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1.
Genet Mol Res ; 14(4): 14979-88, 2015 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-26634459

RESUMO

The leptin gene product is released into the blood stream, passes through the blood-brain barrier, and finds the leptin receptor (LEPR) in the central nervous system. This hormone regulates food intake, hematopoiesis, inflammation, immunity, differentiation, and cell proliferation. The LEPR Gln223Arg polymorphism has been reported to alter receptor function and expression, both of which have been related with prognostics in several tumor types. Furthermore, several studies have shown a relationship between the Gln223Arg polymorphism and tumor development, and its role in oral and oropharyngeal squamous cell carcinoma is now well understood. In this study, 315 DNA samples were used for LEPR Gln223Arg genotyping and 87 primary oral and oropharyngeal squamous cell carcinomas were used for immunohistochemical expression analysis, such that a relationship between these and tumor development and prognosis could be established. Homozygous LEPR Arg223 was found to be associated with a 2-fold reduction in oral and oropharyngeal cancer risk. In contrast, the presence of the Arg223 allele in tumors was associated with worse disease-free and disease-specific survival. Low LEPR expression was found to be an independent risk factor, increasing the risk for lymph node metastasis 4-fold. In conclusion, the Gln223Arg polymorphism and LEPR expression might be valuable markers for oral and oropharyngeal cancer, suggesting that LEPR might serve as a potential target for future therapies.


Assuntos
Biomarcadores Tumorais/genética , Leptina/genética , Neoplasias Bucais/genética , Neoplasias Orofaríngeas/genética , Polimorfismo Genético/genética , Receptores para Leptina/genética , Adulto , Idoso , Alelos , Substituição de Aminoácidos , Barreira Hematoencefálica , Índice de Massa Corporal , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Fatores de Risco
2.
Genet Mol Res ; 14(4): 15123-32, 2015 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-26634475

RESUMO

Five-year survival rates for oral squamous cell carcinoma (OSCC) are 30% and the mortality rate is 50%. Immunohistochemistry panels are used to evaluate proliferation, vascularization, apoptosis, HPV infection, and keratin expression, which are important markers of malignant progression. Keratins are a family of intermediate filaments predominantly expressed in epithelial cells and have an essential role in mechanical support and cytoskeleton formation, which is essential for the structural integrity and stability of the cell. In this study, we analyzed the expressions of keratins 17 and 19 (K17 and K19) by immunohistochemistry in tumoral and non-tumoral tissues from patients with OSCC. The results show that expression of these keratins is higher in tumor tissues compared to non-tumor tissues. Positive K17 expression correlates with lymph node metastasis and multivariate analysis confirmed this relationship, revealing a 6-fold increase in lymph node metastasis when K17 is expressed. We observed a correlation between K17 expression with disease-free survival and disease-specific death in patients who received surgery and radiotherapy. Multivariate analysis revealed that low expression of K17 was an independent marker for early disease relapse and disease-specific death in patients treated with surgery and radiotherapy, with an approximately 4-fold increased risk when compared to high K17 expression. Our results suggest a potential role for K17 and K19 expression profiles as tumor prognostic markers in OSCC patients.


Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/metabolismo , Queratinas/metabolismo , Neoplasias Bucais/metabolismo , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/terapia , Intervalo Livre de Doença , Células Epiteliais/metabolismo , Feminino , Humanos , Metástase Linfática/patologia , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/patologia , Neoplasias Bucais/terapia , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/patologia , Prognóstico
3.
Genet Mol Res ; 14(3): 10515-23, 2015 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-26400283

RESUMO

Death-associated protein 1 (DAP1) is a member of the DAP family. Its expression is associated with cell growth and normal death of the neoplastic cells, regulated by the mammalian target of the rapamycin protein. Activated DAP1 negatively regulates autophagy, which has been associated with the development and progression of several diseases, such as cancer, and with prognosis and survival of diverse tumor types. Therefore, in this study we analyzed DAP1 expression in 54 oral squamous cell carcinoma tumor samples and in 20 non-tumoral margins by immunohistochemistry. The results showed that DAP1 is more frequently expressed in tumor tissues compared with marginal non-tumoral cells. Additionally, high DAP1 expression is associated with a 4-fold increase in the risk of lymph node metastases. Our results suggest that the DAP1 protein can be used as a potential marker of lymph node metastases predisposition, helping define the best therapy for each patient to minimize risk of developing metastases.


Assuntos
Proteínas Reguladoras de Apoptose/genética , Carcinoma de Células Escamosas/genética , Predisposição Genética para Doença , Linfonodos/patologia , Neoplasias Bucais/genética , Adulto , Idoso , Proteínas Reguladoras de Apoptose/metabolismo , Autofagia/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Feminino , Expressão Gênica , Humanos , Imuno-Histoquímica , Linfonodos/metabolismo , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Boca/metabolismo , Boca/patologia , Neoplasias Bucais/metabolismo , Neoplasias Bucais/mortalidade , Neoplasias Bucais/patologia , Estadiamento de Neoplasias , Prognóstico , Risco , Análise de Sobrevida
4.
Genet Mol Res ; 14(3): 11145-53, 2015 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-26400345

RESUMO

Currently, the most important prognostic factor in oral squamous cell carcinoma (OSCC) is the presence of regional lymph node metastases, which correlates with a 50% reduction in life expectancy. We have previously observed that expression of hypoxia genes in the tumor inflammatory infiltrate is statistically related to prognosis in OSCC. FAS and FASL expression levels in OSCC have previously been related to patient survival. The present study analyzed the relationship between FASL expression in the inflammatory infiltrate lymphoid cells and clinical variables, tumor histology, and prognosis of OSCC. Strong FASL expression was significantly associated with lymph node metastases (P = 0.035) and disease-specific death (P = 0.014), but multivariate analysis did not confirm FASL expression as an independent death risk factor (OR = 2.78, 95%CI = 0.81-9.55). Disease-free and disease-specific survival were significantly correlated with FASL expression (P = 0.016 and P = 0.005, respectively). Multivariate analysis revealed that strong FASL expression is an independent marker for earlier disease relapse and disease-specific death, with approximately 2.5-fold increased risk compared with weak expression (HR = 2.24, 95%CI = 1.08-4.65 and HR = 2.49, 95%CI = 1.04-5.99, respectively). Our results suggest a potential role for this expression profile as a tumor prognostic marker in OSCC patients.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Proteína Ligante Fas/metabolismo , Neoplasias Bucais/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/secundário , Feminino , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Linfócitos do Interstício Tumoral/metabolismo , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/mortalidade , Neoplasias Bucais/patologia , Análise Multivariada , Prognóstico , Modelos de Riscos Proporcionais
5.
Lasers Med Sci ; 29(2): 813-21, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23982721

RESUMO

The treatment of muscle injuries is a common practice at rehabilitation centers. Low-level laser therapy (LLLT) has demonstrated positive effects regarding the modulation of the inflammatory response, the enhancement of the tissue repair process and the prevention of fibrosis. The aim of the present study was to evaluate the effects of LLLT on morphological aspects of muscle tissue, collagen remodeling and activity of matrix metalloproteinase 2 (MMP-2) in rat skeletal muscle following acute injury. Wistar rats were divided into five groups: (1) control group (n = 10), (2) sham group (n = 10), (3) LLLT group (n = 30), (4) non-treated injury group (n = 30) and (5) injury + LLLT group (n = 30). Cryoinjury was performed on the belly of the tibialis anterior (TA) muscle. LLLT was performed daily with an AlGaAs laser (780 nm; beam spot of 0.04 cm(2), output power of 40 mW, power density of 1 W/cm(2), energy density of 10 J/cm(2) and 10-s exposure time). Animals were euthanized at 1, 3 and 7 days. The TA muscles were removed and weighed. Morphological aspects were evaluated using H & E staining. The amount and distribution of collagen fibers were evaluated by picrosirius staining. Characterization and activity of MMP-2 were evaluated by zymography and Western blot techniques, respectively. The results revealed that LLLT induced a reduction in inflammatory infiltrate and myonecrosis after 1 day, an increase in the number of blood vessels after 3 and 7 days as well as an increase in the number of immature muscle fibers and MMP-2 gelatinase activity after 7 days. In conclusion, LLLT has a positive effect on the inflammatory process, MMP2 activity and collagen organization and distribution in the repair process of rat skeletal muscle.


Assuntos
Terapia com Luz de Baixa Intensidade/métodos , Músculo Esquelético/lesões , Tíbia/patologia , Animais , Colágeno/metabolismo , Fibrose/radioterapia , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Músculo Esquelético/patologia , Ratos Wistar
6.
BMC Med Genomics ; 3: 14, 2010 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-20441585

RESUMO

BACKGROUND: The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression. METHODS: The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells. RESULTS: We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (ARID4A, CALR, GNB2L1, RNF10, SQSTM1, USP9X) were validated by real time PCR. CONCLUSIONS: A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.


Assuntos
Regulação Neoplásica da Expressão Gênica , Neoplasias Bucais/metabolismo , Proteoma/metabolismo , Anexina A5/metabolismo , Apoptose , Proliferação de Células , Regulação para Baixo , Eletroforese em Gel Bidimensional , Fibroblastos/metabolismo , Genômica , Células Hep G2 , Humanos , Queratinas/metabolismo , Neoplasias Bucais/genética , Hibridização de Ácido Nucleico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Células Estromais/metabolismo , Vimentina/metabolismo
7.
Oral Oncol ; 45(2): 121-6, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18621570

RESUMO

The selection of housekeeping genes is critical for gene expression studies. To address this issue, four candidate housekeeping genes, including several commonly used ones, were investigated in oral squamous cell carcinoma cell lines. A simple quantitative RT-PCR approach was employed by comparing relative expression of the four candidate genes within two cancerous cell lines (HN6 and HN31) and one noncancerous cell line (HaCaT) treated or not with EGF and TGF-beta1. Data were analyzed using ANOVA followed by the NormFinder software program. On this basis, stability of the candidate housekeeping genes was ranked and non statistical differences were found using ANOVA test. On the other hand, the NormFinder was able to show that GAPD and TUBB presented the less variable results, representing appropriated housekeeping genes for the samples and conditions analyzed. In conclusion, this study suggests that the GAPD and the TUBB represent adequate normalizers for gene profiling studies in OSCC cell lines, covering, respectively, high and low expression levels genes.


Assuntos
Carcinoma de Células Escamosas/genética , Perfilação da Expressão Gênica/métodos , Gliceraldeído-3-Fosfato Desidrogenases/genética , Neoplasias Bucais/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Tubulina (Proteína)/genética , Linhagem Celular Tumoral , Humanos , Padrões de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas
8.
Oral Dis ; 14(6): 510-3, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18826382

RESUMO

AIM: To investigate the immunoexpression and diagnostic applicability of human erythrocyte-type glucose transporter protein (GLUT-1) in oral peripheral nerve sheath tumors. MATERIAL AND METHODS: Specimens diagnosed as oral peripheral nerve sheath tumors archived in the Oral Pathology Service of Universidade Federal de Minas Gerais from 1966 to 2006 were evaluated. Thirty-four lesions were included: 15 traumatic neuromas, 11 neurofibromas, four neurilemmomas, and four malignant peripheral nerve sheath tumors (MPNST). One case of neurofibroma was associated with neurofibromatosis type I. Immunohistochemistry for S-100 and GLUT-1 was performed. S-100 was immunopositive in all lesions. RESULTS: Benign lesions were immunopositive for GLUT-1 except in two (18.2%) cases of neurofibromas. In the traumatic neuroma, the perineuriums were immunopositive for GLUT-1. In the neurofibroma, the immunoreactivity was heterogeneous. Immunopositivity was observed at levels of 54.5% in the periphery of the lesion, 9.1% in the center, and 18.2% in both. The neurilemmoma demonstrated immunopositivity in the capsule. One case (25%) of MPNST presented GLUT-1 positive stain in occasional cells distributed homogeneously in all the tumor area. CONCLUSION: GLUT-1 is a useful marker for perineurial cells and should be included in the oral peripheral nerve sheath tumors immunophenotyping thus aiding in the correct diagnosis of these lesions.


Assuntos
Transportador de Glucose Tipo 1/análise , Neoplasias Bucais/patologia , Neoplasias da Bainha Neural/patologia , Biomarcadores Tumorais/análise , Núcleo Celular/patologia , Tecido Conjuntivo/patologia , Diagnóstico Diferencial , Humanos , Imuno-Histoquímica , Imunofenotipagem , Fibras Nervosas/patologia , Neurilemoma/patologia , Neurofibroma/patologia , Neurofibromatose 1/patologia , Neuroma/patologia , Proteínas S100/análise
9.
Oral Dis ; 13(6): 564-9, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17944673

RESUMO

OBJECTIVES: Plasmablastic lymphoma (PBL) of the oral cavity is an aggressive neoplasm derived from B cell, considered to be the second more common among human immunodeficiency virus (HIV)-associated malignancies. As Epstein-Barr virus (EBV) infection has been associated with this neoplasm, the aim of the present study was to assess the presence of EBV in 11 cases of oral HIV-related PBL and investigate the controversial issue of the presence of Human herpesvirus-8 (HHV-8) in these tumors. METHODS: DNA was extracted from nine cases of HIV-associated oral lymphomas, diagnosed as PBL, and genomic material was amplified by polymerase chain reaction to verify the presence of EBV. In situ hybridization (ISH) for EBV was performed in five cases. Immunohistochemical analysis was conducted to confirm previous diagnosis and verify HHV-8 infection. RESULTS: The 11 cases had diagnosis confirmed by immunohistochemical analysis. Only nine cases presented an adequate amount of DNA for analysis, and EBV was detected in seven of them. The five cases tested for EBV viral infection by ISH showed positive signals. All 11 cases were negative for HHV-8. CONCLUSION: The presence of EBV in all cases studied favors a direct role of this virus in the development of HIV-related PBL, and this finding could be considered when dealing with HIV patients.


Assuntos
DNA Viral/análise , Herpesvirus Humano 4/isolamento & purificação , Linfoma Relacionado a AIDS/virologia , Neoplasias Bucais/virologia , Adulto , Infecções por Vírus Epstein-Barr/virologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
10.
Oral Dis ; 12(4): 424-7, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16792730

RESUMO

BACKGROUND: Homeobox are a family of developmental genes involved in morphogenesis and cellular differentiation. Participation of homeobox within normal and malignant tissue has been recently discussed in the literature. OBJECTIVE: To analyze the presence of HOXB13 transcript expression in human minor salivary gland. MATERIAL AND METHODS: Ten-micrometer sections from frozen samples were evaluated employing non-radioactive in situ hybridization technique and HOXB13 mRNA probes. RESULTS: HOXB13 was found to be expressed in ducts and mucous acini but not in serous acini. CONCLUSIONS: RESULTS suggest that HOXB13 transcripts are differently expressed in normal mucous and serous acini, and it may possibly reflect a different role in salivary gland carcinogenesis.


Assuntos
Proteínas de Homeodomínio/biossíntese , Glândulas Salivares Menores/metabolismo , Carcinoma de Células Escamosas/cirurgia , Feminino , Expressão Gênica , Genes Homeobox/fisiologia , Humanos , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/metabolismo , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias das Glândulas Salivares/cirurgia , Análise de Sequência de RNA
11.
Br J Oral Maxillofac Surg ; 42(4): 363-4, 2004 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15225961

RESUMO

We present the case of a 65-year-old woman who had a painless mass in the left buccal mucosa. Histology showed a benign osteolipoma.


Assuntos
Lipoma/patologia , Mucosa Bucal/patologia , Neoplasias Bucais/patologia , Ossificação Heterotópica/patologia , Doenças Raras/patologia , Idoso , Feminino , Humanos
12.
Oral Oncol ; 40(3): 326-31, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-14747065

RESUMO

The expression of G1-phase cell-cycle regulators is commonly deregulated in human malignancies. In the present study, we investigate components of the retinoblastoma (RB) pathway in normal salivary glands (NSG) and in salivary gland tumours (SGT). Samples of NSG, pleomorphic adenoma (PA), adenoid cystic carcinoma (ACC), mucoepidermoid carcinoma (MEC), epithelial-myoepithelial carcinoma (EMC), malignant myoepithelioma (MEM), carcinoma ex pleomorphic adenoma (CEPA), and polymorphous, low-grade adenocarcinoma (PLGA) were examined immunohistochemically using antibodies to cyclin D1, cyclin-dependent kinase 4 (CDK-4), retinoblastoma protein (pRb), CDK inhibitor p16 and transcription factor E2F-1. In normal salivary glands, cyclin D1 and cdk-4 were not expressed in any case while p16 was positively expressed. pRb was abundant and E2F-1 moderately expressed. In tumors, cdk-4 was overexpressed in half of the cases. Most tumour cases showed decreased pRb immunoexpression compared to normal salivary glands. In contrast, expression of p16 and E2F-1 increased. pRb expression was absent in three cases of PA, two of EMC and one of CEPA. One case of MEM and one of PLGA showed no E2F-1 expression. Statistical analyses revealed positive correlations between cyclin D1 and cdk-4, cyclin D1 and E2F-1, cdk-4 and E2F-1, and p16 and E2F-1. The benign and malignant tumours expressed retinoblastoma pathway proteins differently form the normal salivary gland. Our findings suggest that, pRb pathway deregulation in salivary gland neoplasms is unrelated to their biological behaviour.


Assuntos
Proteínas de Ciclo Celular/metabolismo , Proteína do Retinoblastoma/metabolismo , Neoplasias das Glândulas Salivares/metabolismo , Glândulas Salivares/metabolismo , Ciclo Celular , Humanos , Neoplasias das Glândulas Salivares/patologia , Glândulas Salivares/citologia
13.
Int J Paediatr Dent ; 12(6): 446-9, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12452989

RESUMO

The oral findings of 12 patients with Moebius syndrome are described. Facial weakness, hypoplastic upper lip, microstomia, mouth-angle drooping, hypoplasia of mandible, gothic palate, tongue weakness, fissured tongue, tongue atrophy and open bite were found. In addition to describing the oral findings, this report discusses some implications for dental treatment of patients with this syndrome.


Assuntos
Síndrome de Möbius/complicações , Anormalidades da Boca/etiologia , Criança , Pré-Escolar , Cárie Dentária/etiologia , Feminino , Humanos , Masculino , Sialorreia/etiologia
14.
J Forensic Odontostomatol ; 20(1): 6-9, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12085524

RESUMO

Human identification can be accomplished by several technical procedures, especially by the comparative analysis of dental documents. Recent advances in molecular biology have now widened the scope for human identification and defined the protocols for the collection of biological material. Allele patterns of a population are particularly useful and they should be verified since they vary in different populations. We have studied the frequency of the alleles in three STR loci of the Y chromosome (DYS390, DYS391 and DYS393) in a group of Brazilian caucasian subjects. Results presented alleles 21, 22, 23, 24, 25 and 26 in locus DYS390, alleles 8, 9, 10, 11 12 and 13 in STR DYS391 and alleles in STR DYS393 were 11, 12, 13 14 and 15. The highest frequencies were 24 (0.46), 11 (0.37) and 13 (0.45). Each of the three STR systems had a PE (power of exclusion) of 0.6764 (DYS390), 0.5988 (DYS391) and 0.6136 (DYS393), The combination of the three STR systems revealed a PE of 0.9498, suggesting that this data can help in human identification.


Assuntos
Antropologia Forense/métodos , Cromossomo Y , Alelos , Brasil , Mapeamento Cromossômico , Bases de Dados Factuais , Frequência do Gene , Humanos , Masculino , Repetições Minissatélites , Reação em Cadeia da Polimerase
15.
Oral Oncol ; 38(1): 117-9, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11755832

RESUMO

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.


Assuntos
Lipossarcoma/patologia , Neoplasias da Língua/patologia , Idoso , Diagnóstico Diferencial , Humanos , Lipossarcoma/cirurgia , Masculino , Neoplasias da Língua/cirurgia , Resultado do Tratamento
16.
Pesqui Odontol Bras ; 15(4): 314-9, 2001.
Artigo em Português | MEDLINE | ID: mdl-11787320

RESUMO

There are several protocols reported in the literature for the extraction of genomic DNA from formalin-fixed paraffin-embedded samples. Genomic DNA is utilized in molecular analyses, including PCR. This study compares three different methods for the extraction of genomic DNA from formalin-fixed paraffin-embedded (inflammatory fibrous hyperplasia) and non-formalin-fixed (normal oral mucosa) samples: phenol with enzymatic digestion, and silica with and without enzymatic digestion. The amplification of DNA by means of the PCR technique was carried out with primers for the exon 7 of human keratin type 14. Amplicons were analyzed by means of electrophoresis in an 8% polyacrylamide gel with 5% glycerol, followed by silver-staining visualization. The phenol/enzymatic digestion and the silica/enzymatic digestion methods provided amplicons from both tissue samples. The method described is a potential aid in the establishment of the histopathologic diagnosis and in retrospective studies with archival paraffin-embedded samples.


Assuntos
DNA/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Humanos , Técnicas de Amplificação de Ácido Nucleico , Inclusão em Parafina
17.
Dev Biol ; 216(1): 369-81, 1999 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-10588886

RESUMO

Bone morphogenetic protein 4 (Bmp4) is expressed during multiple stages of development of the chicken inner ear. At the otocyst stage, Bmp4 is expressed in each presumptive sensory organ, as well as in the mesenchymal cells surrounding the region of the otocyst that is destined to form the semicircular canals. After the formation of the gross anatomy of the inner ear, Bmp4 expression persists in some sensory organs and restricted domains of the semicircular canals. To address the role of this gene in inner ear development, we blocked BMP4 function(s) by delivering one of its antagonists, Noggin, to the developing inner ear in ovo. Exogenous Noggin was delivered to the developing otocyst by using a replication-competent avian retrovirus encoding the Noggin cDNA (RCAS-N) or implanting beads coated with Noggin protein. Noggin treatment resulted in a variety of phenotypes involving both sensory and nonsensory components of the inner ear. Among the nonsensory structures, the semicircular canals were the most sensitive and the endolymphatic duct and sac most resistant to exogenous Noggin. Noggin affected the proliferation of the primordial canal outpouch, as well as the continual outgrowth of the canal after its formation. In addition, Noggin affected the structural patterning of the cristae, possibly via a decrease of Msx1 and p75NGFR expression. These results suggest that BMP4 and possibly other BMPs are required for multiple phases of inner ear development.


Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Orelha Interna/embriologia , N-Acetilglucosaminiltransferases , Proteínas/genética , Fatores de Transcrição , Animais , Apoptose/efeitos dos fármacos , Vírus do Sarcoma Aviário/genética , Proteína Morfogenética Óssea 4 , Proteínas Morfogenéticas Ósseas/genética , Bromodesoxiuridina , Proteínas de Transporte , Embrião de Galinha , Proteínas de Drosophila , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Proteínas de Homeodomínio/metabolismo , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Proteínas de Insetos/metabolismo , Fator de Transcrição MSX1 , Morfogênese/efeitos dos fármacos , Fenótipo , Proteínas/farmacologia , Receptores de Fator de Crescimento Neural/metabolismo , Canais Semicirculares/embriologia , Transfecção
18.
Development ; 125(1): 11-20, 1998 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9389659

RESUMO

A mature inner ear is a complex labyrinth containing multiple sensory organs and nonsensory structures in a fixed configuration. Any perturbation in the structure of the labyrinth will undoubtedly lead to functional deficits. Therefore, it is important to understand molecularly how and when the position of each inner ear component is determined during development. To address this issue, each axis of the otocyst (embryonic day 2.5, E2.5, stage 16-17) was changed systematically at an age when axial information of the inner ear is predicted to be fixed based on gene expression patterns. Transplanted inner ears were analyzed at E4.5 for gene expression of BMP4 (bone morphogenetic protein), SOHo-1 (sensory organ homeobox-1), Otx1 (cognate of Drosophila orthodenticle gene), p75NGFR (nerve growth factor receptor) and Msx1 (muscle segment homeobox), or at E9 for their gross anatomy and sensory organ formation. Our results showed that axial specification in the chick inner ear occurs later than expected and patterning of sensory organs in the inner ear was first specified along the anterior/posterior (A/P) axis, followed by the dorsal/ventral (D/V) axis. Whereas the A/P axis of the sensory organs was fixed at the time of transplantation, the A/P axis for most non-sensory structures was not and was able to be re-specified according to the new axial information from the host. The D/V axis for the inner ear was not fixed at the time of transplantation. The asynchronous specification of the A/P and D/V axes of the chick inner ear suggests that sensory organ formation is a multi-step phenomenon, rather than a single inductive event.


Assuntos
Proteínas Aviárias , Padronização Corporal , Orelha Interna/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Fatores de Transcrição , Animais , Proteína Morfogenética Óssea 4 , Proteínas Morfogenéticas Ósseas/genética , Diferenciação Celular , Embrião de Galinha , Ducto Coclear/embriologia , Orelha Interna/transplante , Células Ciliadas Auditivas/citologia , Proteínas de Homeodomínio/genética , Imuno-Histoquímica , Hibridização In Situ , Fator de Transcrição MSX1 , Proteínas do Tecido Nervoso/genética , Fatores de Transcrição Otx , Fenótipo , Receptor de Fator de Crescimento Neural , Receptores de Fator de Crescimento Neural/genética , Sáculo e Utrículo/embriologia , Canais Semicirculares/embriologia , Transplante
19.
Appl Opt ; 35(3): 388-99, 1996 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-21069023

RESUMO

A mathematical solution of the wave equation for coaxial fibers having four different refractive-index profiles is presented. The transcendental equations are obtained under LP approximation and calculated for comparison of HE(mn)-mode-dispersion characteristics. Attention is paid to the HE(11) and HE(12) modes because of their importance for modeling directional couplers, and calculations are carried out to obtain dispersion dependence on dimensional parameters of the fibers. The field expressions also are given, and the spatial distributions of the HE(11) and HE(12) modal fields of all coaxial structures for different conditions of propagation are calculated.

20.
Eur Arch Otorhinolaryngol ; 250(2): 105-9, 1993.
Artigo em Inglês | MEDLINE | ID: mdl-7685179

RESUMO

The relationship between the histological grading of malignancy and the expression of vimentin and cytokeratin was studied in 43 cases of oral squamous cell carcinoma. Immunohistochemical analysis was carried out with the avidin-biotin method using monoclonal antibody anti-vimentin, and the peroxidase anti-peroxidase method using the polyclonal antibody anti-cytokeratin. All cases were classified according to the histological malignancy grading system proposed by Anneroth. All of the carcinomas were found to express cytokeratin, while 60.4% expressed vimentin. Vimentin was particularly noted in all tumors scored to have highly malignant cells.


Assuntos
Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/química , Neoplasias Bucais/patologia , Vimentina/análise , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais , Biomarcadores Tumorais , Feminino , Humanos , Técnicas Imunoenzimáticas , Queratinas/análise , Masculino , Pessoa de Meia-Idade , Prognóstico , Vimentina/biossíntese
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