A proposal for reclassification of oral candidosis.
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Acute Disease↗
Biomedical subjects
I Olsen
Publications and source records attributed to I Olsen.
Oral lichen planus is not associated with IgG circulating antibodies to epithelial antigens.
Autoantibodies to a number of epithelial components have previously been described in small groups of patients with lichen planus. Recently a group of antibodies to monkey esophagus have been detected in lichen planus related to hepatitis C virus infection. This study has examined the frequency of serum antiepithelial antibodies in a group of patients with idiopathic oral lichen planus and lichenoid drug reactions. Five of 34 patients with idiopathic lichen planus and two of six patients with lichenoid eruptions had circulating antibodies that gave rise to an antinuclear pattern when examined using epithelial tissue. However, these antibodies were present in only low titer and were not specific to a particular clinical presentation of lichen planus or lichenoid drug reaction. It seems likely therefore that such antibodies do not play an important part in the etiopathogenesis of lichen planus, and their detection is unlikely to be beneficial in the diagnosis of this disease.
Adolescent↗
Immunologic aspects of dermal and oral lichen planus: a review.
There have been many investigations, both experimental and epidemiologic, of the forms of LP affecting the skin and oral mucosae. These studies have provided a varied range of hypotheses to explain not only the factors determining susceptibility to and onset of this disease, but also the immunologic mechanisms leading to the pathosis with which LP is associated. Much progress has been made, especially through in vitro studies, regarding detailed aspects of the immunology of LP. However, data is often conflicting or incomplete. In this review we attempt to bring together the currently available data regarding the immunologic basis of LP.
Animals↗
Fungi in therapy-resistant apical periodontitis.
The occurrence of yeasts in 967 microbiological endodontic samples taken from root canals in persistent endodontic infections was studied. The sampling was done by general practitioners in various parts of Finland from root canal infections which did not respond favourably to standard conservative therapy. The samples were cultivated aerobically on a non-selective enriched horse blood agar medium, on TSBV agar medium in 5% CO2 and anaerobically on horse blood agar medium. Micro-organisms were found in 692 of the samples while 275 showed no growth. Forty-eight fungi were isolated from 47 samples which is 7% of the culture-positive samples. Twenty yeast strains were identified further by their colony morphology, growth and cellular characteristics and patterns of carbohydrate assimilation. All isolates except one belonged to the genus Candida. Candida albicans was the most common species. C. glabrata was found together with C. albicans in one sample. C. guilliermondii, C. inconspicua and Geotrichum candidum were each isolated once. Yeasts were found in pure culture in six samples and together with bacteria in 41 samples. In all the samples except two, the accompanying facultative bacteria were Gram positive. The most frequent of them were alpha- and non-haemolytic Streptococcus species which were found in 31 samples. Anaerobic bacteria were isolated together with yeasts from 12 root canals. They included both Gram positive species such as Peptostreptococcus micros and Gram negative species such as Fusobacterium nucleatum. The regular isolation of yeasts, also in pure culture, indicates that yeasts may have an important role in cases of apical periodontitis persisting after conventional treatment.
Candida↗
Observation of Saccharomyces cerevisiae in blood of patient undergoing root canal treatment.
In this study, an unusual observation of Saccharomyces cerevisiae isolated from an infected root canal and from the blood of a patient undergoing endodontic therapy of a tooth with asymptomatic apical periodontitis is reported. Phenotypic (biochemical tests, antifungal susceptibility tests and SDS-PAGE of cellular proteins) and genetic (ribotyping) methods were used to characterize the strains. By using these methods it was found that the blood and root canal isolates were identical but differed from S. cerevisiae strains of other sources. It was therefore more than likely that the root canal was the source of the blood isolate and that it had been transferred unintentionally to the bloodstream during root canal treatment.
Antifungal Agents↗
Antibodies to epithelial components in oral lichen planus (OLP) associated with hepatitis C virus (HCV) infection.
Oral lichen planus (OLP) is a common chronic inflammatory disorder sometimes associated with hepatitis C virus (HCV) infection. An increased prevalence of autoimmune markers has been reported in patients with HCV infection. The aim of the present study was to determine, by conventional indirect immunofluorescence, the nature and frequency of circulating antibodies to epithelial antigens in the sera of HCV-positive patients who also have OLP. The study comprised four groups: 14 patients with OLP and HCV infection. 14 HCV-seronegative patients with OLP, 21 HCV-seropositive patients without OLP and 18 healthy controls. We found a significant association between the concomitance of OLP and HCV infection and the presence of such antibodies. It is concluded that some patients with HCV-associated OLP may have circulating antibodies to epithelial antigens, although their precise aetiological role in the development of this disease in HCV infection remains unknown.
Antiviral Agents↗
Expression and splicing of the fibronectin gene in healthy and diseased periodontal tissue.
Fibronectin is a major component of the extracellular matrix and is considered to have an important role in chronic inflammatory periodontal disease. The fibronectin gene product has been shown to be subject to alternative splicing in 3 regions, each generating different mRNA transcripts associated specifically with normal adult tissue, embryogenesis, tissue regeneration, and wound healing. In the present study, using the reverse-transcribed polymerase chain reaction to examine splicing profiles of the primary transcript, we found that healthy periodontal tissue expressed all alternatively spliced embryonic isoforms, indicative of the extensive and ongoing rebuilding processes which occur in these tissues. In marked contrast, only the exon-skipped transcripts were generated in tissue from chronic inflammatory periodontal disease patients. The loss of the high molecular weight isoforms in lesional tissues may be due to the excess production of inflammatory mediators in this disease, since we observed that high concentrations of the cytokine IL-1beta caused down-regulation of these transcripts in normal periodontal cells in tissue culture. Moreover, we also demonstrated that growth factors likely to be involved in periodontal regeneration and repair, such as PDGF, IGF-1 and TGF-beta, elicited pronounced upregulation of the embryonic isoforms of fibronectin in these cells. Although the functional activities of the antigens corresponding to the alternatively spliced variants of fibronectin are not yet known, our finding that they are selectively expressed suggests that they have highly specific roles in both periodontal breakdown and repair.
Adult↗
Comparison of lipopolysaccharides from Bacteroides, Porphyromonas, Prevotella, Campylobacter and Wolinella spp. by tricine-SDS-PAGE.
Lipopolysaccharides (LPSs) of 11 bacterial strains from the type species of the genera Bacteroides (B. fragilis), Prevotella (Pr. melaninogenica), Porphyromonas (Po. gingivalis), Campylobacter (C. fetus subsp. fetus), and Wolinella (W. succinogenes), and from the type strains of B. distasonis, B. forsythus, B. ureolyticus, Po. levii, Po. macacae, and C. gracilis, were extracted with hot water-phenol (Westphal method). S-form LPSs, obtained from all organisms, were well resolved with tricine-sodium-dodecyl-sulphate polyacrylamide gel electrophoresis and visualized by silver staining. Lipid A was not stained. Also profiles from LPS of Escherichia coli, serotypes 0111:B4 and 055:B5, could be distinguished. While W. succinogenes showed a relatively short S-form LPS on electrophoregrams, the other bacteria, including B. fragilis, exhibited long-ladder LPSs. Po. gingivalis displayed the largest number of bands and the longest O-chain. The long O-chain of this bacterium may be important for its virulence.
Antigens, Bacterial↗
Establishment of long-term myogenic cultures from patients with Duchenne muscular dystrophy by retroviral transduction of a temperature-sensitive SV40 large T antigen.
We have established long-term human myogenic cultures from adult human skeletal muscle biopsies by infecting primary explant cultures with an amphotropic retroviral construct encoding a temperature-sensitive SV40 large T antigen, tsA58-U19. Infected myoblasts expressed the large T antigen and showed greatly enhanced proliferative capacity when cultured at 33 degrees C, compared with noninfected cells. When the infected cultures were incubated at 39 degrees C, the cells withdrew from cycle, aligned, and fused to form multinucleated myotubes which expressed certain antigens that are similarly expressed in nontransduced differentiating muscle cells. Myogenic clones with greatly increased proliferative capacity were generated, for the first time, from biopsies obtained from Duchenne muscular dystrophy patients as well as from normal, dystrophin-positive individuals. Cell lines produced by this approach may prove valuable for in vitro studies of myogenesis and for investigating the cellular and molecular consequences of inherited muscle diseases.
Antigens, Polyomavirus Transforming↗
Polymerase chain reaction analysis of oestrogen and androgen receptor expression in human gingival and periodontal tissue.
Oestrogen and androgen receptors mediate the effects of their respective hormones by acting as ligand-activated transcription factors that control a wide range of biological processes. In order to determine whether periodontal and gingival tissues could respond to oestrogen and androgen, the specific and highly sensitive reverse-transcribed polymerase chain reaction (PT-PCR) was used to examine the presence of the mRNAs corresponding to these receptors. Expression of the androgen receptor was readily detected in periodontal and gingival tissue and in fibroblasts derived from these tissues, but transcripts for the oestrogen receptor were not detected in these samples. Moreover, treatment of the cultured fibroblasts with the oestrogen diethylstilbesterol (DES) or the androgen dihydrotestosterone (DHT) did not induce the expression of the oestrogen receptor mRNA; nor did the hormones modify the activity of the androgen receptor gene. These results suggest that, while periodontal and gingival tissues are not able to respond directly to oestrogen, they may nevertheless be highly sensitive to the anabolic effects of androgens.
Cells, Cultured↗
Three subtypes of the tet(M) gene identified in bacterial isolates from periodontal pockets.
The tet(M) genes were characterized from 84 isolates of 10 different bacterial species isolated from the periodontal pockets of 16 patients with periodontal disease. A 740 bp polymerase chain reaction product from the hypervariable region of the tet(M) structural gene was cleaved with the restriction enzymes AluI and HinfI. Three different restriction patterns were identified for each of the two enzymes. By DNA sequencing, using a direct solid-phase automated sequencing method, the isolates could be grouped into 3 different clusters of tet(M) subtypes. The internal DNA homology within each subtype was 98-100%; the homology between clusters was 89-94%. Two different subtypes were identified in 9 of 10 bacterial species, and the remaining species had 3 different subtypes. One of the subtypes (M3) was seen mainly in the anaerobic isolates. This subtype was different from all earlier sequenced structural tet(M) genes present in the Genbank. Most patients had two different subtypes of tet(M), and a third subtype was seen in the 3 patients who exhibited the greatest variety of tetracycline-resistant bacterial species. It appears that the presence of one subtype of the tet(M) gene within a patient or bacterial species does not prevent the acquisition of another subtype of the same gene. This study identified a new subtype of the tet(M) gene and grouped it into 3 distinct yet highly homologous genetic subtypes.
Bacterial Proteins↗
Tetracycline resistance in Prevotella isolates from periodontally diseased patients is due to the tet(Q) gene.
Tetracycline-resistance in gram-negative periodontal bacteria is often due to the presence of the tet(Q) gene. In the present study the polymerase chain reaction (PCR) was used to examine 54 isolates of gram-negative anaerobic rods (Prevotella intermedia, Prevotella nigrescens and related or Bacteroides-like species) for the presence of the tet(Q) gene. The isolates were recovered from 42 patients with periodontal disease living in northern Europe and North America. An 814 base-pair segment of the tet(Q) gene was amplified from all 41 isolates resistant to tetracycline with minimal inhibitory concentrations of 4 micrograms/ml and above. The presence of the tet(Q) gene was verified using hybridization with a specific oligonucleotide internal to the amplified region and restriction endonuclease digestion with DdeI. A PCR product of the same size was also amplified from one tetracycline susceptible isolate (minimal inhibitory concentration = 0.5 microgram/ml). However, this isolate and the one isolate that was resistant to tetracycline at 4 micrograms/ml showed a weaker signal than the remaining isolates when hybridized with the internal probe. Typing of the PCR products using restriction endonuclease digests with AluI and HpaII revealed two clusters of distinct electrophoresis patterns, indicating that two different subtypes of the tet(Q) gene were present in this material. A control strain containing the tet(Q) gene from Bacteroides thetaiotaomicron had a different electrophoresis pattern for AluI. This study indicated that subtypes of the tet(Q) gene in tetracycline-resistant gram-negative periodontal bacteria exist both within the same patient and within the same species.
Base Sequence↗
Characterization of new periodontal and endodontic isolates of spirochetes.
Gas chromatographic analysis of cellular fatty acids (CFA), biochemical reactions, electrophoresis of soluble cellular proteins, as well as immunodiffusion were used to discriminate between 16 fresh spirochete isolates from periodontal and endodontic infections. CFA patterns were compared by the Hewlett Packard MIDI library to all available reference strains, and results of the biochemical tests were compared to VPI's records for treponemal strains. The electrophoretograms of soluble proteins of the fresh isolates were compared to those of previously well described strains of Treponema denticola, T. pectinovorum, T. vincentii, T. socranskii subsp. socranskii, T. socranskii subsp. paredis, T. socranskii subsp. buccale, and T. socranskii 04. Immunodiffusion was carried out by using adsorbed polyclonal rabbit antibodies to representative strains of the species mentioned. These methods separated most clinical strains from approved species strains, suggesting that new species had been isolated.
Bacterial Proteins↗
3-hydroxy fatty acids in a lipopolysaccharide-like material from Treponema denticola strain FM.
Treponemes are associated with major oral diseases such as apical and marginal periodontitis. Lipopolysaccharide (LPS) is regarded as an important virulence factor in these diseases. It is unclear whether LPS is present in oral treponemes. Therefore, the present study was undertaken to examine if a common oral treponeme-Treponema denticola-possesses LPS. A modified Westphal method (phenol water-ethanol-hexane extraction) was used to extract LPS-like material from T. denticola, reference strain FM. It was cultivated in prereduced anaerobically sterilized pectin medium. Gas chromatography and gas chromatography-mass spectrometry of the extract detected three hydroxy fatty acids: C3-OH-i-13:0, C3-OH-i-15:0, and C3-OH-16:0 which constituted 12% of its fatty acid content. These acids, commonly regarded as markers of LPS, suggested the presence of LPS in T. denticola.
Biomarkers↗
LFA-3 delta D2: a novel in vivo isoform of lymphocyte function-associated antigen 3.
Lymphocyte function-associated antigen 3 (LFA-3) has previously been described as occurring as two isomers in vivo, a transmembrane (TM) form and a glycosyl phosphatidylinositol (GPI)-linked form, differing only in their membrane anchoring mechanism. A third isoform, LFA-3 delta D2, which has the cytoplasmic tail of TM LFA-3 but a truncated extracellular domain, has been identified in vitro. We report that the LFA-3 delta D2 isoform, identified by RT-PCR analysis and DNA sequencing, is also present in vivo and appears to share a signal sequence with the TM and GPI isoforms. Expression of LFA-3 delta D2 was observed in both normal and diseased human buccal mucosa and gingiva. Thus, while specific functional differences between isoforms remain to be established, our results show that LFA-3 delta D2 is constitutively expressed in vivo, along with the other, previously described, isoforms of LFA-3.
Amino Acid Sequence↗
Capillary zone electrophoresis as a new tool in the chemotaxonomy of oral treponemes.
The existing taxonomy of oral treponemes is not satisfactory. This is due to the fact that these strict anaerobic bacteria are not easily cultivated or differentiated. Therefore, new techniques that can contribute to improved cultivation, classification, and identification of these fastidious organisms should be welcomed. In the present study capillary zone electrophoresis (CZE) was used to distinguish oral treponemes by their metabolic product patterns generated in a liquid medium. To our knowledge this technique has not previously been used in bacterial chemotaxonomy. Reference strains of Treponema denticola, Treponema pectinovorum, Treponema vincentii, Treponema socranskii subspecies buccale and Treponema socranskii subspecies socranskii were cultured anaerobically in duplicate on different days in Pectin medium for 4 days at 37 degrees C under nitrogen atmosphere. Treponemal cells were harvested by centrifugation. Thereafter, their supernatants were filtered through 0.22-micron Millipore filters and subjected to CZE. The resulting electropherograms clearly distinguished T. denticola, T. pectinovorum and T. vincentii. Minor differences were detected between T. socranskii subspecies buccale and T. socranskii subspecies socranskii. Subspecies were clearly different from species. It seems that CZE of culture metabolites, which showed high resolution and good reproducibility, may be a valuable tool in the chemotaxonomy of oral treponemes, even at the subspecies level.
Culture Media↗
A new method for routine isolation of oral treponemes using U-tubes and pectin medium.
Determination of the composition of the oral microflora has traditionally been based on cultivation. Treponemes are prevalent in many oral infections but, unfortunately, are not regularly cultured. In this study a new method was established for routine isolation of oral treponemes from clinical samples. Bacterial samples from 47 periodontal pockets and 4 endodontic infections were incubated anaerobically under nitrogen atmosphere at 37 degrees C in U-tubes containing pectin medium. In the U-tube a 'bacterial sample side' and a 'sterile medium side' were established on separate sides of a membrane filter and an agar plug. Using this method we were able to isolate viable treponemes from all bacterial samples. This was in contrast to previously established methods such as the agar dilution technique, the technique involving the membrane filter placed on the surface of solid agar media and the well in agar plate technique. We believe that the 'U-tube method' is a valuable supplement to previously described techniques in routine isolation of treponemes from clinical samples.
Journal Article↗
Scleroderma-derived human fibroblasts retain abnormal phenotypic and functional characteristics following retroviral transduction with the SV40 tsT antigen.
In this study an amphotropic retrovirus has been used to efficiently transduce normal human (NF) and scleroderma (systemic sclerosis; SSc) dermal fibroblasts (SScF) with a sequence encoding a temperature-sensitive mutant of the SV40 large T antigen (tsA58-U19). From the primary outgrowths of skin explants, cultures were generated whose growth was stringently temperature-dependent. When grown at a low, permissive temperature (35 degrees C), both normal and SSc-transduced cells continuously divided with similar doubling times, whereas at a high, nonpermissive temperature (39.5 degrees C), division of both the NF and SScF cells was rapidly arrested. These cells have been passaged more than 50 times, have the typical morphological appearance of fibroblasts, and have retained an anchorage-dependent phenotype. The transduced normal cells (tsT-NF) synthesized the matrix molecules collagen and fibronectin and expressed phenotypic antigens characteristic of their nontransduced counterparts, including MHC Class I, VLA beta 1 (CD29), Hermes 1 (CD44), VLA-4 alpha (CD49d), ICAM-1 (CD54) and LFA-3 (CD58) and the cell surface ectoenzymes neutral endopeptidase (CD10), aminopeptidase N (CD13), and dipeptidyl peptidase IV (CD26). Analysis of the transduced SSc fibroblasts (tsT-SScF) showed that these cells exhibited certain major features of the SSc pathology, notably the abnormally high synthesis of type I collagen, increased expression of ICAM-1, and depressed levels of CD26. Moreover, these phenotypic characteristics were retained even after prolonged culture in vitro. The tsT-SScF cells also retained their responsiveness to cytokines, since interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) both produced a marked increase in ICAM-1 expression. Our findings show that infection of SScF with the SV40 tsT antigen extends the life span of these cells and does not ablate their abnormal phenotypic and functional characteristics.
Antibodies↗