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Biomedical subjects

Michael Wilson

Publications and source records attributed to Michael Wilson.

At least 37 records · Page 2Linked to original sources
PubMed2004

Mercuric resistance genes in gram-positive oral bacteria.

Mercury-resistant bacteria isolated from the oral cavities of children carried one of two types of merA gene that appear to have evolved from a common ancestor. Streptococcus oralis, Streptococcus mitis and a few other species had merA genes that were very similar to merA of Bacillus cereus strain RC607. Unlike the B. cereus RC607 merA gene, however, the streptococcal merA genes were not carried on Tn5084-like transposons. Instead, comparisons with microbial genomic sequences suggest the merA gene is located on a novel type II transposon. Coagulase-negative staphylococci and Streptococcus parasanguis had identical merA genes that represent a new merA variant.

Adolescent↗
PubMed2004

Lethal photosensitisation of oral bacteria and its potential application in the photodynamic therapy of oral infections.

Chemical antibacterial agents are increasingly being used in prophylactic and therapeutic regimes for dental plaque-related diseases, which are among the most common human infections. As these agents are difficult to maintain at a therapeutic concentration in the oral cavity and can be rendered ineffective by resistance development in the target organisms, there is a need to develop alternative antimicrobial approaches. Bacteria and other microbes can be sensitised to light through prior treatment with a chemical photosensitising agent. Lethal photosensitisation of a wide range of bacteria responsible for caries, periodontal diseases and root canal infections has been demonstrated using red light in conjunction with a number of photosensitisers, including Toluidine Blue, phthalocyanines and chlorins. The advantages of this approach are that bacteria can be eradicated in very short periods of time (seconds or minutes), resistance development in the target bacteria is unlikely and damage to adjacent host tissues and disruption of the normal microflora can be avoided. This approach may be a useful alternative to antibiotics and antiseptics in eliminating cariogenic and periodontopathogenic bacteria from disease lesions and for the disinfection of root canals. Not only would this be of benefit for the treatment of these diseases but, by replacing the antimicrobial agents that are currently used for such purposes, it would help to conserve our dwindling supply of antimicrobial agents that are effective in the treatment of serious systemic infections.

Bacteria↗
PubMed2004

Biofilm-specific surface properties and protein expression in oral Streptococcus sanguis.

OBJECTIVE: Oral streptococci are primary colonisers of the tooth surface and are abundant in dental plaque biofilms. Bacteria growing in these relatively dense, surface-associated communities are phenotypically quite distinct from their planktonic counterparts. The purpose of the present study was to develop a method to investigate biofilm-specific surface protein expression by Streptococcus sanguis to help provide a better understanding of the critical events in plaque development. DESIGN: Biofilm cells were grown on the surface of glass beads in a biofilm device fed with mucin-containing artificial saliva. Planktonic cells were grown in continuous culture at approximately the same growth rate. Surface hydrophobicity of biofilm and planktonic cells was determined by hexadecane partitioning, and expression of streptococcal fibronectin adhesin CshA was determined in ELISA using specific antiserum. Antisera raised to glutaraldehyde-fixed whole biofilm or planktonic grown cells were used to screen an expression library of S. sanguis genomic DNA, and isolated clones were sequenced. RESULTS: Phenotypic analysis of biofilm and planktonic cells confirmed that mode of growth affected surface properties of S. sanguis. Thus, hydrophobicity and CshA expression was significantly elevated in biofilm cells. Library screening with biofilm antiserum yielded 32 recombinant clones representing 21 different S. sanguis proteins involved in adhesion and colonisation, carbohydrate utilisation or bacterial metabolism. In differential analysis of four selected Escherichia coli clones, biofilm antiserum reacted five times stronger than planktonic antiserum with cell-free extracts of clones encoding homologues of CshA and Cna collagen adhesin of Staphylococcus aureus, suggesting that these surface proteins are up-regulated in biofilm cells. In contrast, both antisera reacted equally strongly with cell-free extracts of the remaining two clones (encoding dihydrofolate synthase and an unknown protein). CONCLUSIONS: The method described represents a useful means for determining bacterial protein expression in biofilms based on a combination of molecular and immunological techniques. Surface expression of putative fibronectin and collagen adhesins was up-regulated in biofilm cells.

Bacterial Adhesion↗
PubMed2004

Transfer of blastocysts and morulae on day 5.

OBJECTIVE: To limit the number of embryos transferred and reduce high-order multiple pregnancies without compromising a patient's opportunity to become pregnant. DESIGN: Retrospective, nonrandomized analysis of embryo development and patient outcome when embryos were transferred on day 5. SETTING: Private practice. PATIENT(S): Women undergoing in vitro fertilization (IVF) treatment. INTERVENTION(S): Extend embryo culture to day 5 before embryo transfer (ET) to reduce the number of embryos transferred and minimize high-order multiple pregnancies. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate (CPR), implantation rate (IR), and live born rate (LBR) from expanding (expanded blastocysts and blastocysts) and nonexpanding (early blastocysts and morulae) embryos transferred on day 5. RESULT(S): Approximately 60% of patients had expanding embryos (EXE) on day 5. Forty percent of patients having an ET had transferable non-expanding embryos (NEE). Patients with EXE had higher CPR and LBR compared to patients with NEE. Implantation rate and multiple pregnancy rate (MPR) were also higher for patients with EXE. The miscarriage rate (MCR) for patients with EXE and NEE was not different. Approximately 5.5% of patients did not have an ET, with most (>98%) of the ET failures from patients with <==3 two pronuclei (2PN) embryos. The number of 2PN embryos had an effect on CPR, LBR, MPR, and the number of patients having cryopreservation. CONCLUSION(S): Day 5 ET allows for the selection of embryos with the highest implantation potential as evidenced by acceptable pregnancy rates for patients with either EXE or NEE, without the need to transfer more than two embryos.

Blastocyst↗
PubMed2004

Characterisation of viridans group streptococci with different levels of Tet(M)-mediated tetracycline resistance.

Streptococcus oralis 264-3, Streptococcus mitis 254-1 and S. mitis 264-1, isolated from the oral cavities of two children were each found to carry the tet(M) gene but exhibited different degrees of reduced susceptibility to tetracycline (tetracycline MICs of 2, 8 and 64 mg/L, respectively). The aim of this study was to determine the molecular basis for the different levels of tetracycline resistance (Tc(R)) observed. Escherichia coli HB101 carrying the cloned tet(M) genes exhibited similar levels of tetracycline susceptibility to those observed in the parental streptococcal strains (MICs of 1, 16, and 64 mg/L for tet(M) genes from S. oralis 264-3, S. mitis 254-1 and S. mitis 264-1, respectively). DNA sequencing revealed that S. oralis 264-3 had a tet(M) gene highly homologous to tet(M) carried by Tn916 from Enterococcus faecalis (99.6% identity), while the intermediate- and high-level Tc(R) strains had tet(M) sequences that resembled the tet(M) gene of Tn5251 from Streptococcus pneumoniae (99.3% and 99.4% identity, respectively). No differences were observed in the upstream attenuator structure for each of the strains and differences in reduced tetracycline susceptibilities could be attributed to changes in the deduced amino acid sequences of the Tet(M) proteins.

Adolescent↗
PubMed2004

Neutral density liquid formulations for nematode-based biopesticides.

Entomopathogenic nematodes are widely used as biological insecticides for agricultural and horticultural pests. In the vast majority of cases, commercial products contain nematodes partially dehydrated on to inert solid carriers. Nematode survival in these products is generally poor, they are difficult to handle and are not suitable for use with all nematode species. We have developed a non-viscous, non-adhesive and non-toxic liquid formulation for nematode storage and transport based on neutral density colloidal silica suspensions. Survival and virulence of nematodes stored in this formulation without aeration was superior to nematodes stored in aerated quarter strength Ringer's solution.

Animals↗
PubMed2004

Antibody-directed photodynamic therapy of methicillin resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a world-wide public health problem, causing nosocomial and community-acquired infections. Furthermore, MRSA is increasingly resistant to many conventional antimicrobials, so there is a real need to develop alternative approaches for MRSA decolonization and treatment. Previously, we have demonstrated that MRSA can be killed with an immunoglobulin G (IgG)-SnCe6 conjugate and red light, but effectiveness was dependent on the particular strain and the growth phase. In this investigation, we used an antibody raised against MRSA to make an Ab-SnCe6 conjugate capable of targeting many MRSA strains in all growth phases. To suspensions of important epidemic MRSA, each grown to stationary, lag, or exponential phase, the Ab-SnCe6 conjugate was added and samples exposed to red light. Survivors were then enumerated. This treatment was very effective at killing all the different MRSA strains tested, in all growth phases. The Ab-SnCe6 conjugate was able to kill EMRSA-16 selectively in a mixed suspension of EMRSA-16 and Escherichia coli, and was much better at killing EMRSA-16 than a coagulase-negative staphylococcus, S. epidermidis. These results demonstrate that photodynamic therapy of MRSA is very effective when the photosensitizer is targeted to the pathogen using a suitable antibody and may be a good candidate for a novel treatment of MRSA infections.

Humans↗
PubMed2004

Can thallium-pertechnetate subtraction scanning play a role in the preoperative imaging for minimally invasive parathyroidectomy?

PURPOSE: Preoperative Tc-99m sestamibi scanning can identify candidates for minimally invasive parathyroid surgery. However, a significant number of patients with single gland disease have negative scans and are not considered for the minimally invasive procedure. MATERIALS AND METHODS: To determine if T1-201/Tc-99m sodium pertechnetate subtraction scanning (TPSS) is a viable alternative imaging technique for patients with primary hyperparathyroidism (1 degrees HPTH), we reviewed our experience. The outcomes of 100 consecutive patients with 1 degrees HPTH who underwent preoperative TPSS and parathyroid exploration between 1995 and 2000 at our institution were retrospectively reviewed. RESULTS: The mean preoperative calcium and parathyroid hormone levels were 10.8 mg/dL and 220 pg/mL, respectively. The overall cure rate was 96%. Single gland disease was present in 88%. Of the 100 patients studied, 15 underwent both a Tc-99m sestamibi scan and TPSS, whereas the other 85 had only the TPSS. CONCLUSIONS: The sensitivity and positive predictive value of the TPSS were 73.3% and 90.4%, respectively. In the patients undergoing both TPSS and Tc-99m sestamibi scans, the results concurred in 60%. However, in 20% of the remaining patients, TPSS correctly localized the abnormal parathyroid(s) when Tc-99m sestamibi failed. TPSS has a comparable sensitivity and positive predictive value to Tc-99m sestamibi scanning. In patients with a negative Tc-99m sestamibi scan, TPSS can provide additional localizing information. As a result of the high positive predictive value of TPSS, a single parathyroid gland localized by TPSS alone can then be approached by minimally invasive parathyroidectomy.

Female↗
PubMed2004

Effect of nematodes on rhizosphere colonization by seed-applied bacteria.

There is much interest in the use of seed-applied bacteria for biocontrol and biofertilization, and several commercial products are available. However, many attempts to use this strategy fail because the seed-applied bacteria do not colonize the rhizosphere. Mechanisms of rhizosphere colonization may involve active bacterial movement or passive transport by percolating water or plant roots. Transport by other soil biota is likely to occur, but this area has not been well studied. We hypothesized that interactions with soil nematodes may enhance colonization. To test this hypothesis, a series of microcosm experiments was carried out using two contrasting soils maintained under well-defined physical conditions where transport by mass water flow could not occur. Seed-applied Pseudomonas fluorescens SBW25 was capable of rhizosphere colonization at matric potentials of -10 and -40 kPa in soil without nematodes, but colonization levels were substantially increased by the presence of nematodes. Our results suggest that nematodes can have an important role in rhizosphere colonization by bacteria in soil.

Animals↗
PubMed2004

Characterization of Tn916S, a Tn916-like element containing the tetracycline resistance determinant tet(S).

We have characterized a transferable tetracycline resistance (Tcr) element from a Streptococcus intermedius isolate. The gene responsible for this resistance was identified by PCR and Southern hybridization as tet(S). Furthermore, the genetic support for this determinant was shown to be a conjugative transposon closely related to Tn916. This element has been designated Tn916S.

Base Sequence↗
PubMed2004

Prevalence of periodontal pathogens in dental plaque of children.

Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Tannerella forsythensis have been implicated as the main etiological agents of periodontal disease. The purpose of this work was to estimate the prevalence of these organisms in plaque from children without gingivitis (group 1; n = 65) and from those with gingivitis (group 2; n = 53). Extracted DNA from plaque was subjected to two rounds of PCR targeting the 16S rRNA gene using both universal primers and species-specific primers. The results were as follows: group 1, P. gingivalis, 49%; A. actinomycetemcomitans, 55%; and T. forsythensis, 65%; group 2, P. gingivalis, 47%; A. actinomycetemcomitans, 59%; and T. forsythensis, 45%. T. forsythensis was detected more frequently in children with no gingivitis than in those with gingivitis (P = 0.03). There was no significant difference between the two groups with respect to the presence of P. gingivalis or A. actinomycetemcomitans in either group (P > 0.05). Logistic regression analysis revealed that the odds of a patient having gingivitis were 2.3 times greater in the absence of T. forsythensis. In conclusion, the results of this study have shown that the three pathogens can be detected in the dental plaque of healthy children and of those with gingivitis and that T. forsythensis is associated with dental plaque at sites with no gingivitis.

Aggregatibacter actinomycetemcomitans↗
PubMed2003

Prevalence and identification of tetracycline-resistant oral bacteria in children not receiving antibiotic therapy.

The prevalence of tetracycline-resistant oral bacteria in healthy 4- and 6-year-old children who had not received antibiotics during the 3 months prior to sampling was investigated. Of the 47 children sampled, 46 harboured tetracycline-resistant bacteria. The median proportion of cultivable anaerobic and aerobic oral bacteria resistant to tetracycline was 1.1% and the MIC50 of these was 64 microg ml(-1). The majority (56%) of tetracycline-resistant bacteria were resistant to at least one other antibiotic, usually erythromycin. The most commonly identified tetracycline-resistant bacteria were the oral streptococci (65%), the next most prevalent groups were Veillonella spp. (10%) and Neisseria spp. (9%). The most frequently identified tetracycline resistance determinant was tet(M). The results of this study have shown that tetracycline-resistant oral bacteria were widespread amongst the children studied.

Anti-Bacterial Agents↗
PubMed2003

Oral bacteria resistant to mercury and to antibiotics are present in children with no previous exposure to amalgam restorative materials.

Dental plaque from 76 children without amalgam restorations was screened for bacteria resistant to mercuric chloride. Seventy-one per cent of the children harboured mercury-resistant oral bacteria and the median percentage of the total oral microflora resistant to mercuric chloride was 0.007% (range 0-5.3%). Eighty-seven mercury-resistant bacteria were isolated and 86% of these were streptococci with Streptococcus mitis predominating. Sixty per cent of the mercury-resistant isolates were also resistant to at least one of the four antibiotics tested (penicillin, ampicillin, erythromycin and tetracycline) with resistance to tetracycline (40% of isolates) most frequently encountered.

Anti-Bacterial Agents↗
PubMed2003

Identification of a novel lipase gene mutated in lpd mice with hypertriglyceridemia and associated with dyslipidemia in humans.

Triglyceride (TG) metabolism is crucial for whole body and local energy homeostasis and accumulating evidence suggests an independent association between plasma TG concentration and increased atherosclerosis risk. We previously generated a mouse insertional mutation lpd (lipid defect) whose phenotype included elevated plasma TG and hepatic steatosis. Using shotgun sequencing (approximately 500 kb) and bioinformatics, we have now identified a novel lipase gene lpdl (lpd lipase) within the lpd locus, and demonstrate the genetic disruption of exon 10 of lpdl in the lpd mutant locus. lpdl is highly expressed in the testis and weakly expressed in the liver of 2-week old mice. Human LPDL cDNA was subsequently cloned, and was found to encode a 460AA protein with 71% protein sequence identity to mouse lpdl and approximately 35% identity to other known lipases. We next sequenced the human LPDL gene exons in hypertriglyceridemic subjects and normal controls, and identified seven SNPs within the gene exons and six SNPs in the adjacent introns. Two hypertriglyceridemic subjects were heterozygous for a rare DNA variant, namely 164G>A (C55Y), which was absent from 600 normal chromosomes. Two other coding SNPs were associated with variation in plasma HDL cholesterol in independent normolipidemic populations. Using bioinformatics, we identified another novel lipase designated LPDLR (for 'LPDL related lipase'), which had 44% protein sequence identity with LPDL. Together with the phospholipase gene PSPLA1, LPDL and LPDLR form a new lipase gene subfamily, which is characterized by shortened lid motif. Study of this lipase subfamily may identify novel molecular mechanisms for plasma and/or tissue TG metabolism.

Amino Acid Sequence↗
PubMed2003

Diagnosis and monitoring of hemorrhagic shock during the initial resuscitation of multiple trauma patients: a review.

The initial management of the multiple trauma victim requires evaluation for potential hemorrhage and ongoing monitoring to assess the efficacy of resuscitation and avoid complications related to hemorrhagic shock. A variety of strategies exist to assess circulatory status, including hemodynamic monitoring, tissue perfusion measurement, and use of serum markers of metabolism. We review available technologies used to assess fluid status and tissue perfusion in patients with blood loss or hemorrhagic shock, discuss how these methods can be used effectively and efficiently during initial trauma resuscitation to guide therapy and disposition, and suggest directions for future research to improve outcomes by providing more appropriate and timely care and avoiding unnecessary complications.

Biomarkers↗
PubMed2003

The application of the precautionary principle to the blood system: the Canadian blood system's vCJD donor deferral policy.

The precautionary principle is an influential concept that has been widely used in international treaties and declarations involving the protection of the environment. The principle is now being applied to the development of transfusion policy. In this article, we examine the application of the precautionary principle in the policy process leading to Canada's decision to defer donations from individuals who had traveled to the United Kingdom because of concerns over variant Creutzfeldt-Jakob disease. We found that, although the principle prominently influenced the decision-making process, problems existed with its interpretation. In particular, there was difficulty in balancing the risk prevented by applying a precautionary measure against the risk introduced by the same measure; in this case, the potential for shortages of blood. This dilemma is somewhat unique to the public health sector and will likely recur in future applications of the principle to transfusion policy.

Blood Banks↗
PubMed2003

Light-activated antimicrobial coating for the continuous disinfection of surfaces.

The ability of a photosensitizer-containing cellulose acetate film to kill bacteria was evaluated. Substantial kills were achieved following irradiation of the film with white light for up to 24 hours. Applying a photosensitizer-containing coating to surfaces could reduce the environmental load of pathogens, thus helping to prevent infectious disease transmission.

Anti-Bacterial Agents↗