Effect of ZEB1 Associated with microRNAs on Tumor Stem Cells in Head and Neck Cancer.

Cancer biologists have focused on studying cancer stem cells (CSCs) because of their ability to self-renew and recapitulate tumor heterogeneity, which increases their resistance to chemotherapy and is associated with cancer relapse. Here, we used two approaches to isolate CSCs: the first involved the metabolic enzyme aldehyde dehydrogenase ALDH, and the second involved the three cell surface markers CD44, CD117, and CD133. ALDH cells showed a higher zinc finger E-box binding homeobox 1 (ZEB1) microRNA (miRNA) expression than CD44/CD117/133 triple-positive cells, which overexpressed miRNA 200c-3p: a well-known microRNA ZEB1 inhibitor. We found that ZEB1 inhibition was driven by miR-101-3p, miR-139-5p, miR-144-3p, miR-199b-5p, and miR-200c-3p and that the FaDu Cell Line inhibition occurred at the mRNA level, whereas HN13 did not affect mRNA expression but decreased protein levels. Furthermore, we demonstrated the ability of the ZEB1 inhibitor miRNAs to modulate CSC-related genes, such as TrkB, ALDH, NANOG, and HIF1A, using transfection technology. We showed that ALDH was upregulated upon ZEB1-suppressed miRNA transfection (Mann-Whitney ** p101 = 0.009, t-test ** p139 = 0.009, t-test ** p144 = 0.002, and t-test *** p199 = 0.0006). Overall, our study enabled an improved understanding of the role of ZEB1-suppressed miRNAs in CSC biology.

Occupational stress and work engagement among primary healthcare physicians: a cross-sectional study

Abstract BACKGROUND: Brazil's Family Health Strategy is based on a primary healthcare model, which is considered to have case resolution capacity, with physicians at its center. OBJECTIVES: To evaluate the levels of occupational stress and work engagement among primary healthcare physicians. DESIGN AND SETTING: Cross-sectional study conducted in 2017, in São José do Rio Preto, São Paulo, Brazil. METHODS: A non-probability sample including 32 physicians from family health teams was used. Three self-applied instruments were used: a scale developed by the researchers seeking sociodemographic and professional variables, the Work Stress Scale and the Utrecht Work Engagement Scale. RESULTS: Female professionals (59.4%), permanent employees (56.3%), workload of 40 hours per week (59.4%) and 3-10 years of acting in primary care (68.8%) were more prevalent. Six professionals (19.4%) exhibited significant stress (score ≥ 2.5). The main stressors were lack of prospects for career growth (2.9 ± 1.3), form of task distribution (2.7 ± 1.0), poor training (2.7 ± 1.2) and insufficient time to perform the job (2.6 ± 1.2). Levels of work engagement ranged from 4.3 to 4.6 and were rated as high in all dimensions. Physicians with occupational stress had average levels of work engagement, whereas those without occupational stress had high levels of work commitment. CONCLUSIONS: A notable percentage of the physicians were experiencing occupational stress. The physicians had high levels of work engagement. Occupational stress was negatively correlated with work engagement, and it significantly compromised physicians' levels of work engagement and interfered with their positive relationship with the work environment.

Treatment effects of the EGFR pathway drugs on head and neck cancer stem cells.

(1) Head and neck cancer (HNC) is the sixth most common cancer worldwide and show low survival rates and drug resistance, which can be due to the presence of cancer stem cells (CSCs), a small cell population with metastatic potential, invasion and self-renewal ability. (2) Here, seven tumor cells were sorted as CD44+/CD117+/CD133+ or ALDH+, considered as HNC stem cells (HNCSCs), and as CD44-/CD117-/CD133- or ALDH-, considered non-HNCSCs after both cells sorted criteria was compared to evaluate cell migration, invasion, and colony forming assays. These subpopulations were treated with Cetuximab, Paclitaxel, or a combination of both drugs and evaluated for cell viability. Quantitative PCR and western blot were performed to evaluate EGFR, TRKB, KRAS and HIF-1α gene and protein expression. (3) HNCSCs presented more colonies and appeared to be more sensitive to the drug combination when compared with non-HNCSCs, regardless cells sorted criteria and primary tumor subsite. The EGFR, TRKB, KRAS and HIF-1α genes and proteins were upregulated in CSCs compared with non-HNCSCs, thus explaining the drug resistance. (4) This study contributes to the better development of specific therapeutic protocols based on Cetuximab and Paclitaxel drugs in the treatment of HNC in the presence of CSCs and cell proliferation biomarkers.

Occupational stress and work engagement among primary healthcare physicians: a cross-sectional study.

BACKGROUND: Brazil's Family Health Strategy is based on a primary healthcare model, which is considered to have case resolution capacity, with physicians at its center. OBJECTIVES: To evaluate the levels of occupational stress and work engagement among primary healthcare physicians. DESIGN AND SETTING: Cross-sectional study conducted in 2017, in São José do Rio Preto, São Paulo, Brazil. METHODS: A non-probability sample including 32 physicians from family health teams was used. Three self-applied instruments were used: a scale developed by the researchers seeking sociodemographic and professional variables, the Work Stress Scale and the Utrecht Work Engagement Scale. RESULTS: Female professionals (59.4%), permanent employees (56.3%), workload of 40 hours per week (59.4%) and 3-10 years of acting in primary care (68.8%) were more prevalent. Six professionals (19.4%) exhibited significant stress (score ≥ 2.5). The main stressors were lack of prospects for career growth (2.9 ± 1.3), form of task distribution (2.7 ± 1.0), poor training (2.7 ± 1.2) and insufficient time to perform the job (2.6 ± 1.2). Levels of work engagement ranged from 4.3 to 4.6 and were rated as high in all dimensions. Physicians with occupational stress had average levels of work engagement, whereas those without occupational stress had high levels of work commitment. CONCLUSIONS: A notable percentage of the physicians were experiencing occupational stress. The physicians had high levels of work engagement. Occupational stress was negatively correlated with work engagement, and it significantly compromised physicians' levels of work engagement and interfered with their positive relationship with the work environment.

Relationship between Serotonin-2A Receptor Gene Polymorphism and Wound Healing in Brazilian Patients

Abstract Genetic changes in platelet serotonin receptors (5-HTR2A) impair the initial process of tissue repair, regardless of the triggering factor of the skin wound. Objective was to determine the prevalence of the 102T-C polymorphism in the 5-HTR2A gene in Brazilian patients with and without skin wounds. Cross-sectional case-control study, in which 100 patients were evaluated as Cases Group (subdivided into I-with Chronic Wound and II-with Acute Wound) and 100 individuals as Controls, of both genders. DNA was extracted from leukocytes of peripheral blood and the region that covers the polymorphism was amplified by the molecular techniques Polymerase Chain Reaction/Restriction Fragment Length Polymorphism. The TT genotype was significantly associated with the protective factor against alterations in the healing process of skin wounds (OR: 0.4833; 95%CI: 0.2704-0.8638; p<0.05) in the Control Group. The genotypic analysis between Cases Group (I-Chronic Wound and II-Acute Wound) determined that the TT genotype was significantly associated with the protection factor in Case II (OR: 0.3333; 95%CI: 0.1359-0.8177; p<.005) and the CC genotype was significantly associated with the chance to develop chronic ulcers in the Case I (OR: 6.667; 95%CI: 1.801-24.683; p<0.05). Patients with chronic skin wounds have a higher prevalence of the 102T-C polymorphism in the 5-HTR2A gene, which is associated to alterations in the healing process in this population. There are differences, at the molecular level, in patients, with and without these lesions, and the probable role of the serotonergic system in wound healing.

Anti-EGFR treatment effects on laryngeal cancer stem cells.

Laryngeal cancer (LC) is one of the common head and neck neoplasms and is characterized by resistance to conventional therapy and poor prognosis. This may result from the presence of cancer stem cells (CSCs), which form a small population in tumors with metastatic potential, high invasive capacity, self-renewal, and differentiation. This study aimed to evaluate the effectiveness of 5-fluorouracil and cisplatin individually, as well as the combination of cetuximab and paclitaxel in a CSC subpopulation separated with biomarkers related to tumoral growth (CD44, CD117, and CD133). In addition, expression of TrkB, KRAS, HIF-1α, and VEGF-A genes and proteins related to cell proliferation were evaluated in this subpopulation. The CD44, CD133, and CD117 biomarkers were used to analyze the identification and separation of both subpopulations using FACSAria Fusion. Subpopulations positive for CD44, CD133, and CD117 or lacking these biomarkers were classified as laryngeal cancer stem cells (LCSCs) or laryngeal cancer non-stem cells (non-LCSCs), respectively. Matrigel invasion and colony forming assays were performed to confirm CSC presence. Subpopulations were cultured and exposed to 5-fluorouracil, cisplatin, and cetuximab/paclitaxel drugs for 24 h. Cell proliferation was determined using MTS assay. KRAS and TrkB gene expression levels were evaluated using quantitative real time PCR with TaqMan® Assay in both subpopulations. The non-LCSC subpopulation was considered as the control for relative expression. We found that the LCSC subpopulation demonstrated more resistance to cetuximab and paclitaxel combination chemotherapy when compared with the non-LCSC subpopulation of the cell line. These LCSC subpopulations presented up-regulated expression of KRAS, HIF-1α, and VEGF-A genes and proteins and no TrkB gene expression, but TrkB protein expression was up-regulated in the LC cell line when compared to the non-CSC subpopulation. "In conclusion, the combination of CD44, CD133, and CD117 biomarkers has stem cell properties. Moreover, LCSCs, are capable of resisting treatment and present high KRAS, HIF-1α, and VEGF-A gene expression".

Gene Polymorphisms Involved in Folate Metabolism and DNA Methylation with the Risk of Head and Neck Cancer.

BACKGROUND: Folate is essential for DNA synthesis, repair, and methylation. Polymorphisms in genes associated with folate metabolism may alter these processes and, consequently, modulate cancer development. AIM: We aimed to assess DNMT3B -149C/T (rs2424913), DNMT3B -283T/C (rs6087990), DNMT3B -579G/T (rs2424909), DHFR 19-pb ins/del (rs70991108), SHMT1 1420C/T (rs1979277), and TYMS 28-bp tandem repeat (rs34743033) polymorphisms with risk of head and neck cancer. METHODS: A case-control study was conducted in 1,086 Brazilian individuals. Real-time and conventional polymerase chain reactions-PCR were performed for genotyping the polymorphisms. RESULTS: The single nucleotide polymorphism (SNP), DNMT3B -283T/C, revealed a higher risk of head and neck squamous cell carcinoma (HNSCC) when compared with the C group in the codominant (p < 0.001), dominant (p <0.001), and overdominant (p= 0.001) models for T/C and C/C genotypes. DNMT3B -149C/T and DNMT3B -579G/T revealed no association between groups in any model. The DHFR 19-pb ins/del polymorphism protected against HNSCC development compared to the C group by the codominant (p < 0.001), dominant (p < 0.001), and overdominant (p < 0.001) models. In the TYMS, the 3R/3R genotype had a protective effect against HNSCC development compared with the C group by the recessive models (p= 0.009). In contrast, SHMT1 1420 C/T presented no association between the HNSCC and C groups. DHFR 19-pb ins/del polymorphisms protected against oral cavity cancer (p= 0.003), and only TYMS-28 3R/3R decreased the risk of tumor progression (p= 0.023). In the Kaplan-Meier curve, an association was found between DHFR ins/ins and TYMS -28 3R carriers with respect to relapse-free time; further, DNMT3B -579 T and TYMS-28 2R/2R carriers had longer survival times. CONCLUSION: DNMT3B -283T/C is associated with higher risk, whereas DHFR 19-pb ins/del and TYMS 28 3R/3R protect against head and neck cancer. We also highlighted the association of TYMS 3R/3R genotype carriers with relapse-free cancer protection and survival time.

Glutathione S-transferase Polymorphisms in Head and Neck Squamous Cell Carcinoma Treated with Chemotherapy and/or Radiotherapy.

BACKGROUND/AIM: The Glutathione S-transferases (GSTs) are important carcinogen-metabolizing enzymes. Polymorphisms involved in these enzymes can modulate the development and treatment of head and neck cancer. To investigate the association of GSTs polymorphisms with head and neck cancer and risk factors, clinical-pathological features, and survival time of the patients treated with chemotherapy and/or radiotherapy. METHODS: The GST gene polymorphisms were evaluated in 197 cases and 514 controls by PCR-RFLP-Polymerase Chain Reaction Restriction Fragment Length Polymorphism. RESULTS: The GSTP-313 was associated with a decreased risk for HNSCC (p=0.050). The GSTP1 haplotype analysis revealed a higher frequency of the AC and AT haplotypes in the case group than in the control group (p=0.013 and p=0.019, respectively), and the opposite for G-C haplotype (p = 0.015). Yet, the different combinations between the genotypes were associated with an increased risk of cancer. The study showed no association between the polymorphisms and primary tumor site, clinical-pathological characteristics, treatment (chemotherapy and/or radiotherapy) and survival time of the patients. CONCLUSION: The GST polymorphisms combination showed an increased risk for carcinogenesis, and studies with larger casuistry can contribute to the clarification of the role in individual patient differences for the response to chemotherapy and/or radiotherapy and identify biomarkers of susceptibility.

Polymorphisms in MTHFR, MTR, RFC1 and CßS genes involved in folate metabolism and thyroid cancer: a case-control study.

INTRODUCTION: Polymorphisms in genes coding enzymes involved in folate metabolism may cause alterations in this metabolic pathway and contribute to carcinogenesis, because folate is essential for DNA synthesis, methylation and repair. The objective of this study was to investigate the association of MTHFR 677C>T (rs1801133), MTR 2756A>G (rs1805087), RFC1 80A>G (rs1051266) and CßS 844ins(68) (no rs#) polymorphisms and thyroid cancer development. The association of these polymorphisms with demographic risk factors and clinical histopathological parameters was also evaluated. MATERIAL AND METHODS: The study is a case-control analysis with a total of 462 individuals (151 patients and 311 controls). Polymerase chain reaction-restriction fragment length polymorphism technique was used for genotyping. The χ2 and multiple logistic regression were utilized for statistical analysis. RESULTS: The polymorphisms analysis revealed an association between the MTHFR 677C>T polymorphism (OR = 2.87, 95% CI: 1.50-5.48, p < 0.01, codominant model), (OR = 1.76, 95% CI: 1.18-2.64, p < 0.01, dominant model), (OR = 2.37, 95% CI: 1.28-4.39, p < 0.01, recessive model) and thyroid cancer. RFC1 80A>G polymorphism also was associated with thyroid cancer under recessive mode of inheritance (OR = 1.55; 95% CI: 1.02-2.38; p = 0.04); however, this polymorphism showed Hardy-Weinberg disequilibrium in the control group (χ2 = 24.71, p < 0.001). Furthermore, alcohol (OR = 1.56, 95% CI: 1.36-1.89, p < 0.01) and tobacco consumption (OR = 1.97, 95% CI: 1.28-3.04, p < 0.01) were associated with increased risk for thyroid cancer. The MTR 2756A>G polymorphism showed an association with tumor extent (OR = 2.69, 95% CI: 1.27-5.71, p < 0.01) and aggressiveness (OR = 4.51, 95% CI: 1.67-12.1, p < 0.01). CONCLUSIONS: MTHFR 677C>T is significantly associated with increased risk for thyroid cancer and MTR 2756A>G is associated with tumor extent and aggressiveness. In addition, alcohol and tobacco consumption were associated with increased risk of thyroid cancer. These results may contribute to a better prognosis for thyroid cancer.

Candidate Biomarkers for Oral Squamous Cell Carcinoma: Differential Expression of Oxidative Stress-Related Genes

Background: Alteration in the biotransformation of exogenous compounds can result in production of reactive oxygen species (ROS), which can predispose cells to malignant transformation in the head and neck. This study aimed to evaluate the expression of genes involved in antioxidant metabolism in the oral squamous cell carcinoma (OSCC). Methods: The expression of eighty-four genes was evaluated in OSCC and non-tumor tissues by quantitative real-time polymerase chain reaction using the TaqMan Gene Expression Array. The biological mechanisms related to the differentially expressed genes were investigated using Gene ­ NCBI, KEGG, UNIPROT and REACTOME databases. Results: Twenty-one genes encoding enzymes involved in antioxidant metabolism were differentially expressed in the OSCC case. Four genes (ATOX1, PRDX4, PRNP, and SOD2) were up-regulated, and seventeen (ALOX12, CAT, CSDE1, DHCR24, DUOX1, DUOX2, EPHX2, GLRX2, GPX3, GSR, GSTZ1, MGST3, PRDX1, OXR1, OXSR1, SOD1, and SOD3) were down-regulated. We identified 14 possible novel biomarkers for OSCC. The differentially expressed genes appeared related to important biological processes involved in carcinogenesis, such as inflammation, angiogenesis, apoptosis, genomic instability, invasion, survival, and cell proliferation. Conclusions: Our study identified novel biomarkers which might warrant further investigation regarding OSCC pathogenesis since the altered expression in the genes can modulate biological processes related to oxidative stress and predispose cells to malignant transformation in the oral cavity.

Overexpression of Antiangiogenic Vascular Endothelial Growth Factor Isoform and Splicing Regulatory Factors in Oral, Laryngeal and Pharyngeal Squamous Cell Carcinomas

Background: Overexpression of proangiogenic vascular endothelial growth factor A family VEGFAxxx is associated with tumor growth and metastasis. The role of the alternatively spliced antiangiogenic family VEGFAxxxb is poorly investigated in head and neck squamous cell carcinomas (HNSCCs). The antiangiogenic isoform binds to bevacizumab and its expression level could influence the treatment response and progression-free survival. In this study, the relative expression of VEGFAxxx and VEGFA165b isoforms and splicing regulatory factors genes was investigated in a series of HNSCCs. Methods: VEGFAxxx, VEGFA165b, SRSF6, SRSF5, SRSF1 and SRPK1 gene expression was quantified by quantitative real time PCR in 53 tissue samples obtained by surgery from HNSCC patients. Protein expression was evaluated by immunohistochemistry. Results: VEGFAxxx and VEGFA165b were overexpressed in HNSCCs. Elevated protein expression was also confirmed. However, VEGFA isoforms demonstrated differential expression according to anatomical sites. VEGFAxxx was overexpressed in pharyngeal tumors while the VEGFA165b isoform was up-regulated in oral tumors. The VEGFA165b isoform was also positively correlated with expression of the splicing regulatory genes SRSF1, SRSF6 and SRSF5. Conclusions: We concluded that VEGFAxxx and VEGFA165b isoforms are overexpressed in HNSCCs and the splicing regulatory factors SRSF1, SRSF6, SRSF5 and SRPK1 may contribute to alternative splicing of the VEGFA gene. The findings for the differential expression of the antiangiogenic isoform in HNSCCs could facilitate effective therapeutic strategies for the management of these tumors.

Rhinoseptoplasty in children / Rinosseptoplastia em crianças

Abstract Introduction: Untreated septal and/or nasal pyramid deviation in children should be corrected as soon as possible, because they can result in esthetic or functional problems years later. Objective: To report the surgical experience in treating children with nasal septum and/or nasal pyramid deviation. Methods: Review of medical records of 202 children, 124 (61.4%) males and 78 (38.6%) females, between 4 and 16 years of age (M = 11 years) who underwent rhinoplasty and/or septoplasty in a Pediatric Otolaryngology Service of the Dept. of Otolaryngology and Head and Neck Surgery between January 1994 and January 2010. Results: Septoplasty performed in 157 cases (77.7%); rhinoseptoplasty in 23 cases (11.4%), and rhinoplasty in 22 cases (10.9%). Conclusion: Nasal changes should be corrected in children, in order to provide harmonious growth, and prevent severe sequelae found in mouth breathers.

Resumo Introdução: Desvio do septo e/ou da pirâmide nasal em crianças, se não tratado, pode apresentar problemas estéticos ou funcionais após anos, devendo ser corrigido o quanto antes. Objetivo: Relatar experiência cirúrgica no tratamento de crianças com desvio de septo nasal e/ou pirâmide nasal. Método: Revisão de prontuários de 202 crianças, 124 (61,4%) do gênero masculino e 78 (38,6%) do feminino, entre quatro e 16 anos (M = 11 anos), submetidas a rino e/ou septoplastia de janeiro de 1994 a janeiro de 2010, no Serviço de Otorrinopediatria do Departamento de Otorrinolaringologia e Cirurgia de Cabeça e Pescoço. Resultados: Septoplastia feita em 157 casos (77,7%); rinosseptoplastia em 23 casos (11,4%) e rinoplastia em 22 casos (10,9%). Conclusão: Alterações nasais devem ser corrigidas em crianças, para proporcionar crescimento harmônico e evitar as graves sequelas encontradas no respirador bucal.

Rhinoseptoplasty in children.

INTRODUCTION: Untreated septal and/or nasal pyramid deviation in children should be corrected as soon as possible, because they can result in esthetic or functional problems years later. OBJECTIVE: To report the surgical experience in treating children with nasal septum and/or nasal pyramid deviation. METHODS: Review of medical records of 202 children, 124 (61.4%) males and 78 (38.6%) females, between 4 and 16 years of age (M=11 years) who underwent rhinoplasty and/or septoplasty in a Pediatric Otolaryngology Service of the Dept. of Otolaryngology and Head and Neck Surgery between January 1994 and January 2010. RESULTS: Septoplasty performed in 157 cases (77.7%); rhinoseptoplasty in 23 cases (11.4%), and rhinoplasty in 22 cases (10.9%). CONCLUSION: Nasal changes should be corrected in children, in order to provide harmonious growth, and prevent severe sequelae found in mouth breathers.

Molecular approach of auditory neuropathy / Abordagem molecular da neuropatia auditiva

INTRODUCTION: Mutations in the otoferlin gene are responsible for auditory neuropathy. OBJECTIVE: To investigate the prevalence of mutations in the mutations in the otoferlin gene in patients with and without auditory neuropathy. METHODS: This original cross-sectional case study evaluated 16 index cases with auditory neuropathy, 13 patients with sensorineural hearing loss, and 20 normal-hearing subjects. DNA was extracted from peripheral blood leukocytes, and the mutations in the otoferlin gene sites were amplified by polymerase chain reaction/restriction fragment length polymorphism. RESULTS: The 16 index cases included nine (56%) females and seven (44%) males. The 13 deaf patients comprised seven (54%) males and six (46%) females. Among the 20 normal-hearing subjects, 13 (65%) were males and seven were (35%) females. Thirteen (81%) index cases had wild-type genotype (AA) and three (19%) had the heterozygous AG genotype for IVS8-2A-G (intron 8) mutation. The 5473C-G (exon 44) mutation was found in a heterozygous state (CG) in seven (44%) index cases and nine (56%) had the wild-type allele (CC). Of these mutants, two (25%) were compound heterozygotes for the mutations found in intron 8 and exon 44. All patients with sensorineural hearing loss and normal-hearing individuals did not have mutations (100%). CONCLUSION: There are differences at the molecular level in patients with and without auditory neuropathy. .

INTRODUÇÃO: Mutações no gene da otoferlina (OTOF) são responsáveis pela neuropatia auditiva. OBJETIVO: Investigar a prevalência de mutações no gene OTOF em pacientes com e sem neuropatia auditiva. MÉTODO: Estudo de casos em corte transversal sendo avaliados 16 casos índice com neuropatia auditiva, 13 pacientes com deficiência auditiva sensorioneural (DASN) e 20 indivíduos ouvintes. DNA foi extraído de leucócitos do sangue periférico e regiões do gene OTOF foram analisadas pela técnica PCR-RFLP. RESULTADOS: Dos 16 casos índice, 9 (56%) são do gênero feminino e 7 (44%) do masculino. Dos 13 pacientes com DASN, 7 (54%) são masculinos e 6 (46%) femininos. Dos 20 ouvintes, 13 (65%) são masculinos e 7 (35%) femininos. Treze (81%) casos índice apresentam o genótipo selvagem (AA) e 3 (19%) o genótipo heterozigoto AG para a mutação IVS8-2A-G (intron 8). A mutação 5473C-G (exon 44) foi encontrada em heterozigose (CG) em 7 (44%) dos casos índice e 9 (56%) apresentam o genótipo selvagem (CC). Destes mutantes, dois (25%) são heterozigotos compostos para as mutações encontradas no intron 8 e exon 44. Os pacientes com DASN e os ouvintes não apresentam mutações (100%). CONCLUSÃO: Existem diferenças, ao nível molecular, em pacientes com e sem neuropatia audi tiva. .

Molecular approach of auditory neuropathy.

INTRODUCTION: Mutations in the otoferlin gene are responsible for auditory neuropathy. OBJECTIVE: To investigate the prevalence of mutations in the mutations in the otoferlin gene in patients with and without auditory neuropathy. METHODS: This original cross-sectional case study evaluated 16 index cases with auditory neuropathy, 13 patients with sensorineural hearing loss, and 20 normal-hearing subjects. DNA was extracted from peripheral blood leukocytes, and the mutations in the otoferlin gene sites were amplified by polymerase chain reaction/restriction fragment length polymorphism. RESULTS: The 16 index cases included nine (56%) females and seven (44%) males. The 13 deaf patients comprised seven (54%) males and six (46%) females. Among the 20 normal-hearing subjects, 13 (65%) were males and seven were (35%) females. Thirteen (81%) index cases had wild-type genotype (AA) and three (19%) had the heterozygous AG genotype for IVS8-2A-G (intron 8) mutation. The 5473C-G (exon 44) mutation was found in a heterozygous state (CG) in seven (44%) index cases and nine (56%) had the wild-type allele (CC). Of these mutants, two (25%) were compound heterozygotes for the mutations found in intron 8 and exon 44. All patients with sensorineural hearing loss and normal-hearing individuals did not have mutations (100%). CONCLUSION: There are differences at the molecular level in patients with and without auditory neuropathy.

Gradual approach to refinement of the nasal tip: surgical results / Abordagem gradativa para refinamento da ponta nasal: resultados cirúrgicos

Introduction: The complexity of the nasal tip structures and the impact of surgical maneuvers make the prediction of the final outcome very difficult. Therefore, no single technique is enough to correct the several anatomical presentations, and adequate preoperative planning represents the basis of rhinoplasty. Objective: To present results of rhinoplasty, through the gradual surgical approach to nasal tip definition based on anatomical features, and to evaluate the degree of patient satisfaction after the surgical procedure. Methods: Longitudinal retrospective cohort study of the medical charts of 533 patients of both genders who underwent rhinoplasty from January of 2005 to January of 2012 was performed. Cases were allocated into seven groups: (1) no surgery on nasal tip; (2) interdomal breakup; (3) cephalic trim; (4) domal suture; (5) shield-shaped graft; (6) vertical dome division; (7) replacement of lower lateral cartilages. Results: Group 4 was the most prevalent. The satisfaction rate was 96% and revision surgery occurred in 4% of cases. Conclusion: The protocol used allowed the implementation of a gradual surgical approach to nasal tip definition with the nasal anatomical characteristics, high rate of patient satisfaction with the surgical outcome, and low rate of revision. .

Introdução: A complexidade das estruturas da ponta nasal e o impacto de manobras cirúrgicas sobre o seu suporte dificultam a previsão da forma final da mesma. Devido a isso, nenhuma técnica isolada é suficiente para corrigir adequadamente as numerosas apresentações anatômicas, sendo o planejamento pré-operatório, a base da rinoplastia. Objetivos: Apresentar resultados de rinoplastias, por meio da abordagem cirúrgica gradativa para definição da ponta nasal baseada nas características anatômicas, e avaliar o grau de satisfação dos pacientes após a realização do procedimento cirúrgico. Método: Estudo em coorte histórica longitudinal no qual foram avaliados os prontuários de 533 pacientes de ambos os gêneros submetidos à rinoplastia no período de Janeiro de 2005 a Janeiro de 2012. Os pacientes foram divididos em sete grupos: (1) Nenhuma cirurgia na ponta nasal;( 2) Divulsão interdomal; (3) Ressecção cefálica; (4) Sutura domal; (5) Enxerto em escudo; (6) Divisão vertical dos domus; (7) Reconstrução das cartilagens alares maiores. Resultados: O grupo 4 foi o de maior prevalência. A taxa de satisfação foi de 96% e a revisão cirúrgica ocorreu em 4% dos casos. Conclusão: O protocolo utilizado permitiu a associação da abordagem cirúrgica gradativa para definição da ponta nasal com as características anatômicas nasais, alta taxa de satisfação como resultado cirúrgico e baixa taxa de revisão. .

Gradual approach to refinement of the nasal tip: surgical results.

INTRODUCTION: The complexity of the nasal tip structures and the impact of surgical maneuvers make the prediction of the final outcome very difficult. Therefore, no single technique is enough to correct the several anatomical presentations, and adequate preoperative planning represents the basis of rhinoplasty. OBJECTIVE: To present results of rhinoplasty, through the gradual surgical approach to nasal tip definition based on anatomical features, and to evaluate the degree of patient satisfaction after the surgical procedure. METHODS: Longitudinal retrospective cohort study of the medical charts of 533 patients of both genders who underwent rhinoplasty from January of 2005 to January of 2012 was performed. Cases were allocated into seven groups: (1) no surgery on nasal tip; (2) interdomal breakup; (3) cephalic trim; (4) domal suture; (5) shield-shaped graft; (6) vertical dome division; (7) replacement of lower lateral cartilages. RESULTS: Group 4 was the most prevalent. The satisfaction rate was 96% and revision surgery occurred in 4% of cases. CONCLUSION: The protocol used allowed the implementation of a gradual surgical approach to nasal tip definition with the nasal anatomical characteristics, high rate of patient satisfaction with the surgical outcome, and low rate of revision.

Inflammation and cancer: role of annexin A1 and FPR2/ALX in proliferation and metastasis in human laryngeal squamous cell carcinoma.

The anti-inflammatory protein annexin A1 (ANXA1) has been associated with cancer progression and metastasis, suggesting its role in regulating tumor cell proliferation. We investigated the mechanism of ANXA1 interaction with formylated peptide receptor 2 (FPR2/ALX) in control, peritumoral and tumor larynx tissue samples from 20 patients, to quantitate the neutrophils and mast cells, and to evaluate the protein expression and co-localization of ANXA1/FPR2 in these inflammatory cells and laryngeal squamous cells by immunocytochemistry. In addition, we performed in vitro experiments to further investigate the functional role of ANXA1/FPR2 in the proliferation and metastasis of Hep-2 cells, a cell line from larynx epidermoid carcinoma, after treatment with ANXA1(2-26) (annexin A1 N-terminal-derived peptide), Boc2 (antagonist of FPR) and/or dexamethasone. Under these treatments, the level of Hep-2 cell proliferation, pro-inflammatory cytokines, ANXA1/FPR2 co-localization, and the prostaglandin signalling were analyzed using ELISA, immunocytochemistry and real-time PCR. An influx of neutrophils and degranulated mast cells was detected in tumor samples. In these inflammatory cells of peritumoral and tumor samples, ANXA1/FPR2 expression was markedly exacerbated, however, in laryngeal carcinoma cells, this expression was down-regulated. ANXA1(2-26) treatment reduced the proliferation of the Hep-2 cells, an effect that was blocked by Boc2, and up-regulated ANXA1/FPR2 expression. ANXA1(2-26) treatment also reduced the levels of pro-inflammatory cytokines and affected the expression of metalloproteinases and EP receptors, which are involved in the prostaglandin signalling. Overall, this study identified potential roles for the molecular mechanism of the ANXA1/FPR2 interaction in laryngeal cancer, including its relationship with the prostaglandin pathway, providing promising starting points for future research. ANXA1 may contribute to the regulation of tumor growth and metastasis through paracrine mechanisms that are mediated by FPR2/ALX. These data may lead to new biological targets for therapeutic intervention in human laryngeal cancer.

Gene expression profile of 5-fluorouracil metabolic enzymes in laryngeal cancer cell line: predictive parameters for response to 5-fluorouracil-based chemotherapy.

BACKGROUND: 5-fluorouracil (5-FU) is an antifolate chemotherapeutic that has become established in many therapeutic regimes, but sensitivity variations and development of resistance are common problems that limit the efficiency of the treatments. Inter-individual variations to 5-FU outcome have been attributed to different expression profiles of genes related to folate metabolism. METHODS: To elucidate the mechanisms of variations to 5-FU outcome, the authors investigated MTHFR, DHFR, TYMS and SLC19A1 folate genes expression for 5-FU response in laryngeal cancer cell line (Hep-2). Concentrations of 10, 50, and 100 ng/mL of 5-FU chemotherapeutic were added separately in Hep-2 cell line for 24 hours at 37 °C. Cell sensibility was evaluated with fluorescein isothiocyanate (FITC) label Bcl-2 by flow cytometry. The real-time quantitative PCR (qPCR) technique was performed for quantification of gene expression using TaqMan(®) Gene Expression Assay. ANOVA and Bonferroni's post hoc tests were utilized to statistical analysis. RESULTS: The numbers of viable Hep-2 cells with 10, 50, and 100 ng/mL concentrations of 5-FU chemotherapy were 15.87, 28.3 and 68.9%, respectively. Statistical analysis showed significant association between control group and increased expression for TYMS gene in cells treated with 100 ng/mL/5-FU chemotherapy (P<0.05). CONCLUSIONS: The authors found association between the highest 5-FU dose chemotherapy and increased expression levels for TYMS folate gene in laryngeal cancer cell line. Although these experiments were performed in vitro, the results suggest that genetic factors are thought to play an important role in drug metabolism and may be useful for predicting treatment outcomes.