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Biomedical subjects

A J Potts

Publications and source records attributed to A J Potts.

At least 19 recordsLinked to original sources

Strain persistence of invasive Candida albicans in chronic hyperplastic candidosis that underwent malignant change.

OBJECTIVES: The aim of this study was to assess persistence and tissue invasion of Candida albicans strains isolated from a 65 year-old patient with chronic hyperplastic candidosis (CHC), that subsequently developed into squamous cell carcinoma (SCC). MATERIALS AND METHODS: C. albicans (n=7) were recovered from the oral cavity of the patient over seven years. Confirmation of CHC and SCC in this patient was achieved by histopathological examination of incisional biopsy tissue. DNA fingerprinting was performed on the seven isolates from the CHC patient together with a further eight isolates from patients with normal oral mucosa (n=2), chronic atrophic candidosis (n=1), SCC (n=1) and CHC (n=4). Genotyping involved the use of inter-repeat PCR using the eukaryotic repeat primer 1251. Characterisation of the tissue invasive abilities of the isolates was achieved by infecting a commercially available reconstituted human oral epithelium (RHE; SkinEthic, Nice, France). After 24 h, C. albicans tissue invasion was assessed by histopathological examination. RESULTS: DNA fingerprinting demonstrated strain persistence of C. albicans in the CHC patient over a seven year period despite provision of systemic antifungal therapy. The strain of C. albicans isolated from this patient was categorised as a high invader within the RHE compared to other isolates. CONCLUSIONS: Candidal strain persistence was evident in a patient with CHC over seven years. This persistence may be due to incomplete eradication from the oral cavity following antifungal therapy or subsequent recolonisation from other body sites or separate exogenous sources. The demonstration of enhanced in vitro tissue invasion by this particular strain may, in part, explain the progression to carcinoma.

Aged↗

PCR fingerprinting of Candida albicans associated with chronic hyperplastic candidosis and other oral conditions.

The purpose of this study was to genotype strains of Candida albicans to determine whether specific types were associated with chronic hyperplastic candidosis (CHC). A total of 67 candidal isolates from CHC patients (n = 17) and from patients with other oral conditions (n = 21) were genotyped by PCR fingerprinting employing two interrepeat primer combinations (1245 and 1246 primers or 1251 primer) and a single minisatellite-specific M13 primer. The most suitable primer for fingerprint analysis was found to be primer 1251, yielding well-resolved banding patterns. For the 67 isolates tested, PCR fingerprinting delineated 25 (1245 and 1246 primers), 27 (1251 primer), and 25 (M13 primer) profiles. The majority of C. albicans isolates from multiple sites within the mouth produced identical profiles (six out of nine subjects examined). For patients for whom a series of longitudinal isolates was available, strain persistence for up to 7 years was evident for five out of eight individuals, despite episodes of antifungal therapy. Computer-assisted comparison of the interrepeat PCR fingerprints identified seven distinct profiles that were shared among isolates from different individuals. However, no association was evident among isolates of C. albicans from specific clinical conditions. Eight isolates that were initially identified as C. albicans but having atypical PCR profiles were later confirmed as Candida dubliniensis. In conclusion, the genotypic data do not indicate clonal restriction of C. albicans with respect to CHC. Furthermore, these results have demonstrated that in the majority of individuals, colonizing populations of C. albicans are clonal in nature and exhibit strain persistence.

Candida albicans↗

Identification of Candida dubliniensis, based on ribosomal DNA sequence analysis.

Differentiation of Candida albicans and the recently described C. dubliniensis has proven difficult due to the high degree of phenotypic similarity of these species. The present study examines sequence variations in the ribosomal DNA (rDNA) intergenic transcribed spacer (ITS) regions of C. albicans (n = 5) and C. dubliniensis (n = 7) strains, with a view to identifying sequence differences that would enable consistent differentiation of these two species by restriction fragment length polymorphism (RFLP) analysis. The ITS1 and ITS2 regions, together with the entire 5.8S rRNA gene of the strains, were amplified by the polymerase chain reaction (PCR), using primers ITS1 and ITS4, PCR products from both C. albicans and C. dubliniensis were of similar size (around 540 bp); however, sequence analysis revealed over 20 consistent base differences between the products of the two species. On the basis of sequence variation, the restriction enzyme MspA1 I was selected and used to differentiate the PCR products of C. albicans and C. dubliniensis by RFLP analysis. MspA1 I yielded two discernible fragments from C. albicans PCR products, whilst those from C. dubliniensis appeared undigested, thereby providing an approach to differentiate the two species.

Candida↗

Sensitivity of Seescan TV image analysis in discriminating between normal, dysplastic and malignant oral smears.

OBJECTIVE: Smears from premalignant and malignant lesions may contain large proportions of normal cells together with atypical cells; that could reduce the sensitivity of cytologic diagnosis. The present study assessed the performance of the Seescan TV image analysis system (TVIAS) in distinguishing between normal, premalignant and malignant oral smears. The sensitivity of Seescan TVIAS was tested using both white and monochromatic light. STUDY DESIGN: Nuclear area (NA) and corrected integrated optical density (IOD) of 50 Feulgen-stained nuclei were measured in smears collected from normal oral mucosa (n = 6), lesions displaying epithelial dysplasia (n = 5) and invasive squamous cell carcinoma (n = 5) using a Seescan TVIAS with both white and monochromatic light. RESULTS: There was a significant increase (P < .001) in mean IOD for nuclei in smears from dysplastic lesions and carcinomas as compared with normal smears. For smears from carcinomas, the mean NA was significantly elevated as compared with dysplastic (P < .001) and normal smears (P < .01). Mean NA for dysplastic smears was significantly reduced as compared with normal smears. While all smears from premalignant and malignant lesions contained mostly normal nuclei, a significant proportion of abnormal nuclei was identified in each smear. CONCLUSION: Although oral smears contain large amounts of normal cells, the Seescan TVIAS could successfully identify dysplastic and malignant cells on the basis of both IOD and NA values with or without the use of a monochromatic filter.

Carcinoma, Squamous Cell↗

Adherence of Candida albicans to oral epithelial cells differentiated by Papanicolaou staining.

OBJECTIVE: To examine the relative adherence of Candida albicans to oral epithelial cells differentiated by Papanicolaou staining. METHODS: Oral epithelial cells were collected from 10 healthy adults (five male, five female) and counted. Equal volumes of oral epithelial cells and candida were mixed and incubated. The epithelial cells from this mix were collected by filtration through 10 microns polycarbonate membrane filters. Cells retained on the membrane filters were stained with crystal violet followed by Papanicolaou stain. The number of yeast attached to each of 100 red, orange, and green staining oral epithelial cells was determined by direct microscopic examination. RESULTS: C albicans had a higher level of adherence (p < 0.001) to red staining oral epithelial cells (mean (SD) number of candida attached to 100 oral epithelial cells 562 (159)) than to cells staining either orange (105 (47)) or green (161 (66)). CONCLUSIONS: Oral epithelial cell variability for candidal adherence is confirmed. The technique provides an opportunity to examine the relation between oral epithelial cell type and oral candidosis in specific groups, such as tobacco smokers, where increased epithelial cell keratinisation and candidal colonisation has been reported.

Adult↗

Immunocytochemical detection of Candida albicans in formalin fixed, paraffin embedded material.

AIM: To assess the ability of the commercially available monoclonal antibody 1B12 (BioGenex, San Ramon, USA) to identify C albicans in formalin fixed, paraffin wax embedded material (FFPE). METHODS: Broth cultures of 20 strains of seven Candida species were resuspended in 4% agarose blocks, fixed in formalin for 24 hours, and embedded in paraffin wax. In addition, 16 blocks of FFPE tissue known to contain periodic acid-Schiff positive fungal hyphae were examined. Antigen retrieval involved microwave treatment of specimens in citrate buffer (0.01 M; pH 6.5) before addition of 1B12 antibody for 24 hours. Bound antibody was subsequently detected using a biotinylated link antibody and a peroxidase conjugated streptavidin. RESULTS: Only C albicans strains were 1B12 positive in the agarose blocks. All FFPE tissue blocks were found to contain 1B12 positive hyphal structures, indicating the presence of C albicans. CONCLUSIONS: The ability to identify candida organisms penetrating the lesional tissue in cases of chronic hyperplastic candidosis will help to clarify the role of individual Candida spp in this important form of oral candidosis.

Antibodies, Monoclonal↗

A novel technique for assessment of adherence of Candida albicans to solid surfaces.

A novel approach for the assessment of adherence of Candida albicans to translucent acrylic material is described. The method uses the inverted microscope to visualise yeast adhering to acrylic surfaces while the test material remains immersed in buffer. Adherent cells were not subjected to surface tension forces that can occur during drying processes, so that an even distribution of yeast with no aggregation occurred. The process of counting attached yeast was subsequently performed without difficulty. From the 11 C albicans isolates examined, two groups were evident with respect to acrylic adherence: one group of four isolates with an adherence level of 400 yeast/mm2 acrylic, and one group of seven isolates with adherence levels of 1000 yeast/mm2 acrylic.

Acrylic Resins↗

Opinions of dental consultants on the standard of referral letters in dentistry.

AIM: To ascertain the views of dental consultants on the relative importance of a range of topics relevant to specialist referral. SUBJECTS: 200 randomly selected dental consultants working in the UK in 1995. MAIN OUTCOME MEASURES: Data items which referral letters should contain; standard of referral letters; appropriate reasons for referral; how referrals could be improved; should restrictions be placed on specialist referrals. RESULTS: 161 replies were received. Overall there was only slight variation between specialities with regard to data items, appropriateness of referral and standard of referral letter. The perceived standard of referral letters was adequate or better on 76% of occasions; 21% were deemed to be of a poor standard; 2% were described as appalling. CONCLUSIONS: A Section 63 course on how to refer competently could be of benefit. Consultants were generally not in favour of restricting referrals to them.

Consultants↗

Depressive symptoms and alterations in sucrose taste perception: cognitive bias or a true change in sensitivity?

Previous studies have reported elevated taste thresholds in depressed subjects, but those studies did not control for changes in response bias. The current study used signal detection analyses to address this shortcoming. Sucrose detection thresholds were measured (1) in subjects with high and low Hamilton Depression Rating Scale (HAM-D) scores who did not meet standard criteria for current Major Depressive Episode (MDE); and (2) in subjects who did fulfil standard criteria for MDE. Subjects with low HAM-D scores produced significantly more false alarms than the other two groups, but taste sensitivity, as indexed by d', did not vary significantly across groups. These results suggest that changes in response bias underlie previously reported increases in sucrose taste thresholds in depressed subjects.

Adult↗

Characterisation of the inflammatory cell infiltrate in chronic hyperplastic candidosis of the oral mucosa.

The inflammatory cell infiltrate in biopsy material of chronic hyperplastic candidosis (CHC) from the oral mucosa was characterised using immunocytochemical techniques. Nine specimens were stained for human kappa and lambda immunoglobulin light chains, CD68 antigen (macrophages), lysozyme (macrophages, granulocytes), CD3 antigen (T-lymphocytes), CD20 antigen (B-lymphocytes) and leucocyte common antigen (LCA). In addition, these and a further 13 specimens were also examined for immunoglobulin (Ig)-containing cells (IgA, IgG and IgM). The density of the infiltrate varied considerably between cases; T-lymphocytes were the dominant cell type (53.9%), with fewer B-lymphocytes (8.2%) and macrophages (14.2%). Many Ig-containing cells were seen, and although IgG-containing cells predominated, (60.8%, SD +/- 9.0) there was a high proportion of IgA-containing cells (36.7%, SD +/- 9.1) with few IgM-containing cells (2.5%, SD +/- 3.0). Many neutrophils, together with smaller numbers of T-lymphocytes and macrophages, were seen in the epithelium. It is suggested that mucosal defence to Candida infection involves a cell-mediated reaction in which there is recruitment of macrophages and local production of immunoglobulin with a prominent IgA component.

Adult↗

Identification of Candida species in formalin fixed, paraffin wax embedded oral mucosa by sequencing of ribosomal DNA.

Aim-To identify Candida species in formalin fixed, paraffin wax embedded tissue by sequencing candidal rDNA.Methods-Target rDNA sequences were amplified by polymerase chain reaction (PCR) from fresh isolates of Candida and from 18 preserved oral mucosal tissue samples (16 cases of chronic hyperplastic candidiasis and two fibroepithelial polyps), shown histologically to contain Candida. Identification of Candida species within tissue was based on a comparison of the rDNA sequences obtained with those from the fresh isolates of Candida and those present in the GenBank database.Results-The PCR products obtained from 12 of the 18 tissue specimens studied were characteristic of Candida albicans. In two of these cases a second, larger PCR product was obtained and these sequences were characteristic of Candida glabrata.Conclusions-Candidal DNA was amplified successfully from formalin fixed, paraffin wax embedded tissue. Sequencing of the PCR product enabled identification of the Candida species present.

Journal Article↗

Orofacial radiological manifestations of systemic sclerosis.

OBJECTIVE: To ascertain the orofacial radiological manifestations of systemic sclerosis. METHODS: Twenty-one patients with known systemic sclerosis underwent panoramic, and where appropriate periapical, radiography parotid sialography and a labial gland biopsy. RESULTS: The prevalence of periodontal ligament space widening was 33%, bone resorption 9.5% and abnormal parotid gland sialographic findings 43%. Root resorption affected 4 (33%) of the 12 dentate patients. CONCLUSIONS: Orofacial radiological manifestations occur commonly in systemic sclerosis. Root resorption has not previously been reported.

Adult↗

Identification of Candida species by PCR and restriction fragment length polymorphism analysis of intergenic spacer regions of ribosomal DNA.

The PCR was used to amplify a targeted region of the ribosomal DNA from 84 Candida isolates. Unique product sizes were obtained for Candida guilliermondii, Candida (Torulopsis) glabrata, and Candida pseudotropicalis. Isolates of Candida albicans, Candida tropicalis, Candida stellatoidea, Candida parapsilosis, and Candida krusei could be identified following restriction digestion of the PCR products.

Base Sequence↗

Oral presentation of metastatic calcinosis cutis.

An 80-year-old man presented with a hard swelling in the lower labial sulcus. Radiological investigation showed a mineralized mass on the labial aspect of an edentulous mandible which on excision, demonstrated the histological features of calcinosis cutis. No other sites seemed to be affected by calcific deposits in this patient, who appears to be the oldest reported case of metastatic calcinosis, and the first intra-oral case, to date. The different forms of calcinosis cutis are highlighted.

Aged↗

Proteolytic cleavage of the integrin beta 4 subunit.

The integrin beta 4 subunit often undergoes proteolytic cleavage within its long cytoplasmic tail to yield a characteristic protein pattern of 205, 165, and 125 kDa. The results in this study suggest that beta 4 cleavage often occurs during or after cell lysis, where it was readily inhibitable by calcium chelators (EDTA, EGTA) and inhibitors of cysteine proteases (E64c, leupeptin). The cleavage of beta 4 is catalyzed by a calpain-like enzyme because (i) it requires calcium, (ii) it is mimicked by purified milli-calpain, and (iii) it is inhibited by several calpain inhibitors including the calpain-specific inhibitor calpastatin. Within intact cells, cleavage of beta 4 was cell type-specific and observed only when the cells were made permeable to calcium. Substantial cell viability was retained during beta 4 cleavage induced by ionomycin plus calcium, indicating that cleavage within intact cells was not necessarily a consequence of cell death. However, manipulations of cells including suspension, synchronization, and stimulation with serum, phorbol esters, or other agents all failed to induce cleavage, suggesting that if cleavage is physiologically relevant, it is not easily duplicated in vitro. Analysis of multiple cell types showed a wide variation in beta 4 sensitivity to proteolytic cleavage, suggesting that this process might be differentially regulated depending on the cellular environment.

Antigens, Surface↗

Plasma cell populations in labial salivary glands from patients with and without Sjögren's syndrome.

Plasma cells expressing IgG, IgA and IgM were quantified in labial salivary glands from patients with Sjögren's syndrome (n = 25) and compared with glands from patients with a variety of systemic diseases (n = 32) and normal individuals (n = 15). Based on qualitative and quantitative analysis, glands from the systemic disease group were divided into normal histology (n = 24) and non-specific inflammation (n = 8) groups. There were no significant differences in cell densities or Ig class proportions between histologically normal glands from patients and those from normal volunteers. Total immunocyte densities were significantly increased in sialadenitis (P < 0.025) and Sjögren's syndrome (P < 0.001) compared with normal histology glands. In both the sialadenitis and Sjögren's syndrome groups there were significant increases in IgG and IgM cell densities (IgG, P < 0.006; IgM, P < 0.001) and proportions (IgG, P < 0.05; IgM, P < 0.001). There were no significant differences in immunocyte densities or proportions between the sialadenitis and Sjögren's syndrome groups except for a lower percentage proportion of IgA cells in the latter (P < 0.038). In all groups the total and individual Ig-class cell densities showed significant positive correlations with extent of leucocyte infiltration (P < 0.01) and negative correlations between IgA and IgG and/or IgM cell proportions. Analysis of the plasma cell data alone and in combination with quantifiable histological parameters failed to yield specific or sensitive diagnostic information. The results suggest that changes in glandular plasma cell populations in Sjögren's syndrome are non-specific.

Adult↗

Knowledge-based design of removable partial dentures using direct manipulation and critiquing.

A knowledge-based system for designing removable partial dentures is described in which graphical representations of denture components are manipulated directly by the user to build the required denture design. The arch form of individual patients may be depicted by linear movement and rotation of the icons representing teeth. The size, shape and position of many denture components are determined dynamically to conform to the shape of abutment teeth and to the juxtaposition of other elements. Expert clinical knowledge is used to analyse the developing design and suggest alternate approaches.

Computer-Aided Design↗

Intra-oral salivary gland tumours in the West Midlands.

Intra-oral salivary gland tumours reported from the West Midlands over the years 1975-88 were examined as to a number of epidemiological variables; the question as to their increasing incidence was also addressed. The findings were largely in agreement with those of most previous authors in that most of the tumours occurred in the palate, but whereas most of these were benign, a large proportion of those in other sites were malignant. However, firm conclusions are difficult to draw since there is much disagreement over reporting criteria. An international workshop and comprehensive tumour registries are required in order to determine more accurate data and conclusions. Similar cooperation is necessary in order to substantiate or refute the suggestion that the incidence of salivary gland tumours is increasing.

Adenocarcinoma↗