Abstracts of scientific papers presented at the forty-sixth annual meeting of the American Academy of Oral Pathology

Abstracts of scientific papers presented at the forty-sixth annual meeting of the American Acad.emy of Oral Pathology THE BIRTH OF ORAL PATHOLOGY: PART I, FIRST DENTAL JOURNAL REPORTS OF BENIGN ORAL TUMORS AND CYSTS, 1839-1859. Bouquot J, Lense E. West Virginia University, Morgantown, W. Vu. Oral pathology is traditionally presumed to have its origins in the 1930s and 194Os, perhaps commencing with Bunting’s Textbook of Oral Pathology, Thomas’ opus, Oral Pathology, or the first issues of the Archives of Clinical Oral Pathology, or Oral Surgery. The first Professorship of “Dental Pathology”, however, was established with the Baltimore School of Dental Medicine in 1840, and the first text dedicated to oral pathology was published by Thomas Bond in 1848. It appears that the mid-19th century was the time of the true birth and early development of oral pathology. A fascination for pathologic processes was an integral part of organized or “modern” dentistry at its inception from 1830 to 1860, and half of all scientific articles in the first volumes of the earliest dental journals were related to pathology of the mouth and jaws. The first comprehensive U.S. dentistry text, Bell’s 1829 classic, dealt extensively with pathologic processes, including first reports of numerous oral lesions. Because of an American dominance in dentistry, references in early American dental journals are presumed to be among the first references to a variety of oral pathologic entities. The purpose of the present paper is to present such references. It represents a. review of early concepts of benign oral tumors and cysts resulting from a page-by-page reading of all papers published in all volumes of all journals published in English from 1839 to 1860.

GIANT CELL ARTERITIS. Uthman A. State University of New York, Bufsalo, N.Y. Giant cell arteritis affecting the temporal or maxillary artery is a rare but an important cause of facial pain, and an early diagnosis is of vital importance in avoiding disastrous consequences. The disease is defined as a focal granulomatous inflammation of arteries of medium and small size that affects principally the cranial vessels, especially temporal arteries, and usually occurs in older Caucasion women. The prevalence of temporal arteritis (TA) increases with age and it reaches an incidence of 850 per 100,000 population

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aged 80 and older. Classically TA patients present with headache, tenderness of temporal artery, facial pain, and later visual loss. Additional systemic manifestations are anemia, fever, a significant rise in erythrocyte sedimention rate, and in about 10% to 20% of cases the patient also suffers from jaw claudication. The most serious complication of TA is sudden blindness caused by opthalmic arteritis. An early diagnosis and treatment by steroids will avoid blindness and other consequences. A biopsy of the temporal artery is conclusive; however, in 40% of cases, the biopsies are negative, even in patients with classic manifestations of TA. In such cases, one must assume that the lesions were focal and were missed on biopsy. The cases presented today, attended the orofacial pain clinic of Oral Medicine Department with a medical and a dental referral for TMD problems. They presented with some of the classical features attributed to this most common of vasculitides. Diagnoses were confirmed through blood analysis, and steroid treatment completely eliminated their pain and prevented other consequences.

CINNAMON-INDUCED Bernstein M, Miller Louisvilly, Ky.

CONTACT STOMATITIS. R, Gould A. U. of Louisville,

A clinicopathologic entity characterized clinically as a white or red/white mucosal lesion and histologically as a lichenoid mucositis with prominent perivasculitis is described. In a review of 32 histopathologic cases with this microscopic appearance, 24 patients gave a positive history of cinnamon use. Eleven of these cases were seen clinically by the oral pathologist. In 19 cinnamon-using patients, discontinuance of cinnamon resulted in prompt regression of the lesions that had usually been present for months and had been refractory to other treatment. The reappearance of lesions was experienced by the two patients who were rechallenged with cinnamon. The additional five patients have been too recently accessioned at the time of this writing to evaluate their response. Cinnamon-induced stomatitis venenata should be considered in patients with symptomatic red and white mucosal lesions that otherwise might resemble lichen planus, candidiasis, snuff-dipper’s keratosis, cheek biting, galvanism, contact allergy, or premalignant mucosal disease. Alternatively, when the pathologist accessions histologic lesions that show lichenoid mucositis with perivasculitis, it may be appropriate to include a note regarding possible cinnamon causes along with the usual comments regarding lichen planus and lupus erythematosus.

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ICO (NEURALGIA-lN~UCiN~ STEONECROSIS): RADIOGRA ANCE OF THE “INVISIBLE” OSTEOMYELITIS. Bouquot J, Roberts A. West Virginia University,

Morgantown,

W. Va.

The various radiographic appearances of a newly identified form of jawbone osteomyelitis, neuralgia-inducing cavitational osteonecrosis (NICQ), are presented. NICO lesions have been found almost exclusively in patients with facial neuralgias, and curettage of NICO lesions has produced long-term and permanent pain reduction in such patients. Radiographic appearances may include one or all of the following: poorly demarcated, nonexpansile radiolucencies; moth-eaten radiolucencies; soap-bubble radiolucencies; radiopague flecks and stippling; cotton-wool radiopacities; groundglass radiopacities; “rat’s nest” radiopacities; vertical, irregular remnants of lamina dura in extraction sites; remnants of horizontal interradicular trabeculae in edentulous bone; unremodeled sockets in extraction sites; focal destruction of the inferior alveolar canal or maxillary sinus walls. Radiographic appearances can be so subtle as to be virtually unidentifiable without considerable diagnostic experience. A few NICO lesions have been identified before the onset of pain, and future pain sites can be predicted in some cases via radiographic evaluation. NICO lesions may or may not present with a positive technetium 99 scan, CAT scan, or MRI, and several patients have demonstrated negative jawbone filling on angiography.

NTITY: NEUROEPlTHELlAL HAMARTOMA. Mesa M, Baden E, Grodjesk J. University of

Medicine and Dentistry of New Jersey, Newark and Jersey City, N.J. The epithelium of the gingiva has a broad-spectrum ation potential. This is documented by the occurrence

differentiof hamarto-

mas, choristomas, and benign odontogenic tumors of the gingiva. A heretofore undescribed neuroepithelial lesion is reported in a 24year-old woman presenting clinically as a linear “exostosis” along the lingual gingival margins extending from the first to the third right mandibular molar. Light microscopic examination included hematoxylin-eosin, Masson trichrome, and Bodian method stains. Immunohistochemical studies for S- 100 protein, epithelial membrane antigen (EMA), neuron-specific enolase (NSE), musclespecific actin (MSA), and polyclonal cytokeratins were performed using the avidin-biotin complex technique. Light microscopy revealed a lesion in the lamina propria consisting of two components, namely, epithelial islands of well-differentiated squamous cells with occasional microcystic changes and focal keratinization; and numerous nervelike organoid structures, which were found in close apposition to the epithelium. Histochemistry and immunochemistry confirmed the neural nature of the latter component, which was S-100 protein and NSE positive, whereas it was negative for MSA. The epithelial islands exhibited positivity for EMA and poiyclonal cytokeratins. The review of the literature failed to reveal an identical lesion. The organ of Chievitz, although presenting collision structures of epithelial islands and nerves, has not been reported in this location. In the skin, a similar pattern has been de-

scribed in a lesion reported as “‘neurofollicular propose the term “neuroepithelial hamartoma”

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hamartoma.” We for this lesion.

NEVUS SEBACEUS OF JAQASSOHN: SIS OF 41 CASES. Fowler C, Houston

Hall USAF Medical Center, Lackland Base, San Antonio, Tex.

G. Wilford

Air Force

Nevussebaceus of Jadassohn (NSJ), also referred to as organoid nevus, is a hamartoma of the skin that most frequently occurs on the scalp, face, or neck. It is usually present at or appears shortly after birth, but may not become noticeable until puberty when it acquires a verrucous, or papillomatous appearance. In adulthood, a variety of benign or malignant skin appendage tumors may develop secondarily within lesions of NSJ. In this study of 41 cases of NSJ retrieved from the files of the department of pathology at Wilford Hall United States Air Force Medical Center, most patients presented for treatment in the second decade of life. Males outnumbered females by a ratio of 2.2/ 1. Twenty-eight (68.2%) of the patients were white. The scalp was the most common location (20 cases), followed by the face (15 cases), periauricular area (4 cases), neck (1 case), and shoulder (1 case). Ten (24.4%) of the patients had developed secondary tumors in their lesions with syringocystadenoma papilliferum (4 cases) and basal cell carcinoma (2 cases) being the most common. One patient had linear nevus sebaceus syndrome with extensive lesions of the scalp and face, and oral involvement. Pathologic material from staged excisions revealed plexiform neurofibromas associated with this patient’s NSJ although no other findings specific for neurofibromatosis were identified. In addition, this patient developed an ameloblastic fibro-odontoma of the anterior maxilla, which is interesting when it is considered as perhaps another component of his overall hamartomatous syndrome.

ORAL KAPOSI’S SARCOMA: A IQ-YEAR RETR SPECTIVE HISTOPATHOLOGIC S 1991). Regezi JA, MacPhail LA, Daniels TE, et al.

University of California,

San Francisco.

Biopsies of 120 oral Kaposi’s sarcomas (KS) submitted to the UCSF oral pathology service from 198 1 to 199 1 were reviewed to describe clinical-pathologic features and to search for unrecognized cases of bacillary angiomatosis, a recently described bacterial skin disease that mimics KS. Clinically, lesions were purple/blue/red in color except one that was mucosal colored; elevated/nodular 69 or macular 26. Most frequently biopsied sites were palate (61/120, 50%) and gingiva (34/120, 28%). Ten patients had multiple oral lesions and 25 also had cutaneous KS. Pain and ulceration were infrequently described. Histologically, the specimens could be divided into two types: small well-delineated lesions (31/120, 26%) and larger infiltrative lesions (89/120; 74%). The small, predominantly macular, lesions were characterized by inconspicuous patches of spindle cells containing ill-defined vascular spaces. They were found in the midsubmucosa and did not involve the lamina propria. Larger lesions exhibited a diffuse, infiltrative pattern of spindle cells. Slitlike and frequently (54/89,61%) angular branching or open vascular channels characterized this type of KS. Nu-

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clear atypia and mitoses were less common in small than large lesions (16% vs 38% and 26% vs 61% respectively). Red blood cell count extravasation (94 and 100%) hemosiderin (42 and 47%), and hyaline globules (45 and 45%) were seen in near equal frequency in both small and large lesia’ns. No cases of bacillary angiomatosis were found. Two pathologic patterns emerge for oral Kaposi’s sarcoma; small macular KS colmposed of patches of spindle cells with undeveloped vascular channels, and larger infiltrative KS composed of spindle cells with atypical vascular spaces. Supported by POl-DE07946.

EOSINOPHILIC ULCER OF THE ORAL MUCOSA: IMMUNOHISTOCHEMISTRY AND REPORT OF 38 NEW CASES. El-Mofty S, Wick M, Miller A. Wash-

ington University, St. Louis, and Temple University, Philadelphia. Eosinophilic ulcer of the oral mucosa (EUOM), also known as traumatic eosinophilic granuloma, is diagnostically challenging lesion for both clinician and pathologist. It commonly presents as a lingual ulcer that could be mistaken for squamous cell carcinoma. Histologically it shows mononuclear cell infiltrate that might be misdiagnosed as lymphoma. Trauma is believed to play a role in its cause, however, the exact pathogenesis is not known. The purpose of this study was twofold: first, to review the demographic, clinical, and histopathologic features of 38 new cases of EUOM, and second, using immunohistochemical stains, to characterize the mononuclear cell infiltrate. Thirty-eight previously unreported cases of EUOM were retrieved from the files of Washington University and Temple University oral pathology sections. Representative cases were selected for immunohistochemical study and were stained with the following marker antibodies: LCA, L26, MB2, UCHLl, MTI, KPl, Mac387,SlOO, Factor VIII,andUlex.Thepatient ages ranged from 6 to 88 years with an average of 56.6 years. The female:male ratio was 1.5. The majority of the lesions occurred either on the tongue or buccal mucosa. Only 19 cases presented as ulcers. A history of trauma was given in seven cases. Histologically a dense mononuclear cell infiltrate was present in a richly vascular stroma. Eosinophils were present in all lesions but varied in their density. The mononuclear cell infiltrate was composed of lymphocytes, plasma cells, histiocytes, and mast cells. Immunostains showed that the lymphocytic infiltrate was predominantly T cells, and the histiocytes were non-Langerhans’ antigen presenting-type cells. Few phagocytic macrophages were present. Although trauma might be an etiologic factosr, this study shows that cell-mediated immunity

is a likely

pathogenetic

mechanism

for EUOM.

SMOKELESS TOBACCO-ASSOCIATED EPITHELIAL AND LANGERHANS’ CELL CHANGES. Daniels TE, Hansen LS, Greenspan JS, et al. University of

California,

San Francisco.

We studied specimens ‘of smokeless tobacco (ST)-associated oral mucosal lesions from professional baseball players (95% from snuff users, 5% from chewing-tobacco users) to assess lesional histopathology and Langerhans’ cell (LC) density and antigen expression. We observed four histopathologic types of epithelial

601

change in 138 specimens: hyperparakeratosis, hyperorthokeratosis, pale surface staining, and basal cell hyperplasia; all were benign. The type of change was associated with the type of ST used, but not with the duration (years) or amount (hours per day) of ST use. The thickness of the hyperkeratotic layer in a specimen correlated directly with the amount of ST use (p = 0.005). In the 17 lesions (from snuff users) studied by immunohistochemical analysis with four monoclonal antibodies (T6, HLA-DR, -DP, -DQ), there were fewer LC in all lesion specimens than in their autologous control specimens. The average reduction in LC was 58% (range, 3% to 95%). Specimens with all types of epithelial change showed similar reductions in LC. There were no significant differences in the numbers of LC expressing any of the four antigens, in either lesion or control specimens. Our data indicate that snuff causes a greater variety and severity of epithelial change than does chewing tobacco, and that snuff reduces the number of LC at its site of contact with the oral mucosa. This may focally alter immune surveillance in oral mucous membranes. NIDR grant PO1 DE08547.

CARTILAGE IN THE WALLS OF ODONTOGENIC KERATOCYSTS. Kratochvil F, Brannon R. Armed

Forces Institute of Pathology, Washington, D.C., and Naval Dental School, Bethesda, Md. There have been a number of publications in the past that pertained to metaplasia of the epithelial lining of different types of odontogenic cysts. Very few references addressed metaplastic changes that might be induced in the fibrous connective tissue of cyst walls by the odontogenic cystic lining. Cartilage in the wall of an odontogenic cyst has only once been reported as a chondroma in the wall of an odontogenic keratocyst (OKC). We are reporting four cases of OKCs that demonstrate cartilage within the fibrous connective tissue walls (three cases from the files of the AFIP and one case from Emory University). These observations would tend to suggest that OK.Cs exert a heretofore unreported inductive effect.

SEARCH FOR CORRELATION OF RADON LEVELS AND INCIDENCE OF SALIVARY GLAND TUMORS. Miller AS, Harwick RD, Alfaro-Miranda M, Sundararajan M. Temple University, Philadelphia. A number of scientific studies have linked exposure to ionizing radiation to the development of salivary gland tumors in human beings and laboratory animals. The EPA has reported Pennsylvania radon readings to be the highest in the United States. Studies were undertaken to determine if there is a statistical correlation between radon levels and incidence of salivary gland tumors. In Part I of the study, the incidence of minor salivary gland tumors accessioned by Temple University, Emory University and University of Southern California from 1986 to 88 were correlated with average radon levels in the three loci using standard statistical analyses. In Part II, the occurrence of malignant salivary gland was obtained for each of the 67 counties in Pennsylvania for 1986 to 88 and correlated statistically with radon levels and population figures in each of those counties. Results. In Part I we could not demonstrate a statistical correlation between radon levels and incidence of minor salivary gland tumors when the Philadelphia metropoli-

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November tan area was compared with Los Angeles and Atlanta. In Part II, no correlation was found between the incidence of malignant saiivary gland and radon levels in Pennsylvania. Conclusions. Even though Oral Pathology Laboratory at Temple University accessions greater numbers of salivary gland tumors annually than the two laboratories compared in this study, the percentage of salivary gland tumors to total accessions is not significantly different and cannot be correlated with radon levels in the three geographic areas. There was no correlation between radon levels and populationbased malignant salivary glands in Pennsylvania.

MULTIPLE CARCINOMAS OF THE MOUTH AND THROAT: THE 54-YEAR EXPERlENCE OF AN ENTIRE U.S. COMMUNITY. Bouquot J, Weiland L, Kurland L. West Virginia University, Morgantown, W. Vu. and Mayo Clinic, Rochester, Minn. Hospital-based investigations have concluded that as few as 1% and as many as 40% of mouth and throat cancer patients develop additional primary carcinomas of these mucosal sites. Such a wide variation probably reflects different patient referral patterns of individual hospitals more than real differences in risk, but no population-based data have been available for comparison. The present study is the first to identify all cases of oral, pharyngeal, and laryngeal carcinoma in a well-delineated population to determine the relative frequency and characteristics of multiple cancers in an unbiased patient sample. Medical records of Rochester, Minn. residents diagnosed with these cancers between 1935 and 1988 were retrieved from all primary-, secondary-, and tertiary-care hospitals where Rochester residents may have gone for care. Of 292 primary mucosal carcinomas of the upper aerodigestive tract (excluding lip vermilion) identified in 259 residents, 25 lesions in 22 persons were additional primary carcinomas of those sites (39 of 339 cancers if lip vermilion is included). Thus multiple primaries occurred in only 8.5% of upper aerodigestive tract carcinoma patients; and 12%, 8%, and 6%, respectively, of laryngeal, oral and pharyngeal carcinomas were second or third primary carcinomas. Also, 58% (n = 14) were synchronously diagnosed. Clinicopathologic data will be presented.

UTERINE LEIOMYOSARCOMA METASTATIC TO LOWER LIP. Ailen C, Mallery S, Sebastian R, Marsh W. The OhioState University, Columbus, andSpringfield, Ohio. Metastatic lesions involving the oral soft tissues are relatively uncommon, and such lesions involving the lower lip specifically are rare. A recent review of 422 oral metastatic deposits showed only three located in the lower lip. In addition, most metastases of the oral region are carcinomas rather than sarcomas. The purpose of this report is to describe a case of metastatic uterine leiomyosarcoma to the lower lip that proved to be the initial manifestation of a recurrent malignant condition. Case report: A 65-year-old white woman presented to her oral surgeon with an 0.8 cm nontender mass in her lower lip, to the right of the midline. Clinical impression was mucocele, and the lesion was excised. Medical history was significant for acute myelocytic leu-

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kemia (1983), which was in remission, and a hysterectomy (1989) for uterine leiomyosarcoma that was believed to be confined to the wall of the uterus. Histologic examination of the lip lesion showed a well-circumscribed collection of spindle-shaped cells with a central area of necrosis. The lesional cells exhibited eosinophilic cytoplasm and pleomorphic nuclei with occasional mitotic figures. Immunohistochemistry showed that the lesional cells were positive for muscle-specific actin, and negative for S-l00 and cytokeratin. Comparison with the previously resected uterine lesion showed similarity. Further evaluation showed metastatic deposits on the chest roentgenogram and elevated alkaline phosphatase activity. Abdominal CT also demonstrated bilateral nodules suggestive of metastatic disease. The patient is currently being treated by medical oncology.

LATERAL RADICULAR INFLAMMATORY LESION ASSOCIATED WITH VITAL TEETH EXHIBITING ECCENTRIC DENS INVAGINATUS. Damm D, Harris J, Drummond J, White D. University of Kentucky, Lexington. The classic coronal dens invaginatus presents with a surface invagination that is lined by enamel and is positioned over the underlying pulp tissue. A less well-recognized pattern does occur in which the invagination perforates the lateral aspect of the root. This has been described by Oehlers and designated Type 3 dens invaginatus. Two very similar examples of this type are presented. In both cases, the developmental abnormality resulted in the creation of a periodontal abscess overlying the radicular opening of the invagination. In the first case, thorough curettage of the area was performed, and this was followed by endodontics of the vital tooth. Neither attempt was successful at resolving the problem. The source of the infection was discovered only after extraction of the tooth. In the second case, the defect was recognized, the base of the invagination was exposed, and the tooth was recontoured to eliminate the entire invagination, while maintaining vitality of the tooth. Radicular forms of dens invaginatus have been reported and must be separated from this coronal variant. The radicular variants can be lined by enamel but do not communicate with the surface and are not associated with inflammatory lesions unless recession exposes the radicular opening of the invagination. Recognition of this unusual variation of coronal dens invaginatus is important to provide a prompt diagnosis with appropriate therapy in these persistent lateral radicular inflammatory lesions.

ANAPLASTIC LARGE-CELL Ki-1 PHOMA P SENTING AS A MANDIBULAR S. Hicks Nichols CM, Flaitz CM, Luna MA. Bering Denta; Clinic, University of Texas Health Science Center Dental Branch, M.D. Anderson Cancer Center, and Baylor College of Medicine, Houston. HIV-infected persons have an increased risk for development of high-grade, non-Hodgkin’s lymphomas. Anaplastic large-Gel! Ki-1 lymphoma is a recently described lymphoid neoplasm character-

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ized by cellular

pleomorphism,

a sinusoidal

growth

pattern

and

Ki-1 (CD30) reactivity. This type of lymphoma may be mistaken for metastatic carcinoma, melanoma, or malignant histiocytosis. Although persons with AIDS frequently present with nonHodgkin’s lymphoma at extranodal sites, the oral cavity and mandible, in particular, are unusual sites. A 35year-old, HIV-positive homosexual white man presented with a mandibular 3 X 5 cm indurated mass with an irregular mucosal surface of 2 months duration. A panoramic radiograph showed extensive involvement of the underlying bone. Microscopic examination of the incisional biopsy showed large, pleomorphic, cohesive tumor cells arranged in nests and cords with minimal epidermitropism. Occasional tumor giant cells and atypical mitotic figures were present. Immunocytochemical studies were positive for Ki-1 (CD30), HD37(CDl9) and L26(CD20); and negative for LCA(CD45), UCHL-l(CD45RO), EMA, Keratin, and Desmin. In situ hybridization for EBV-DNA was positive with tumor cells. Flow cytometry on paraffin, formalin-fixed tissue revealed tetraploidy and a G2 + M of 16.5%, characteristic of high-grade lymphomas. The patient underwent a chemotherapy protocol for lymphoma with rapid resolution of the intraoral mass. Intraoral presentation of rapidly enlarging, soft tissue masses may represent a high-grade non-Hodgkin’s lymphoma in HIV-infected persons. Although rare, anaplastic large-cell Ki-1 lymphoma should be considered and requires immunocytochemical study to eliminate the possibility of other AIDS-associated malignant conditions.

CELLULAR FETAL RHABDOMYOMA OF THE BUCCAL MUCOSA. Lense E, Bycroft El, Vitt M, Nieusma

G, Bouquot J. West Virginia town, W. Va.

University,

Morgan-

Although rhabdomyosarcoma is the most common soft tissue malignancy of childhood, rhabdoyomas are much less common and account for less than 2% of all striated muscle tumors. Extracardisc rhabdomyomas are subdivided by clinical and morphologic features into three categories: genital, adult, and fetal. Fetal rhabdomyomas occur principally in the head and neck area of male children younger than 3 years old. Only a handful of oral cases have been noted in the literature. We report a case of a cellular fetal rhabdoyoma occuring in the buccal mucosa of a 2-year-old boy. He presented with a 3 X 2 cm movable mass that was clinically thought to represent a salivary gland tumor. On surgical dissection, the tumor was found to consist of a main body with finger-like projections and to be somewhat adherent to the mucosal surface. Histologically, the tumor was characterized by loosely arranged striated muscle fibers and bundles, interspersed with plump mesenchymal cells that exhibit some variability in nuclear size and shape. However, mitotic activity was rare, and necrosis was absent. Cellular differentiation at the periphery of the tumor made distinction from surrounding muscle difficult. A comparison of immunohismchemical staining patterns between tumor muscle fibers and adjacent normal muscle was made with the use of myoglobin, desmin, and vimentin. Although the fetal rhabdoyoma bears little resemblance to the adult form, its similarity to the embryonal rhabdomyosarcoma makes the recognition of this rare tumor important,

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PYOSTOMATITISVEGETANSANDORALCROHN’S DISEASE: AN UNUSUAL CLINICAL ASSOCIATION

Ficarra G, Cicchi P, Amorosi A, Piluso S. University of Florence and USL 10/D, Florence, Italy. Pyostomatitis vegetans (PV) is a rare eruption of the oral cavity characterized by tiny yellow pustules coating the surface of the friable eroded mucosa. It is considered a marker for inflammatory bowel disease. Oral features of Crohn’s disease (CD) include ulcers, lip fissuring, cobblestone plaques, angular cheilitis, polypoid tag lesions, and perioral erythema. We describe a 45-year-old woman with a 6-month history of painful sores in her mouth, diarrhea, weight loss, and cutaneous lesions. Oral examination showed cobblestones plaques on the tongue and friable vegetating pustules on the labial commissures. Staphylococcus epidermis was isolated from the pustules. Laboratory studies revealed eosinophilia, and low hemoglobin, and zinc level (40 pg/dL). Histologic study of the labial lesions revealed hyperplastic epithelium with intraepithelial clefts containing eosinophils, and neutrophils. Tongue lesions showed chronic inflammation with noncaseating granulomas. Colonoscopy and biopsy demonstrated intestinal CD. P\J lesions regressed after oral zinc supplementation. Prednisone treatment (60 mg/day) resulted in healing of the tongue lesions. In our patient, PV appeared to be related to zinc deficiency caused by malabsorption. The pathogenetic relationship between PV and CD will be discussed.

INTRALESIONAL VINBLASTINE TREATMENT OF INTRAORAL KAPOSI’S SARCOMA IN AIDS PATIENTS. Nichols CM, Flaitz CM, Hicks MJ. Bering

Dental Clinic, University of Texas Health Science Center Dental Branch, and Baylor College of Medicine, Houston. Kaposi’s sarcoma (KS) is the most frequent malignant neoplasm in persons with AIDS, occurring in 15% to 20% of all AIDS patients and in >30% of homosexual, HIV-infected men. The purpose of this study was to evaluate the effects of intralesional vinblastine administration on intraoral KS in homosexual males with AIDS. Eighty-two KS lesions in 24 persons were treated. Most frequent lesion sites were: hard palate (35%), soft palate (17%), maxillary facial (12%) and mandibular facial (10%) gingivae, and maxillary tuberosity (6%). Lesion area ranged from 0.1 to 35 cm2 (mean 4.2 cm2). Complete clinical resolution was found in 69.6% of lesions up to 56 weeks after the last vinblastine treatment (mean, 15.2 weeks). Partial response (66% reduction in mean lesion area) occurred with the remaining 30.4% of lesions. Intralesional vinblastine dosage administered was 0.1 mg/cm2, with a mean of 2.4 treatments (range 1 to 6). The overall recurrence rate (RR) was 37.8%, with a mean recurrence time of 15.6 weeks after the last vinblastine treatment. Recurrence rates varied with clinical appearance. Recurrence rates by lesion type were: 60% for purple mixed macular/ nodular lesions; 40% for nodular lesions; 39% for purple macular lesions; 9% for red macular lesions, and 0% for red mixed macular/nodular lesions. Sixty-five percent of recurrences coincided with discontinuing AZT. Postoperative complications included:

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November pain (83%), mucosal ulceration (21%), and transient paresthesia (17%). Intralesional vinblastine administration results in clinical resolution in a substantial number of intraoral Kaposi’s sarcoma lesions. The clinical appearance of the lesion and changes in overall management of HIV infection may be prognostic factors in predicting recurrences. Monitoring for possible retreatment of recurrent lesions and treatment of new lesions is necessary.

SMOKELESS-TOBACCO EXT REPLICATION OF HERPES SI 1 (HSV-1) IN CULTURED HU LlAL CELLS Murrah VA, Gilchrist EP, Moyer MP. Departments of Pathology and Surgery, The University of Texas Health Science Center at San Antonio. The high prevalence of both tobacco use within the population and the number of persons carrying latent herpes simplex virus type 1 (HSV-1) suggests the opportunity for synergism between these agents as cocarcinogens. A prerequisite for HSV-mediated cell transformation appears to be prevention of host cell lysis. Smokeless-tobacco extracts have recently been shown to inhibit HSVmediated lysis in nonhuman cell lines. In this study, cultured human oral epithelial cells were infected with wild type HSV-1 pretreated with and exposed periodically to both 2% extracts of loose leaf, moist, and dry snuffs and to purified tobacco-specific carcinogens; 4-(N-methyl-N-nitrosamino)-l-(3-pyridinyl)-l-butanone [NNK] at 1.0 rig/ml, nitrosonornicotine [NNN] at 1.0 rig/ml, and benzo (a) pyrene [B(a)P] at 0.25 rig/ml. Parameters measured included onset and time course of cytopathic effects, percentage of cultures undergoing active virus production, and concentration of virus released into the media over time. Results showed both inhibition of HSV-mediated cell lysis and inhibition of viral replication by all types of tobacco extract. Tobacco-specific carcinogens, NNK, NNN and B(a)P did not significantly alter the natural history of HSV-I infection in the concentrations used. In summary, this is the first time that both inhibition of lyric effects of HSV-1 infection and inhibition of virus replication by tobacco extracts have been demonstrated in human oral epithelial cells. This is also the first time that such an effect has been demonstrated using the wild type virus in the absence of prior attenuation procedures, such as UV-irradiation or heat inactivation. This work therefore contributes significant evidence to support the hypothesis that HSV-1 and smokeless tobacco have the capability of acting as synergistic agents in the development of oral cancer. This work was supported by an American Cancer Society Clinical Oncology Career Development Award, an ACS Institutional Cancer Research Grant and a grant from the Smokeless Tobacco Research Council.

IMAGE AND FLOW CYTOMETRIC DNA ANALYSlS OF PRIMARY AND RECURRENT AMELOBLASTOMA AND AMELOBLASTIC CARCINOMA OF THE JAWS. Muller S, Cohen C, De ose P, Waldron C. Emory University, Atlanta, Ga. Ameloblastomas, the second most common odontogenic tumor, behave as benign locally aggressive tumors. Ameloblastic carcinomas, on the other hand, show histologic features of malignancy and

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1992

may metastasize. By image cytometric analysis of Feulgen-stained sections from decalcified, formalin-fixed paraffin-embedded tumors; we have compared nuclear DNA content of 22 ameloblastomas (17 primary lesions and 5 recurrent cases) and 5 ameloblastic carcinomas. At least 200 nuclei from each case were selected and analyzed by a pathologist with the use of the CAS 200 system (Cell Analysis Systems, Elmhurst, Ill.). Of the 22 ameloblastomas, 17 (77%) were diploid (< 2SN) whereas 5 were hyperdiploid (> 2.5N). No significant differences in ploidy were identified between primary and recurrent ameloblastomas or among plexiform, follicular, and acanthomatous types of ameloblastoma. One plexiform unicystic and one granular cell ameloblastoma were diploid and aneuploid respectively. Of the five ameloblastic carcinomas, four (80%) were aneuploid (two hyperdiploid and two multiploid); one (20%) was diploid. Among the ameloblastic carcinomas, ploidy was not significantly correlated with the incidence of metastasis. Flow cytometry, with the use of a modified Hedley’s technique, was performed on three carcinomas and 100% concordance between image and flow cytometric data was seen. Although aneuploidy is significantly more common in ameloblastic carcinomas than ameloblastoma (<.OOl), malignant potential does not a!ways correlate with DNA aneuploidy.

COMPARISON OF CYTOPATH HAIRY LEUKOPLAKIA WITH IN TION FOR EPSTEIN-BARR VIRUS. Greenspan JS, Regezi JA, Greenspan D, De Souza YG, Daniels TE. University of California San Francisco. It has been recently observed that the cytopathic changes associated with hairy leukoplakia (HL) correlate strongly with ultrastructural evidence of intra-keratinocyte herpes-type viral particles. Because in situ hybridization is considered to be the most accurate method for confirmation of Epstein-Barr virus (EBV) induced HL, this investigation evaluates the compatibility of histopathologic findings, which many believe to be diagnostic, with in situ hybridization results in 60 patients with clinically suggestive HL. Hematoxylin and eosin-stained (H and E) sections were seviewed independently by three oral pathologists without knowledge of in situ results. The presence of keratinocyte nuclear inclusions or homogenization, believed to be specific for EBV in these lesions, were used as indicators of infection. Cytoplasmic changes were evaluated separately. Results showed that with in situ hybridization, 48 cases were positive and 12 were negative. When the two methods were compared, pathologist concurrence ranged from 8 3% to 92%. False negatives ranged from 6% to 19%, and false positives ranged from 8% to 25%. Cytoplasmic ballooning, homogenization, and perinuclear clearing were evident in all cases of in situ confirmed HL; however, these changes were also noted in 75% (9,’ 12) of the in situ negative cases. Most cases of HL exhibited botb nuclear homogenization and inclusions, though the former was more constant. Nuclear chromatin was typically marginated along the nuclear membrane. Nuclear changes were evident in mid- and upper-level keratinocytes. Frequently, these alterations were subtle and evident in few cells. It is concluded that there is generally good correlation between nuclear morphology and in situ hybridization for EBV. In the appropriate clinical setting, the finding of

Volume Number

Abstracts

74 5

nuclear inclusions or homogenization may be diagnostic. However, because of false negatives and false positives, H and E cytopathology should not be considered a substitute for in situ hybridization. Supported by POl-DE07956

AMPLIFICATION OF c-erbB-2 ONCOGENE IS ASSOCIATED WITH MALIGNANTCHANGES IN PLEOMORPHIC ADENOMAS. Vigneswaran N, Cohen C, Miiller S, DeRose P. Emory University, Atlanta, Ga. The biologic behavior of malignant salivary gland neoplasms are sometimes difficult to predict by their histopathologic features alone. The oncoprotein c-erbB-2 (c-erbB-2) sharing close similarity with epidermal growth factor receptor, is frequently expressed in adenocarcinomas and correlates with poor prognosis in breast cancer. Cathepsin-D (Cath-D) is a lysosomal enzyme and it& presence in tumors is related to their proliferative and/or infiltrative nature. CA 15-3 is a high mw. mucin-like glycoprotein produced by mammary carcinomas. Anti-microbial substances like lactoferrin (Lf) and lysozyme (Ly) are synthesized by normal salivary gland and their expression in salivary gland tumors is reported to reflect their well-differentiated nature with better prognosis. Six pleomorphic adenomas (PA), 3 carcinomas ex-PA (CAEX-PA), 5 adenoid cystic carcinomas (ACC), 3 acinic cell carcinomas (AC), 6 mucoepidermoid carcinomas (MEC), 3 polymorphous low-grade adenocarcinomas (PLAC), and 2 adenoca.rcinomas not otherwise specified (AC-NOS) were immunostained with antibodies (AB) to c-erbB-2 (mono- and polyclonal) Cath-D, CA 15-3, Lf and Ly. Strong membranous staining for c-erbB-2 was seen in malignant cells of CAEX-PA with both monoclonal and polyclonal ABs whereas normal salivary gland tissue and all other tumors were immunonegative. Infiltrating cells of all malignant tumors demonstrated strong cytoplasmic: positivity for Cath-D except for one PLAC and a low grade MEC. Focal and weak reactivity for Cath-D was also noted in ductal cells of normal salivary gland and PA. Strong reactivity for CA 15-3 was observed in all cases of ACC, AC, 5/6 MEC, AC-NOS, CAEX-PA, whereas all three PLAC remained negative. Weak lunninal staining for CA 15-3 was found in

605

The study of tumor markers is one major emphasis in the study of carcinogenesis. Placental form of glutathione S-transferase (GST-P) is one of the most reliable tumor markers in hepatocarcinogenesis. This study was designed to study the usefulness of GST-P as a tumor marker for oral carcinogenesis. The expression of GST-P with the use of antirat liver GST-P antibody was investigated in hamster buccal pouch mucosa treated with 0.5% dimethylbenz(a)anthracene (DMBA) biweekly for 12 weeks. This study for the first time showed that the antirat liver GST-P antibody is applicable to the HBPM model and that DI#BA treatment induced GST-P positive foci. These foci were randomly distributed and frequently involved the hyperplastic and dysplastic segments of the epithelium, as well as tumors. The control pouches, untreated or treated with mineral oil biweekly, showed no GST-P positive foci. The results showed that GST-P may be a useful tumor marker for oral carcinogenesis. Further study is needed to explore the kinetics of these GST-P positive foci and the value of GST-P as a tumor marker in oral carcinogenesis.

BIOENERGETIC AND THIOL CHARACTERIZATION OF HUMAN MICROVASCULAR ENDOTHELIAL CELLS (MVEs) Mallery SR, Lantry LE, Stephens RE. Ohio State University, Columbus, Ohio. The regulation of angiogenesis, a process that involves the microvasculature, is integral in both physiologic (e.g., fetal development) and pathologic (e.g., tumor angiogenesis) processes. Previous studies from our laboratory have shown an association between cellular glutathione (GSH) status, redox state NAD(P)H/ NAD(P)+, and cell cycle kinetics. The cytoprotective role of GSH was also of interest, because the microvasculature is the primary site for cellular adhesions and blood-tissue exchange. The purpose of this study was to isolate, culture, and initiate biochemical characterization of human MVEs. The choriocapillaris was selected as the tissue primary site because of its unique anatomy, which permits separation of the microvascular bed. GSH levels were determined by a kinetic, dual beam spectrophotometric assay. Significantly higher levels of GSH were present in MVEs relative to hu-

ductal cells of normal salivary gland and PA. Focal expression of both Lf and Ly was detected in both PA and salivary gland carcinomas except for PLAC. Malignant change in PA is associated with the amplification of c-erbB-2 oncogene; this gene does not appear to play any significant role in the evolution of other malignant salivary gland tumors. High grade malignant salivary gland neoplasms overexpress Cath-D and CA 15-3. Reactive patterns of Ly and Ly in salivary gland tumors do not correlate with their biologic behavior. Thus detection of c-erbB-2, Cath-D and Ca 15-3 in salivary gland tumors may be useful in predicting their malignant nature and prognosis, in the absence of recognizable histopathologic patterns.

man keratinocytes (HK), (GSH) as nmol/mg protein, MVEs = 60.8 +3.6,x1 = 6,HK = 33.0 + 6.2,n = lO.HPLCbioenergetic profiles demonstrated that relative to HK, MVEs contain appreciably greater levels of high energy phosphates (ATP, GTP, UTP), as well as higher levels of reducing equivalents (NADH&NADPH). Because the in vivo MVE environment may be rich in reactive oxygen species, because of phagocytic cell interactions, the cytoprotective role of GSH peroxidase may be beneficial to cell survival. Therefore the biochemical characteristics of MVEs may reflect cellular adaptations that have developed as a result of the unique environment and function of the microvasculature.

DEVELOPMENT OF PLACENTAL GLUTATHIONE S-TRANSFERASE (GST-P) STAINED FOCI DURING HAMSTER BUCCAL POUCH MUCOSA CARCINOGENESIS. Zhang L, Mock D, Cameron R. University

KERATIN PROTEIN PROFILE IN PREMALIGNANT LESIONS AND SQUAMOUS CELL CARCINOMA OF THE HUMAN ORAL MUCOSA. Lin LM, Tsang SH, Chen YK. Kaohsiung Medical College, Department of Oral Pathology, Kaohsiung, Taiwan, Republic of

of British Columbia, Vancouver, and University Toronto, Toronto, Ontario.

of

China.

606

Abstracts

ORALSU~O

O~L~LIMED

O~~LPATHO~

November The aim of the present study is to compare the keratin pattern of the premalignant and malignant oral Lesions with normal oral mucosa by electrophoretic (SDS-PAGE) technique and immunohistochemical (PAP) method with the use of AEl and AE3 antibodies. Biopsy specimens including 21 leukoplakia with and without epithelial dysplasia; 14 verrucous hyperplasia (VH), 9 verrucous carcinoma (VC), 26 squamous cell carcinoma (SCC) (18WD, 7MD, 2PD) were used. We have seen a whole epithelial staining of AEl and suprabasal staining of AE3 in normal oral epithelium. Compared with normal oral mucosa, AEl and AE3 staining pattern were almost preserved in lesions of leukoplakia without dysplasia and VH. Absence of AEl staining in the basal layer was noted in leukoplakia with dysplasia suggesting some relationship with the malignant potential. Though epithelial dysplasia is accepted to precede carcinoma, not all dysplastic lesions progress to become carcinoma when left untreated. Also, carcinoma may occur in lesions without epithelial dysplasia. Hence, lack of AEl staining in basal cells in some lesions of leukoplakia without dysplasia may imply a malignant potential. There seemed to a progressive loss of AEl and AE3 immunostained keratin transition from well to poorly differentiated SCC. The basic pattern of keratin separated by SDS-PAGE, typical for normal nucosa, was still recognizable in leukoplakia and VH. Increasing frequency of LMW keratin expression and loss of normally expressed keratin were occurred in both VC and SCC. This altered keratin expression was not associated with the severity of gross or microscopic features observed in tumor tissues. Keratin pearls formation and individual cell keratinization, histopathologically noted in WD SCC did not parallel the altered expression of HMW keratin. This may be because the aberrant expression of HMW keratin and loss of normally expressed keratin are related to changes of corresponding mRNA or message translation. Thus changes in HMW keratin may not consistently be reflected with the state of differentiation observed at microscopic level. Therefore further study of the roles of mRNA plays is warranted.

THE RELATIONSHIP OF FLOW CYTOMETRIC ANALYSIS AND NUCLEOLAR QRGANIZER REGION ENUMERATION IN ARCHIVAL ORAL PREMALIGNANT LESIONS. Kahn M, Mincer H, Dockter M, Hermann J. University of Tennessee, Memphis. Studies of benign and malignant tumors using either flow cytometric analysis (FCA) or nucleolar organizer region-associated proteins stained by an argyrophil method (AgNOR) have found that either technique can demonstrate the increased cellular proliferation observed in malignant tumors. In FCA this is reflected in increased DNA index (DI), S-phase (S), and total proliferative index (TPI) whereas in AgNOR an increase in mean number of NORs (MNOR) has been observed. Although both have been used individually in assessing the histopathologic nature of various human tumors, very few studies have investigated the relationship between the two techniques in a single series. Therefore we examined a series of 36 formalin-fixed paraffin-embedded tissues of the oral mucosa (five controls, four epithelial hyperplasias, nine mild dysplasias, nine moderate dysplasias, six severe dysplasias, and three squamous cell carcinomas) to assess if AgNOR aided in the

1992

subjective light microscopic diagnosis (LMD) of oral mucosal dysplasias and if the parameters recorded for the specimens (age, race, gender, site, LMD, DI, TPI, S, and MNOR) exhibited any positive correlation. FCA was performed based on a modified procedure of Hedley, et al. whereas AgNOR was accomplished by the silver nitrate colloid staining method of Cracker, et al. A positive correlation was observed between DI and MNOR (r = 0.434;~ < 0.012), and between TPI and S (r = 0.774;~ < 0.0001) holding gender and race constant. Additionally, the MNOR increased linearly with increasing grade of dysplasia with one exception. The results indicated that there is positive correlation between FCA and AgNOR and that AgNOR may be a diagnostic aid in grading oral mucosal dysplasias.

COMPARATIVE EFFECTS OF TPA HAMSTER CHEEK POUCH MUCOSA. Singh B, Hall

J, McCoy B. Medical Ga.

College of Georgia, Augusta,

We have previously reported the effects of tetradecanoyl-phorbol-13-acetate (TPA) and sn-1.2-dioctanoylglycerol (DiC8) on hamster cheek epithelium implicating the role of protein kinase C (PKC) in cell proliferation. The objective of this investigation was to compare the effects of the tumor promoter TPA with its nonpromoter congener 12-deoxyphorbol-13-0-phenylacetate (DOPP) using anti-inflammatory agent fluocinolone acetonide (FA). For this purpose the animals were divided into five groups (5 animals/ group) and treated as follows: Group I, 5 pg TPA; Group Ii, TPA + 5 pg FA; Group HI, 5 pg DOPP; Group IV, DOPP + 5 pg FA; and Group V, 0.25 ml acetone that served as a control. The animals were sacrificed 72 hours after injection with vinblastine sulfate to monitor mitosis. The tissues were fixed in neutral buffered formalin, processed, and stained with hematoxylin and eosin. The epithelial mitotis (MI) were reflected per 1 mm of the basement membrane. As compared with the acetone controls (MI 1 .I + O.l), both TPA and DOPP significantly enhanced epithelial mitosis activity (MI 4.6 rf- 0.4 and 3.9 I 0.3 respectively). FA markedly reduced epithelial mitosis as well as inflammation and hyperplasia induced by these agents. In conclusion, these data show that DOPP has some selective activities in common with TPA. Because DOPP only partially activates PKC in heterogenous mixtures it is plausible that other parameters such as PKC isozymes and induction of dark (stem?) cells might be involved for complete tumor promotion.

IMMUNOCYTOCHEMICAL ANALYSIS QF GALACTOSYLTRANSFERASE IN ORAL SQUAMOUS CELL CARCINOMAS. Tajima Y, Yokose S, Utsumi N.

Meikai pan.

University School of Dentistry, Saitama, Ja-

The purpose of this study was to evaluate the histochemical and cytochemical expression of galactosyltransferase (GT) in oral squamous cell carcinomas (SCCa). Fifteen cases of intraoral SCCa (6 tongue; 4 gingiva; 4 floor of the mouth; 1 buccal mucosa) were examined. The tissues for light microscopic analysis were prepared from formalin-fixed and paraffin-embedded materials. For electron

Volume Number

Abstracts

74 5

microscopy, tissue samples from one floor of the mouth case and one tongue case were fixed in 2%~ glutaraldehyde-2% paraformaldehyde in phosphate-buffered saline (pH 7.2) and embedded in Lowicryl K4M. For detection of the enzyme protein, murine monoclonal antibody against human GT (supplied by Dr. S. Nozawa, Keio University, Japan) was used at dilutions of 1:3OOO, 1:4000, and 1:5000. For the immunostaining, the ABC and protein A-gold methods were applied for light and electron microscopy, respectively. GT displayed a distinct distribution in the carcinoma cells infiltrating into the neighboring tissue. The immunoreactivity was seen intracellularly as fine granular forms, and was demonstrated in seven of the fifteen cases examined. Immunoelectronmicroscopy revealed a preferential labeling of the gold particles in the trans-Golgi complexes. No significant reaction was detected in the plasma membrane. The results provide evidence that the squamous carcinoma cells present a highly promoted glycosylation pattern than that seen in their normal counterpart.

UPREGULATION OF EPITHELIAL IN INFLAMMATORY DISEASES COSA. Eversole LR, Christensen

University of California

CALPROTECTIN OF ORAL MUR, Miyasaki K.

Los Angeles.

Leukocyte calprotectin is an extragranular cytosolic calcium binding protein, belonging lo the S-100 family. It consists of alpha and beta polypeptide chains that exist as homo and heterodimers. Biologically, calprotectin exhibits profound antibacterial and antifungal effects even from necrocytes. Monoclonal Ab MAC 387 (Dakopatts) reacts with the heterodimer and has been used as a leukocyte marker. This antibody has also been found to react with keratinocytes from normal oral epithelium, yet is only expressed in skin when inflammation is present. Epithelial and leukocyte calprotectins show homology by isoelectric focusing and on immunoblots. In this investigation, we evaluated for the presence of keratinocyte calprotectin by immunoperoxidase and immunotluorescence methods with mAb MAC 387 in inflammatory processes of oral mucosa in which both immunopathologic and infectious mechanisms were extant. Regardless of the disease mechanism, epithelial calprotectin is significantly upregulated in inflamed mucosa. This molecule may provide a biochemical antimicrobial adjunctive function to the phyysical barrier of oral epithelium.

AMYLASE EXPRESSION IN ACINIC CELL CARCINOMA: COMPARISON WITH PAROTID TUMORS IN v-Ha-ras TRANSGENIC MICE AS A MODEL FOR HUMAN ACINIC CELL CARCINOMA. Dardick I, Burford-Mason A, Mackay A. Departments of Pathology and Otolaryngology, University of Toronto

and Conacher Head and Neck Cancer Research Unit, The Toronto Hospital, Toronto, Ontario, Canada. Animal models are an essential requirement for investigating human salivary gland tumors (HSGT). Male transgenic mice with the mouse mammary tumor virus (mMTV)/v-Ha-ras transgene develop parotid tumors. Histologically these are adenocarcinomas that can show cytoplasmic secretory granules and ultrastructurally have the rough endoplasmic reticulum and zymogen-type granules

607

characteristic of acinar cell differentiation. These parotid tumors appear to closely resemble human acinic cell carcinomas (ACCs). To assess whether amylase expression might serve as an another marker for comparison with human ACC, immunocytochemistry for amylase was performed on 22 human ACCs and compared with 14 ACCs occurring in seven male mMTV/v-Ha-ras mice. Whether in human or mouse ACCs, amylase was always focal and limited to 10% or less of the tumor cells. Amylase was detected in only 18% and 29% of human and mouse tumors, respectively. Zymogen-type granules were detected ultrastructurally in both systems but may not contain amylase because staining of mouse parotid glands with tumor rnicrofoci suggests repression of amylase expression is an early event. Mouse AACs, the first in vivo animal model for a HSGT, parallel the human neoplasm in amylase expression and this emphasizes the suitability of this animal model for studying the developmental

and biologic

processes in the human

ACCs.

RELATIONSHIP BETWEEN HYPHAL CONCENTRATION AND RAT TONGUE LESION INDUCTION. Copete M, Allen C, Bradway S. The Ohio State Univer-

sity, Columbus,

Ohio.

In the rat model of oral candidiasis, several strains of C. albicans exhibit hyphal penetration of the keratin layer of dorsal tongue mucosa, but only some strains appear to be able to induce the inflammatory infiltrate and proliferative epithelial changes associated with lesion formation. Some investigators have proposed that lesion formation in human beings is dependent on a threshold effect, that is, colonization by a minimal concentration of hyphae. Others feel that only certain C. albicuns isolates are capable of lesion induction, perhaps because of stimulation of the skin-associated lymphoid tissue with strain-specific antigens. The purpose of this study was to determine the anatomic location and concentration (hyphae/linear mm) of hyphae in dorsal tongue mucosa of rats inoculated either with candidal strains that produce lesions (Group I) or strains that penetrate keratin, but do not produce lesions (Group II). Paraffin-embedded tongues from rats, inoculated with the same number of either Group I (n = 16) or Group II (n = 11) organisms for 20 weeks, were step-serial sectioned and stained with periodic acid-Schiff stain. The number of hyphae/linear mm involving the posterior one third of the dorsal tongue mucosa were counted by light microscopy using an ocular micrometer. No attempt was made to delineate between the hyphae of individual organisms; rather, the number of hyphae in a given linear unit of tissue was used to provide a relative indication of the “hyphal mass” present in the mucosal tissue. Preliminary results showed that C. ulbicuns in Group I was confined to the dorsal tongue lesions that primarily affected the giant conical papillae. Group II exhibited no clinically or histologically demonstrable dorsal tongue lesions and exhibited a slightly more diffuse distribution of hyphae with concentrations of organisms just posterior to the giant conical papillae. There appeared to be no significant difference in hyphal concentration between Group I and Group II dorsal tongue mucosa. These preliminary observations suggest that hyphal concentration is probably not the primary etiologic determinant in dorsal tongue lesion induction in the rat.

60% Abstracts

ORALSURGORAL

MEDORAL

November

QOUBLE IMMUNOSTAINING OF MINOR SALlVARY GLAND NEOPLASMS. Vincent SD, Hammond HL, Roegler C. University of Iowa, Iowa City, Iowa. Some benign and malignant salivary gland neoplasms are composed of both epithelial and myoepithelial cells. Past efforts to evaluate for both cell types have included light and electron microscopy as well as immunologic staining techniques. We have used a dual immunologic staining technique that provides distinctive light microscopic marking of two different surface antigens on one surgical section. To mark myoepithelial cells, we used alpha smooth muscle actin (Biogenex). For epithelial cells we used high or low molecular weight cytokeratin (Biogenex). Internal and external controls consisted of normal surface ora! mucosa and minor salivary glands. The procedure involves the initial application of primary monoclonal mouse anti-ASMA, a biotintylated link, streptavidin label and diaminibenzidine/chromogen solution. Remaining uncovered primary anti-ASMA antibodies are blocked. The second primary antibody, mouse monoclonal anticytokeratin is then applied, followed by linkage, streptavidin conjugated alkaline phosphatase label, and a naphthol phosphate/fast red substrate/chromogen solution. Internal staining controls consisted of normal salivary gland acini, blood vessels, and surface stratified squamous epithelium. Evaluation of benign and malignant minor salivary gland neoplasms revealed evidence of both epithelial and myoepithelial cell surface antigens in varying proportions with the possible exception of canalicular and trabecular adenomas and some mucoepidermoid carcinomas that showed little evidence of myoepithelial surface antigens. This staining technique can be used for other neoplasms when evaluation of several cellular antigens on a single section can be of diagnostic value.

HISTOPATHOLOGIC EVALUATION OF PERSISTENT HIV-ASSOCIATED INTRAORAL ULCERATIONS WITH CLINICAL CORRELATION. Flaitz CM, Nichols CM, Hicks MJ. Bering Dental Clinic, University of Texas Health Science Center Dental Branch, and Baylor College of Medicine, Houston. Intraoral ulcerations present a diagnostic challenge in HIV-infected persons because of the multiplicity of etiologic factors. The purpose of this study was to describe clinical, histopathologic, and immunocytochemical characteristics of persistent, intraoral ulcerations in HIV-infected persons. Thirteen homosexual men and 1 IVDA woman (mean, CD4 = 23) with an average of 2.5 ulcers/ patient at initial exam were included in the study. After therapy with antiviral agents or corticosteroids, a single persistent ulcer (mean duration, 4 weeks) from each patient was biopsied. Lesion area ranged from 0.4 to 13.5 cm2 (mean, 2.9 cm2). Most frequent sites were buccal and labial mucosae and tonsillar pillar region. At time of biopsy, 65% of HIV-infected patients were receiving antiretroviral agents, and 57% were receiving acyclovir. Tissue sections were stained with H&E, PAP, PAS, GMS and AFB. Immunocytochemical stains for CMV and HSV were performed. The lesions were compared with 14 consecutive, nonspecific intraoral ulcers from our biopsy service. With HIV-associated ulcers, 42.8% of lesions were due to combined CMV and HSV infection. CMV

P.4ThoL

1992

infection, alone, was found in 21.8% of lesions. Aphtbous-like ?esions accounted for 35.7% of lesions. Candidal organisms were associated with 60% of aphthous-like ulcers. HSV was associated with CMV infection; however, no cases of HSV infection alone were identified. With nonspecific control ulcers, no infectious agents were identified. PAP correlated well with immunocytochemical detection of CMV and may be useful for screening purposes in suspected CMV infections. A substantial proportion of persistent, HIV-associated ulcers are due to either CMV infection alone, or in combination with HSV, which respond to ganciclovir therapy.

SINUSITIS. Warnock 6, Watkins G, Bianchi D. Naval Dental School, National Naval Medical Center, Bethesda, Md. ALLERGIC

FUNGAL

Although sinusitis is a common disorder, fungal sinusitis has previously been regarded as rare, particularly in immunocompetent patients. Increased recognition of allergic fungal sinusitis over the past decade reflects growing awareness of this entity both clinically and histologically. Originally described as aspergillus sinusitis, the majority of cases are now attributed to Bipolaris (formerly classified as Drechslera), a Dematiaceous fungi. Patients are usually immunocompetent with persistent unilateral or pansinusitis, nasal polyposis, and sinus opacification that are refractory to medical management. Bony expansion, occasionally with erosion, is common, suggesting a neoplastic process. The histologic hallmark is the presence of “allergic mucin” containing abundant aggregations of eosinophils and CharcotLeyden crystals. Fungal elements are minimal, usually seen in the mutinous pools and seldom show frank tissue invasion. The paucity of fungi requires diligent searching and special fungal stains. Presently treatment is surgical debridement with systemic and or topical nasal steroid therapy. Four cases of allergic sinusitis are presented, emphasizing: histologic features,

clinical features, pertinent laboratory tests, and controversies in treatment modalities.

ASSOCIATION BETWEEN PERIODONTAL EASE SEVERITY AND LOW HUMAN BI WEIGHT. Boyd DL, Collins JG, Arnold RR, Offenbather S. University of North CaroEina, Chapel Hill, N. C. Certain gram-negative infections have been clearly associated with an increased risk for low birth weight (LBW) in humans and in animal models. These infections do not invade the fetal-placenta unit but are believed to elicit a maternal cytokine response that indirectly has deleterious effects on fetal growth Previous studies in our laboratory have demonstrated that either an intravenous challenge with lipopolysaccharide (LPS) isolated from P. gingivalis oi a subcutaneous infection with P. gingivalis in a chamber model can markedly retard fetal growth in the pregnant hamster. These data lead us to test the hypothesis that periodontal disease may be a risk factor for human LBW. We conducted a case/control cohort study of 24 pairs of mothers in which a mother with a LBW newborn (case, < 2.0 kg) was matched to a normal birth weight (NBW) mother (control, > 2.0 kg). Matching was performed on 16 covariables that are known to influence pregnancy outcome, including

Volume Number

Abstracts

74 5

age, race, tobacco, alcohol and drug usage, as well as nutritional and socioeconomic status. All cases and controls had moderate to severe periodontitis with an overall mean attachment loss (ALOSS) of 3.05 mm/site, with no significant differences in mean ALOSS comparing cases to controls. Within the NBW patients there was no relationship between the newborn birth weight and the mean maternal ALOSS. However, within the LBW patients there was a significantly positive correlation between maternal mean ALOSS and decreasing birth weight at p = 0.016. In this group, an increased mean severity of 0.5 mm/site resulted in a decreased birth weight of 635 gm. Although the increased risk for LBW if periodontitis is present or absent cannot be estimated from this periodontally diseased population, these pilot data clearly demonstrate that severe periodontal disease may be a risk factor for LBW in certain susceptible patients. Supported by NICHD grant #ROlHD26652.

THE CYTOBRUSH ORAL CYTOLOGY.

PLUS CELL

COLLECTOR

IN

Jones A, Pink F, Sandow P, Baughman R, Migliolrati C, Stewart C. University of Florida

College

of Dentistry,

Gainesville.

The Cytobrush tongue depressor

609

Plus Cell Collector was compared with a wooden during oral cytologic smears. Factors evaluated

were degree of patient discomfort, clinician ease of use, and ability of pathologist to make an accurate cytopathologic diagnosis. Fifty patients were randomly divided into five groups (vesiculo-bullous/ ulcerative, erythroplakia, candidiasis, squamous cell carcinoma, normal controls). One cytologic smear was performed on every patient for both the cytobrush and tongue depressor. In questionable cases, an incisional biopsy was performed to establish a definitive diagnosis. Two-factor ANOVA analysis with Tukey HSD multiple comparison revealed a significant difference in pain between controls and pathology cases (p = O.Ol), no significant (N.S.) difference in pain between devise used (p = 0.72), and N.S. interaction was shown (p = 0.83). Nonparametric Kruskal-Wallis analysis revealed the cytobrush was significantly more convenient and provided a thinner cytologic smear. The same method revealed no significance difference in diagnosis ability, in distribution of cells, and in quantity of cells, with both methods proving adequate. In support of our hypothesis, the cytobrush elicited no significant difference in pain, was more convenient to the clinician, and allowed an accurate cytopathologic diagnosis when compared with the wooden tongue depressor.

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