Master of Science in Biomedical Sciences / Department of Clinical Sciences / Mary Lynn Higginbotham / This study evaluated sixteen canine nasal carcinoma and five normal nasal epithelium samples for expression and phosphorylation of known targets of toceranib [vascular endothelial growth factor receptor-2 (VEGR2), platelet derived growth factor alpha (PDGFR-[alpha]), platelet derived growth factor receptor beta (PDGFR-[beta]), and stem cell factor receptor (c-KIT)] and epidermal growth factor receptor 1 (EGFR1) using immunohistochemistry, RT-PCR and a receptor tyrosine kinase (RTK) phosphorylation panel. Protein for VEGFR2 was expressed in neoplastic cells of all carcinomas, PDGFR-[alpha] was noted in 15/16, whereas PDGFR-[beta] was detected in 3/16 samples, but showed primarily stromal staining. Protein expression for c-KIT was present in 4/16 and EGFR1 was noted in 14/16 samples. Normal tissue showed variable protein expression of the RTKs. Messenger RNA for VEGFR2, PDGFR-[beta], and c-KIT were noted in all samples. Messenger RNA for PDGFR-[alpha] and EGFR1 were detected in 15/16 samples. All normal nasal tissue detected messenger RNA for all RTKs of interest. Constitutive phosphorylation of VEGFR2, PDGFR-[alpha], PDGFR-[beta] and c-KIT was not observed in any carcinoma or normal nasal sample, but phosphorylation of EGFR1 was noted in 10/16 carcinoma and 3/5 normal samples. The absence of major phosphorylated RTK targets of toceranib suggests the clinical effect of toceranib may occur through inhibition of alternative and currently unidentified RTK pathways in canine nasal carcinomas. The observed protein and message expression and phosphorylation of EGFR1 in the nasal carcinoma samples merits further inquiry into EGFR1 as a therapeutic target for this cancer.

The nasal turbinates of ungulates are complex bony scrolls within the nasal cavity. These intricate bony plates, covered by moist epithelium, provide a large surface area that facilitates a countercurrent exchange of both water and heat between turbinal lining and respired air. Given their functional importance, maxilloturbinate size and structural dimensions may vary among species of different body sizes, activity levels and from different habitats, and may also serve as a predictor of the ability of the species to cope with high temperatures or limiting water resources. This is the first study to measure nasal turbinate surface area in ungulates using high-resolution computed tomography (CT) scanning. Heads of eleven South African bovid species and one European bovid species were successfully scanned and surface area measurements made using stereological techniques. These species included Cape grysbok, springbuck, bushbuck, blesbok, impala, mountain reedbuck, fallow deer, kudu, nyala, gemsbok and blue wildebeest; and represent species of different body sizes, from a range of habitats and with different water dependencies and predator avoidance strategies. The total maxilloturbinate surface area increased with body size for all study species. The surface areas of the nasal turbinates varied rostrocaudally, with the highest surface area occurring approximately midway along the length of the maxilloturbinate bones. The Cape grysbok stood out as having a nasal turbinate surface area of 12.77 cm2/kg, which was lower than the observed trend, the reason for this not being clear from these data. Phylogenetic independent analyses showed that log body mass and water dependence had a significant effect on nasal turbinate surface area, with habitat, distributional range and anti-predator behaviour having no effect. Subsequent phylogenetic species comparisons showed that structural variations in nasal turbinate surface area were phylogeny based due to the close genetic relatedness of the study species, and not associated with any environmental factors. The environmentally linked results for water dependency need further investigation in future studies of larger sample sizes and a broader range of species. Changes in climatic conditions may impact on a species‟ activity patterns, with individuals being forced to make behavioural modifications rather than physiological or anatomical adjustments. However, there is no clear evidence to indicate large differences in nasal turbinate surface area in relation to water dependence. This subsequently rules out the use of nasal turbinate surface area as a predictor to which and how ungulate species will respond to increasing global temperatures.

Lung cancer surpasses all other causes of cancer-related deaths worldwide. Gene-expression microarrays have shown that differences in the cytologically normal bronchial airway can distinguish between patients with and without lung cancer. In research reported here, we have used microRNA expression in bronchial epithelium and gene expression in nasal epithelium to advance biological understanding of the lung-cancer "field of injury" and develop new biomarkers for lung cancer diagnosis. MicroRNAs are known to mediate the airway response to tobacco smoke exposure but their role in the lung-cancer-associated field of injury was previously unknown. Microarrays can measure microRNA expression; however, they are probe-based and limited to detecting annotated microRNAs. MicroRNA sequencing, on the other hand, allows the identification of novel microRNAs that may play important biological roles. We have used microRNA sequencing to discover novel microRNAs in the bronchial epithelium. One of the predicted microRNAs, now known as miR-4423, is associated with lung cancer and airway development. This finding demonstrates for the first time a microRNA expression change associated with the lung-cancer field of injury and microRNA mediation of gene expression changes within that field. The National Lung Screening Trial showed that screening high-risk smokers using CT scans decreases lung-cancer-associated mortality. Nodules were detected in over 20% of participants; however, the overwhelming majority of screening-detected nodules were non-malignant. We therefore need biomarkers to determine which screening-detected nodules are benign and do not require further invasive testing. Given that the lung-cancer-associated field of injury extends to the bronchial epithelium, our group hypothesized that the field of injury may extend farther up in the airway. Using gene expression microarrays, we have identified a nasal epithelium gene-expression signature associated with lung cancer. Using samples from the bronchial epithelium and the nasal epithelium, we have established that there is a common lung-cancer-associated gene-expression signature throughout the airway. In addition, we have developed a nasal epithelium gene-expression biomarker for lung cancer together with a clinico-genomic classifier that includes both clinical factors and gene expression. Our data suggests that gene expression profiling in nasal epithelium might serve as a non-invasive approach for lung cancer diagnosis and screening

The acute phase response (APR) has been shown to alter expression and activity of biotransformation enzymes, such as the phase I cytochromes P450 and phase II glutathione s-transferases (GSTs). The cytochromes P450 and GSTs are expressed abundantly and colocalized to non-neuronal cells of the olfactory mucosa. Previous studies indicate that olfactory cytochromes P450 expression and activity is altered during periods of localized inflammation and infection. Little is understood, however, about the influence of the APR on olfactory GST enzymes. This study investigated effects of the APR on olfactory GST isozymes expression and activity in mouse olfactory mucosa after 24-hr treatment with the acute phase inducer, polyinosinic: polycytidylic acid (polyIC). Western blot analysis using antibodies directed against specific GST isoforms α (A1-1), μ (M1-1), and π (P1-1) demonstrated that their expression was unaltered by polyIC treatment. In contrast, olfactory P450 2E1 expression was significantly decreased. Enzymatic activity of the olfactory GSTs toward the general substrate, 1-chloro-2,4-dinitrobenzene (CDNB) was unchanged during the APR. Analysis of olfactory glutathione content during the APR showed that it was also unaffected by polyIC. The insensitivity of these olfactory GST isoforms during the APR may play a significant role toward limiting the impact of infection and inflammation on the olfactory system.

Background and objective Dexmedetomidine is characterized with effects of sedation, analgesia, amnesia and lack of respiratory depression. Hence, it should be suitable for awake fibreoptic intubation (AFOI). Methods We enrolled 30 oral cancer patients with limited mouth openings who were undergoing AFOI for elective surgery. Patients were randomly allocated into two groups; the Dex group (nU16) that received dexmedetomidine (1.0mgkg-1) infusion and the Control group (nU14) that received fentanyl (1.0mgkg-1) infusion. Main outcomes were evaluated by grading scores presenting conditions for nasal intubation and postintubation. Other analysed parameters included airway obstruction, haemodynamic changes, consumption time for intubation, amnesia level and satisfaction. Results Intubation score (1-5) representing condition for nasal intubation was significantly better in the Dex group [2(1-3)] than in the Control group [3(2-5)] (PU0.001). Postintubation score (1-3) representing tolerance to intubation also showed more favourable results in the Dex group[1(1-3)] than intheControl group[2(2-3)] (PU0.002). The Dex group showed significantly reduced haemodynamic response to intubation than the Control group. Incidence requiring temporary haemodynamic support was higher in the Dex group but not of significance. Both levels of amnesia and satisfaction score were significant in the Dex group. Other analysed parameters such as consumption time for intubation, airway obstruction score and postoperative adverse events did not differ significantly. Conclusion Combination of dexmedetomidine loading with topical anaesthesia provides significant benefit for AFOI in intubation condition, patient tolerance, haemodynamic response, amnesia and satisfaction. Dexmedetomidine is effective for AFOI in anticipated difficult airway with only minor and temporary haemodynamic adverse effects.

In search of evidence for or against the endothermic dinosaur hypothesis, a recent study by Ruben et al. (1996) revealed that endotherms tend to have larger nasal cross-sectional areas than ectotherms of the same mass. The reason offered for this observation was that larger nasal passages are needed to house the complex respiratory turbinates possessed by endotherms. Whales were excluded from the study on the grounds that they have no nasal turbinates. In the present study, the cross-sectional area of the nasal passages of nine species of Odontoceti were measured by the use of latex casts. The regression of log cross-sectional area vs. log mass yielded the same line for the whales of the current study as for the endotherms of the previous study. Alternative explanations for the large nasal cross-sectional area of endotherms are sought.

Objetivos: Determinar e comparar os valores de nasalância em amostras de fala na presença e ausência da turbulência nasal (TN) e da hipernasalidade. Métodos: Foram analisadas um total de 288 amostras de fala com fonemas de alta e baixa pressão, sendo que, 110 foram excluídas por apresentarem articulação compensatória (AC) ou disfonia. Cinco (5) juízes avaliaram as 178 amostras (110 + 20%) para indicar a presença e ausência da TN e da hipernasalidade. Resultados: As palavras e frases contendo fonemas líquidos (baixa pressão) foram julgados, por todos os juízes, com ausência de TN, e não foi encontrada diferença significante entre as amostras analisadas (lalá, lalá olhou a lua, e frases LO). No caso das 5 amostras com fonemas de alta pressão (papai, bebê, papai pediu pipoca, o bebê babou, frases PO) observou-se que somente nas frases PO a diferença foi estatisticamente significante (p=0,02) para o fator hipernasalidade. Conclusão: Comparando os valores de nasalância na presença e ausência da TN podemos concluir que a nasalância apresentou-se elevada em todas as palavras e frases contendo fonemas orais de alta pressão, porém a diferenças encontradas não foram estatisticamente significantes. Comparando os valores de nasalância na presença e ausência da hipernasalidade notamos que os valores apresentaram-se elevados nas palavras e frases contendo fonemas orais sonoros de alta pressão e nas frases LO. / Objective:The objective of this study was to determine and to compare nasalance scores at the presence and absence of nasal turbulence (NT) and hypernasality. Methods: Nasometric scores and audio recordings were obtained simultaneously from 30 participants with operated unilateral cleft lip and palate during production of speech samples with low and high pressure phonemes. From a sample of 288 recording, 110 were excluded when rated with presence of compensatory articulation (CA) or dysphonia by a group of 5 judges and the remaining 110 sample were classified according to the presence or absence of NT and hypernasality and their mean nasalance scores were calculated. Results: Intra-rater agreement between the judges ranged from regular to perfect and inter-rater agreement ranged from moderate to substancial. The words and phrases with low pressure phonemes (lalá, lalá olhou a lua e low pressure phrases) were rated by all judges with absence of NT. All nasalance scores for low pressure phonemes were found to be suggestive of the presence of hypernasality for the samples rated as hypernasal. There was no significant difference between the nasalance scores for all low pressure samples analysed (lalá, lalá olhou a lua and other low pressure phrases). The nasalance scores for high pressure phonemes (papai, bebê, papai pediu pipoca, o bebê babou, phrases high pressure) were found to be suggestive of the presence of hypernasality for the samples rated as hypernasal and also when the samples were rated with presence of NT. There was significant difference between the nasalance scores only for the high pressure phrases (p=0,02). Conclusion: While statistically significant higher nasalance scores were found only for the longer speech sample of high pressure phrases, elevated nasalance scores were observed for all samples rated with presence hypernasality or presence of NT suggesting the importance, during clinical practice, of carefully interpreting nasalance scores at the presence of turbulence.

O órgão vômeronasal (OVN), é uma estrutura que detecta feromônios, emitindo sinais que modulam o comportamento social e reprodutivo. Possui células-tronco que se dividem e migram para substituir neurônios ao longo da vida. O objetivo deste estudo foi isolar e caracterizar as células derivadas do órgão vômeronasal de coelhos da raça Nova Zelândia e testar seu potencial terapêutico no tratamento da ablação da via vômeronasal. Utilizou-se 10 coelhos machos com 120 dias, sendo 9 submetidos a ablação do OVN e 1 utilizado para coleta de material para cultivo celular. Foram testados três meios de cultivo DMEM High Glucose, DMEM/F12 e MEM Alfa. No cultivo celular observou-se maior confluência e crescimento quando utilizado o DMEM High Glucose, confirmado pelos ensaios de MTT e Azul de Trypan. Na imunocitoquímica observou-se PCNA+, OCT4+, Nanog+, GFAP+, Vimentina+, Nestin+, Stro-1+, B-Tubulina+, CK-18+, CD73+, CD90+, CD105+, CD34-, CD117- e CD45-. Na citometria de fluxo foi observado PCNA+, OCT-3/4+, Nanog+, GFAP+, Vimentina+, Nestin+, Stro-1+, B-Tubulina+, CK-18+, CD73+, CD90+, CD105+, CD34- e CD45-. Na análise molecular foi possível observar a expressão de CD73, CD105, Oct-4, Nestina, Vimentina e GAPDH. Funcionalmente as células se diferenciaram em adipócitos, osteócitos e condrócitos e não possuem potencial tumorigênico em camundongos Balb-cnu/nu. A análise hormonal demonstrou que nos animais tratados após 7 e 14 dias não houveram diferenças significativas entre os grupos tratados e controles. No grupo após 21 dias notou-se uma diferença de 50% nos níveis hormonais no grupo tratado em relação ao grupo controle. Na análise da expressão para eGFP observou-se que os animais tratados após 7 dias as células injetadas formavam aglomerados celulares na região subjacente ao epitélio sensitivo. Após 14 dias continuavam a circundar o epitélio neurosensorial e após 21 dias observou-se pouca expressão de células localizadas apenas na região subjacente ao epitélio neurosensorial. A linhagem celular derivada do órgão vômeronasal possui uma população de células progenitoras, que foram visualizadas no foco da lesão, demonstrando sua capacidade de migração, o que a torna uma boa fonte para terapia celular em ablação da via vômeronasal. / The vômeronasal organ (VNO) is a structure that detects pheromones, emits signals that modulate social and reproductive behavior. It has stem cells that divide and migrate to replace neurons throughout life. The aim of this study was to isolate and characterize cells derived from the vômeronasal organ from rabbits New Zealand and test their therapeutic potential in the treatment of ablation of the vômeronasal pathway. 10 male rabbits with 120 days were submitted to the ablation of VNO and 2 controls animals used to collect material for cell culture. Three different culture media DMEM High Glucose, Alpha MEM and DMEM/F12 were tested. In cell culture observed greater confluence and growth when used DMEM High Glucose, confirmed by MTT assay and Trypan Blue. In immunocytochemistry observed PCNA + OCT4 +, Nanog +, GFAP + Vimentin +, Nestin +, Stro-1 + B-tubulin +, CK-18 +, CD73 +, CD90 +, CD105 +, CD34-, CD117-and CD45-. In flow cytometry was observed PCNA + OCT-3/4 + Nanog +, GFAP + Vimentin +, Nestin +, Stro-1 + B-tubulin +, CK-18 +, CD73 +, CD90 +, CD105 +, CD34-and CD45-. Molecular analysis was possible to observe the expression of CD73, CD105, Oct-4, Nestin, vimentin and GAPDH. Functionally, the cells differentiate into adipocytes, osteocytes and chondrocytes have not Balb-cnu/nu tumorigenic potential in mice. Hormonal analysis demonstrated that animals treated after 7 days and 14 days there were no significant differences between treated groups and controls. In group after 21 days noticed a 50% difference in hormone levels in the treated group compared to the control group. In expression analysis for eGFP was observed that the treated animals after 7 the injected cells formed cell clusters in the underlying sensory epithelium region. After 14 days continued to surround the neurosensory epithelium and after 21 days there was little expression in the cells located just behind the neurosensory epithelium region. The cell line derived from the vômeronasal organ has a population of progenitor cells that were visualized in the lesion focus, demonstrating their ability to migrate, which makes it a good source for cell therapy in ablation of the vômeronasal pathway.

O objetivo neste trabalho foi de analisar a geometria nasal, a nasofaringe e a orofaringe, em crianças respiradoras orais com indicação de cirurgia desobstrutiva das vias aéreas superiores, e verificar a existência de uma possível influência direta da respiração na determinação do tipo facial. Foram avaliadas 657 crianças e, dentre elas, foram selecionadas 75, que se submeteram a avaliação otorrinolaringológica por meio de exames clínico, de radiografia cavum e/ou nasofibroscopia, e com as quais ficaram evidenciadas as obstruções das vias aéreas superiores com indicação cirúrgica. Ao final 41 crianças (21 do gênero masculino e 20 do feminino), entre 6,16 e 14,66 anos de idade, brasileiras, leucodermas, sem história de tratamento ortodôntico ou otorrinolaringológico cirúrgico (remoção de tonsilas faringeanas, palatinas, ou estruturas internas da cavidade nasal), aceitaram participar do estudo. Foram obtidas telerradiografias em norma lateral para a determinação do tipo facial e realizado o exame de rinometria acústica para a obtenção da área transversal mínima da cavidade nasal. A análise estatística dos dados (ANOVA, Razão de Verossimilhanças, análises de variâncias com medidas repetidas com dois fatores, comparações múltiplas de Bonferroni, teste Kruskal- Wallis; com nível de significância de 5%) e a interpretação dos resultados obtidos não mostraram diferença estatisticamente significativa na área total em MCA1 e MCA2 entre os tipos faciais (p > 0,05). Além disso, a obstrução das tonsilas faringeanas e palatinas avaliadas separadamente ou combinadas não variou estatisticamente segundo os tipos faciais (p = 0,582 para tonsila faringeana e p = 0,733 para tonsila palatina; e p = 0,925 quando combinadas). Conclui-se que, no presente estudo, não foi encontrada evidência de que a obstrução das vias aéreas superiores tenha influência determinante na definição do tipo facial. / The objective of this paper was to analyze the nasal geometry, the nasopharynx and the oropharynx in oral breathing children with indication for surgery clearence upper airway, in order to verify the existence of a possible direct influence of breathing in the determination of the facial type. A group of 657 children was evaluated and, among them, 75 were selected for otorhinolaryngologic evaluation through clinical exams, cavum radiography and/or nasal endoscopy , when the obstruction of the upper airway with surgical indication has been confirmed. From this subgroup, 41 children agreed to participate in the study (21 males: 20 females), with ages between 6,16 and 14,66 years, brazilians, whites, with no history of orthodontic treatment nor otorhinolaryngologic surgery (removal of the pharyngeal or palatine tonsils or internal structures of the nasal cavity). Radiographs were taken in lateral norm for the determination of the facial type and the acoustic rhinometry exam was performed for the determination of the nasal minimal transversal area. The statistical analyses of the data were made with the variance analysis (ANOVA), likelihood ratio test, analyses of variance with measures repeated with two factors, Bonferroni multiple comparisons, and Kruskal-Wallis test. The significance level was chosen as 0.05. No statistically significant difference was detected in the total area in MCA1 and MCA2 between the facial types. Besides that, the obstruction of the pharyngeal or palatine tonsils, taken separately or in combination, did not vary statistically according to the facial pattern (p = 0.582 for pharyngeal tonsil; p = 0.733 for palatine tonsil; and p = 0.925 when combined). Therefore, in the present study, no evidence has been found that the obstruction of the superior airway has determinant influence in the determination of the facial type.

Objetivo: Determinar as áreas seccionais e os volumes de segmentos específicos da cavidade nasal, incluindo a nasofaringe, de indivíduos sem evidências de obstrução nasal, e os mesmos volumes na produção da fala, com vistas ao estabelecimento de valores de referência. Modelo: Análise prospectiva. Local de Execução: Laboratório de Fisiologia, HRAC-USP. Participantes: 30 voluntários caucasianos, com idade entre 18 e 30 anos, sendo 14 homens e 16 mulheres. Intervenções/Variáveis: Foram avaliadas, por rinometria acústica, áreas de secção transversa nos segmentos correspondentes à válvula nasal (AST1), região anterior da concha nasal inferior (AST2) e região posterior da concha nasal inferior (AST3) e volumes, nos segmentos correspondentes à região da válvula nasal (V1), dos cornetos (V2) e da nasofaringe (V3). As medidas foram feitas antes e após a descongestão nasal, no repouso (suspensão voluntária da respiração) e durante a atividade velofaríngea (produção contínua e silenciosa do fonema /f/). Resultados: Os valores médios (±DP) das áreas (em cm2) e volumes (em cm3) obtidos no repouso, foram: AST1=0,54±0,13, AST2=0,98±0,31, AST3=1,42±0,44, V1=1,68±0,32, V2=3,98±1,12 e V3=18,93±3,51. Após a DN os valores foram significantemente maiores (p<0,05). Na fala, os volumes foram os seguintes: V1=1,80±0,22; V2=4,22±1,20; V3=15,32±5,40. O valor de V3 na fala foi significantemente menor que o observado na ausência de atividade velofaríngea (p<0,05). Conclusão: A comparação das áreas seccionais e dos volumes obtidos com os relatados na literatura validam seu uso como valores de referência, assim como o emprego da rinometria acústica como método de avaliação objetiva da geometria nasal e da atividade velofaríngea na fala. / Objective: To determine cross-sectional areas and volumes of specific segments of the nasal cavity, including the nasopharyngeal region, of individuals without evidences of nasal obstruction, and the same volumes during speech, with the purpose of establishing reference values. Model: Prospective analysis. Setting: Laboratory of Physiology, HRAC-USP. Participants: 30 caucasians volunteers, aged between 18 and 30 years (14 males and 16 females). Intervention/Variables: Cross-sectional areas corresponding to the nasal valve (CSA1), anterior edge of the inferior turbinate (CSA2) and posterior edge of the inferior turbinate (CSA3), and volumes corresponding to the nasal valve (V1), turbinate (V2) and nasopharyngeal regions (V3) were evaluated. Measurements were performed by acoustic rhinometry before and after nasal decongestion, at rest (voluntary interruption of respiration) and during velopharyngeal activity (silent continuous production of the sound /f/). Results: Mean values (±SD) of areas (in cm2) and volumes (in cm3) were the following: CSA1=0,54±0,13, CSA2=0,98±0,31, CSA3=1,42±0,44, V1=1,68±0,32, V2=3,98±1,12 and V3=18,93±3,51. After decongestion, the values were significantly larger (p<0,05). During speech, volumes were the following: V1=1,80±0,22; V2=4,22±1,20; V3=15,32±5,40. Nasopharyngeal volume (V3) measured during speech was significantly smaller than at rest (p<0,05). Conclusion: The comparison of the nasal cross-sectional areas and volumes obtained with those reported in the literature validate their use as reference values and the use of acoustic rhinometry to assess nasal geometry and velopharyngeal activity during speech.