Analysis of multivitamin supplements and polymorphisms in the folic acid metabolic pathway enzymes in mothers of nonsyndromic cleft lip-palate individuals

Orientador: Ricardo Della ColettaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: A fissura labial e/ou palatina (FL/P) não-sindrômica é uma malformação congênita do lábio e/ou palato com alta frequência na população brasileira. A etiologia das fissuras é complexa e conta com a participação de fatores genéticos e ambientais. Inúmeros estudos demonstraram que variantes polimórficas das enzimas relacionadas ao metabolismo do ácido fólico podem ser importantes fatores de risco materno para o nascimento de uma criança FL/P não-sindrômica. O objetivo deste estudo foi estudar a influência do consumo de suplementos vitamínicos durante o primeiro trimestre de gravidez e comparar a frequência alélica e genotípica de 4 genes (MTHFR, MTHFD1, MTR e RFC1) que codificam enzimas da via metabólica do ácido fólico entre mães de indivíduos portadores de FL/P não-sindrômicas (grupo experimental) e mães de indivíduos clinicamente normais (grupo controle). Amostras de DNA de 184 mães do grupo controle e de 106 mães do grupo experimental foram genotipadas por reação em cadeia da polimerase associada à análise de polimorfismo de fragmentos de restrição enzimática (PCR-RFLP). A ausência de suplemento vitamínico durante o primeiro trimestre de gravidez aumentou de forma discreta (aproximadamente em 0,4 vezes) o risco de uma mulher ter um filho com FL/P não-sindrômica. Dos 15 polimorfismos analisados neste estudo, 2 apresentaram diferenças entre os grupos. No polimorfismo rs2274976 do gene MTHFR, o alelo A e o genótipo GA ocorreram em uma frequência significantemente maior no grupo experimental que no grupo controle (p<0,000001), aumentando em aproximadamente 6 vezes o risco de uma mãe ter um filho com FL/P não-sindrômica. O genótipo AA no lócus polimórfico rs2236225 do gene MTHFD1 foi significantemente mais prevalente no grupo experimental comparado com o grupo controle (p=0,02). A presença deste genótipo aumentou em aproximadamente 2 vezes o risco de uma mãe ter um filho com FL/P não-sindrômica. Análise multivariada demonstrou que estes fatores contribuíram de maneira independente para a etiologia das FL/P não-sindrômicas. O presente estudo demonstra que os polimorfismos rs2274976 do gene MTHFR e rs2236225 do gene MTHFD1 e a suplementação vitamínica durante o primeiro trimestre de gravidez estão associados ao desenvolvimento de FL/P não-sindrômicas na população brasileira. Este estudo corrobora com evidências prévias que demonstraram a influência de fatores ambientais e genéticos na etiopatogenia das FL/P não-sindrômicas.Abstract: Nonsyndromic cleft lip with or without cleft palate (CL/P) is a congenital malformation of the lip and/or palate with elevating frequency in the Brazilian population. The etiology of the nonsyndromic CL/P is complex and both environmental and genetic factors play important roles. Several studies demonstrated that polymorphisms in the folic acid metabolic enzymes may be important maternal risk factor for the birth of a child with nonsyndromic CL/P. The aim of this study was to determine the influence of the multivitamin supplements during the first trimester of pregnancy and to compare the allele and genotypic frequencies of 4 genes (MTHFR, MTHFD1, MTR and RFC1) that encode enzymes of the acid folic metabolic pathway between mothers of nonsyndromic CL/P patients (experimental group) and mothers of clinically normal children (control group). DNA samples from 184 mothers of the control group and from 106 mothers of the experimental group were genotyped by polymerase chain reaction associated with reaction fragment length polymorphism (PCR-RFLP). The lack of multivitamin supplementation during the pregnancy first trimester increased in approximately 0.4-fold the maternal risk of a nonsyndromic CL/P child. Two out of 15 polymorphisms showed differences between groups. In rs2274976 MTHFR polymorphism, allele A and genotype GA occurred in a significantly higher frequency on experimental group when compared to control group (p<0.000001), rising in approximately 6 times the risk of a mother giving birth to a nonsyndromic CL/P child. Genotype AA in the rs2236225 MTHFD1 polymorphic locus was significantly more prevalent in experimental group than in control group (p=0.02). This genotype raised in approximately twice the risk of a mother giving birth to a nonsyndromic CL/P child. Multivariate analysis demonstrated that those factors contributed in an independent manner to nonsyndromic CL/P etiology. The present study shows that rs2274976 MTHFR and rs2236225 MTHFD1 polymorphisms, as well as the multivitamin supplementation during the first trimester of pregnancy, are associated with the development of nonsyndromic CL/P in the Brazilian population. This study corroborates with previous evidences demonstrating the influence of environmental and genetic factor on etiopathogenesis of the nonsyndromic CL/P.MestradoPatologiaMestre em Estomatopatologi

Suppurative minor salivary gland sialolithiasis

Sialolithiasis is a common nonneoplastic disease of the major salivary glands that often affects the submandibular glands. Minor salivary gland involvement by sialolithiasis is uncommon, with only 273 cases reported. A long clinical history, acute symptoms, and mucopurulent discharge are unusual features of these cases. Herein, we report the case of a 63-year-old woman who complained of symptomatic nodular swelling of the buccal mucosa associated with purulent discharge for several days. The clinical history lasted 15 years, with episodes of asymptomatic non-suppurative swelling in the same area. The patient underwent surgical excision. The microscopic examination revealed chronic nonspecific sialadenitis associated with psammomatous calcifications, confirming minor salivary gland sialolithiasis. After 3 years of follow-up, the patient was free of symptoms. Patients with sialolithiasis are usually asymptomatic; however, swelling, pain, and fistula may be present in rare cases. The presence of purulent exudate should lead to the differential diagnosis of stomatitis glandularis, a rare inflammatory condition affecting the minor salivary glands. Sialolithiasis and stomatitis glandularis should be considered in the clinical differential diagnosis of symptomatic suppurative nodular swelling affecting the oral mucosa, and histopathological analysis is necessary for the diagnosis

Myofibroblasts in the stroma of oral cancer promote tumorigenesis via secretion of activin A

SummaryMyofibroblasts are essential during wound healing and are often found in the stroma of oral squamous cell carcinomas (OSCC). Although the molecular mechanisms by which myofibroblasts influence OSCC remain largely unknown, previous studies demonstrated that presence of myofibroblast in OSCC stroma is an important risk factor of patient’s shortened survival. Here we showed that some growth factors are produced in higher levels by tumor-associated myofibroblasts compared to tumor-associated fibroblasts, including activin A. Myofibroblast-conditioned media containing activin A significantly increased OSCC cell proliferation and tumor volume, whereas down-regulation of activin A in the conditioned media decreased proliferation. In addition, myofibroblasts induced in vitro invasion of OSCC cells, which was accompanied by an increased production of matrix metalloproteinases (MMP). In vivo, a significant correlation between presence of myofibroblasts and activities of MMP-2 and MMP-9 was observed in OSCC samples. However, blockage of activin A synthesis by myofibroblasts did not affect invasion and MMP production by OSCC cells. Together, our data demonstrate that activin A is required for the proliferative effects of myofibroblasts on OSCC cells. We conclude that myofibroblasts in the stroma of OSCC may influence proliferation and invasion, resulting in more aggressive tumor

Chronic ulcerative stomatitis : a systematic review of the clinical and microscopic features

the purpose of this study was to perform a systematic review regarding clinical and histopathological characteristics, immunopathological findings, and treatment for chronic ulcerative stomatitis (CUS).  articles in English, published from January 1962 up to November 2017, assessing clinical and immunological features, treatment, and follow-up of patientes with CUS, were retrieved from three databases (PubMed, Cochrane Library and SCOPUS). A manual literature search was also conducted. A total of 12 studies met inclusion criteria, therefore, were analyzed in this review. CUS shares similiar clinical and microscopic features to those found in oral lichen planus (OLP) and oral lichenoid lesions (OLL). Hence, direct immunofluorescence (DIF) is indispensable to define a final diagnosis. Due to the poor sample availability in the current literature, it is not possible to accurately confirm the prevalence and features of CUS. in order to better evaluate this condition?s findings, further studies with a greater amount of similar immune-mediated diseases should be performed

Photobiomodulation in the treatment of xerostomia associated with hyposalivation in a pediatric patient with systemic scleroderma

Scleroderma is a rare autoimmune disease characterized by excessive collagen production. The oral manifestations of the patient with scleroderma can include microstomia, xerostomia, and changes in the resorption teeth. We report the case of a 7-year-old female patient diagnosed with systemic scleroderma where photobiomodulation therapy was used to treat xerostomia associated with hyposalivation. She attended a pediatric clinic and presented with dry and rigid facial skin, trismus, xerostomia, malocclusion, and difficulty swallowing. Stimulated salivary flow was assessed before, during, and after treatment. Photobiomodulation therapy was conducted at four points at the sublingual glands with 660 nm, 100 mW, and 0.8 J/cm2 to each point; eight points at the parotid glands; and six points at the submandibular glands with 808 nm, 100 mW, and 0.8 J/cm2 for 8 seconds at each point. After this therapy, an increase in salivary flow, remission of the xerostomia, and an improvement in mastication and swallowing were observed. Photobiomodulation therapy was effective in controlling xerostomia in this pediatric patient, resulting in increased salivary flow and an improvement in her quality of life

Intraoral lipoma with degenerative changes mimicking atypical lipomatous tumor: an immunohistochemical study

Lipomas are mesenchymal neoplasms relatively uncommon in the oral cavity. Lipomas can exhibit histopathological features mimicking atypical lipomatous tumors (ALT) or dysplastic lipoma (DL) in the presence of degenerative changes. Relevantly, immunohistochemistry assists in the correct diagnosis. Herein, we present the case of a 54-year-old male with a sessile nodule located on the dorsum of the tongue. The histopathological analysis showed a diffuse, non-circumscribed adipocytic proliferation constituted by cells of variable size containing cytoplasmic vacuoles and displaced nuclei, some resembling lipoblasts supported by fibrous connective tissue stroma. By immunohistochemistry, tumor cells were positive for vimentin, S100, FASN, CD10, and p16. Rb expression was intact. Moreover, CD34, p53, MDM2, and CDK4 were negative. After 2-year of follow-up, no alteration or recurrence was observed. In conclusion, MDM2, CDK4, p53, and Rb immunomarkers can be used reliably to differentiate benign lipoma with degenerative changes from ALT and DL

Post-radiotherapy recurrence of conventional oral squamous cell carcinoma showing sarcomatoid components: an immunohistochemical study

Spindle cell squamous cell carcinoma (SpSCC) is a rare biphasic malignant neoplasm, uncommonly affecting the oral cavity. The SpSCC diagnosis is difficult, especially when it exhibits inconspicuous morphology, inadequate tissue sampling, or association with an exuberant inflammatory reaction. Post-radiotherapy recurrent SpSCC occurring at the same site of conventional SCC is a rare phenomenon. A 59-year-old man was complained of “painful injury on the tongue” with 20 days of duration. He reported smoking and alcohol consumption. Medical history revealed conventional SCC on the tongue treated with surgery and radiotherapy 10 years ago. Intraoral examination showed a polypoid lesion with ulcerated areas, measuring 3 cm in diameter, on the tongue and floor of the mouth, at the same site of previous conventional SCC. The microscopical analysis showed small foci of carcinomatous component admixed with an exuberant inflammatory reaction. Immunohistochemistry highlighted the sarcomatoid component. Both malignant components were positive for EMA, CD138, p40 (deltaNp63), p63, and p53. Moreover, CK AE1/AE3 evidenced the carcinomatous component, whereas vimentin stained the sarcomatoid component. The Ki-67 was &gt;10%. The current case emphasizes the importance of immunohistochemistry in the differential diagnosis of SpSCC from mimics and documents a rare complication of Ionizing Radiation

EBV-negative lymphoepithelial-like carcinoma of the lower lip

Lymphoepithelial-like carcinoma (LEC) is a rare malignant neoplasm, which can be associated with Epstein-Barr virus (EBV) infection. Histologically, LEC is an undifferentiated carcinoma with an intermixed reactive lymphoplasmacytic infiltrate. LEC appears to be an uncommon tumor type of lip carcinoma. An 82-year-old white woman presented a lesion on her lower lip that developed over the last year. The lesion was characterized by ulceration with flat edges, hardened base, painful, and absence of regional lymphadenopathy. Microscopical analysis evidenced an intense inflammatory infiltrate, composed of lymphoplasmacytic cells, associated with scarce pleomorphic epithelial cells. Immunohistochemistry highlighted the LEC cells with strong expression of pan-CK AE1/AE3, EMA, p63, and p53. CD138 was also faintly positive. Ki-67 was &gt;85%. In situ hybridization analysis did not show evidence of EBV. A diagnostic of EBV-negative LEC was made. We present an uncommon type of lip carcinoma, which can represent a diagnostic challenge for clinicians and pathologists

Contribution of polymorphisms in genes associated with craniofacial development to the risk of nonsyndromic cleft lip and/or palate in the Brazilian population

Background and Objective: Nonsyndromic cleft lip and/or palate (NSCL/P) is a complex disease associated with both genetic and environmental factors. One strategy for identifying of possible NSCL/P genetic causes is to evaluate polymorphic variants in genes involved in the craniofacial development. Design: We carried out a case-control analysis of 13 single nucleotide polymorphisms in 9 genes related to craniofacial development, including TBX1, PVRL1, MID1, RUNX2, TP63, TGFB3, MSX1, MYH9 and JAG2 , in 367 patients with NSCL/P and 413 unaffected controls from Brazil to determine their association with NSCL/P. Results: Four out of 13 polymorphisms (rs28649236 and rs4819522 of TBX1, rs7940667 of PVRL1 and rs1057744 of JAG2 ) were presented in our population. Comparisons of allele and genotype frequencies revealed that the G variant allele and the AG/GG genotypes of TBX1 rs28649236 occurred in a frequency significantly higher in controls than in the NSCL/P group (OR: 0.41; 95% CI: 0.25-0.67; p=0.0002). The frequencies of rs4819522, rs7940667 and rs1057744 minor alleles and genotypes were similar between control and NSCL/P group, without significant differences. No significant associations among cleft types and polymorphisms were observed. Conclusion: The study suggests for the first time evidences to an association of the G allele of TBX1 rs28649236 polymorphism and NSCL/P

Cathepsin K Is Present in Invasive Oral Tongue Squamous Cell Carcinoma In Vivo and In Vitro

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