Substance P regulates the expression of matrix metalloproteinases and tissue inhibitors of metalloproteinase in cultured human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Substance P may play a role in the pathogenesis of periodontal disease; however, its mechanisms of modulation are not clear. This study evaluated the effect of two concentrations of Substance P on the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in cultured human gingival fibroblasts. MATERIAL AND METHODS: Fibroblasts were stimulated for 48 h with 10(-4) or 10(-9) m Substance P; untreated fibroblasts served as controls. The expression of MMP-1, -2, -3, -7 and -11 and of TIMP-1 and -2 was evaluated using real-time polymerase chain reaction and western blotting. RESULTS: There was a significant, concentration-dependent stimulatory effect of Substance P on MMP-1, -2, -3 and -7 and TIMP-2 gene expression (p < 0.05), and a probable effect on MMP-11 (p = 0.06). At the higher concentration (10(-4) m Substance P), MMP-1, -2, -3, -7 and -11 and TIMP-2 showed the greatest up-regulation; at the lower concentration (10(-9) m Substance P), MMP-1, -3 and -7 and TIMP-2 exhibited diminished up-regulation, with MMP-2 and -11 showing down-regulation (p < 0.05). Expression of TIMP-1 was not affected by Substance P (p > 0.05). Western blotting confirmed that Substance P up-regulated MMP-1, -2, -3 and -11 and TIMP-2. MMP-1, -3 and -11 and TIMP-2 showed greater up-regulation at the higher Substance P concentration and diminished up-regulation at the lower concentration. MMP-2 was up-regulated to a similar degree at both Substance P concentrations. CONCLUSION: In gingival fibroblast cells, Substance P at the higher concentration (10(-4) m) induced greater up-regulation of MMP-1, -3 and -11 and TIMP-2 expression, but at the lower concentration (10(-9) m) induced diminished up-regulation, which may represent a mechanism for modulating periodontal breakdown.

Peroxiredoxin I is differentially expressed in multiple myelomas and in plasmablastic lymphomas.

BACKGROUND: Plasmablastic lymphoma (PBL) and multiple myeloma (MM) are B cell-derived malignancies that share many morphologic and immunophenotypic traits, making the differential diagnosis particularly complicated. We have recently demonstrated that peroxiredoxin I (PrdxI) is expressed in plasma cells but not in B lymphocytes, suggesting that its expression is development-associated. AIM: To analyze PrdxI expression in PBL and in MM in order to study its utilization as an additional diagnostic molecular tool. METHODS AND RESULTS: Eight cases of PBL and nine of MM were studied by immunohistochemistry. We have demonstrated that PrdxI expression is closely connected with the immunoglobulin production capacity of the cells, which means high in MM, but absent in PBL cases, except one, wherein few cells were stained. CONCLUSIONS: We hypothesize PrdxI as a component of the unfolded protein response (UPR), an adaptive pathway essential for plasma cell differentiation. As we have not detected immunoglobulin in our PBL cases, we suggest that UPR was not activated in the cells, accounting for the impediment of the developmental process, and for the inhibition of PrdxI expression observed. PrdxI could be considered an additional plasma cell functional marker and could also be speculated as a therapeutic target in the treatment of MM.

Effect of a topical antimicrobial paste on healing after extraction of molars in HIV positive patients: randomised controlled clinical trial.

AIM: To assess the efficacy of a medicated dressing in the socket on the healing after dental extractions in HIV positive patients. PATIENTS AND METHODS: Forty HIV positive patients with CD4 counts below 200 cells/mm(3) who were to have molar teeth extracted were randomly allocated to have the socket dressed with a medicated paste (Rifocort) with camphorated parachlorophenol and iodoform) and sutured (treated group), or merely sutured (control group). RESULTS: The sockets in the treated group healed more quickly than in the control group.

Basal cell adenocarcinoma of the palate with squamous metaplasia.

Basal cell adenocarcinoma is a rare salivary gland tumour, especially in minor glands. The clinical, histological, and immunohistochemical features of a case involving the palate are described. Formalin fixed, paraffin embedded sections of the tumour were examined in haematoxylin and eosin (H&E) sections and also using immunostaining for cytokeratins 7, 8, 13, 14, 18, 19, vimentin, muscle specific actin (HHF35), and laminin. H&E sections showed that the tumour was composed mainly of basaloid cells and a striking feature was the presence of squamous metaplasia. Neural invasion was also conspicuous. Immunohistochemical reactions indicated that cytokeratin 14 was expressed by all tumour cells and vimentin by all cells except those in the areas of squamous metaplasia. The remaining cytokeratins and actin were present in some of the tumour cells, while laminin showed discreet positivity around cell arrangements. The foci of squamous metaplasia and the immunohistochemical findings are helpful in distinguishing basal cell adenocarcinoma from other salivary gland tumours which show basaloid cells.

Application of immunohistochemistry to the diagnosis of salivary gland tumors.

A panel of antibodies composed of the cytokeratins (CKs), vimentin, and actin was applied to 114 minor salivary gland tumors to evaluate its diagnostic value. The results revealed that luminal cells of intercalated duct-like structures, such as those seen in pleomorphic adenoma, basal cell adenoma, adenoid cystic carcinoma, and epithelial-myoepithelial carcinoma, expressed CKs 7, 8, 14, and 19. The outer cells of these structures exhibited vimentin or vimentin plus muscle-specific actin, but rarely CK14, which is seen particularly in pleomorphic adenoma, in the tubular type of basal cell adenoma, and seldom in the tubular type of adenoid cystic carcinoma. Modified myoepithelial cells of pleomorphic adenoma and myoepithelioma exhibited a variable immunoprofile. CKs 7 and 8 were also observed in acinar cell adenocarcinoma and polymorphous low-grade adenocarcinoma with vimentin in the latter. CK13 was expressed only by canalicular adenoma and mucoepidermoid carcinoma cells. This study showed that the panel of antibodies employed is effective in distinguishing among salivary gland tumors.

Study of minor salivary gland mucoepidermoid carcinoma differentiation based on immunohistochemical expression of cytokeratins, vimentin and muscle-specific actin.

The expression of cytokeratins (CKs) 7,8,10,13,14,18,19, vimentin and muscle-specific actin (MSA) was investigated in 17 mucoepidermoid carcinomas (MEC) by the streptavidin-biotin technique. The results revealed that CKs 7, 8 and 18 were positive for intermediate, luminal columnar and mucous cells. For epidermoid cells, the expression was heterogeneous and discrete. The reaction with CK19 was similar to that seen for the above CKs, except for the fact that mucous cells were negative. CK 14 was preferentially expressed in the intermediate cells localised in basal, parabasal and epidermoid cells. CK13 was localised in intermediate, epidermoid and luminal columnar cells. In stratified epithelium, CK13 was expressed in intermediate cells and negative in basal cells. These findings were more expressive in cystic areas of the tumours. CK10 was negative for all the cases studied. MSA was positive only in stromal elements, and only two cases of CME were heterogeneously positive for vimentin. The result obtained showed that the immunoprofile of MEC, for the studied antigens, is similar to that exhibited by the excretory duct of normal salivary glands.

Effect of N-CAM on in vitro invasion of human adenoid cystic carcinoma cells.

Adenoid cystic carcinoma of salivary glands is characterised by aggressive behaviour, high rate of local recurrences, neurotropism and late metastasis. In a previous work we demonstrated that adenoid cystic carcinoma cultured cells (CAC2 cells) expressed N-CAM. It was suggested that this expression, modulated by extracellular matrix, would be correlated to cell movement. The aim of our study was to verify whether CAC2 cells presented invasion capacity. Moreover, we tested whether the neural adhesion molecule (N-CAM) would participate in this process. CAC2 cells were either previously treated, or not (control), with a monoclonal antibody against N-CAM. Invasion assays were carried out using a modified Boyden chamber (Transwell chamber). CAC2 cells (10(5)) were dispensed into Transwell upper chamber on the top of Matrigel coated filter. The cells that invaded the filters in the first 8 h were counted under light microscopy, yielding data for the invasion rates (%). Control CAC2 cells presented an invasion rate of 5.28+/-0.04%. The invasion rate raised to 6.53+/-0.2% when N-CAM was blocked with monoclonal antibody. N-CAM impaired the adenoid cystic carcinoma cell invasion in vitro. Therefore, we suggest an anti-invasive role for N-CAM in adenoid cystic carcinoma.

The role of basement membrane proteins on the expression of neural cell adhesion molecule (N-CAM) in an adenoid cystic carcinoma cell line.

We investigated the presence of neural cell adhesion molecule (N-CAM) in the adenoid cystic carcinoma cell line CAC2 using immunofluorescence microscopy. Additionally, we analysed the role of laminin and type IV collagen in N-CAM expression. We demonstrated that cultured adenoid cystic carcinoma cells express N-CAM. Control cells presented a scattered N-CAM expression on cell membrane, and type IV collagen had no effect in N-CAM distribution. CAC2 cells grown on laminin-coated coverslips expressed N-CAM concentrated on cell lamellipodia suggesting relationship with migratory activity. Our results showed that cultured adenoid cystic carcinoma cells express N-CAM and this expression is modulated by extracellular matrix.

p53 In biopsies of oral squamous cell carcinoma. A comparative study with a malignancy grading system.

The relationship between the histological grading of malignancy and p53 protein expression was studied in 40 biopsies of oral squamous cell carcinomas. An immunohistochemical analysis was carried out using the streptavidin method preceded by a treatment with citric acid in a microwave oven. All cases were classified according to the histological malignancy grading system proposed by Anneroth et al. (Scandinavian Journal of Dental Research 1987, 95, 229-249). The expression of p53 was found in 62.5% of the carcinomas studied. Positivity of p53 staining showed a correlation with the histological grade of malignancy and with the degree of keratinisation, nuclear polymorphism and number of mitoses.

Immunohistochemical Mdm2 expression in minor salivary gland tumours and its relationship to p53 gene status.

Mdm2 protein is a cellular regulator of p53 protein activity. Minor salivary gland tumours were investigated for immunohistochemical expression of Mdm2 protein and for p53 gene status. Formalin-fixed sections were submitted to monoclonal antibody anti-Mdm2 through use of the streptavidin-biotin method. Nuclear immunoreactivity was scored 1 (0-25% nuclei positive), 2 (26-50%), 3 (51-75%) and 4 (> 75%). The scores found were: PLGA = 1-4; ACC = 3 and 4; ACA = 2 and 4; PA = 3. Genomic DNA of p53 gene exons 5-8 was examined by polymerase chain reaction and no alterations were detected. The strong immunohistochemical Mdm2 expression may represent an alternative mechanism to the development of salivary gland tumours.

Immunohistochemical distinction of high-grade mucoepidermoid carcinoma and epidermoid carcinoma of the parotid region.

The correct diagnosis of high-grade mucoepidermoid (MEC), which is composed of solid islands of intermediate and squamous cells, may be challenging, due to its similarity to other tumours, mainly with squamous cell carcinoma (SCC). The present report employed immunohistochemical technique against different cytokeratins (CKs), in order to differentiate these two entities. : Six high-grade MEC and six SCC of the parotid region, retrieved from the files of both Oral Pathology Department of the School of Dentistry of University of São Paulo and Pathology Department of A.C. Camargo Hospital, were submitted immunohistochemical technique against Cks 7,8, 10, 13, 14 and 19. : High-grade MEC was positive for Cks 7, 8, 13, 14 and 19. The cases of SCC showed strong positivity for CK14, and CK10 was present only in focal areas. Our results highlight the use of CKs (especially CK14) to differentiate high-grade MEC and SCC.

CD30+ lymphocytes in chronic gingivitis from HIV-positive patients: a pilot study.

Th2 type lymphocytes are characterized by high expression of CD30 glycoprotein. Increased serum levels of CD30 and Th2 IL-4 producing T-cells are found during AIDS progression. Since HIV-positive patients are more susceptible to periodontal disease, quantitative analysis of positive cells for the CD30 receptor in chronic gingivitis of both HIV-infected and non-infected patients (NSG) would help to clarify the immunoregulation of HIV-associated periodontal diseases. The purpose of this study was to evaluate CD30+ lymphocytes in gingival biopsies from sites exhibiting chronic gingivitis on HIV-positive patients (CG-HIV) and NSG. A biotin-streptavidin amplified system was used for identification of the CD30 receptor. The results demonstrated increased proportions of Th2 cells in CG-HIV as compared to NSG. Additional studies are necessary to understand the importance of these cells to the biological activity or inactivity of the disease.

The use of subcutaneously implanted araldite disks for microstructural studies of inflammatory macrophages.

Subcutaneously implanted epoxy resin disks can be used instead of glass coverslips in mice to recruit inflammatory macrophages. Implanted araldite disks exhibited large numbers of macrophages on both surfaces which could be observed conveniently by light and phase contrast microscopy. Most importantly, the disks can be embedded directly in an araldite mixture for the preparation of thin sections for electron microscope examination.

Healing of the displaced condylar process fracture: an experimental study.

Healing of the displaced condylar process fracture was analysed by means of an experimental model using adults rats. Displaced fractures of the right condylar process were achieved under general anaesthesia. Histological data initially demonstrated neutrophilic exudation along the articular capsule and adjacent muscle fibres. One week later, devitalized bone areas at the fracture site, as well as proliferation of cartilaginous and osseous tissue were observed. Subsequently, exuberant callus formation and a decrease in the inflammatory process occurred. After 3 months, the condylar process presented characteristics of normality and was centralized into the temporal fossa, with interposition of the articular disc. These results indicate that experimentally induced, displaced condylar process fractures heal by callus formation with simultaneous repositioning of the condyle.

Extranodal follicular dendritic cell sarcoma of the palate.

Follicular dendritic cell tumors are uncommon and usually occur in lymph nodes. We report the case of a follicular dendritic cell tumor that occurred in the palate of a 14-year-old boy and manifested itself as a nodular mass. Histologically, the neoplasm consisted of spindle-shaped or oval-shaped cells with eosinophilic cytoplasms and nuclei with delicate, dispersed chromatin. The lesional cells were principally arranged in diffuse, fascicular patterns with vaguely whorled or storiform areas. Focal multinucleate tumor giant cells and lymphocytes were observed throughout the neoplasm. Immunohistochemically, tumor cells were positive for the follicular dendritic cell markers CD21, CD35, and CD23 and for S-100 protein, CD68, and muscle-specific actin. Tumor cells were negative for LCA, CD20, EMA, CK (AE1/AE3), HMB45, and CD34. Lymphocytes were positive for LCA and CD45RO. Although follicular dendritic cell sarcoma is a very uncommon tumor, it should be included in the differential diagnosis of tumors in this location.

Intraosseous leiomyoma of the mandible.

An intraosseous leiomyoma arising within the mandible was diagnosed in a 24-year-old woman. Clinically, a nodular swelling of the lower border of the mandible was noted. Radiographically, a unilocular ellipsoid radiolucency, loss of the lower border of the mandible, and tooth resorption were observed. The mitotic count was 0.4 per 10 high-power fields, which supported the benign nature of the tumor. The clinical and histologic parameters for distinguishing between benign and malignant smooth muscle neoplasms are discussed.

Langerhans cell histiocytosis: recurrent lesions affecting mandible in a 10-year-old patient.

Hand-Schuller-Christian disease is a multifocal variant of eosinophilic granuloma, characterised by the classical triad of bony lesions, exophthalmos and diabetes insipidus. This case relates recurrent Langerhans' cell histiocytosis lesions presented as destruction of periodontal support associated with diabetes in a 10-year-old patient. Medical history suggests that the case represents a case of Hand-Schuller Christian disease.

Diagnosis of HIV infection using saliva samples: comparative study between adults and children.

The use of saliva samples to detect the anti-HIV antibody has been described recently as an interesting alternative in relation to blood samples. We have analyzed 172 saliva samples from HIV positive and negative patients assisted at CASP, 108 of them adults and 64 children, in order to assess the specificity and sensitivity of the GACELISA test. A specificity was found of 85% and 80% respectively for the adult and children groups and sensitivity of 97.7% and 100% respectively for the adult and children groups. The statistical analysis revealed no differences in specificity and in sensitivity between these groups. We concluded that GACELISA test is useful for children.

Accuracy assessment of the axial images obtained from cone beam computed tomography.

OBJECTIVE: The aim of this study was to evaluate accuracy of linear measurements assessed from axial tomograms and the influence of the use of different protocols in two cone beam CT (CBCT) units. METHODS: A cylinder object consisting of Nylon® (Day Brazil, Sao Paulo, Brazil) with radiopaque markers was radiographically examined applying different protocols from NewTom 3G(TM) (Quantitative Radiology s.r.l, Verona, Veneto, Italy) and i-CAT(TM) (Imaging Sciences International, Hatfield, PA) units. Horizontal (A-B) and vertical (C-D) distances were assessed from axial tomograms and measured using a digital calliper that provided the gold standard for actual values. RESULTS: There were differences when considering acquisition protocols to each CBCT unit. Concerning all analysed protocols from i-CAT(TM) and Newtom 3G(TM), both A-B and C-D distances presented underestimated values. Measurements of the axial images obtained from NewTom 3G(TM) (6 inch 0.16 mm and 9 inch 0.25 mm) were similar to the ones obtained from i-CAT(TM) (13 cm 20 s 0.3 mm, 13 cm 20 s 0.4 mm and 13 cm 40 s 0.25 mm). CONCLUSION: The use of different protocols from CBCT machines influences linear measurements assessed from axial images. Linear distances were underestimated in both equipments. Our findings suggest that the best protocol for the i-CAT(TM) is 13 cm 20 s 0.3 mm and for the NewTom 3G(TM), the use of 6 inch or 9 inch is recommended.