Programmable bio-nanochip-based cytologic testing of oral potentially malignant disorders in Fanconi anemia.

Fanconi anemia (FA) is caused by mutations of DNA repair genes. The risk of oral squamous cell carcinoma (OSCC) among FA patients is 800-folds higher than in the general population. Early detection of OSCC, preferably at it precursor stage, is critical in FA patients to improve their survival. In an ongoing clinical trial, we are evaluating the effectiveness of the programmable bio-nanochip (p-BNC)-based oral cytology test in diagnosing oral potentially malignant disorders (OPMD) in non-FA patients. We used this test to compare cytomorphometric and molecular biomarkers in OSCC cell lines derived from FA and non-FA patients to brush biopsy samples of a FA patient with OPMD and normal mucosa of healthy volunteers. Our data showed that expression patterns of molecular biomarkers were not notably different between sporadic and FA-OSCC cell lines. The p-BNC assay revealed significant differences in cytometric parameters and biomarker MCM2 expression between cytobrush samples of the FA patient and cytobrush samples of normal oral mucosa obtained from healthy volunteers. Microscopic examination of the FA patient's OPMD confirmed the presence of dysplasia. Our pilot data suggests that the p-BNC brush biopsy test recognized dysplastic oral epithelial cells in a brush biopsy sample of a FA patient.

A retrospective study of Enterococcus faecalis infective endocarditis: comparison of clinical characteristics and outcomes associated with treatment.

Introduction: A synergistic antibiotic combination of a penicillin and gentamicin (AG) or ceftriaxone (AC) is used in the management of Enterococcus faecalis infective endocarditis (EFIE). We compare the treatment outcomes between AG and AC, including low and high dose ceftriaxone (1 and 2 g 12 hourly). Methods: A retrospective cohort study of patients treated for EFIE at single tertiary centre (2012-2019). Outcome measures examined were 90- and 180-day mortality, treatment associated adverse events and relapse of bacteraemia (within 1 year). Results: 39 patients were enrolled [61.6% given (AC) (n = 24), 24% received ACL (n = 10) and 34% received ACN (n = 14)], 38.4% received AG (n = 15). We noted a difference in the mortality outcomes at 90 and 180 days between those treated with AG and AC overall (6.7% and 33.3%, respectively) although this did not reach statistical significance (P = 0.114, P = 0.061). No significant difference was noted between these groups in incidence of relapsed bacteraemia with two cases noted in the AC cohort (8.3%, 2/24) and none observed (0/15) in the AG cohort (P = 0.662, P = 0.414). A greater number of adverse events was observed in the AG group (11/15, 73.3%) compared to the overall AC group (6/24, 25.0%) (P = 0.009), with no difference between the high and low dose ceftriaxone groups (P = 0.05). Conclusion: Combination treatment of EFIE with AC is associated with a reduced number of adverse events in comparison to AG groups. Although increased mortality was observed in the AC group, this did not reach statistical significance, and reflects the greater comorbidities and reduced capacity for surgical source control in this cohort.

Nuclear F-actin Cytology in Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma.

Oral cavity cancer has a low 5-y survival rate, but outcomes improve when the disease is detected early. Cytology is a less invasive method to assess oral potentially malignant disorders relative to the gold-standard scalpel biopsy and histopathology. In this report, we aimed to determine the utility of cytological signatures, including nuclear F-actin cell phenotypes, for classifying the entire spectrum of oral epithelial dysplasia and oral squamous cell carcinoma. We enrolled subjects with oral potentially malignant disorders, subjects with previously diagnosed malignant lesions, and healthy volunteers without lesions and obtained brush cytology specimens and matched scalpel biopsies from 486 subjects. Histopathological assessment of the scalpel biopsy specimens classified lesions into 6 categories. Brush cytology specimens were analyzed by machine learning classifiers trained to identify relevant cytological features. Multimodal diagnostic models were developed using cytology results, lesion characteristics, and risk factors. Squamous cells with nuclear F-actin staining were associated with early disease (i.e., lower proportions in benign lesions than in more severe lesions), whereas small round parabasal-like cells and leukocytes were associated with late disease (i.e., higher proportions in severe dysplasia and carcinoma than in less severe lesions). Lesions with the impression of oral lichen planus were unlikely to be either dysplastic or malignant. Cytological features substantially improved upon lesion appearance and risk factors in predicting squamous cell carcinoma. Diagnostic models accurately discriminated early and late disease with AUCs (95% CI) of 0.82 (0.77 to 0.87) and 0.93 (0.88 to 0.97), respectively. The cytological features identified here have the potential to improve screening and surveillance of the entire spectrum of oral potentially malignant disorders in multiple care settings.

Large Draining Focal Fibrous Hyperplasia Secondary to Periapical Granuloma.

Periapical granuloma is a pathological diagnosis associated clinically and radiographically with a nonvital tooth and a periapical radiolucency, respectively. It is frequently seen as a sequela of long-standing pulpal necrosis. Often times, a draining fistula is observed near the nonvital tooth. We report an unusual case of a large draining focal fibrous hyperplasia in association with a large periapical granuloma treated at our clinic. The diagnosis was made by the clinical presentation, radiologic and histopathologic findings.

Cytokeratin expression in corneal endothelium in the iridocorneal endothelial syndrome.

The immunocytologic characteristics of two formalin-fixed, paraffin-embedded corneas from patients with the iridocorneal endothelial (ICE) syndrome and unaffected control corneas were studied. Binding of polyclonal antisera to Factor VIII, S-100 protein, involucrin, neuron specific enolase (NSE), and the lectins peanut agglutinin and Ulex europaeus agglutinin-1 was performed using the standard peroxidase-anti-peroxidase method. We detected reactive patterns of monoclonal antibodies to cytokeratins (34BE12 is a 56-58 kD mouse IgG reactive to stratified epithelia; Pkk1 is a 44-54 kD mouse IgG reactive to simple epithelia; and KL1 is a 55-57 kD mouse IgG reactive to epidermis and simple epithelia) using the standard avidin-biotin complex method. Staining properties were similar for the polyclonal antisera, lectins, NSE, and chromogranin in corneas with ICE syndrome and in the controls. However, the cytokeratins 34BE12, Pkk1, and KL1 were detected in the endothelium of the corneas with the ICE syndrome but not in the controls. These findings suggest that various cytokeratins are expressed in the corneal endothelium in the ICE syndrome that are not expressed in unaffected corneal endothelium.

Expression patterns of epithelial differentiation antigens and lectin-binding sites in ameloblastomas: a comparison with basal cell carcinomas.

Whether the peripheral ameloblastoma (PA) and intraoral basal cell carcinoma (BCC) are two different clinical entities or essentially the same lesion still remains unresolved. The immunophenotypes of neoplastic cells of peripheral and intraosseous ameloblastomas, ameloblastic carcinomas, and BCCs were studied using a panel of monoclonal/polyclonal antibodies and lectins. The major cytokeratins (CKs) of neoplastic cells of ameloblastomas were CKs 5 and 14, whereas co-expression of CKs 8, 18, and 19 was observed in the cells of the stellate reticulum-like areas. Metaplastic squamous and keratinizing cells found in follicular and acanthomatous variants of ameloblastomas expressed CKs 1 and 10, involucrin, and binding sites for the lectins Ulex europeaus agglutinin I and Helix pomatia agglutinin. beta 2-Microglobulin was uniformly negative in all cases of ameloblastomas and ameloblastic carcinomas studied. Cutaneous BCCs also demonstrated similar reactive patterns with the above-mentioned antigens. The most striking feature is the presence of a peritumorous band-like peanut agglutinin staining found in both BCCs and PAs but not in intraosseous ameloblastomas. This unique peanut agglutinin staining pattern of PA may be diagnostically useful for its histopathologic distinction from an intraosseous ameloblastoma that has infiltrated the soft tissue. The neoplastic cells of ameloblastomas express markers of less-differentiated epithelial cells. Despite differences in epithelial origins, PAs are tumors analogous to cutaneous BCCs.

Variable expression of cathepsin B and D correlates with highly invasive and metastatic phenotype of oral cancer.

The expression levels of cathepsins B, D, and L in oral cancer surgical specimens were determined using immunocytochemical analysis. Cathepsins B and D are frequently overexpressed in squamous cell carcinomas, whereas their overexpression was less frequent in verrucous carcinoma and basaloid squamous cell carcinomas. Elevated level of cathepsin B in oral carcinomas was significantly associated with advanced tumor stage (P < .05) and poor histologic malignancy grade (P < .001). Increased expression of cathepsin D correlated significantly with the presence of metastasis (P < .05), poor histologic malignancy grade (P < .001), and high proliferation rate (P < .05). Cathepsin L was less frequently overexpressed in oral cancers than cathepsin B and D. These findings indicate that there is a strong cause/effect relationship between the expression levels of cathepsin B and D in oral cancers and their local invasive and metastatic growth patterns. Thus, cathepsins B and D are useful prognostic markers as well as promising gene therapy targets for oral cancer.

Are differences in filaggrin expression suitable for discriminating benign, premalignant and malignant skin lesions? An immunohistochemical study.

The differential diagnosis of benign and malignant skin lesions may pose considerable difficulties. Therefore, 137 formalin-fixed paraffin-embedded biopsies of various benign, premalignant and malignant skin tumours were examined immunohistochemically for the presence of filaggrin using the monoclonal anti-filaggrin antibody AKHl. The proliferating cells of both benign and malignant tumours were negative for filaggrin. Most of the benign tumours exhibited an intense filaggrin-positive granular layer identical to the adjacent healthy skin whereas benign conditions such as irritated seborrhoeic keratoses and clear cell acanthomas were completely devoid of filaggrin. Keratoacanthomas revealed an inconsistent staining pattern for filaggrin. Premalignant and malignant lesions including basal cell carcinomas were either focally positive or completely negative for filaggrin. The absence or presence of filaggrin in these lesions did not correspond to the degree of dysplasia. Moreover, none of these conditions gave a staining pattern for filaggrin to be used for the histological differential diagnosis of these skin lesions.

Cathepsin-D and tumor associated antigen DF3 in salivary gland neoplasia. Differential diagnostic and prognostic applications.

Salivary gland tumors pose considerable difficulty in diagnostic and prognostic assessment based on the histopathologic features alone. Cathepsin-D is overexpressed in cancer cells where its concentration in the primary tumor is correlated with increased risk of metastasis. DF3 antigen is a tumor associated glycoprotein that is specific for malignant cells of glandular origin. We examined the distribution patterns of cathepsin-D and DF3 antigens in benign (n = 11) and malignant (n = 44) salivary gland tumors of various histologic types. The frequency of cathepsin-D expression is significantly increased (p < 0.001) in salivary gland carcinomas compared to benign mixed tumors (BMT). High levels of cathepsin-D expression was frequent in carcinomas ex BMT, mucoepidermoid carcinomas, poorly differentiated adenocarcinomas-NOS and adenoid cystic carcinomas (ACC). Acinic cell carcinomas and polymorphous low-grade adenocarcinomas were mostly negative. Intense cytoplasmic staining for DF3 antigen was noted in the tumor cells of mucoepidermoid carcinomas, carcinomas ex BMT and poorly differentiated adenocarcinomas-NOS whereas other types of salivary gland carcinomas exhibited either negative or only focal membrane staining. The noted differences in the reactive patterns of cathepsin-D and DF3 antigen among various histologic types of salivary gland carcinomas may have differential diagnostic and prognostic applications.

Aggressive osteoblastoma of the mandible closely simulating calcifying epithelial odontogenic tumor. Report of two cases with unusual histopathologic findings.

Aggressive osteoblastoma is a rare bone-forming neoplasm composed of prominent epithelioid cells that demonstrate locally invasive growth with a high rate of recurrence but no metastatic potential. Clinical, radiographic and pathologic features of mandibular aggressive osteoblastoma in a 21-year-old African-American male and a 12-year-old Caucasian female are presented. Both tumors were resected with wide surgical margins and neither patient had adjuvant radiation or chemotherapy. The patients showed no evidence of local recurrence or distant spread either clinically or radiographically after two years of follow-up. These tumors were composed of solid sheets of pleomorphic epithelioid cells, eosinophilic amorphous osteoid with foci of calcification, which closely simulated amyloid. Differentiation of this tumor from histologically similar calcifying epithelial odontogenic tumor and low-grade osteosarcoma proved difficult. Immunohistochemical study with osteocalcin confirmed the osteoblastic nature of these epithelioid cells.

A comparative lectin histochemical study of major and minor salivary glands with special reference to the labial glands.

There is debate about the nature of the secretory cells in labial salivary glands. To characterize their basophilic acini the binding patterns of different lectins were examined in these glands (n = 30) and compared with those of major salivary glands (n = 10) and palatal salivary glands (n = 20). Binding in formalin-fixed paraffin-embedded sections was revealed using either anti-lectin antibodies and the peroxidase-antiperoxidase method or biotinylated lectins and the avidin biotin method. Binding of peanut agglutinin was seen in all basophilic acini of labial glands, whereas serous acini of major glands were completely negative. Focal binding of soybean agglutinin was seen in all basophilic acini of labial glands in addition to diffuse binding in mucous acini of all salivary glands, whereas serous acini were mostly negative. Conversely, there was binding of Ulex europaeus agglutinin I in all mucous and serous acini of all glands except for the basophilic acini of labial glands. Bandeiraea simplicifolia agglutinin I and Helix pomatia agglutinin bound to all basophilic and most mucous acini of labial glands, and to most mucous acini of major salivary glands, whereas only a few serous acini of major glands were reactive with both lectins. Pretreatment with neuraminidase yielded binding sites for peanut agglutinin in most of the serous acini of major glands and mucous acini of labial glands, and increased B. simplicifolia agglutinin I and H. pomatia agglutinin binding in the serous acini of major glands. Thus the different lectin binding patterns, particularly the differences in the reaction patterns of peanut, soybean and U. europaeus agglutinin I, showed that the basophilic acini of labial glands reacted more like mucous acini.

Ameloblastic fibrosarcoma in a bull.

This report describes a malignant odontogenic neoplasm in a 7-year-old bull. The mass, involving the right mandible, was locally invasive and destructive. Histologically, it consisted of islands and cords of benign odontogenic epithelium, entrapped in a population of malignant mesenchymal cells. These morphological features are characteristic of ameloblastic fibrosarcoma in man, an odontogenic tumour not previously described in animals.

Tobacco use and cancer. A reappraisal.

With approximately six million users, smokeless tobacco has received considerable scrutiny as a risk factor for oral cancer. We review the relationship between smokeless tobacco use, keratosis, and oral cancer. Several features of smokeless tobacco keratosis, including the natural history, clinical presentation, and biologic behavior, differentiate it from other leukoplakias that exhibit greater malignant potential. Previous research has demonstrated that the relative risk of oral cancer with smokeless tobacco use is 4.2, about half of the risk from smoking (relative risk = 10 to 15). Mortality data from populations with sustained high-frequency smokeless tobacco use do not support the mistaken prediction of an epidemic of oral cancer with increasing smokeless tobacco use. In fact, the risks of smokeless tobacco use compare so favorably with those of smoking that smokers who switch to smokeless tobacco reduce their risks for all tobacco-related illnesses including oral cancer. Although some criticize this proposal as less than an ideal solution for the nation's smokers, full adoption of this strategy would eventually save over 400,000 lives each year.

Spontaneous bilateral quadriceps tendon rupture.

Spontaneous bilateral quadriceps tendon ruptures are uncommon. We present a 30-year-old man with end-stage renal failure, who sustained this injury, and subsequently had surgical repair of both tendons on separate occasions. He has since regained full range of movement of both knees.