Isolated nonsyndromic cleft palate: multicenter epidemiological study in the Brazil.

BACKGROUND: Nonsyndromic orofacial clefts (NSOC) are the craniofacial most common congenital malformations. There are evidences that the nonsyndromic cleft palate (NSCP) development differs from other NSOC. However, most of the publications treat NSCP without considering that information. Furthermore, few studies focus on NSCP. The aim of this study was to describe epidemiological findings of patients with isolated NSCP in Brazil. METHODS: In this cross-sectional multicenter study, four reference Centers for treatment in three different Brazilian states was investigated. Data were obtained from clinical records of patients, between November 2021 and June 2022. Researched variables were sociodemographic, clinical characteristics and pregnancy and family history. Pearson's chi-square and ANOVA One-way tests were used for associations. RESULTS: Majority were female (58.1%), white (60.7%) with incomplete NSCP (61.2%). There was an association between complete NSCP and a positive history of medical problems during pregnancy (p = 0.016; 27.9%; OR: 1.94; 1.12-3.35). Systemic alterations were perceived in 40.6% of the sample with odds ratio for development of the complete type (OR: 1.21; 0.74-1.97). Higher OR was visualized in medication use during pregnancy (OR: 1.35; 0.76-2.37) and positive family history of oral cleft (OR: 1.44; 0.80-2.55). Dental and surgical care was associated with higher age groups (p < 0.050). CONCLUSIONS: NSCP was most prevalent in white skin color female. Complete NSCP is associated with medical problems during pregnancy. Medication use during pregnancy and positive family history of oral cleft increase the chance of developing complete NSCP.

Overexpression of heat-shock protein 47 impacts survival of patients with oral squamous cell carcinoma.

BACKGROUND: The expression of heat-shock protein 47 (HSP47) has been linked to collagen synthesis control and implicated in fibrotic disorders, but more recent studies have demonstrated its role in solid tumors. In this study, we explored the prognostic impact of HSP47 in oral squamous cell carcinomas (OSCC) and determined the in vitro effects of its loss-of-function on viability, proliferation, migration, invasion, and resistance to cisplatin of OSCC cells. METHODS: The HSP47 expression in tumor samples was assessed by immunohistochemistry in two independent cohorts totaling 339 patients with OSCC, and protein levels were associated with clinicopathological features and survival outcomes. The OSCC cell lines HSC3 and SCC9 were transduced with lentivirus expressing short hairpin RNA to stably silence HSP47 and used in assays to measure cellular viability, proliferation, migration, and invasion. RESULTS: HSP47 was overexpressed in OSCC samples, and its overexpression was significantly and independently associated with poor disease-specific survival and shortened disease-free survival in both OSCC cohorts. The knockdown of HSP47 showed no effects on cell viability or cisplatin sensitivity, but impaired significantly proliferation, migration, and invasion of OSCC cells, with stronger effects on SCC9 cells. CONCLUSION: Our results show a significant prognostic impact of HSP47 overexpression in OSCC and reveal that HSP47 inhibition impairs the proliferation, migration, and invasion of OSCC cells. HSP47 may represent a potential therapeutic target for OSCC.

Prognostic significance of the neural invasion in oral squamous cell carcinoma.

BACKGROUND: Although nerve involvement can predict recurrence and prognosis in oral squamous cell carcinomas, there still have controversies and limitations regarding the standardization for its detection. In this study, we explore the impact of neural invasion in oral squamous cell carcinomas prognosis, comparing intraneural invasion (tumor cells inside nerve structure) and perineural invasion (cells involving the nerve, but not invading its sheath). METHODS: Surgical slides stained with hematoxylin and eosin from 235 patients with oral squamous cell carcinomas were carefully verified for the presence of intraneural invasion and perineural invasion. The location in the tumor (intratumoral vs. peritumoral) and number of foci (unifocal or multifocal) were also explored. Survival analyses for cancer-specific survival and disease-free survival were performed with Cox proportional model. RESULTS: Neural invasion was identified in 74 cases, 64.9% displayed intraneural invasion and 35.1% displayed perineural invasion. Univariate analysis revealed a significantly poorer cancer-specific survival, but not disease-free survival, in patients with intraneural invasion, in contrast to cases with perineural invasion that did not achieve significant association with both cancer-specific survival and disease-free survival. Further analyses revealed that the location in the tumor and number of foci had little impact on discriminatory ability of intraneural invasion. Multivariate analysis confirmed that intraneural invasion is significantly and independently associated with poor cancer-specific survival (hazard ratio: 2.50, 95% CI: 1.31-3.79, p = 0.003). CONCLUSION: This study provides evidence that intraneural invasion, but not perineural invasion, is a relevant predictor of survival in patients with oral squamous cell carcinomas, suggesting that its association with other clinical and pathological prognostic factors should be consider in determining the optimal treatment protocol and prognosis of these patients.

MutSα expression predicts a lower disease-free survival in malignant salivary gland tumors: an immunohistochemical study

Ten cases of normal salivary gland (NSG) and 92 of SGT (54 benign and 38 malignant) were retrieved. Immunohistochemistry was performed for hMSH2, hMSH3, hMSH6. Scanned slides were digitally analyzed based on the percentage of positive cells with nuclear staining. Cases were further classified in MutSαhigh and MutSβhigh based on hMSH2-hMSH6 and hMSH3-hMSH6 expression, respectively. Results: hMSH3 expression was lower in malignant SGT compared to NSG and benign cases. Adenoid cystic carcinoma (ACC) cases with perineural invasion presented a lower percentage of hMSH3 positive cells. hMSH6 was downregulated in both benign and malignant SGT compared to NSG. Malignant SGT cases with MutSαhigh expression had lower disease-free survival compared to MutSαlow cases. A 10.26-fold increased risk of presenting local recurrence was observed. Conclusions: Our findings suggest that a lack of hMSH3 protein function is associated with a more aggressive phenotype (malignancy and perineural invasion) and that MutSα overexpression predicts a poor clinical outcome in malignant SGT.(AU)

Brazilian Multiethnic Association Study of Genetic Variant Interactions among FOS, CASP8, MMP2 and CRISPLD2 in the Risk of Nonsyndromic Cleft Lip with or without Cleft Palate.

Associations of CRISPLD2 (cysteine-rich secretory protein LCCL domain containing 2) and genes belonging to its activation pathway, including FOS (Fos proto-oncogene), CASP8 (caspase 8) and MMP2 (matrix metalloproteinase 2), with nonsyndromic orofacial cleft risk, have been reported, but the results are yet unclear. The aim of this study was to evaluate single nucleotide polymorphisms (SNPs) in FOS, CASP8 and MMP2 and to determine their SNP-SNP interactions with CRISPLD2 variants in the risk of nonsyndromic cleft lip with or without cleft palate (NSCL±P) in the Brazilian population. The SNPs rs1046117 (FOS), rs3769825 (CASP8) and rs243836 (MMP2) were genotyped using TaqMan allelic discrimination assays in a case-control sample containing 801 NSCL±P patients (233 nonsyndromic cleft lip only (NSCLO) and 568 nonsyndromic cleft lip and palate (NSCLP)) and 881 healthy controls via logistic regression analysis adjusted for the effects of sex and genomic ancestry proportions with a multiple comparison p value set at ≤0.01. SNP-SNP interactions with rs1546124, rs8061351, rs2326398 and rs4783099 in CRISPLD2 were performed with the model-based multifactor dimensionality reduction test complemented with a 1000 permutation-based strategy. Although the association between FOS rs1046117 and risk of NSCL±P reached only nominal p values, NSCLO risk was significantly higher in carriers of the FOS rs1046117 C allele (OR: 1.28, 95% CI: 1.10-1.64, p = 0.004), TC heterozygous genotype (OR: 1.59, 95% CI: 1.16-2.18, p = 0.003), and in the dominant model (OR: 1.50, 95% CI: 1.10-2.02, p = 0.007). Individually, no significant associations between cleft risk and the SNPs in CASP8 and MMP2 were observed. SNP-SNP interactions involving CRISPLD2 variants and rs1046117 (FOS), rs3769825 (CASP8) and rs243836 (MMP2) yielded several significant p values, mostly driven by FOS rs1046117 and CASP8 rs3769825 in NSCL±P, FOS rs1046117 in NSCLO and CRISPLD2 rs8061351 in NSCLP. Our study is the first in the Brazilian population to reveal the association of FOS rs1046117 with NSCLO risk, and to support that CRISPLD2, CASP8, FOS and MMP2 interactions may be related to the pathogenesis of this common craniofacial malformation.

Stress induced phosphoprotein 1 overexpression controls proliferation, migration and invasion and is associated with poor survival in oral squamous cell carcinoma.

Objective: Although there have been remarkable achievements in the molecular landscape of oral squamous cell carcinoma (OSCC) in recent years, bringing advances in the understanding of its pathogenesis, development and progression, little has been applied in the prognosis and choosing the optimal treatment. In this study, we explored the influence of the stress induced phosphoprotein 1 (STIP1), which is frequently reported to be highly expressed in many cancers, in OSCCs. Methods: STIP1 expression was assessed in the TCGA database and in two independent cohorts by immunohistochemistry. Knockdown strategy was applied in OSCC cell lines to determine the impact of STIP1 on viability, proliferation, migration and invasion. The zebrafish model was applied for studying tumor formation and metastasis in vivo. The association of STIP1 and miR-218-5p was explored by bioinformatics and mimics transfection. Results: STIP1 was highly expressed in OSCCs and significantly associated with shortened survival and higher risk of recurrence. STIP1 down-regulation decreased proliferation, migration and invasion of tumor cells, and reduced the number of metastases in the Zebrafish model. STIP1 and miR-218-5p were inversely expressed, and the transfection of miR-218-5p mimics into OSCC cells decreased STIP1 levels as well as proliferation, migration and invasion. Conclusion: Our findings show that STIP1 overexpression, which is inversely associated with miR-218-5p levels, contributes to OSCC aggressiveness by controlling proliferation, migration and invasion and is a determinant of poor prognosis.

Estudo Epidemiológico do Câncer Infantojuvenil no Hospital de Câncer de Cascavel Uopeccan entre os Anos 2000 e 2014 / Epidemiological Study of Childhood Cancer in Cascavel Cancer Hospital Uopeccan among 2000 and 2014 / Estudio Epidemiológico del Cáncer Infantil en el Hospital Oncológico de Cascavel Uopeccan entre 2000 y 2014

Introdução: Estudos epidemiológicos de base hospitalar sobre o câncer infantojuvenil são importantes para mostrar o perfil dos pacientes assistidos pelo serviço. Objetivo: Avaliar o perfil clínico-epidemiológico e a sobrevida d e pacientes na faixa etária de 0-19 anos atendidos no Hospital do Câncer de Cascavel Uopeccan (2000-2014). Método: Estudo transversal com avaliação de prontuários para os seguintes desfechos: sexo, idade, cor/raça, outras patologias, histórico familiar de câncer, domicílio, tipo da neoplasia, estadiamento, tratamento, metástases, recidivas, situação do paciente ao final da pesquisa. A estatística descritiva e os testes qui-quadrado e Kaplan-Meier foram aplicados. Resultados: Observou-se maior frequência para meninos (55,2%), faixa etária de 1-4 anos (36,3%), brancos (87%), domicílio urbano (81,6%), leucemia (35,8%) e quimioterapia (50,2%). Ocorreu metástase em 16,41% e recidiva em 22,38%. Não havia relato de histórico familiar de câncer em 47% dos prontuários. Outras patologias foram negadas em 58,9%. Ao final, 55,2% estavam vivos e sem doença. Houve associação estatisticamente significava entre menores de 10 anos com tumores renais e neuroblastoma; maiores de 10 anos com linfomas e neoplasias epiteliais malignas; e entre a situação atual do paciente com metástase, recidivas e estadiamento. Conclusão: Os pacientes analisados na presente pesquisa eram na maioria leucêmicos, do sexo masculino e faixa etária de 1-4 anos. A sobrevida global e a livre de doença foram, respectivamente, de 70,3% e 71,63%

Introduction: Hospital-based epidemiological studies on childhood cancer are important to show the profile of patients cared by the service. Objective: To evaluate the characteristics of cancer patients aged 0-19 years at the Cascavel Cancer Hospital Uopeccan (2000-2014). Method: Cross-sectional study that evaluated medical charts for the following outcomes: gender, age, color/race, comorbidities, family history of cancer, household, cancer type, staging, treatment, metastasis, recurrences, patient's status at the end of study. Descriptive statistics, chi-square and Kaplan-Meier were applied. Results: Boys were more prevalent (55.2%), age range from 1 to 4 years (36.3%), White (87%), urban household (81.6%), with leukemia (35.83%) and in chemotherapy (50.2%). Metastasis occurred in 16.41% and recurrence in 22.38%. There was no report of family history of cancer in 47% of the charts. Other pathologies were denied in 58.9%. In the end, 55.2% were alive and disease-free. There was a statistically significant association between boys younger than 10 years old with renal tumors and neuroblastoma and older than 10 years with lymphomas and malignant epithelial neoplasms and between the current status of the patient with metastasis, relapses, and staging. Conclusion: The patients analyzed in this study were mostly leukemic, males and aged 1-4 years. Global and disease-free survival were, respectively, 70.3% and 71.63%

Introducción: Los estudios epidemiológicos hospitalarios sobre cáncer infantil son importantes para mostrar el perfil de los pacientes atendidos por el servicio. Objetivo: Evaluar las características de los pacientes oncológicos de 0-19 años atendidos en el Hospital do Cáncer de Cascavel Uopeccan (2000-2014). Método: Estudio transversal que evaluó historias clínicas para los siguientes resultados: sexo, edad, color/raza, comorbilidades, antecedentes familiares de cáncer, domicilio, tipo de cáncer, estadificación, tratamiento, metástasis, recurrencias, situación del paciente al final de la investigación. Se aplicó estadística descriptiva, chi-cuadrado y Kaplan-Meier. Resultados: Fueron más prevalentes: niños (55,22%), grupo de edad 1 a 4 años (36,32%), blancos (87,06%), hogares urbanos (81,59%), leucemia (35,83) %) y quimioterapia (50,25%). Hubo metástasis en 16,41% y recidiva en 22,38%. No hubo informes de antecedentes familiares de cáncer en 47% de los pacientes. Se negaron comorbilidades en 58,91%. Al final, 55,23% estaban vivos y sin enfermedad. Hubo asociación estadística entre menores de 10 años con tumores renales y neuroblastoma; mayores 10 años con linfomas y neoplasias epiteliales malignas; e entre la situación del paciente con metástasis, recaídas y estadificación. Conclusión: Los pacientes analizados en esta investigación eran en su mayoría leucémicos, varones y de 1 a 4 años. La supervivencia global y libre de enfermedad fueron, respectivamente, 70,3% y 71,63%

Exploring GRHL3 polymorphisms and SNP-SNP interactions in the risk of non-syndromic oral clefts in the Brazilian population.

OBJECTIVE: To investigate the association of single-nucleotide polymorphisms (SNP) in grainyhead-like 3 (GRHL3) and to verify its possible interactions with others genes responsible for craniofacial development in the risk of non-syndromic oral cleft (NSOC). METHODS: Applying TaqMan allelic discrimination assays, we evaluated GRHL3 SNPs (rs10903078, rs41268753, and rs4648975) in an ancestry-structured case-control sample composed of 1,127 Brazilian participants [272 non-syndromic cleft palate only (NSCPO), 242 non-syndromic cleft lip only (NSCLO), 319 non-syndromic cleft lip and palate (NSCLP), and 294 healthy controls]. Additionally, SNP-SNP interactions of GRHL3 and previously reported variants in FAM49A, FOXE1, NTN1, and VAX1 were verified in non-syndromic cleft lip with or without cleft palate (NSCL ± P). To eliminate false-positive associations, Bonferroni correction or 1,000 permutation method was applied. RESULTS: The multiple logistic regression analysis showed that the CC genotype of rs10903078 (p = .03) and the haplotype C-C formed by the SNPs rs10903078 and rs41268753 (p = .04) were associated with NSCLO, but the p-values did not withstand Bonferroni correction. However, SNP-SNP test revealed significant interactions between GRHL3 SNPs and FAM49A (rs7552), FOXE1 (rs3758249), VAX1 (rs7078160 and rs751231), and NTN1 (rs9891446). CONCLUSIONS: Our results confirm the importance of GRHL3 and its interactions with previously NSOC-associated genes, including FAM49A, FOXE1, NTN1, and VAX1, in the pathogenesis of NSOC in the Brazilian population.

Patterns of dental anomalies in patients with nonsyndromic oral cleft

Aim: To characterize the patterns of dental anomalies (DA) in the mixed and permanent dentitions of patients with nonsyndromic oral cleft (NSOC). Methods: This cross-sectional, observational, case-control study included 173 patients, 61 with mixed dentition (NSOC=29 and control=32) and 112 with permanent dentition (NSOC=57 and control=55). All subjected were submitted to clinical and radiographic examination. Dental anomalies of eruption, number, size and shape outside the cleft area were considered. Results: Although there was no statistical significance among patients with mixed dentition, dental agenesis was the anomaly more common in this group. In patients with permanent dentition, a higher prevalence of DA in NSOC group compared to control group was observed (p=0.02). Gyroversion and dental agenesis were the DA more frequently observed in the permanent dentition and the second premolar was the tooth more affected (p=0.003). Mandible and the left side were more involved, and dental agenesis was more frequently found in patients with unilateral cleft lip with or without cleft palate (NSCL±P). Conclusion: Our findings show a higher frequency of DA in NSOCs than in the control group in patients with permanent dentition, mainly due to a higher occurrence of agenesis of second premolars in patients with unilateral NSCL±P

Odontogenic Keratocysts as First Manifestation of Nevoid Basal Cell Carcinoma Syndrome: Surgical Management and Immunohistochemical Analysis.

Nevoid Basal Cell Carcinoma Syndrome (NBCCS), also known as Gorlin syndrome, is a rare autosomal dominant disorder, with no gender predilection. Individuals with NBCCS are commonly diagnosed between 17 and 35 years old and can present multiple basal cell carcinomas scattered throughout the body, presence of recurrent and early-onset odontogenic keratocysts (OKCs) and skeletal abnormalities. This article describes a case of a 13-year-old white boy who referred complaining of facial asymmetry. Extraoral examination revealed volumetric increase displacing the nasal ala from the right side and extended to the zygomatic bone. The intraoral evaluation showed mixed dentition with moderate degree of malocclusion. In addition, bilateral vestibular fornix swelling was observed in the upper canine region. An increase in volume was also detected on the hard palate on the right side. Computed tomography revealed multiple hypodense lesions with cystic appearance. The aspiration was positive, with a yellowish aspirate of serous consistency of all lesions. Given the numerous lesions, it was decided to decompress them for posterior enucleation procedures. In addition to other manifestations, the patient was diagnosed with NBCCS. Although common, the occurrence of OKCs in pediatric patients, especially in multiple lesions, is highly indicative of NBCCS, and its investigation should be considered, even in the absence of other signs of this syndrome. Synchronous decompression was satisfactory and can be used in similar cases of multiple cystic lesions.

Immunoexpression of hoxb7 and hoxb9 in salivary gland tumours.

BACKGROUND: Salivary gland carcinomas are uncommon neoplasms and the identification of new prognostic indicators could improve their management. HOXB7 and HOXB9 are members of the class I homeobox-containing genes important for normal embryogenesis and that are dysregulated in several human neoplasms. This study investigated HOXB7 and HOXB9 expressions in salivary gland tumourigenesis, their correlation with neoplastic proliferative and angiogenic features and their importance as prognostic markers. METHODS: A hundred and fifty salivary gland tumours were organized in tissue microarray and expressions of CD105, Ki67, HOXB7 and HOXB9 were determined through immunohistochemistry. Reactions were quantified and correlated with clinicopathological parameters. RESULTS: In normal glands, HOXB7 was found in basal cells, whereas HOXB9 was seen in serous acinar and scattered ductal cells. Malignancies exhibited an increased vascular density, proliferative index, HOXB7 and HOXB9 expressions when compared with pleomorphic adenoma and Warthin's tumour. Significant correlation was found between HOXB7 and CD105 (P = 0.004) in adenoid cystic carcinomas, and HOXB7 higher expression significantly correlated with the presence of paresthesia (P = 0.02). No marker exhibited a significant association with survival rates (P > 0.05). CONCLUSION: HOXB7 and HOXB9 were expressed in normal salivary gland and were present in benign and malignant tumours derived from these structures, and HOXB7 significantly correlated with neoangiogenesis in AdCC. These findings suggest that both proteins might play a role in salivary gland tumourigenesis, but they were not significant prognostic determinants in this sample.

Epstein-Barr virus (EBV)--associated posttransplant lymphoproliferative disorder appearing as mandibular gingival ulcers.

Posttransplant lymphoproliferative disorders (PTLDs) comprise a spectrum of complications that affect immunocompromised patients following hematopoietic stem cell transplantation or solid organ transplantation. Its incidence varies depending on the transplanted organ, occurring in approximately 2.3% of kidney transplantations. A 31-year-old woman was referred to the Dental Clinic of the State University of Western Paraná for evaluation of an oral lesion. Her medical history revealed a previous diagnosis of hypertension, Epstein-Barr virus (EBV) seropositivity, and kidney transplantation 12 years prior. She was under standard immunosuppressive therapy. Intraoral examination identified a gingival necrotic lesion with extension to the posterior right lower alveolar bone. An incisional biopsy was performed. Histologic examination showed lymphocytic proliferation of cells with small and hyperchromatic nuclei, atypical mitosis, and cells with large and pale nuclei showing prominent nucleoli permeating connective tissue, muscle fibers, and adipocytes. Correlation of clinical, histologic, and immunohistochemical findings led to a diagnosis of polymorphic EBV-associated PTLD rich in B and T cells.

Vascular endothelial growth factor immunoexpression is increased in malignant salivary gland tumors.

Salivary gland tumors (SGTs) consist of a heterogeneous group of lesions accounting for 3% to 10% of all head and neck neoplasms. Little is known about their angiogenic properties, and despite vascular endothelial growth factor (VEGF) has been previously studied in these lesions, further investigations are warranted to better determine its clinical and prognostic significance. In the current study, a total of 132 formalin-fixed, paraffin-embedded SGTs were organized in tissue microarray blocks and submitted to immunohistochemistry against VEGF protein. Slides were scanned and immunoreactions analyzed using Pixelcount V9 algorithm (Aperio Technologies Inc, Vista, CA, USA). Clinical and follow-up data were retrieved from patients' medical charts. Tumors included 50 cases of pleomorphic adenoma, 32 mucoepidermoid carcinomas, 30 adenocarcinomas not otherwise specified, and 20 adenoid cystic carcinomas. A slight male preponderance was found (1.1:1.0), with a mean age of 47.5 years. Parotid gland was the most affected location. Vascular endothelial growth factor expression was found in the cytoplasm of all cases analyzed with variable intensity, proving to be overexpressed in malignant tumors if compared with pleomorphic adenoma. A significant correlation of VEGF reactivity was found only with age, showing no further significant associations. Age and presence of paresthesia were the only features that predicted a lower specific survival rate under univariate and multivariate analyses. Log-rank test evidenced VEGF high expression as a potential determinant of reduced survival, although a statistical significance could not be reached. Hence, considering VEGF overexpression in malignant tumors and its potential association with a lower survival rate, this protein might be associated with SGTs pathogenesis and aggressiveness.

Clinicopathologic analysis of 493 cases of salivary gland tumors in a Southern Brazilian population.

OBJECTIVE: The aim of this study was to determine the distribution and demographic features of salivary gland tumors (SGTs) in a large Brazilian population. STUDY DESIGN: A total of 493 cases of SGTs diagnosed between 2001 and 2011 from a general pathology laboratory and an oral pathology service were reviewed with respect to their clinicopathologic features. RESULTS: A total of 369 tumors were benign and 124 were malignant. The mean age of patients with benign tumors was 46.3 years and that of patients with malignancies was 54.0 years. The parotid gland was the most common location (42.3%). Pleomorphic adenoma (PA) and Warthin's tumor were the most common benign neoplasias, whereas mucoepidermoid carcinoma (MEC) and adenocarcinoma, not otherwise specified, were the most frequent malignancies. CONCLUSIONS: The present data confirm that PA and MEC are the most common benign and malignant SGTs. However, it is important to consider that differences in tumor types may be influenced by whether a tumor derives from a medical or a dental service.