Biomedical subjects

A Gilmour

Publications and source records attributed to A Gilmour.

At least 19 recordsLinked to original sources

PubMed2000

The growth and resistance to sodium hypochlorite of Listeria monocytogenes in a steady-state multispecies biofilm.

A constant-depth film fermenter (CDFF) was used to culture a steady-state multispecies biofilm consisting of one strain each of Listeria monocytogenes, Pseudomonas fragi and Staphylococcus xylosus. These bacteria were initially grown together in a conventional chemostat to achieve a steady state before being inoculated into the CDFF over an 18-h period. A dilute tryptone soya broth (TSB) medium was supplied to the CDFF and the biofilm allowed to develop over a 28-d period. This mature biofilm was then subjected to increasing levels of sodium hypochlorite solution to measure any antimicrobial effect. The three organisms were seen to reach a steady state after 6 d in the chemostat before being transferred to the CDFF where the mature multispecies biofilm reached steady state at 17 d. Listeria monocytogenes in both planktonic and biofilm growth stabilized at 1. 8 and 1.5%, respectively, of the total plate counts, while Ps. fragi and Staph. xylosus were the predominant organisms in the biofilm at 59% and 39.5%, respectively, of the total microbial population. Steady-state biofilms in the CDFF were exposed to increasing strengths of sodium hypochlorite; 200, 500 and 1000 p.p.m. free chlorine, but a substantial two-log cycle drop in bacterial numbers was only achieved at 1000 p.p.m. free chlorine. In planktonic culture all three organisms were completely eliminated when exposed to 10 p.p.m. free chlorine for a 30-s period.

Biofilms↗

PubMed1999

Adherence of Listeria monocytogenes strains to stainless steel coupons.

An assay was developed to measure the number of Listeria monocytogenes cells adhering to stainless steel, and was used to investigate the adherence of 111 strains of the organism, which included representatives with respect to serotype, carriage of plasmids, source and persistence in the food processing environment. Growth and adherence curves of four L. monocytogenes strains over 48 h were obtained. While the growth curves of all four micro-organisms were seen to reach similar levels at stationary phase, there was still substantial variation among the adherence curves. In addition, a scatter-graph of growth vs adherence counts at 24 h showed poor correlation. These factors indicated that interstrain variation in adherence at stationary phase is due to factor(s) intrinsic to each strain of L. monocytogenes. Persistent strains were found to adhere in significantly greater numbers than sporadic strains, and variation was also found among serotypes, with serotype 1/2c showing significantly greater adherence than serotypes 1/2a and 4b; 4b strains were significantly higher than those of 1/2a strains. No significant difference was found between strains according to source or plasmid carriage.

Bacterial Adhesion↗

PubMed1999

A proinflammatory role for IL-18 in rheumatoid arthritis.

IL-18 is a novel cytokine with pleiotropic activities critical to the development of T-helper 1 (Th1) responses. We detected IL-18 mRNA and protein within rheumatoid arthritis (RA) synovial tissues in significantly higher levels than in osteoarthritis controls. Similarly, IL-18 receptor expression was detected on synovial lymphocytes and macrophages. Together with IL-12 or IL-15, IL-18 induced significant IFN-gamma production by synovial tissues in vitro. IL-18 independently promoted GM-CSF and nitric oxide production, and it induced significant TNF-alpha synthesis by CD14(+) macrophages in synovial cultures; the latter effect was potentiated by IL-12 or IL-15. TNF-alpha and IFN-gamma synthesis was suppressed by IL-10 and TGF-beta. IL-18 production in primary synovial cultures and purified synovial fibroblasts was, in turn, upregulated by TNF-alpha and IL-1beta, suggesting that monokine expression can feed back to promote Th1 cell development in synovial membrane. Finally, IL-18 administration to collagen/incomplete Freund's adjuvant-immunized DBA/1 mice facilitated the development of an erosive, inflammatory arthritis, suggesting that IL-18 can be proinflammatory in vivo. Together, these data indicate that synergistic combinations of IL-18, IL-12, and IL-15 may be of importance in sustaining both Th1 responses and monokine production in RA.

Animals↗

PubMed1997

The effect of high hydrostatic pressure on Listeria monocytogenes in phosphate-buffered saline and model food systems.

Three strains of Listeria monocytogenes (NCTC 1194, a poultry isolate and the Scott A strain) were exposed to a range of pressures (300, 350, 375, 400 and 450 MPa) in 10 mmol 1(-1) phosphate-buffered saline (PBS) at pH 7.0 for up to 30 min at ambient temperature. Generally, increasing the magnitude and duration of compression resulted in increasing levels of inactivation, although the inactivation kinetics varied depending on the strain and pressure applied. The three strains also exhibited a wide variation in their resistance to high pressure. The resistance of the three strains to high pressure (375 MPa) was also assessed in a series of model food systems containing one of each of the three main food constituents: protein (1, 2, 5 and 8% w/v bovine serum albumin in PBS), carbohydrate (1, 2, 5 and 10% w/v glucose in PBS) and lipid (olive oil (30% v/v) in PBS emulsion). Overall, increasing the concentrations of bovine serum albumin (BSA) and glucose in the suspending medium resulted in decreasing levels of inactivation of all three strains; however, the minimum concentration of BSA and glucose required to increase survival to a level greater than that observed in PBS alone varied depending on the strain and on the duration of the treatment. The survival of all three strains was greater in the olive oil/PBS emulsion than in PBS alone at all treatment times.

Animals↗

PubMed1997

The effect of high hydrostatic pressure on the activity of intracellular enzymes of Listeria monocytogenes.

The effect of high hydrostatic pressure (100-550 MPa, 15 min, ambient temperature) on the activity of 13 metabolic enzymes produced by all three strains of Listeria monocytogenes (NCTC 11994, a poultry isolate and Scott A) was examined using gel electrophoresis. The enzymes assayed exhibited a wide variation in barotolerance. The pressure resistance of each particular enzyme was not dependent on the strain from which it was derived. This would seem to indicate that these enzymes were not a determining factor in relation to previously observed differences in the overall pressure resistance of the three strains.

Hydrostatic Pressure↗

PubMed1996

Multilocus enzyme electrophoresis for characterization of Listeria monocytogenes isolates: results of an international comparative study.

Multilocus enzyme electrophoresis (MEE) is a standard technique that is used to elucidate the epidemiology of a variety of bacterial species. Recently, the method has been employed by several laboratories for investigations of clinical and foodborne isolates of Listeria monocytogenes. To assess the sensitivity and reproducibility of MEE in characterising L. monocytogenes isolates for epidemiological purposes and, ultimately, to agree on a standard protocol, seven laboratories participated in a blinded study of 80 strains. The strain collection included both epidemiologically related and unrelated isolates. Each laboratory used its own protocol for MEE. The number of enzymes that were assayed by the laboratories ranged from 8 to 23, and the total number of identified electrophoretic types (ETs) varied between 14 and 25. Of the II pairs of duplicate strains, the number of pairs recognised as identical by the seven laboratories ranged from 3 to 10 (median = 8). From 10 to 18 (median = 15) of the 22 groups of epidemiological related strains were recognised as homogeneous by the different laboratories. The discriminatory power of the method, calculated using Simpson's index of diversity for 69 strains (80 strains minus the 11 duplicates), ranged from 0.827 to 0.925. This relatively low discriminatory power is a consequence of a somewhat low genetic diversity of L. monocytogenes compared to other bacterial species. Efforts should be pursued to standardise the method in order to improve the intra- and inter-laboratory reproducibility.

Alleles↗

PubMed1996

DNA fingerprinting of benign and malignant breast lesions.

Paired blood and breast tissue samples from 96 patients undergoing surgical excision of a breast lesion were subjected to DNA fingerprint analysis using the minisatellite probes 33.6 and 33.15. The 'fingerprints' of the blood and breast DNA were compared. DNA fingerprint changes seen were classified as band additions, band deletions or changes in band intensity. Significantly more DNA fingerprint changes were seen in malignant than in benign lesions (probe 33.6, P<0.01; probe 33.15, P=0.01; both probes, P = 0.04). The frequency of the changes detected in the breast cancers was unrelated to tumour grade or the presence of invasion. DNA ploidy was measured in 70 of the breast carcinomas. DNA fingerprint changes were seen in 41 of these, but flow cytometry detected aneuploidy in only 29 tumours (P=0.04). The DNA fingerprinting technique is unlikely to be useful in differentiating between benign and malignant breast lesions as changes were detected in one of the fibroadenomas studied. However, it may be a useful technique in screening tumour DNA for abnormalities suitable for more detailed analysis.

Breast Diseases↗

PubMed1996

Characterization of Listeria monocytogenes isolates by esterase electrophoresis.

Multiple bands of alpha-naphthyl-propionyl esterase (alpha NPE) activity observed following starch gel electrophoresis of cell extracts allowed 219 Listeria monocytogenes isolates from milk, nondairy foods, and clinical and veterinary sources to be assigned to 17 different alpha NPE types. Multilocus enzyme electrophoresis (MEE) analysis was used to obtain electrophoretic mobility data for alpha NPEs and nine other metabolic enzymes; on the basis of these data, the 219 strains were assigned to 59 electrophoretic types. Each of the methods separated the strains into two main groups, and there was extensive agreement on assignment of the strains to the groups. Although the method is less discriminatory than MEE, the usefulness of alpha NPE electrophoresis alone as a rapid, simple typing method for L. monocytogenes for certain purposes is indicated by agreement among alpha NPE, MEE, and restriction fragment length polymorphism types as molecular markers for persistent and transient L. monocytogenes strains found in sequential raw-milk and nondairy food samples obtained from different processors.

Animals↗

PubMed1995

The influence of fluoride on the adsorption of proteoglycans and glycosaminoglycans to hydroxyapatite.

Proteoglycans and their spatial arms, the glycosaminoglycans, are known to interact with hydroxyapatite, and are considered to have a role in the regulation of mineralization. This study investigates the interactive mechanisms, with particular attention directed at the influence of fluoride on the adsorption process. Proteoglycans and glycosaminoglycans were adsorbed to hydroxyapatite in the presence of fluoride (1-20 ppm range). The adsorbates included a chondroitin 4-sulfate-rich proteoglycan extracted from rat incisor dentine, hyaluronan, chondroitin 4-sulfate, and dermatan sulfate. The order of glycosaminoglycan in decreasing affinity for hydroxyapatite was chondroitin 4-sulfate, dermatan sulfate, and hyaluronan, and the individual glycosaminoglycans showed different responses to the presence of fluoride. Graded increases in fluoride (1-4 ppm) led to 5-40% reduction of glycosaminoglycan adsorption to hydroxyapatite. The proteoglycan showed less affinity for hydroxyapatite, and demonstrated a reduction in adsorption of up to 22% with 20 ppm fluoride. The inhibitory effect of fluoride indicated an electrostatic mechanism, presumably via the calcium sites in the hydroxyapatite lattice.

Adsorption↗

PubMed1995

Impedance as an alternative to MPN enumeration of coliforms in pasteurized milks.

Samples (900) of pasteurized whole, semi-skimmed and skimmed milk were subjected to conventional enumeration of coliforms by a nine-tube most probable number (MPN) technique, and impedance enumeration, in parallel. Regression analysis of the positive samples (98) showed that impedance enumeration was at least as accurate as the MPN method but results were obtained faster, with all testing being completed in 20 h, rather than 48 h. Consumable requirements, and staffing levels, were also much less with the impedance system. The impedance method could therefore beneficially replace the conventional method.

Animals↗

PubMed1995

Characterization of Listeria monocytogenes isolated from poultry products and from the poultry-processing environment by random amplification of polymorphic DNA and multilocus enzyme electrophoresis.

A total of 289 Listeria monocytogenes strains isolated from a poultry-processing environment and poultry products over a 6-month period were characterized by random amplification of polymorphic DNA, (RAPD) to pinpoint sources of contamination within the plant and gain some measure of the persistence of individual genotypes within this environment. Eighteen RAPD profiles (A through R) were identified within this group, with 64% (184 of 289) of all strains displaying a single RAPD profile, RAPD type A. This genotype was more prevalent in the raw-poultry-processing environment, where, although its origin within this environment appeared to be the incoming birds, it was also widespread on food contact surfaces, floors, and drains. This was the only genotype which persisted throughout the entire 6-month period, and it and RAPD type B were the only two genotypes found in both the raw- and cooked-poultry-processing environments. L. monocytogenes strains isolated from cooked poultry products and the cooked-poultry-processing environment up to 1 year later (17 strains) contained only RAPD types A and B, highlighting the potential which exists for persistent strains to cross-contaminate foods processed in that environment. The other genotypes (C through R) occurred more sporadically, suggesting varied sources of contamination. These were confined to either the raw- or the cooked-poultry-processing environment and were relatively short-lived. Further characterization of a selection of RAPD type A strains, together with strains of RAPD types B through R, was carried out by multilocus enzyme electrophoresis. Strains of RAPD type A contained two electrophoretic types, one of which was serotype 1/2a and the other was 1/2c.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

The effects of softwater acclimation on respiratory gas transfer in the rainbow trout Oncorhynchus mykiss

Gill O2 uptake, CO2 excretion, ventilation and blood respiratory/acidbase variables were evaluated in control and softwater-acclimated trout (Oncorhynchus mykiss) to test the hypothesis that gill chloride cell (CC) proliferation, elicited by 2 weeks of softwater exposure, impairs the diffusion of respiratory gases across the gill. The proliferation of CCs in softwater fish was verified using light microscopy, and its impact on respiratory gas transfer was assessed in vivo by continuous monitoring of arterial blood PO2 (PaO2), PCO2 (PaCO2) and pH (pHa) using an extracorporeal blood circulation under conditions of normoxia and graded hypoxia [water PO2 (PwO2) was lowered from 20.0 kPa to 5.3 kPa within 20 min]. During normoxia, ventilation frequency was significantly higher in the softwater trout (78±4 versus 57±4 breaths min-1; mean ± s.e.m.), while ventilation amplitude was similar in both groups (1.01.1 cm opercular displacement). PaCO2 and plasma HCO3- concentration were significantly lower in the softwater fish and the blood acidbase status was characterized by a mixed respiratory alkalosis and metabolic acidosis such that blood pH was not statistically different between the two groups. CO2 excretion (2.52.8 mmol kg-1 h-1) and O2 uptake rates (2.35.1 mmol kg-1 h-1), as measured during normoxia, were unaffected by acclimation to soft water. During hypoxia, ventilation frequency and amplitude increased in the control trout, whereas only ventilation amplitude increased in the softwater-acclimated fish. The rate of PaO2 reduction during hypoxia was significantly greater in the softwater fish (0.84±0.06 versus 0.65±0.06 kPa PaO2 kPa-1 PwO2) and, at the most severe level of hypoxia (PwO2=5.3 kPa), PaO2 was significantly lower in the softwater fish. The rate of PaCO2 reduction (caused by hyperventilation) was significantly lower in the softwater-acclimated fish (0.002±0.001 versus 0.005±0.001 kPa PaCO2 kPa-1 PwO2; mean ± s.e.m.; P<0.06) and, indeed, was not statistically different from zero. Blood pH did not change significantly during hypoxia in either group but, through much of the hypoxic period (715 kPa PwO2), pHa was statistically lower in the softwater-acclimated fish. These results demonstrate that exposure of trout to soft water for 2 weeks is associated with proliferation of lamellar CCs and impaired branchial gas transfer. Hyperventilation was identified as a compensatory physiological adjustment.

Journal Article↗

PubMed1994

A non-isotopic DNA hybridisation assay for the identification of Staphylococcus aureus isolated from foods.

A digoxygenin-labelled total genomic DNA probe was used to identify Staphylococcus aureus. Isolation and identification of organisms was possible in less than 4 days. Identification alone could be completed in less than 2 days, compared with over 5 days for identification of isolates by multipoint inoculation. The probe showed excellent discrimination of S. aureus from other staphylococci and from a wide range of bacteria commonly associated with milk and meat. The effectiveness of this probe was tested against cultural isolation of staphylococci in milk using Baird-Parker agar followed by identification using multipoint inoculation and API Staph. The probe gave comparable results to the conventional methods and, for large sample numbers, offered lower cost and greater ease of use.

Animals↗

PubMed1994

Some factors inhibiting amplification of the Staphylococcus aureus enterotoxin C1 gene (sec+) by PCR.

PCR amplification of the sec+ gene for staphylococcal enterotoxin C1 (SEC1) can be achieved from as little as 10 fg total genomic DNA (equivalent to less than 10 cells) using two nested primer pairs. The presence of bacterial cells, particularly thermonuclease-producing staphylococci, and the thermonuclease enzyme (TNase) itself, were found to be factors which individually and together reduced the sensitivity of PCR amplification.

DNA, Bacterial↗

PubMed1994

Incidence of Listeria spp. and Listeria monocytogenes in a poultry processing environment and in poultry products and their rapid confirmation by multiplex PCR.

The incidence of Listeria spp. and Listeria monocytogenes in a poultry processing plant and in raw and cooked poultry products was determined over a 6-month period. Within the raw and cooked poultry processing environments, 46% (36 of 79) and 29% (51 of 173) of the samples contained Listeria spp. while 26% (21 of 79) and 15% (27 of 173) contained L. monocytogenes, respectively. Various sites within the processing environment were found to be consistently positive for L. monocytogenes throughout the entire sampling period. Of the raw and cooked products tested, 91% (53 of 58) and 8% (8 of 96) were found to contain Listeria spp. while 59% (34 of 58) and 0% (0 of 96) contained L. monocytogenes, respectively. Although L. monocytogenes was not detected in the cooked products examined, the presence of other Listeria spp. highlights the potential which exists for postprocessing contamination. Multiplex PCR proved to be a convenient and time-saving technique for rapid confirmation of Listeria spp. and L. monocytogenes in a single reaction.

Animals↗

PubMed1994

Application of multilocus enzyme electrophoresis and restriction fragment length polymorphism analysis to the typing of Listeria monocytogenes strains isolated from raw milk, nondairy foods, and clinical and veterinary sources.

The powerful discriminatory typing capabilities of multilocus enzyme electrophoresis and restriction fragment length polymorphism analysis were applied to Listeria monocytogenes strains from raw milk, nondairy foods, and clinical and veterinary sources. The raw milk and nondairy food strains were sequential isolates obtained over a year-long period from a number of different producers and manufacturers. Results obtained by the two typing methods were in substantial agreement and showed that both raw milk and nondairy foods frequently contain recurrent L. monocytogenes strains, thus suggesting that the presence of these organisms in such commodities often arises because of contamination from within their respective processing environments. Most recurrent strains were serogroup 1/2, with only one instance of recurrent serogroup 4 strains. Some recurrent L. monocytogenes strains, including the serogroup 4 strains, were found by analysis of multilocus enzyme electrophoresis results to be closely related to clinical and veterinary strains, thus suggesting that strains adapted for survival in the food-processing environment retain their potential for pathogenicity.

Animals↗

PubMed1993

Occurrence and characteristics of Listeria in foods produced in Northern Ireland.

The incidence of L. monocytogenes and other Listeria species was determined for 513 food samples produced in Northern Ireland. Selected L. monocytogenes isolates were typed using MEE and RFLP analysis. The overall incidence of Listeria in the foods examined was 35% (Listeria monocytogenes 18.3%). The incidence of Listeria and L. monocytogenes in four different food categories (graded I-IV according to decreasing extent of processing) being: I, 11.1% Listeria and 4.7% L. monocytogenes; II, 27.1% Listeria and 12.2% L. monocytogenes; III, 89.1% Listeria and 50% L. monocytogenes and IV, 100% Listeria and 100% L. monocytogenes. Within food categories I and II, the incidence of Listeria on occasions varied markedly between similar products produced by different processors. The most frequently isolated species was L. innocua, followed by L. monocytogenes, L. seeligeri and L. welshimeri. The L. monocytogenes isolates were predominantly serogroup 1. A modified USDA method was the most productive of four enrichment procedures investigated. Over the one-year duration of the survey, a distinctive Listeria microflora could be discerned in products from certain processors and was confirmed in some cases by MEE and RFLP typing of L. monocytogenes isolates.

DNA, Bacterial↗

PubMed1993

Oestrogen receptor expression in ductal carcinoma in situ of the breast: relationship to flow cytometric analysis of DNA and expression of the c-erbB-2 oncoprotein.

The expression of oestrogen receptor protein (ER) was examined in 151 cases of symptomatic or screening detected pure ductal carcinoma in situ (DCIS) of the breast by immunocytochemical assay (ERICA), in formalin-fixed paraffin-embedded tissue, with the monoclonal antibody H 222 (Abbott). Forty-eight tumours (31.8%) of cases were ER positive. Twenty-seven (17.9%) of cases showed high level ER expression and 21 (13.9%) of cases showed low level ER immunoreactivity. Significant associations of positive tumour ER immunoreactivity and non-comedo architecture chi 2 = 6.76; (d.f. = 1): P < 0.001, small cell size chi 2 = 4.49; (d.f. = 1): P = 0.034, higher S-phase fraction chi 2 = 4.71; (d.f. = 1): P = 0.03 and lack of c-erbB-2 protein overexpression chi 2 = 7.96; (d.f. = 1): P < 0.01 were identified. No significant associations of ER expression and patient age, histological grade of necrosis in DCIS, or DNA ploidy were found. ER expression is detectable in less than one third of symptomatic and screening detected cases of DCIS, implying that endocrine therapy of DCIS may be a more appropriate form of management for morphological subtypes of DCIS which show higher rates of oestrogen receptor expression, particularly those of non-comedo and small cell type.

Adult↗