Oral hairy leukoplakia: clinicopathologic features, pathogenesis, diagnosis, and clinical significance

Oral hairy leukoplakia (OHL) is a lesion frequently, although not exclusively, observed in patients infected by human immunodeficiency viruses (HIV). OHL is clinically characterized by bilateral, often elevated, white patches of the lateral borders and dorsum of the tongue. Histologically, there is profound acanthosis, sometimes with koilocytic changes, and a lack of a notable inflammatory infiltrate. The koilocytic changes are due to intense replication of Epstein-Barr virus (EBV), while epithelial hyperplasia and acanthosis are likely to result from the combined action of the EBV-encoded proteins, latent membrane protein-1, and antiapoptotic BHRF1. How OHL is initiated and whether it develops after EBV reactivation from latency or superinfection remain unresolved; nevertheless, definitive diagnosis requires the demonstration of EBV replicating vegetatively in histological or cytological specimens. In patients with HIV infection, the development of OHL may herald severe HIV disease and the rapid onset of AIDS, but despite its title, OHL is not regarded as premalignant and is unlikely to give rise to oral squamous cell carcinoma.     

Verotoxins induce apoptosis in human renal tubular epithelium derived cells

We have developed an animal model to study human delayed-type hypersensitivity reactions. Previous studies in humans have shown after tuberculin injection the presence of a mononuclear cell infiltration, with almost no eosinophils, associated with a preferential Th-1-type cytokine profile. Human skin graft obtained from tuberculin-reactive donors was grafted onto the back of severe combined immunodeficient mice. After healing, mice were reconstituted intraperitoneally with peripheral mononuclear cells. Tuberculin and diluent were injected intradermally, and skin biopsies were performed 72 hours later. Skin grafts were divided into two parts, one for immunohistochemistry and one for in situ hybridization studies. Immunohistochemistry was performed on cryostat sections using the alkaline phosphatase anti-alkaline phosphatase technique. In the tuberculin-injected sites as compared with the diluent-injected sites, there were significant increases in the number of CD45+ pan leukocytes and CD4+, CD8+, CD45RO+ T cells but not in CD68+ monocytes/macrophages and EG2 or MBP+ eosinophils. The activation markers CD25 and HLA-DR were up-regulated in the tuberculin-injected sites. In situ hybridization was performed using 35S-labeled riboprobes for interleukin (IL)-2, interferon (IFN)-gamma, IL-4, and IL-5. After tuberculin injection, a preferential Th-1-type cytokine profile was observed with significant increases in the numbers of IL-2 and IFN-gamma mRNA-expressing cells. These results are similar to those reported after tuberculin-induced delayed-type hypersensitivity in humans, suggesting that this model might be useful to study cutaneous inflammatory reaction.     

Detection of Epstein-Barr virus in oral scrapes in HIV infection, in hairy leukoplakia, and in healthy non-HIV-infected people

Epstein Barr virus (EBV) has been implicated in the pathogenesis of oral hairy leukoplakia (OHL). EBV is normally detected by lesional biopsy. The objectives of this study were to examine oral scrapes containing squamous epithelial cells (squames) from HIV-infected people with and without clinical lesions of OHL, and from healthy non-HIV-infected controls, for EBV-DNA by the polymerase chain reaction (PCR). EBV-DNA was detected in 65% of HIV-infected people and 20% of healthy HIV-negative controls but in HIV-infected individuals it was found as frequently in those without OHL as in those with. Moreover, EBV-DNA was not detected in all HIV-infected individuals, nor in all OHL. The results suggest that the presence or absence of detectable EBV-DNA in oral scrapes, though a guide, cannot be regarded as absolutely reliable in the diagnosis or exclusion of OHL.     

Oral cancer risk in relation to sexual history and evidence of human papillomavirus infection

BACKGROUND: Experimental models and analyses of human tumors suggest that oncogenic, sexually transmittable human papillomaviruses (HPVs) are etiologic factors in the development of oral squamous cell carcinoma (SCC). We conducted a population-based, case-control study to determine whether the risk of this cancer is related to HPV infection and sexual history factors. METHODS: Case subjects (n = 284) were 18-65-year-old residents of three counties in western Washington State who were newly diagnosed with oral SCC from 1990 through 1995. Control subjects (n = 477) similar in age and sex were selected from the general population. Serum samples were tested for HPV type 16 capsid antibodies. Exfoliated oral tissue collected from case and control subjects and tumor tissue from case subjects were tested for HPV DNA. Odds ratios (ORs) were calculated after adjusting for age, sex, cigarette smoking, and alcohol consumption. RESULTS: Among males only, oral SCC risk increased with self-reported decreasing age at first intercourse, increasing number of sex partners, and a history of genital warts. Approximately 26% of the tumors in case subjects contained HPV DNA; 16.5% of the tumors contained HPV type 16 DNA. The prevalence of oncogenic HPV types in exfoliated oral tissue was similar in case and control subjects. The ORs for HPV type 16 capsid seropositivity were 2.3 (95% confidence interval [CI] = 1.6-3.3) for all oral SCCs and 6.8 (95% CI = 3.0-15.2) for oral SCCs containing HPV type 16 DNA. The joint association of cigarette smoking and HPV type 16 capsid seropositivity with oral SCC (OR = 8.5; 95% CI = 5.1-14.4) was stronger than predicted from the sum of individual associations with current smoking (OR = 3.2; 95% CI = 2.0-5.2) and seropositivity (OR = 1.7; 95% CI = 1.1-2.6). CONCLUSIONS: HPV type 16 infection may contribute to the development of a small proportion of oral SCCs in this population, most likely in combination with cigarette smoking.     

Secretion by parafollicular cells beginning at birth: ultrastructural evidence from developing canine thyroid

The ultrastructure of developing canine parafollicular cells has been examined. The hypothesis that secretion is relatively inactive prior to birth but very active at and following birth was tested. Parafollicular cells develop and accumulate characteristic secretory granules prior to birth. However, there is little or no evidence of exocytosis at this time. At birth and during the neonatal period, but not in adult thyroids, signs of exocytosis of granular contents by parafollicular cells are abundant. Just prior to the expected date of birth and before evidence of exocytosis appears, parafollicular cells accumulate intracisternal granules within rough endoplasmic reticulum. These observations are consistent with the view that parafollicular cells first become actively secretory around the time of birth and are more active at this time than in early fetal or later adult life.     

Localization of Merkel cells at hairless and hairy human skin sites using keratin 18

Merkel cells are neurosecretory cells of the skin with epithelial features such as desmosomes and expression of keratins 8, 18, 19, and 20. Merkel cells are scarcely distributed in adult human skin. Although they are present in hair follicles, their density is higher at hairless anatomic sites such as palms and soles. These cells are often innervated by sensory nerve fibers and are thought to be specialized mechanosensory skin receptor cells. However, their precise origin and function are not clearly established. The aim of this study was to localize Merkel cells in human hairless and hairy skin by immunohistochemistry with antibodies Ks18.174 and Ks19.1 directed against keratins 18 and 19, respectively. In glabrous skin of palm and sole, Merkel cells have been localized at the bottom of the rete ridges, in the epidermal basal layer. To study Merkel cell distribution at hairy anatomic sites, we have chosen breast skin, a tissue containing small hair follicles typical of those covering most of the body's surface. Merkel cells were present in the interfollicular epidermis. In hair follicles, they have been identified in the isthmus region.     

Isolation and characterization of basal cells from human upper respiratory epithelium

Cellular pathways of normal and reparative differentiation of upper airway epithelium are not well understood. Of the three main cell types, basal and secretory cells are known to divide, while ciliated cells are considered terminally differentiated. Several investigations support the role of the basal cell as a progenitor cell type, but others suggest that the secretory cell can regenerate a complete mucocilliary epithelium. Thus, lineage relationships within renewing adult epithelia are still unclear. Understanding the pathways involved in upper airway epithelial cell differentiation is critical for studying injury and repair mechanisms and for developing clinical strategies for tracheal reconstruction. We undertook the current studies to determine the integrin profile of isolated human upper airway basal cells. Respiratory epithelial cells (REC) were isolated by elastase digestion, stained with FITC-labeled Griffonia simplicifolia isolectin B4 (GSI-B4), and sorted by flow cytometry. Approximately 80% of the lectin-positive cells were basal cells, as determined by morphology and cytokeratin staining. These cells expressed integrins alpha 1, alpha 2, alpha 3, alpha 5, alpha v beta 5, beta 1, beta 3, and alpha 6 beta 4, by immunohistochemistry. This is the first report to identify the integrin profile of isolated human upper airway basal cells. These basal cells could be maintained on type I collagen for at least 7 days, where they became partially confluent and retained expression of cytokeratins 5 and 14. Availability of pure populations of basal cells should permit investigations of their role in both normal and maladaptive repair of adult upper airway epithelium.     

Cyclic ultrastructural changes in ewe uterine tube (oviduct) infundibular epithelium

Ultrastructural features of the uterine tube (oviduct) infundibulum of ewes have been studied, with special reference to cyclic changes in the ciliated and the secretory cells. Tissue from the uterine tube infundibulum was taken from 12 Rambouillet crossbreed ewes which were killed at intervals (days 1 (or estrus), 3, 9, 10, 12, and 16) throughout the estrous cycle. The presence of cilia was demonstrated throughout the estrous cycle, and true degeneration or loss of cilia was not apparent at any phase of the cycle. Presence of fibrous granules, which are supposedly related to basal body replication, was demonstrated in the apical cytoplasm of ciliated cells on day 1 of the estrous cycle. Small ciliary buds were especially present on day 1, indicating active formation of cilia during the follicular phase of the cycle. The presence of fibrous granules, basal bodies, and ciliary buds at estrus indicates that ciliogenesis in the ewe uterine tube is stimulated by high levels of endogenous estrogen. Rootlets were observed both during the follicular and the luteal phases of the cycle. The rootlets were about 1 mum long, and their fine structure indicates that they might function as anchoring structures for the motile cilia. The most striking feature during estrus was the occurrence of glycogen granules in the cytoplasm of ciliated and secretory cells. These granules were in the apical cytoplasm and basal region of some epithelial cells. They were minimal or absent during the luteal phase of the estrous cycle. The presence of electron-dense glycogen particles was clearly demonstrated within basal bodies. Possibly the glycogen within the basal bodies functions as a source of energy for ciliary movement and the cytoplasmic glycogen as nourishment for the ovum. The secretory cells also showed characteristic cytologic changes which were correlated with the phase of the estrous cycle. Maximal secretory cell differentiation was apparent during the follicular phase, at which time these cells were characterized by well-developed rough endoplasmic reticulum, numerous ribosomes, and secretory granules of varied size, shape, and density. A most remarkable feature of the granules was their membranous structure, consisting of concentric lamellae of equal dimensions. Typical extrusion of secretory granules into the tubal lumen was apparent during the follicular and the luteal phases of the estrous cycle. Cytoplasmic projections containing nuclei protruded into the tubal lumen and some were free in the lumen, especially during the luteal phase of the estrous cycle. The presence of a well-developed endoplasmic reticulum and numerous secretory granules during estrus indicate that secretion in the ewe uterine tube is presumably under the control of circulating high plasma concentrations of estrogen.     

Nucleoli in human pseudostratified columnar epithelium cells. (Microscopic classification of nucleoli)

The human pseudostratified epithelium was investigated to provide more information on the incidence of main nucleolar types in its epithelial cells. The compact nucleoli or nucleoli with nucleolonemas investigated by light microscopy were usually present in epithelial cells with highly basophilic cytoplasm. The absence of these nucleoli and the presence of ring-shaped nucleoli and/or micronucleoli (reflecting the decrease or inhibition of the RNA synthesis) were observed in epithelial cells which were characterized by the diminished or disappeared basophilic properties of the cytoplasm. These observations indicating the gradual inhibition of the nucleolar (ribosomal) RNA suggest that similar maturation processes may occur in investigated human pseudostratified columnar epithelium which are known for cell lines of the mesenchymal origin.     

Observation of ultrastructural changes in cultured retinal pigment epithelium following exposure to blue light

BACKGROUND: The retina can be damaged by light even when levels of energy are well below the threshold for thermal damage, and the experimental damage of the retinal pigment epithelium (RPE) may be induced more easily by blue light than by longer wavelengths of visible light. The present study demonstrates the ultrastructural damage produced by exposure to blue light in cultured RPE. METHODS: Long-Evans rats were enucleated 8-10 days after birth for primary culture. One week after seeding, the monolayer culture of RPE cells was exposed to a cool blue light (wavelength = 440 +/- 10 nm) for 36 h (12 h/day, 3 days) at 2.0 mW/cm2. Transmission electron microscopy was used to compare the exposed RPE with the control. The entire experiment was repeated 3 times independently. RESULTS: The cytoplasm of the exposed RPE exhibited degenerative changes, such as large whorls of membrane, lamellar whorls and whorled inclusions. CONCLUSION: The RPE cells can be damaged directly by blue light after excluding the possible influence of phagosomes. This primary culture of RPE can also serve as an in vitro model for the study of light damage to the RPE.     

Macromolecular synthesis and ultrastructural changes induced in human larynx carcinoma cells following photodynamic therapy in vitro

Human larynx carcinoma cells (HEp2) were sensitized with different concentrations of Hematoporphyrin and irradiated with a He-Ne laser at different fluences. The degree of PDT-effects were estimated by two parameters: a) macromolecular synthesis and b) observations using electron microscopy. All experiments were evaluated after 68 hr at 37 degrees C. The results showed that PDT exposure of HEp2 cells is characterized by: 1) inhibition of macromolecular synthesis and 2) different cellular and subcellular lesions. Summing up, these studies indicate the existence of a strong correlation between different PDT exposures and the degree of biochemical and ultrastructural changes in human larynx carcinoma cells in vitro.     

Force and mobility in the ageing human tongue

Auditory training means different things to different people. In this brief paper, an attempt is made to review the various aims of and claims made for auditory training, and to reference some of the evidence which bears on the validity of these claims.     

Three-dimensional appearance of Langerhans cells in human gingival epithelium as revealed by confocal laser scanning microscopy

Despite the reducing exposure to allogeneic blood in cardiac surgery, most of patients with anemia still require allogeneic blood. In this study, we have attempted to harvest the blood from cardiac patients with baseline hemoglobin levels below 11.0 g/dl using recombinant human erythropoietin (rHuEPO). 29 anemic patients undergoing cardiac surgery at our hospital between January 1994 and March 1997 were divided into two groups: 3 weeks' treatment with recombinant human erythropoietin (rHuEPO) and blood donation (group 1, n = 15) and iron supplementation alone (group 2, n = 14). There were no statistically significant differences among the two groups in patients characteristic and surgical data. No serious adverse events after phlebotomy were apparent in patients donating autologous blood. Patients in group 1 had significantly higher hemoglobin levels than patients in group 2 at 7 days before operation. The number of reticulocytes were increased at just before operation in group 1, whereas group 2 showed no significant increase. The estimated hemoglobin increase in group 1 were higher at 7 days and just before operation. In 75% of group 1, allogeneic blood transfusion could be avoided, while all patients in group 2 received allogeneic blood transfusion. This study suggests that the combination of rHuEPO administration and autologous blood donation would be beneficial for anemic patients in elective cardiac surgery. The use of rHuEPO should not be restricted to anemic patients.     

Interferon alpha and intracytoplasmic free calcium in hairy cell leukemia cells

Hairy cell leukemia (HCL) is a B-cell tumor affecting the pre-plasma stage of B cell differentiation. One of the most striking characteristics of this disease is its remarkable responsiveness to alpha-interferon (IFN-alpha) therapy. Interferons constitute a heterologous family of multifunctional cytokines displaying anti-viral, anti-proliferative and immunoregulatory properties. These activities have been extensively studied in hairy cells, but the mechanism of action of IFN-alpha in hairy cell leukemia remains unknown. Our approach to investigate the mode action of IFN-alpha in HCL has been to identify abnormalities which occur in these tumor cells and then to ascertain whether these abnormalities can be rectified by IFN-alpha treatment. A high level of free Ca2+ in the cytoplasm of hairy cells was identified. Increases in cytosolic Ca2+ are believed to be a pivotal signal in regulating cell proliferation, cell differentiation and cell death. These high Ca2+ levels in hairy cells could be reduced upon treatment with IFN-alpha either in vitro or in vivo, probably acting by reducing Ca2+ influx into the leukemic cells. Moreover, the effect of IFN-alpha on [Ca2+]i seems to be correlated with down-regulation of CD20 phosphorylation, a B cell specific phosphoprotein involved in Ca2+ influx across the plasma membrane. The possible origins and implications of Ca2+ deregulation and the possible mechanisms or sites of action of IFN-alpha in tumor cells from HCL are explored in this review.     

Vibrotactile and temperature sensory interaction in the human tongue

Guinea-pig dermal scar was shown to contain type III collagen, and, from densitometric analysis of gel electrophoretograms, it was shown to have a higher concentration than the surrounding dermis. This finding is consistent with the 'embryonic' nature of newly formed dermal wound tissue, reflected in increased hydroxylation of collagen lysine and the presence of dihydroxylysinonorleucine (after reduction) as the major cross-link.     

Beta-carotene concentration in buccal mucosal cells with and without dysplastic oral leukoplakia after long-term beta-carotene supplementation in male smokers

OBJECTIVE: To measure the beta-carotene concentration in buccal mucosal cells in smoking men who had received long-term beta-carotene (BC) supplementation in a controlled trial. To assess the association of cellular BC on the prevalence of dysplasia in oral leukoplakia. DESIGN: An end-of-trial examination of a part of subjects in the Alpha-Tocopherol, Beta Carotene Cancer Prevention Study. SUBJECTS AND METHODS: 343 men who for 5-7 years had received BC (20 mg/d) or alpha-tocopherol (AT) (50 mg/d), or both of these or placebo. BC concentration of buccal mucosal cells was compared in the subjects with BC supplementation (n = 173) to that of those without it (n = 170). Oral mucosae were examined clinically and lesions showing leukoplakia histopathologically. RESULTS: Mean (s.d.) BC concentration in buccal mucosal cells was 7.7 (10.3)mg/kg protein in the subjects who received BC compared to 1.1 (1.7) mg/kg protein in those who did not. The BC concentration in the cells of supplemented subjects correlated with their serum BC levels (P < 0.001). AT supplementation had no effect on BC concentration nor was daily amount of smoking statistically significantly associated with the BC concentration in buccal cells. Altogether 17 subjects showed oral leukoplakia, 7 had dysplasia. In these 7 subjects, the BC concentration in buccal mucosal cells did not differ statistically significantly compared to subjects with only hyperkeratosis (n = 10) (F-test, P = 0.74). CONCLUSIONS: After long-term BC supplementation, BC concentration in oral mucosal cells was 7-fold greater than without supplementation. There was no evidence to support an association between cellular BC concentration and precancerous lesions among the few subjects having them in their oral mucosae.     

Porphyromonas gingivalis invades human pocket epithelium in vitro

The present study examined the adhesive and invasive potential of Porphyromonas gingivalis interacting with human pocket epithelium in vitro. Pocket epithelial tissue, obtained during periodontal surgery of patients with advanced periodontal disease, generated a stratified epithelium in culture. P. gingivalis strains W50 and FDC 381 (laboratory strains), OMGS 712, 1439, 1738, 1739 and 1743 (clinical isolates) as well as Escherichia coli strain HB101 (non-adhering control) were tested with respect to epithelial adhesion and invasion. Adhesion was quantitated by scintillation spectrometry after incubation of radiolabeled bacteria with epithelial cells. The invasive ability of P. gingivalis was measured by means of an antibiotic protection assay. The epithelial multilayers were infected with the test and control strains and subsequently incubated with an antibiotic mixture (metronidazole 0.1 mg/ml and gentamicin 0.5 mg/ml). The number of internalized bacteria surviving the antibiotic treatment was assessed after plating lyzed epithelial cells on culture media. All tested P. gingivalis strains adhered to and entered pocket epithelial cells. However, considerable variation in their adhesive and invasive potential was observed. E. coli strain HB101 did not adhere or invade. Transmission electron microscopy revealed that internalization of P. gingivalis was preceded by formation of microvilli and coated pits on the epithelial cell surfaces. Intracellular bacteria were most frequently surrounded by endosomal membranes; however, bacteria devoid of such membranes were also seen. Release of outer membrane vesicles (blebs) by internalized P. gingivalis was observed. These results support and extend previous work from this laboratory which demonstrated invasion of a human oral epithelial cell-line (KB) by P. gingivalis.     

DNA synthesis in cells of mouse corneal epithelium]

Method of autoradiography was used in order to study the kinetics of population of the cornea epithelium cells of mice. Intervals of different duration were found to exist in the DNA synthesis within the limits of S-period of one mitotic cycle. On the basis of personal and literature data a hypothesis has been put forward of a successive pattern of replication in the cells of eukaryots according to which synthesis of a fragment of the DNA daughter thread (or a chromosome subunit) occurs at each moment in a restricted site of a single matrix thread of DNA (matrix chromosome subunit). No DNA synthesis takes place at this moment in the complementary site of the second matrix thread (matrix chromosome subunit), the fragment (chromosome subunit) of one matrix thread being somewhat larger than the complementary fragment (chromosome subunit of the other matrix thread.