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Diagnosis of tuberculosis in pediatric patients remains challenging due to inherent difficulties associated with obtaining respiratory samples for molecular and culture-based testing. To address this, recent studies have highlighted the utility of tongue swabs to detect Mycobacterium tuberculosis genomic DNA in the oral epithelia of tuberculosis infected adults. It is unknown whether tongue swabs have similar utility for diagnosis of childhood tuberculosis and if the presence of DNA in these swabs was associated with whole bacilli. We therefore sought to conduct a preliminary assessment of the utility of tongue swabs to detect tubercle bacilli and their associated genetic material in young children. For this, we recruited hospitalized children with clinically diagnosed tuberculosis (n = 26) or lower respiratory tract infection (LRTI, n = 9). These categories were blinded for downstream laboratory tests, which included PCR, spoligotyping, smear microscopy, and culture. Mtb genomic DNA was detected by PCR only in clinically diagnosed TB cases [11/26 (31.4%)] and not in cases with LRTI. Of these, 5/11 [45.5%] were associated with a spoligotype. Spoligotyping also detected an additional six specimens that were negative by PCR. Using smear microscopy, 19/26 [73.1%] and 4/9 [44.4] were Mtb positive in the tuberculosis or LRTI categories respectively. We noted positive results on all three tests in 5/26 [19.2%] in the tuberculosis category and 0/9 in the LRTI category. All specimens were culture negative. Collectively, these preliminary data present a compelling case for broader testing of tongue swabs to diagnose tuberculosis in children where obtaining standard sputum specimens is not easy.

Phenolic compounds (PC) are linked to astringency sensation. Astringency studies typically use simple models, with pure PC and/or proteins, far from what is likely to occur in the oral cavity. Different oral models have been developed here, comprising different oral epithelia (buccal mucosa (TR146) and tongue (HSC-3)) and other main oral constituents (human saliva and mucosal pellicle). These models, were used to study the interaction with two PC extracts, one rich in flavanols (a green tea extract) and one rich in anthocyanins (a red wine extract). It was observed that within a family of PC, the PC seem to have a similar binding to both TR146 and HSC-3 cell lines. When the oral constituents occur altogether, flavanols showed a higher interaction, driven by the salivary proteins. Conversely, anthocyanins showed a lower interaction when the oral constituents occur altogether, having a higher interaction only with oral cells. Epigallocatechin gallate, epicatechin gallate, epigallocatechin-3-O(3-O-methyl) gallate were the flavanols with the highest interaction. For the studied anthocyanins (delphinidin-3-glucoside, peonidin-3-glucoside, petunidin-3-glucoside and malvidin-3-glucoside), there was not a marked difference on their interaction ability. Overall, the results support that the different oral constituents can have a different function at different phases of food (PC) intake. These differences can be related to the perception of different astringency sub-qualities.

The present study was conducted to assess the applicability of liquid-based cytology (LBC) using an innovative oral brush, Orcellex. Fifty healthy volunteers were recruited. From each subject, four samples were collected using “Orcellex” from apparently normal oral mucosal sites. A plastic spatula was also used to obtain an additional sample. Data on the tolerability and acceptability of the Orcellex were collected from the subjects, together with assessments of the adequacy of LBC slide preparations for cellularity, preparation quality, and the types of cells observed. The Orcellex brush was well accepted by the volunteers, who reported relatively little pain. Orcellex brush LBC preparations were of good quality in terms of cell morphology and staining, with a clean background. Only two smears (2/200; 1%) were found to be inadequate due to low cellularity. All of the plastic spatula LBC preparations were inadequate. Representative cells from all layers of the different oral epithelia examined were documented. Oral liquid-based cytology using the Orcellex brush may have considerable potential for early detection of oral cancer and precancer.

Objectives Evaluating structural changes in oral epithelium can assist with the diagnosis of cancerous lesions. Two-dimensional (2D) non-invasive optical coherence tomography (OCT) is an established technique for this purpose. The objective of this study was to develop and test the diagnostic accuracy of a three-dimensional (3D) evaluation method. Methods The oral lip mucosa of 10 healthy volunteers was scanned using an 870-nm spectral-domain OCT device (SD-OCT) with enhanced depth imaging (EDI). Four raters semi-automatically segmented the epithelial layer twice. Thus, eighty 3D datasets were created and analyzed for epithelial thickness. To provide a reference standard for comparison, the raters took cross-sectional 2D measurements at representative sites. The correlation between the 2D and 3D measurements, as well as intra- and inter-rater reliability, were analyzed using intraclass correlation coefficients (ICC). Results Mean epithelial thickness was 280 ± 64μm (range 178–500 μm) and 268 ± 49μm (range 163–425 μm) for the 2D and 3D analysis, respectively. The inter-modality correlation of the thickness values was good (ICC: 0.76 [0.626–0.846]), indicating that 3D analysis of epithelial thickness provides valid results. Intra-rater and inter-rater reliability were good (3D analysis) and excellent (2D analysis), suggesting high reproducibility. Conclusions Diagnostic accuracy was high for the developed 3D analysis of oral epithelia using non-invasive, radiation-free OCT imaging. Clinical significance This new 3D technique could potentially be used to improve time-efficiency and quality in the diagnosis of epithelial lesions compared with the 2D reference standard., oral epithelium, semi-automatic segmentation, optical coherence tomography, thickness measurement.

The oral cavity provides an entrance to the alimentary tract to serve as a protective barrier against harmful environmental stimuli. The oral mucosa is susceptible to injury because of its location; nonetheless, it has faster wound healing than the skin and less scar formation. However, the molecular pathways regulating this wound healing are unclear. Here, we show that transient receptor potential vanilloid 3 (TRPV3), a thermosensitive Ca(2+)-permeable channel, is more highly expressed in murine oral epithelia than in the skin by quantitative RT-PCR. We found that temperatures above 33°C activated TRPV3 and promoted oral epithelial cell proliferation. The proliferation rate in the oral epithelia of TRPV3 knockout (TRPV3KO) mice was less than that of wild-type (WT) mice. We investigated the contribution of TRPV3 to wound healing using a molar tooth extraction model and found that oral wound closure was delayed in TRPV3KO mice compared with that in WT mice. TRPV3 mRNA was up-regulated in wounded tissues, suggesting that TRPV3 may contribute to oral wound repair. We identified TRPV3 as an essential receptor in heat-induced oral epithelia proliferation and wound healing. Our findings suggest that TRPV3 activation could be a potential therapeutic target for wound healing in skin and oral mucosa.

Tastes are senses resulting from the activation of taste cells distributed in oral epithelia. Sweet, umami, bitter, sour, and salty tastes are called the five “basic” tastes, but why five, and why these five? In this review, we dissect the peripheral gustatory system in vertebrates from molecular and cellular perspectives. Recent behavioral and molecular genetic studies have revealed the nature of functional taste receptors and cells and show that different taste qualities are accounted for by the activation of different subsets of taste cells. Based on this concept, the diversity of basic tastes should be defined by the diversity of taste cells in taste buds, which varies among species.

The present study was designed to evaluate whether chronic exposure to a volcanically active environment might result in genotoxic and cytotoxic effects in human oral epithelial cells. A study group of 120 individuals inhabiting a volcanically active environment (exposed group; Furnas village) and a reference group of 122 individuals inhabiting a village without manifestations of volcanic activity (Santo António village) were examined in this study. Individuals from Furnas village inhabit a volcanically active environment marked by several degassing manifestations, including fumarolic fields, thermal and cold CO2 springs and soil diffuse degassing areas. For each individual, 1000 buccal epithelial cells were analyzed for the frequency of micronucleated cells (MNC) and the frequency of cells with other nuclear anomalies (ONA: pyknosis, karyolysis and karyorrhexis), by using the micronucleus assay. Information on life-style factors and an informed consent were obtained from each participant. The frequencies of MNC and of ONA per 1000 cells in the exposed group (4.3‰ and 23.5‰, respectively) were significantly higher than in the reference group (1.7‰ and 7.7‰, respectively). The risk of having a high frequency of MNC or ONA was 2.4 and 3.1-fold higher in exposed individuals compared to reference group. The analyzed confounding factors (age, gender, smoking status, alcohol consumption, and mouthwash use) did not show any significant association with the frequency of MNC or ONA. The higher risk of DNA damage in individuals inhabiting a volcanically active environment is for the first time clearly highlighted with this biomonitoring study. Given that MNC in oral epithelia are recognized as a predictive biomarker of cancer risk within a population of healthy subjects, these findings could contribute to explain the high incidence rates of lip, oral cavity and pharynx cancers previously referred for Furnas village inhabitants. Keywords: Buccal exfoliated cells, Micronuclei, Genotoxic, Volcanism

While human immunodeficiency virus (HIV) transmission through the adult oral route is rare, mother-to-child transmission (MTCT) through the neonatal/infant oral and/or gastrointestinal route is common. To study the mechanisms of cell-free and cell-associated HIV transmission across adult oral and neonatal/infant oral/intestinal epithelia, we established ex vivo organ tissue model systems of adult and fetal origin. Given the similarity of neonatal and fetal oral epithelia with respect to epithelial stratification and density of HIV-susceptible immune cells, we used fetal oral the epithelium as a model for neonatal/infant oral epithelium. We found that cell-free HIV traversed fetal oral and intestinal epithelia and infected HIV-susceptible CD4 + T lymphocytes, Langerhans/dendritic cells, and macrophages. To study the penetration of cell-associated virus into fetal oral and intestinal epithelia, HIV-infected macrophages and lymphocytes were added to the surfaces of fetal oral and intestinal epithelia. HIV-infected macrophages, but not lymphocytes, transmigrated across fetal oral epithelia. HIV-infected macrophages and, to a lesser extent, lymphocytes transmigrated across fetal intestinal epithelia. In contrast to the fetal oral/intestinal epithelia, cell-free HIV transmigration through adult oral epithelia was inefficient and virions did not infect intraepithelial and subepithelial HIV-susceptible cells. In addition, HIV-infected macrophages and lymphocytes did not transmigrate through intact adult oral epithelia. Transmigration of cell-free and cell-associated HIV across the fetal oral/intestinal mucosal epithelium may serve as an initial mechanism for HIV MTCT.

The possibility of obtaining transplantable oral epithelia opens new perspectives for oral treatments. Most of them are surgical, resulting in mucosal failures. As reconstructive material this in vitro epithelia would be also useful for other parts of the human body. Many researchers still use controversial methods; therefore it was evaluated and compared the efficiency of the enzymatic and direct explant methods to obtain oral keratinocytes. To this project oral epithelia fragments were used. This work compared: time needed for cell obtainment, best cell amount, life-span and epithelia forming cell capacity. The results showed the possibility to obtain keratinocytes from a small oral fragment and we could verify the advantages and peculiar restrictions. We concluded that under our conditions the enzymatic method showed the best results: in the cells obtaining time needed, cell amount and life-span. Both methods showed the same capacity to form in vitro epithelia.

By light microscopy, the dysplastic oral epithelia due to the neoplastic processes are similar to epithelial changes due to the inflammatory processes. Scanning electron microscopy may elucidate the different surface changes between the two. The aim of this study was to examine the surface appearances of the dysplastic oral epithelia adjacent to oral squamous cell carcinoma to see if there are any surface changes. A total of 2 specimens, one specimen from each patient with oral squamous cell carcinoma, were used for this study. Each specimen was divided in two. One half was prepared for light microscopy and the other half was prepared for scanning electron microscopy. Light microscopically, the epithelia showed mild dysplasia. By scanning electron microscopy, the keratinized cells showed irregular microridges surrounding pits, which were variable and irregular in size and shape, and the nonkeratinized cells showed parallel microridges with irregularly widened intervals between each microridge. Irregular, broad, and partly swollen microridges and irregular short, stubby surface projections were also seen. The oral epithelia adjacent to oral squamous cell carcinoma showed mild dysplasia light microscopically but appeared abnormal by scanning electron microscopy. The abnormal epithelial cells showed pleomorphism, irregular and disoriented microridges, and abnormal surface microstructures.

The temperature of taste stimuli can modulate gustatory processing. Perceptual data indicate that the adapted temperature of oral epithelia also influences gustation, although little is known about the neural basis of this effect. Here, we electrophysiologically recorded orosensory responses (spikes) to 25°C (cool) and 35°C (warm) solutions of sucrose (0.1 and 0.3 M), NaCl (0.004, 0.1, and 0.3 M), and water from taste-sensitive neurons in the nucleus of the solitary tract in mice under varied thermal adaptation of oral epithelia. Conditions included presentation of taste stimuli isothermal to adaptation temperatures of 25°C (constant cooling) and 35°C (constant warming), delivery of 25°C stimuli following 35°C adaptation (relative cooling), and presentation of 35°C stimuli following 25°C adaptation (relative warming). Responses to sucrose in sucrose-oriented cells ( n = 15) were enhanced under the constant and relative warming conditions compared with constant cooling, where contiguous cooling across adaptation and stimulus periods induced the lowest and longest latency responses to sucrose. Yet compared with constant warming, cooling sucrose following warm adaptation (relative cooling) only marginally reduced activity to 0.1 M sucrose and did not alter responses to 0.3 M sucrose. Thus, warmth adaptation counteracted the attenuation in sucrose activity associated with stimulus cooling. Analysis of sodium-oriented ( n = 25) neurons revealed adaptation to cool water, and cooling taste solutions enhanced unit firing to 0.004 M (perithreshold) NaCl, whereas warmth adaptation and stimulus warming could facilitate activity to 0.3 M NaCl. The concentration dependence of this thermal effect may reflect a dual effect of temperature on the sodium reception mechanism that drives sodium-oriented cells.

To study cell kinetics of rat gingival (GE), sulcular (SE) and junctional (JE) epithelia in the steady-state and after application of mechanical pressure.Elastic bands were inserted between first and second maxillary molars of 8-week-old male rats, which were labelled with H(3) TdR and killed in groups of six to seven animals together with equal-sized groups of labelled control animals at intervals between 1 and 168 h. Autoradiographs were used to determine epithelial cell proliferation on the pressure side of M1 by calculating the percentage of (3)H TdR-labelled cells (PLC) in the basal (BL) and suprabasal (SL) layers of GE, SE and JE and to estimate median cell cycle (MCC) duration of BL cells by plotting mean and median grain counts against time.(3)H TdR-labelled cells were present in SL of SE and JE 1-12h after isotope injection suggesting that the BL might be not the only source of progenitor cells for JE as they might also be derived through migration from adjacent SE. Application of pressure significantly (ANOVA, P0.05) reduced PLC in BL of GE, SE and JE indicating a decrease of cell proliferation after 1-12h in response to pressure. In steady-state, the MCC durations of BL cells of GE, SE and JE were 39, 14 and 9h, respectively. After application of pressure, they increased significantly (chi(2)-test, P0.05) to 48, 44 and 34 h, respectively.Sustained pressure may lead to reduction of proliferative activity of these epithelia inducing slower progression of progenitor cells through the cell cycle.

Background: Dysplasia is an important feature of leukoplakia. Because agreement among oral pathologists is poor regarding lesional diagnosis, silver stainable nucleolar organizer regions (AgNORs) as replicatory markers may have a place in objectively characterizing dysplasia in tissue specimens. Methods: We studied 41 normal oral epithelia, 51 oral leukoplakia (26 dysplastic, 25 non-dysplastic), and 51 cases of squamous cell carcinoma specimens for their mean AgNOR counts. Results: Mean AgNOR counts increased gradually from normal epithelium to non-dysplastic to dysplastic leukoplakia to squamous cell carcinoma. Using ROC analysis, we determined a mean AgNOR count cut-point (2.37) that can be used to distinguish between dysplastic and non-dysplastic leukoplakia. The test had a sensitivity of 75% and specificity of 83% with area under the curve being 88%. Conclusions: Mean AgNOR count could be a valuable criterion for defining objective parameters for diagnosis/determination of dysplasia distinguishing between dysplastic and non-dysplastic leukoplakia.

The stratified squamous epithelium of the oral mucosa is characterized by significant regional variations mainly with regard to differences in the degree of keratinization. The oral mucosa is also divided into lining, masticatory, and specialized types reflecting the functions in different areas. The permeability barrier in the oral mucosa is mainly a result of membrane-coating granules composed of various lipids that differs between nonkeratinized and keratinized regions of the oral mucosa. Another feature of the environment of the oral cavity is saliva, which contains mucins that lubricate and protect the oral mucosa. Filaggrin is believed to function as a matrix protein supporting the aggregation of keratin filaments, and it has been demonstrated in especially keratinized oral mucosa like the palate. Filaggrin and its metabolites may contribute to difference in the functions and properties of the oral epithelia. Likewise, filaggrin gene (FLG) mutations and altered distribution of filaggrin/profilaggrin in oral mucosa may play a role in disorders affecting the oral epithelia, including oral contact allergy.

Human papilloma viruses (HPV) infection is a significant risk factor for uterine cervical carcinoma. Many previous studies have also demonstrated the presence of HPVs in oral epithelia tissue. However, the role of HPV infection in oral squamous cell carcinoma (OSCC) is still controversy. The present study is to determine the frequency and type of HPV in OSCCs and oral pre-cancer lesions. Methods: DNA samples were collected from 51 OSCCs, 46 oral pre-cancer lesions and 90 normal control specimens by cytobrushings. Nested polymerase chain reaction (PCR), and gene-chip are used to identify multiple HPV types in our samples. Results: The positive rates of overall HPV types (14/46, P=0.0216, OR=2.844, CI=1.186-6.816) and of low- risk types (9/46, P=0.0096, OR=5.529, CI=1.597-19.14) are significantly higher in oral pre-cancer lesions than in control samples. The prevalence of high-risk type (11/51, P=0.0420, OR=2.819, CI=1.051-7.558) is significantly higher in OSCCs than in control but of overall types (13/51, P=0.1066, OR=2.244, CI=0.9266- 5.337) is not to reach the statistical significance.Conclusion: The high-risk HPV may play a role in OSCCs progression and the low-risk ones may associate with the oral pre-cancer lesions.

Background: The aim of this study was to develop and characterize standardized in vitro three-dimensional organotypic models of human junctional epithelium (JE) and sulcular epithelium (SE). Methods Organotypic models were constructed by growing human normal gingival keratinocytes on top of collagen matrices populated with gingival fibroblasts (GF) or periodontal ligament fibroblasts (PLF). Tissues obtained were harvested at different time points and assessed for epithelial morphology, proliferation (Ki67), expression of JE-specific markers (ODAM and FDC-SP), cytokeratins (CK), transglutaminase, filaggrin, and basement membrane proteins (collagen IV and laminin1). Results: The epithelial component in 3- and 5-day organotypics showed limited differentiation and expressed Ki-67, ODAM, FDC-SP, CK 8, 13, 16, 19, and transglutaminase in a similar fashion to control JE samples. PLF supported better than GF expression of CK19 and suprabasal proliferation, although statistically significant only at day 5. Basement membrane proteins started to be deposited only from day 5. The rate of proliferating cells as well as the percentage of CK19-expressing cells decreased significantly in 7- and 9-day cultures. Day 7 organotypics presented higher number of epithelial cell layers, proliferating cells in suprabasal layers, and CK expression pattern similar to SE. Conclusion: Both time in culture and fibroblast type had impact on epithelial phenotype. Five-day cultures with PLF are suggested as JE models, 7-day cultures with PLF or GF as SE models, while 9-day cultures with GF as gingival epithelium (GE) models. Such standard, reproducible models represent useful tools to study periodontal bacteria–host interactions in vitro. draft

In humans, high concentrations of CO2, as found in carbonated beverages, evoke a mixture of sensations that include a stinging or pungent quality. The stinging sensation is thought to originate with the activation of nociceptors, which innervate the respiratory, nasal, and oral epithelia. The molecular basis for this sensation is unknown. Here we show that CO2 specifically activates a subpopulation of trigeminal neurons that express TRPA1, a mustard oil- and cinnamaldehyde-sensitive channel, and that these responses are dependent on a functional TRPA1 gene. TRPA1 is sufficient to mediate responses to CO2 as TRPA1 channels expressed in HEK-293 cells, but not TRPV1 channels, were activated by bath-applied CO2. CO2 can diffuse into cells and produce intracellular acidification, which could gate TRPA1 channels. Consistent with this mechanism, TRPA1 channels in excised patches were activated in a dose-dependent manner by intracellular protons. We conclude that TRPA1, by sensing intracellular acidification, constitutes an important component of the nociceptive response to CO2.

角化程度によるヒト口腔粘膜での上皮細胞表面糖鎖構造の発現変化を, レクチン組織化学的に検索した。粘膜の角化様式は, HE染色およびポーリー反応から,(a) 非角化,(b) 錯角化および (c) 正角化型に分類された。非角化および正角化型上皮では, 皮膚表皮と同様のレクチン結合性を上皮細胞は示した。しかしながら, 錯角化型では, PNA染色性が錯角化層の肥厚に伴い, 消失する傾向にあり, 過錯角化では完全に消失した。さらに同型上皮では, 表皮あるいは他の粘膜上皮において染色性を認めないDBAが, PNAの消失に伴い出現した。以上の結果は, 口腔粘膜上皮表面糖鎖構造の発現様式に角化程度が関与することを示し, 時にPNAとDBAの染色性の変化は, 錯角化形成過程の特徴的な現象であることが示唆された。

The oral cavity provides an entrance to the alimentary tract to serve as a protective barrier against harmful environmental stimuli. The oral mucosa is susceptible to injury because of its location; nonetheless, it has faster wound healing than the skin and less scar formation. However, the molecular pathways regulating this wound healing are unclear. Here, we show that transient receptor potential vanilloid 3 (TRPV3), a thermosensitive Ca-permeable channel, is more highly expressed in murine oral epithelia than in the skin by quantitative RT-PCR. We found that temperatures above 33°C activated TRPV3 and promoted oral epithelial cell proliferation. The proliferation rate in the oral epithelia of TRPV3 knockout (TRPV3KO) mice was less than that of wild-type (WT) mice. We investigated the contribution of TRPV3 to wound healing using a molar tooth extraction model and found that oral wound closure was delayed in TRPV3KO mice compared with that in WT mice. TRPV3 mRNA was up-regulated in wounded tissues, suggesting that TRPV3 may contribute to oral wound repair. We identified TRPV3 as an essential receptor in heat-induced oral epithelia proliferation and wound healing. Our findings suggest that TRPV3 activation could be a potential therapeutic target for wound healing in skin and oral mucosa.—Aijima, R., Wang, B., Takao, T., Mihara, H., Kashio, M., Ohsaki, Y., Zhang, J.-Q., Mizuno, A., Suzuki, M., Yamashita, Y., Masuko, S., Goto, M., Tominaga, M., Kido, M. A. The thermosensitive TRPV3 channel contributes to rapid wound healing in oral epithelia. FASEB J. 29, 182–192 (2015). www.fasebj.org

The micronucleus (MN) assay in exfoliated buccal cells is a useful and minimally invasive method for monitoring genetic damage in humans. This overview has concluded that although MN assay in buccal cells has been used since the 1980s to demonstrate cytogenetic effects of environmental and occupational exposures, lifestyle factors, dietary deficiencies, and different diseases, important knowledge gaps remain about the characteristics of micronuclei and other nuclear abnormalities, the basic biology explaining the appearance of various cell types in buccal mucosa samples and effects of diverse staining procedures and scoring criteria in laboratories around the world. To address these uncertainties, the human micronucleus project (HUMN; see http://www.humn.org) has initiated a new international validation project for the buccal cell MN assay similar to that previously performed using human lymphocytes. Future research should explore sources of variability in the assay (e.g. between laboratories and scorers, as well as inter- and intra-individual differences in subjects), and resolve key technical issues, such as the method of buccal cell staining, optimal criteria for classification of normal and degenerated cells and for scoring micronuclei and other abnormalities. The harmonization and standardization of the buccal MN assay will allow more reliable comparison of the data among human populations and laboratories, evaluation of the assay's performance, and consolidation of its world-wide use for biomonitoring of DNA damage.

The Goal: The goal of this study was to determine the effect of psychoactive substances (drugs) on the presence and frequency of oral Candida species and Candida dubliniensis. Materials and methods: For the purpose of achieving the set goals, we chose a sample. Sixty bed-ridden patients from the Institute for Alcoholism and Other Addictions in Sarajevo Canton, both males and females between 18 and 60 years of age, were included in the research and assigned to two different groups (alcohol addicts and opiate addicts). After extensive anamnesis and a clinical examination, samples of oral epithelia were taken for microbiological identification. Two confirmatory methods were used for the identification of Candida species: the blastesis test and cultivation in a chromatophilic medium (Chrom agar). A yeast assimilation test (API test) was used for the identification of non-albicans Candida. A separate test was used to identify Candida dubliniensis (PAL agar). Results: The results of the microbiological analysis confirmed the frequency of Candida albicans (43%) in psychoactive substance addicts, as well as an increase in non-albicans Candida regardless of the type of addiction (34%). The presence of Candida dubliniensis was proven in psychoactive substance addicts (23%) and it was confirmed that the frequency of bacterial adherence of Candida dubliniensis is directly proportional to the duration of the drug-addiction. Conclusion: The abuse of psychoactive substances has an effect on the frequency of albicans and non-albicans species of oral Candida. Based on the findings, we have concluded that psychoactive substances (opiates and alcohol) lead to an increase in oral Candida dubliniensis regardless of the type of addictions.

Covalently bound lipids have been identified and compared in keratinizing porcine epithelia including epidermis and oral epithelium from palate and gingiva. Stratum corneum was isolated by tryptic digestion, and after extensive extraction of lipids using a series of chloroform-methanol mixtures, the residual tissue was subjected to alkaline hydrolysis to release covalently bound lipids. The lipids so released were analyzed by quantitative thin-layer chromatography. Stratum corncum from each of the three anatomical sites contained ω-hydroxyceramides, ω-hydroxyacids and fatty acids. In epidermal stratum corneum the total covalently bound lipids represented 2.4% of the dry weight of the tissue, but in the oral epithelia this figure was consistently lower: 0.24% in palatal stratum corneum and 0.20% in gingival stratum corneum. Transmission electron microscopy before and after lipid extraction confirms the presence of a lipid envelope in epidermal stratum corneum and demonstrates the absence of this structure in oral stratum corneum.

Epithelial structural proteins, the keratins and keratin-associated proteins, are useful as markers of differentiation because their expression is both region-specific and differentiation-specific. In general, basal cells in all stratified oral epithelia express similar keratins, while the suprabasal cells express a specific set of markers indicating commitment to a distinct program of differentiation. Critical factors in the regulation of epithelial protein expression are now under investigation. The promoter regions of keratin genes are being characterized to determine what sequences within the genes are responsible for differential expression. One important extracellular factor that influences epithelial protein expression is retinol (vitamin A), which exerts its effects via a group of nuclear receptor proteins that may also be expressed in a region-specific manner. These molecular biological approaches enhance our understanding of the mechanisms regulating differentiation of oral epithelia and its regional complexity.

The composition and proliferation of skin from the ear, tail and footpad and oral mucosa from the palate, cheek and tongue of adult mice were examined. The thickness of the nucleated cell layer of skin snowed an approximately two-fold variation; a similar range was found between that of oral mucosae but that was considerably thicker than skin. No direct correlation between epithelial thickness and the number of nucleated cells was observed. Proliferative activity, assessed following the administration of vinblastine sulphate, and turnover of the epithelium showed a broad range of activities but more rapid in the oral epithelia than in the epidermis, suggesting a relationship between functional stress and proliferative activity. The criteria used clearly distinguish between morphologically different epithelia and should prove useful in examining experimentally produced changes in epithelial histodifferentiation.

Portions of clinically healthy non-keratinized and keratinized oral epithelia were removed from adult vervet monkeys (Cercopithecus aethiops). The epithelium was separated from the underlying lamina propria by trypsin digestion, following which the epithelial cells were separated from the various epithelial layers. These were examined with the light and scanning electron microscope. Cells from non-keratinized epithelia have surface microplications while those from keratinized epithelia show microvilli and pits. Two distinct types of cell are therefore present. It is suggested that the different surface appearances are related to different types of mechanical adhesion between the cells.

Basal cell extensions have frequently been observed in a variety of odontogenic lesions as well as in the fibrous epulis (fibrous hyperplasia of the gingiva). In order to determine the frequency of these extensions in normal human oral mucosa, a study using autopsy and surgical material was conducted. The results indicated that a specific type of rete ridge morphology exists for gingiva, including the presence of thin double stranded extensions which may be related to the functional role of the gingiva or to the fact that the odontogenic apparatus is derived from surface epithelium in this region in the developing embryo, or both. Similar extensions have not been found in other oral epithelia.

The permeability of skin and oral mucosa to various compounds has been measured but the actual pathways substances take in traversing the epithelia have not been identified. In this study, radiolabelled cholesterol, ethanol or water were placed on the surface of porcine skin, keratinized gingiva, or nonkeratinized floor of mouth mucosa, and incubated at 37 degrees C for 2 h. The tissue was then snap-frozen, and sectioned in a cryostat, picked up on precoated slides and exposed at -20 degrees C for 40 days for light microscopic autoradiography. Some tissues were freeze-dried and directly embedded in low viscosity resin and prepared for electron microscopic autoradiography. Examination of autoradiographs revealed silver grains over, or adjacent to, intercellular spaces. Counts of the grains over the extra- and intracellular compartments were made in random light and electron microscope fields. For all compounds and tissue regions, there were significantly more (p less than 0.05) grains over the intercellular spaces than over the cells. The results indicate that the intercellular compartment is the predominant route for compounds moving across the superficial barrier layer of epidermis and oral epithelia. The nature of the intercellular material is, thus, a primary determinant of epithelial permeability.

A system has been developed for examining connective tissue influence on keratinization of normal and leukoplakic human oral epithelia. Paired biopsy specimens of normal and leukoplakic mucosa were left intact or were incubated in 1 mM EDTA to permit epithelial-connective tissue separation and various recombinations of the epithelium and connective tissue. The tissues were transplanting to subdermal sites of "nude" (nu/nu) mice and were protected by polyethylene capsules. 44% of implants performed were recovered apparently vital. Preexisting structure was retained after epithelial-connective tissue separation and self-recombination. Cross-recombined specimens showed changes in patterns of keratinization including changes of keratinized leukoplakic epithelium to a non-keratinized type.

Male rats in four different age groups of 2, 9, 19 and 27 months were injected with tritiated thymidine and sacrificed 45 min later. Autoradiographic sections from the palatal and the lingual epithelia were studied. The number of basal cells per mm of length, the number of labeled cells in the basal layer, and in the suprabasal area, were counted. It was found that the progenitor cell population was not limited to the basal layer alone. Thus, the number of basal cells did not coincide with the number of progenitor cells. This study deals with changes in the oral epithelia of the rat at two age periods. One, between 2 and 9 months, is a decrease of the synthesis index while the progenitor size stays constant associated with a change from a rapid growing animal to a mature one. The second, between 9 and 27 months, is a decrease in the progenitor population and an increase in the synthesis index.

An accumulation of inflammatory cells in the tissue adjacent to the newly established dentogingival junction of erupting teeth has been reported by several investigators. In order to elucidate the role played by bacteria in the induction of this presumed defence reaction against irritants from the oral cavity, the first and second molars were studied in seven germfree rats aged 14 to 20 days and in age-matched control animals. After previous demineralisation and embedding in paraffin the molars on one side were serially sectioned sagittally. Inspection of the molar areas revealed no differences in the stage of eruption between the two groups. Also, the general histologic picture was almost identical in the two groups. Before fusion of the dental and oral epithelia, only the ordinary cells of loose connective tissue were present. As soon as the molar appeared in the oral cavity, both germfree and control animals regularly showed some accumulation of inflammatory cells in the vicinity of the epithelial confluence. The greatest number of cells as well as an absolute dominance of leucocytes was noted at the onset of the reaction. Later, lymphocytes became frequent, but then the total number of inflammatory cells, and the proportion of lymphocytes, was smaller in the germfree animals than in the controls. At this stage the numbers of inflammatory cells also varied considerably in different areas around each molar.