was unrelated to A. felis. R. henselae is the proposed species of a newly recognized member of the family Rickettsiaceae, which is a pathogen that may be encountered in immunocompromised or immunocompetent patients. Prolonged fever with bacteremia or vascular proliferative lesions are clinical manifestations of the agent.

Adult↗

Legionella lansingensis sp. nov. isolated from a patient with pneumonia and underlying chronic lymphocytic leukemia.

A Legionella-like organism, strain 1677-MI-H, was isolated from the bronchoscopy washings of a patient with pneumonia who had a 2-year history of progressive, chronic lymphocytic leukemia. The growth characteristics, cellular fatty acids, and ubiquinone content of the isolate were consistent with those for Legionella spp. The isolate was serologically distinct in the slide agglutination test with absorbed antisera. DNA hybridization studies showed that strain 1677-MI-H (ATCC 49751) represents a new Legionella species which is named Legionella lansingensis.

Agglutination Tests↗

Biological intercomparisons of neutron beams used for radiotherapy generated by p(+)-->Be in hospital-based cyclotrons.

The new generation of hospital-based neutron therapy facilities involve cyclotrons using protons on beryllium. The spectrum of neutrons produced includes a large and variable proportion of low-energy neutrons that are poorly penetrating but biologically effective. Cells cultured in vitro were used to compare the three US facilities at Seattle, M.D. Anderson and UCLA, together with the UK facility at Clatterbridge. Cyclotrons were compared within a given experiment on the same day using cells from a common suspension. Among the three US facilities, the relative potency factor at a depth of 25 mm differs by about 11%, with Seattle the least and UCLA the most biologically effective. Clatterbridge was compared directly with M.D. Anderson and found to be less effective by about 5%; it has a slightly lower biological effectiveness than any of the US facilities. There is evidence for an increased biological effectiveness in the build-up region, which reduces the effective skin sparing potential. There is not much difference in build-up between the three US facilities. Using the proton-on-beryllium neutron production process results in a wide spectrum of neutrons with a large but variable low-energy component. The biological effectiveness of the beam depends on target design and thickness as well as the design of the collimating system. Consequently the biological effectiveness of neutron beams generated by this process must be assessed on an individual basis. It cannot be assumed that because cyclotrons have similar accelerating energies that the relative biological effectiveness will be the same.

Animals↗

The dose-rate effect in interstitial brachytherapy: a controversy resolved.

Low-dose-rate interstitial implants generally involve a range of dose rates varying from about 0.3 to 1.5 Gy/h. It has been a matter of some debate as to whether such a variation in dose rate has any impact on clinical results, i.e. whether implant treatment time needs to be taken into account when prescribing the treatment dose. In particular, Paterson and Ellis both published data based on clinical experience suggesting that the prescribed dose should vary with overall treatment time, whereas the Paris school suggested that isoeffect dose should not be varied for treatment times between 3 and 8 days. Experimental radiobiological data obtained in vitro and in vivo imply a significant change in biological effectiveness over this range of dose rates with a greater variation for normal-tissue late effects than for tumour control. We show, based on radiological considerations and model calculations, that the disagreement between the two schools in their recommendations for generating isoeffect doses for different treatment times was a result of two factors. First, the Paterson recommendation was unequivocally based on matching limiting late effects, whereas the Paris system recommendations were based on an analysis of a combination of late and early effects. Second, the Paterson recommendations were based on a dosimetry system in which the dose rate does not change greatly with tumour volume, whereas the Paris recommendations were based on a dosimetric system in which dose rate is correlated with tumour volume, which in turn is correlated with changes in both tumour control and, especially, necrosis.

Animals↗

Conditions for the equivalence of continuous to pulsed low dose rate brachytherapy.

Low dose rate interstitial brachytherapy is extremely useful for those tumors that are accessible for an implant, while the introduction of remote afterloaders has eliminated exposure to nursing personnel. Currently, such machines require an inventory of many sources which are loaded into catheters implanted in the tumor and kept in place during treatment. A significant simplification of such machines would be possible in a pulsed mode, with a single source moving under computer control through the catheters. Assuming that the treatment time and average dose rate are kept unchanged, the question addressed is to find those combinations of radiation pulse widths and frequencies that would be functionally equivalent to a continuous irradiation. The linear-quadratic formalism was used to reanalyze published low dose-rate studies on cells of human origin to obtain 36 parameter sets [alpha, beta, T1/2], where T1/2 is the half time for sublethal damage repair. These data are consistent with those for human tumors. For each parameter set, those combinations of pulse width and frequency were calculated that would yield a functionally equivalent cell survival. For a regimen of 30 Gy in 60 hr, a pulse width of 10 min with a period between pulses of 1 hr would be appropriate for all the cell lines considered. Similar results were found for other possible time/dose combinations. For late effects, a 1-hr period between 10-min pulses might produce up to a 2% increase in late-effect probability, which is probably acceptable for the small volumes irradiated in interstitial brachytherapy.

Brachytherapy↗

The dose-rate effect revisited: radiobiological considerations of importance in radiotherapy.

A wide range of dose-rates have been used in radiation biology and radiation therapy, extending from a few cGy per day to hundreds of Gy in a fraction of a second. The dose-rate range of importance in radiotherapy extends from about 0.1 Gy/hr to several Gy/min. In this range, the fraction of cells killed by a given dose decreases as the dose-rate is reduced, principally because of the repair of sub-lethal damage. In some cell lines, an inverse dose-rate effect is observed where, over a narrow range of dose-rates, the effectiveness of a given dose increases with decreasing dose-rate if cells move through the cycle and are arrested in G2, which is a radiosensitive phase. In recent years data have accumulated for cells of human origin. About 40 data sets have been analyzed for values of the survival curve parameters and the rate of repair of sub-lethal damage. These data have been used to address three questions of relevance to radiotherapy. (1) The proposal to use pulsed rather than continuous irradiation in interstitial brachytherapy. (2) The equivalence of high dose-rate and low dose-rate intracavitary treatments for carcinoma of the cervix. (3) An analysis of equivalent doses for a range of dose-rates in interstitial implants.

Brachytherapy↗

Fractionated high dose-rate versus low dose-rate regimens for intracavitary brachytherapy of the cervix: equivalent regimens for combined brachytherapy and external irradiation.

The conventional treatment for carcinoma of the uterine cervix is a combination of external teletherapy and low dose-rate (LDR) intracavitary brachytherapy. Recently, however, there has been an increasing trend toward the use of high dose-rate (HDR) brachytherapy, in combination with external irradiation. The question is addressed of designing HDR treatments that will produce equivalent results to the more conventional protocols. We argue that for the unique case of radiotherapeutic treatment of carcinoma of the cervix, the criterion for producing an equivalent treatment should be based on the matching of early, not late, effects. In essence, this is because the dose to the tissues at risk for late effects is usually significantly smaller than the prescribed dose. When this effect is factored in with the different shape of dose-response curves for early and late effects, we conclude that, in the majority of cases, late effects will be no worse in a HDR regimen than a LDR regimen, provided that the corresponding doses have been matched to produce equal early effects. We provide a formalism whereby equivalent protocols can be designed for combined "external + HDR brachytherapy" regimens to match current "external + LDR brachytherapy" schedules. Using extensive basic radiobiological in vitro data derived from various cells of human origin, we provide specific examples of equivalent "external + HDR brachytherapy" regimens for 23 current "external + LDR brachytherapy" commonly-used schedules.

Brachytherapy↗

Fractionated regimens for stereotactic radiotherapy of recurrent tumors in the brain.

Radiosurgery (single-fraction stereotactic radiotherapy) was initially developed to treat non-malignant arteriovenous malformations, but there is growing interest in its use for the treatment of recurrent brain tumors. We suggest that there are sound reasons to expect improved results for tumor radiotherapy, in terms of late effects, if a fractionated regimen is used. At present, no published guidelines are available for choosing appropriate doses for fractionated regimens. We present two sets of guidelines, based on experimentally derived radiobiological parameters: first, we estimate gamma-ray doses which, if delivered in various numbers of fractions, should produce equivalent early effects to 70 Gy of 125I X rays delivered at low dose rate; this latter regimen is currently used in RTOG interstitial brachytherapy trials. Second, we estimate doses for multi-fractioned stereotactic radiotherapy which may be advantageous alternatives to particular doses of single-fractioned radiosurgical therapy. As the appropriate hardware is available, the use of fractionated stereotactic radiotherapy deserves serious consideration for the treatment of recurrent tumors in the brain.

Brain Neoplasms↗

Multilocus enzyme analysis of the genus Aeromonas and its use for species identification.

A total of 153 Aeromonas strains were analyzed by multilocus enzyme electrophoresis. All 11 genetic loci were polymorphic with three to 14 alleles per locus (average 6.55). The genetic diversity of each locus varied between 0.095 for indophenol oxidase and 0.881 for the fast variety of malic enzyme. Cluster analysis of the 122 enzyme types revealed a good correlation with taxonomic groupings as determined by DNA-DNA hybridization. In conjunction with biochemical analysis, as few as two enzymes may be sufficient for the identification of all species in the genus Aeromonas.

Aeromonas↗

Biochemical and genetic characterization of autoagglutinating phenotypes of Aeromonas species associated with invasive and noninvasive disease.

The genetic characteristics and biochemical and structural properties of a number of autoagglutinating (AA) strains of Aeromonas associated with invasive and noninvasive disease in humans and infections in animals and from environmental sources were investigated. Of 27 strains analyzed by multilocus enzyme typing and DNA hybridization studies, 25 (93%) were confirmed to belong to either hybridization group 1 (phenospecies and genospecies Aeromonas hydrophila) or 8 (phenospecies Aeromonas sobria; genospecies Aeromonas veronii). Further analysis of 19 of these strains indicated that four major groups could be identified on the basis of serologic and surface characteristics, protein and lipopolysaccharide composition, and virulence properties; these groupings held true regardless of the site of isolation or disease process involved. The major AA+ group identified was serogroup O:11, whose strains possessed an S layer, were resistant to the bactericidal activity of normal serum, and were pathogenic in mice. The results suggest a set of useful phenotypic and structural markers for identification of specific subsets of mesophilic Aeromonas involved in a wide range of infections in the animal kingdom.

Aeromonas↗

Taxonomy and pathogenicity of Erwinia cacticida sp. nov.

A total of 108 pectolytic, soft-rotting Erwinia strains were collected from 11 types of cacti growing in Arizona, Texas, northern Mexico, and Australia between 1958 and 1989. Four strains were collected from soils beneath or close to naturally rotting saguaro cacti. Collectively, these strains caused soft rots of saguaro, organ pipe, and senita cacti, Opuntia (cactus) fruits and pads, tomato fruits, and potato slices, but only occasionally caused soft rots of slices of carrot roots. A numerical cluster analysis showed that 98 of the 112 strains formed a uniform group (cluster 1A) that was distinguished from other pectolytic erwinias by an API 20E code of 1205131, by negative reactions in API 50CHE tests for L-arabinose, myo-inositol, D-cellobiose, melibiose, and D-raffinose, and, in supplemental tests, by positive reactions for malonate and growth at 43 degrees C. The average levels of DNA relatedness of 22 cluster 1A strains to the proposed type strain (strain 1-12) as determined by the hydroxyapatite method were 88% in 60 degrees C reactions (with 1% divergence within related sequences) and 87% in 75 degrees C reactions. The levels of relatedness to the type strains of other Erwinia spp. were less than or equal to 38% in 75 degrees C reactions. Cluster 1A strains also had a characteristic cellular fatty acid profile containing cyclo-(11,12)-nonadecanoic acid (C19:0 Cyclo C11-12) and missing tridecanoic acid (C13:0), heptadecanoic acid (C17:0), and cis-9-heptadecenoic acid (C17:1 CIS 9), which separated them from other pectolytic erwinias. Collectively, these data indicate that the members of cluster 1A are members of a new species, which we name Erwinia cacticida. Three cactus strains in cluster 1B appear to represent a second new species that is closely related to E. cacticida; these strains are designated E. cacticida-like pending the availability of additional strains for testing. The remaining cactus strains (in cluster 4) have the physiological, DNA, and fatty acid profiles of Erwinia carotovora.

DNA, Bacterial↗

Assay for beta-glucuronidase in species of the genus Escherichia and its applications for drinking-water analysis.

Recently, Escherichia species other than Escherichia coli have been isolated from potable water. Environmental isolates as well as clinical isolates of E. adecarboxylata, E. blattae, E. fergusonii, E. hermannii, and E. vulneris were assayed for the enzyme beta-glucuronidase by using EC MUG medium and the Colilert system. None of the isolates were positive for the enzyme by either method.

Escherichia↗

16S rRNA sequences of Bartonella bacilliformis and cat scratch disease bacillus reveal phylogenetic relationships with the alpha-2 subgroup of the class Proteobacteria.

The primary structures of 16S rRNAs of Bartonella bacilliformis, an isolate of the cat scratch disease (CSD) bacillus, and a strain phenotypically similar to the CSD bacillus were determined by reverse transcriptase sequencing. These microorganisms were found to be members of the alpha-2 subgroup of the class Proteobacteria. The sequence from B. bacilliformis was most closely related to the rRNA of Rochalimaea quintana (91.7% homology), the etiologic agent of trench fever. The sequence from the isolate of the CSD bacillus showed the greatest homology with Brucella abortus (89.7%) and, when compared with oligonucleotide catalog data, formed a cluster with Rhodopseudomonas palustris, Pseudomonas carboxidovorans, Nitrobacter species, and Bradyrhizobium species. The 16S rRNA sequence was also determined for the Cleveland Clinic isolate, which was previously shown to be phenotypically similar to and approximately 30% related, by DNA hybridization, to the CSD bacillus. The Cleveland Clinic isolate was isolated from a patient not diagnosed with CSD. The rRNAs from these bacteria exhibited 98.2% homology, confirming that this isolate is a second species in the same genus as the CSD bacillus. Our data suggest that neither B. bacilliformis nor the CSD bacillus is the etiologic agent of bacillary epithelioid angiomatosis.

Alphaproteobacteria↗