Assessment of dental and periodontal indices and Streptococcus mutans virulence in fragile X syndrome patients.

INTRODUCTION: Fragile X syndrome (FXS) is the most common cause of hereditary genetic disorder in a single gene characterised by intellectual disability. Behavioural features such as autism, hyperactivity and anxiety disorder may be present. Biofilm development and pathogenicity of Streptococcus mutans may be altered because FXS renders the dental approach and oral hygiene more complex. OBJECTIVES: The purpose of this study was to compare the levels of transcripts for VicRK and CovR of S. mutans isolated from FXS patients with the levels of transcripts for VicRK and CovR of standard strain ATCC, using a quantitative polymerase chain reaction (qPCR). METHODS: The caries experience index was assessed by the International Caries Detection and Assessment System (ICDAS), Periodontal Condition Index (PCI) and Invasive Dental Treatment Need Index (INI). RESULTS: The clinical index findings revealed a high rate of caries cavities and bleeding on probing of FXS patients. When VicRK and CovR transcript levels were compared with the reference strain, Fragile X patients were found to have significantly higher values. CONCLUSION: The present study demonstrated that FXS patients have more adverse clinical conditions, with increased biofilm accumulation and virulence. When combined with behavioural abnormalities, these patients become even more vulnerable to dental caries.

Spatiotemporal Dynamics and Natural Mortality Factors of Myzus persicae (Sulzer) (Hemiptera: Aphididae) in Bell Pepper Crops.

Pest populations are mostly regulated by climate, intra- and interspecific competition, natural enemies, and host plant quality. Myzus persicae (Sulzer) (Hemiptera: Aphididae) is a widely adapted aphid in the agroecosystems and is one of the main bell pepper pests. In the present study, we determined the spatial and temporal dynamics and the regulatory factors of M. persicae populations in bell pepper crops. The number of aphids and their natural enemies were evaluated during 2 years in four commercial bell pepper fields. Myzus persicae density data were related to temperature, rainfall, and natural enemies by multiple regression analysis and were then submitted to geostatistical analysis. The density of M. persicae was higher during the plant's reproductive growth stage. Rainfall, Chrysoperla spp., and Toxomerus spp. regulate M. persicae populations. Initial infestations of this pest occur along the edges of the fields and subsequently spread towards the center. This information is useful for integrated management programs aimed at anticipating periods of higher abundance of M. persicae and identifying arthropods to be prioritized in biological control.

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.

Lymphatic vascular density and lymphangiogenesis during tumour progression of carcinoma ex pleomorphic adenoma.

AIMS: To assess lymphatic vascular density (LVD) and lymph vessel endothelial proliferation in a series of carcinoma ex pleomorphic adenoma (CXPA) that represents the tumour in the different carcinogenesis phases and tumour progression. METHODS: In 8 cases of early CXPA (intracapsular and minimally invasive tumours), 8 of advanced CXPA (widely invasive tumours) and 10 of pleomorphic adenoma (PA) without malignant transformation, lymphatic vessels and proliferating cells were detected using the antibodies D2-40 and Ki-67 respectively. RESULTS: Comparing early tumours with advanced ones, LVD was not significantly different at the tumour margin. In contrast, regarding intratumoural lymphatics, PA without malignant transformation and early CXPA contained rare, if any, lymph vessels, whereas in widely invasive carcinomas they were more numerous. However, neither intratumoural nor peritumoural LVD were increased in comparison to adjacent normal salivary gland tissue. In no case did dual immunohistochemistry using D2-40 and the cell proliferation marker Ki-67 reveal the existence of proliferating lymphatics. Carcinomatous emboli were found in peritumoural as well as in intratumoural lymphatics only in advanced CXPA without myoepithelial differentiation. CONCLUSION: In CXPA, the lymphatic network is mainly composed of pre-existing lymphatics which are rare in tumours that have not infiltrated outside the confines of the original PA. In the widely invasive CXPA, intratumoural as well as peritumoural lymphatics are a conduit for carcinoma cells, but in carcinomas with myoepithelial differentiation, the neoplastic cells seem to have a lower invasion capacity.

Polymorphous low grade adenocarcinoma presenting an uncommon radiographic aspect.

The aim of this study was to present clinical, histological and immunohistochemical aspects of a polymorphous low grade adenocarcinoma occurring in the mandible. A radiolucent tumour, located in the right mandible, was removed from a 40-year-old woman. Radiographic and CT exams revealed that the lesion expanded bucco-lingual cortical plates and presented an irregular scalloping of the bone. The surrounding lining mucosa was intact. The patient underwent total surgical removal of the lesion with an intraoperative biopsy. Histological diagnosis was polymorphous low-grade adenocarcinoma confirmed by immunohistochemical study. One-year follow up was uneventful. The accurate diagnosis of lesions presenting unusual clinical aspects, as the one presented here, is critical for correctly handling treatment.

Expression of integrin subunits alpha2, alpha3, alpha5, alphav, beta1, beta3 and beta4 in different histological types of ameloblastoma compared with dental germ, dental lamina and adult lining epithelium.

OBJECTIVE: To analyze integrin expression and distribution in different histological types of ameloblastoma, compared with dental germ, dental lamina and adult lining epithelium. MATERIALS AND METHODS: Three-micrometer sections from paraffin-embedded specimens were evaluated employing a streptavidin-biotin immunohistochemical method and anti-integrin alpha2, alpha3, alpha5, alphav, beta1, beta3 and beta4 antibodies. RESULTS: All integrins were present in all specimens, exhibiting different patterns. In follicular ameloblastoma, the integrin staining was stronger in the periphery while integrin alpha2 was not present in the central cells. Acanthomatous ameloblastoma showed a similar pattern, with positive staining for integrins alpha3, alpha5, alphav, beta1 and beta4 in the metaplastic cells. In the unicystic, integrin staining was uniform except for integrins alpha5 and beta3 which showed weaker staining in the upper layers. In the plexiform ameloblastoma, dental germ and lamina integrin staining was uniform. In the adult lining epithelium, staining for integrins alpha2, alpha5 and beta4 was confined to the basal layer, while integrins alphav and beta3 were present in the basal and parabasal, with integrins alpha3 and beta1 in the upper layers. CONCLUSION: Acanthomatous, follicular and unicystic ameloblastomas showed integrin staining patterns similar to the adult lining epithelium while the plexiform ameloblastoma was similar to the dental germ and lamina.

Sialadenoma papilliferum: immunohistochemical study.

Sialadenoma papilliferum (SP) is a rare benign tumour of salivary gland origin, which has been included among the ductal papillomas in the latest classification of tumours by the World Health Organisation. Two SP from the minor salivary gland of the palate of middle age patients were presented and studied by immunohistochemical. Our results showed presence of cytokeratins (CKs) 13, 14, 7, 8, 19 and absence of vimentin and smooth muscle actin. This immunoprofile is similar to the excretory duct of salivary gland.

Cytokeratins in epithelia of odontogenic neoplasms.

Neoplasms and tumours related to the odontogenic apparatus may be composed only of epithelial tissue or epithelial tissue associated with odontogenic ectomesenchyme. The immunohistochemical detection of different cytokeratins (CKs) polypeptides and vimentin has made it easier to explain the histogenesis of many epithelial diseases. The present study aimed to describe the immunohistochemical expression of cytokeratins 7, 8, 10, 13, 14, 18, 19 and vimentin in the epithelial components of the dental germ and of five types of odontogenic tumours. The results were compared and histogenesis discussed. All cells of the dental germ were positive for CK14, except for the preameloblasts and secreting ameloblasts, in which CK14 was gradually replaced by CK19. CK7 was especially expressed in the cells of the Hertwig root sheath and the stellate reticulum. The dental lamina was the only structure to express CK13. The reduced epithelium of the enamel organ contained CK14 and occasionally CK13. Cells similar to the stellate reticulum, present in the ameloblastoma and in the ameloblastic fibroma, were positive for CK13, which indicates a nature other than that of the stellate reticulum of the normal dental germ. The expression of CK14 and the ultrastructural aspects of the adenomatoid odontogenic tumour probably indicated its origin in the reduced dental epithelium. Calcifying odontogenic epithelial tumour is thought to be composed of primordial cells due to the expression of vimentin. Odontomas exhibited an immunohistochemical profile similar to that of the dental germ. In conclusion, the typical IF of odontogenic epithelium was CK14, while CK8, 10 and 18 were absent. Cytokeratins 13 and 19 labelled squamous differentiation or epithelial cells near the surface epithelium, and CK7 had variable expression.

Salivary duct carcinoma: cytokeratin 14 as a marker of in-situ intraductal growth.

AIMS: The aim of this study was to determine the immunoprofile of salivary duct carcinoma and to differentiate intraductal growth from invasive growth. METHODS AND RESULTS: We applied a panel of antibodies (cytokeratins 7, 8, 13, 14, 19, vimentin and alpha-smooth muscle actin) in five cases of salivary duct carcinoma. This panel is currently used for identification of different components of salivary gland tumours in our laboratory. All tumour cells were positive for cytokeratins 7 and 8. Few neoplastic structures expressed cytokeratin 14 in cells surrounding tumour islands. CONCLUSION: The finding of cytokeratin 14 was important to confirm the in-situ intraductal growth, which probably characterizes this low-grade neoplasm.

Well-differentiated liposarcoma of the tongue.

Intraoral liposarcomas are rare, with most reported cases being of the myxoid histological type. We present a well-differentiated liposarcoma of the tongue, in a 65-year-old man. The tumour presented lipoblasts in various stages of differentiation, lipocytes in different sizes and shapes, mesenchymal and signet-ring cells. Lipoma, spindle-cell lipoma, myxoma, hibernoma, angiolipoma, fibrolipoma, pseudosarcomatous faciitis and malignant hysticytoma were considered in the diagnosis process. The patient was treated surgically and so far is free of disease.

Clinicopathologic features of 54 cases of paradental cyst.

OBJECTIVE: The aim of the study was to evaluate the clinicopathologic features of 54 paradental cysts. METHOD AND MATERIALS: The cases were retrieved from the files of the Department of Oral Pathology of the University of São Paulo, in Brazil. Only case files that included complete data, radiographs, and clinical history were used for the study. RESULTS: Most of the cysts were adjacent to a partially erupted mandibular third molar; only two cysts were adjacent to mandibular second molars, and one cyst was adjacent to a first molar. Women were affected more often than were men (33:21), and most of the patients were in their second or third decades. In most cases, the affected tooth had a history of one or more episodes of pericoronitis. Histologically, these cysts presented the typical features of an inflammatory cyst, having a lining epithelium, which apparently resulted from a union between reduced enamel epithelium and oral epithelium. CONCLUSION: Development of the paradental cyst could be avoided if episodes of pericoronitis were properly treated, because the cyst is usually related to pericoronitis.

Ultrastructural aspects of connective tissue in hereditary gingival fibromatosis.

OBJECTIVE: The purpose of this study was to analyze the ultrastructure of gingival connective tissue from patients in one family affected by hereditary gingival fibromatosis (HGF). STUDY DESIGN: Electron microscopic examination was performed with gingival tissue from 10 patients from a Brazilian family with 132 members. Fifty of 96 persons at risk for this disorder were affected, which is consistent with an autosomal dominant pattern of inheritance. RESULTS: The extracellular matrix showed flocculent material and collagen fibrils with structural abnormalities and variation in diameter. Increased numbers of oxytalan fibers were identified; however, elastic fibers were rare in the analyzed areas. CONCLUSIONS: The structural alterations found in HGF appear similar to those described in certain other heritable collagen disorders, suggesting that HGF should be included in the group of hereditary diseases in which connective tissue alterations have a distinct pattern, in contrast to reactive fibrotic gingival enlargements with no genetic component.

Collagenous fibroma (desmoplastic fibroblastoma) of the palate: a case report.

Collagenous fibroma is an uncommon benign soft tissue lesion that has a wide anatomic distribution. We describe a case of a collagenous fibroma that appeared in the left soft and hard palate of a 37-year-old woman as a 5.0-cm solitary, firm nodule. Microscopically, it was composed of stellate or spindle-shaped cells embedded in hypovascular fibrous stroma. Entrapment of fat was focally identified at the edges. Mitotic figures and tumor necrosis were absent. Tumor cells were immunopositive for vimentin, and a few cells were positive for alpha-smooth muscle actin. Tumor extracellular matrix was immunopositive for type I and type III collagen, as well as for fibronectin. These findings satisfied the diagnostic criteria for collagenous fibroma (desmoplastic fibroblastoma). This case, to our knowledge, represents the first report of this tumor in the mouth. The differential diagnosis of fibrous lesions of the mouth is discussed.

Pulp response and cytotoxicity evaluation of 2 dentin bonding agents.

OBJECTIVE: This study evaluated the biocompatibility of two dentin bonding agents (Clearfil Liner Bond 2 and Scotchbond Multi-Purpose) applied in human dental pulps and cell cultures. METHOD AND MATERIALS: In vivo: Twenty human third molars that were scheduled for extraction were used. After cavity preparation, pulp exposure was achieved with a carbide bur. Hemorrhage control was obtained with saline solution. In 16 teeth, adhesive pulp capping was performed and the cavities were sealed with resin composite. In the control group (n = 4), pulps were capped with Ca(OH)2 and the cavities were sealed with IRM. Teeth were extracted 30 or 90 days following treatment and prepared for histological examination and bacterial detection. In vitro: materials were applied in Petri dishes, where NIH-3T3 cells were plated. The cells were counted 2, 4, and 6 days after plating to obtain the growth curves and to determine cell viability. All data were submitted to statistical analysis. RESULTS: In vivo: Dentin bridge formation was seen in all teeth capped with Ca(OH)2, without an inflammatory response. Mild inflammatory responses and dentin bridge formation after 90 days were observed in 50% of specimens treated with Liner Bond 2. Pulps treated with Scotchbond Multi-Purpose presented mild to severe inflammatory response, and no mineralized tissue formation was detected. Bacteria were not disclosed in any specimen. In vitro: The cytotoxicity was similar between the two bonding agents, and both had statistically higher cytotoxic effects (P < 0.002) than Ca(OH)2. CONCLUSION: Ca(OH)2 produced pulp healing in all teeth and exhibited lower cytotoxic effects than both adhesive systems; however, pulp healing was also observed under Liner Bond 2.

Rotated subcondylar process fracture in the growing animal: an experimental study in rats.

Unilateral, rotated subcondylar fractures were surgically induced in young rats under general anaesthesia. A decrease in body weight was observed in the fractured animals, with recovery at the end of the experimental period. A tendency of deviation of the mandible to the fracture side was noticed on axial radiographs, but without significant difference between experimental and sham-operated animals. Histological data initially demonstrated acute inflammation along the articular capsule and adjacent muscle fibres. Within 2 weeks, exuberant callus formation occurred. Neither devitalized bone ends nor intra-articular granulation tissue were seen. After 1 month, the temporomandibular joint presented characteristics of normality, with the condylar process centralized into the temporal fossa, interposed by the articular disc. These results were preserved after 3 months. We concluded that rotational subcondylar fractures in young rats heal by callus formation, with simultaneous and prompt repositioning of the condyle.

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.

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.

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.

Effect of freeze-dried bone and a dextran agglutinant on the healing of defects in the rat mandible.

The effects of inorganic, freeze-dried bone, alone or mixed with a dextran agglutinant, on the repair of experimentally created critical-size bone defects were evaluated in the rat mandible. Histological data demonstrated a progressive resorption of the freeze-dried bone particles, without osseous proliferation into the defect, and a posterior foreign-body reaction. These results indicate that the freeze-dried bone did not induce bony healing in the critical-size defect and that the addition of dextran agglutinant did not change the tissue response. Dextran may have other applications as a biocompatible, resorbable agglutinant.

PCNA/AgNOR and Ki-67/AgNOR double staining in oral squamous cell carcinoma.

This study was performed on oral squamous cell carcinomas (OSCC) in order to investigate the relation between the number of interphase silver-stained nucleolar organizer regions (AgNORs) and the immunolabeling of proliferation-associated markers, using antibodies to Ki-67 and proliferating cell nuclear antigen (PCNA). Fifteen consecutive cases of oral squamous cell carcinoma were used and a double staining technique was performed in order to quantify the number of NORs in PCNA-positive and -negative cells as well as in Ki-67-positive and -negative cells. Our results showed a higher mean number of AgNORs in PCNA- and Ki-67-positive cells than in PCNA- and Ki-67-negative cells. We concluded that there is an association between cell proliferation and AgNOR score in OSCC.