Impact of soft contact lens edge design and mid-peripheral lens shape on the epithelium and its indentation with lens mobility

Purpose. To evaluate the influence of soft contact lens midperipheral shape profile and edge design on the apparent epithelial thickness and indentation of the ocular surface with lens movement. Methods. Four soft contact lens designs comprising of two different plano midperipheral shape profiles and two edge designs (chiseled and knife edge) of silicone-hydrogel material were examined in 26 subjects aged 24.7 ± 4.6 years, each worn bilaterally in randomized order. Lens movement was imaged enface on insertion, at 2 and 4 hours with a high-speed, high-resolution camera simultaneous to the cross-section of the edge of the contact lens interaction with the ocular surface captured using optical coherence tomography (OCT) nasally, temporally, and inferiorly. Optical imaging distortions were individually corrected for by imaging the apparent distortion of a glass slide surface by the removed lens. Results. Apparent epithelial thickness varied with edge position (P < 0.001). When distortion was corrected for, epithelial indentation decreased with time after insertion (P = 0.010), changed after a blink (P < 0.001), and varied with position on the lens edge (P < 0.001), with the latter being affected by midperipheral lens shape profile and edge design. Horizontal and vertical lens movement did not change with time postinsertion. Vertical motion was affected by midperipheral lens shape profile (P < 0.001) and edge design (P < 0.001). Lens movement was associated with physiologic epithelium thickness for lens midperipheral shape profile and edge designs. Conclusions. Dynamic OCT coupled with high-resolution video demonstrated that soft contact lens movement and image-corrected ocular surface indentation were influenced by both lens edge design and midperipheral lens shape profiles. © 2013 The Association for Research in Vision and Ophthalmology, Inc.

Avaliação da pressão inspiratória nasal sniff na população obesa

Introdução: a avaliação da pressão inspiratória nasal (SNIP) é considerada uma manobra complementar da Pressão Inspiratória Máxima estática (PImax) em várias condições clínicas, porém não há relatos na obesidade. Por outro lado, a obesidade tem um importante impacto nos músculos respiratórios especialmente com maiores gordura abdominal o que provavelmente pode ser detectado na avaliação da SNIP que mensura mais precisamente a pressão diafragmática. Objetivo: analisar em obesos a relação entre SNIP e variáveis respiratórias e marcadores de adiposidade. Material e Método: num estudo transversal um total de 92 obesos (38.3±10.2 anos) sem história de doença respiratória ou cardíaca diagnosticada. Foram avaliados na espirometria (capacidade vital forçada-CVF; volume expiratório forçado no primeiro segundo-VEF1; volume de reserva expiratório-VRE) e pressões respiratórias estática (PImax, PEmax e SNIP) e dinâmica (ventilação voluntária máxima-VVM). Sendo considerados os marcadores de adiposidade: índice de adiposidade corporal-IAC; índice de massa corporal-IMC e circunferências do quadril (CQ), cintura (CC) e pescoço (CP). Resultados: 65 obesos mórbidos (IMC=50.8±8.1Kg/m2) e 27 obesos não mórbidos (IMC=35.6±2.7Kg/m2) foram homogêneos (p>0.05) na SNIP (99.1±24.5cmH2O, 87% do predito) e PImax (107.3±26.4cmH2O, 109% do predito). Existe correlação (r=0.5) entre SNIP e PImax somente no grupo de obesos mórbidos. De acordo com as correlações houve associação entre variáveis respiratórias (CVF r=0.48; VEF1 r=0.54; e VVM r=0.54), valores antropométricos (idade r=-0.44) e SNIP somente para os obesos mórbidos. Esses achados foram certificados quando também comparados a quantidade de gordura ao redor do pescoço (CP≥43cm). O modelo de regressão linear stepwise mostrou que a VVM parece ser o melhor preditor para explicar a SNIP nos obesos mórbidos. Nestes obesos a SNIP foi levemente mais baixa (87%predito) que os valores esperados para indivíduos brasileiros saudáveis. Conclusão: em obesos mórbidos a SNIP é moderadamente relacionada a PImax. A SNIP parece ser mais relacionada a VVM que à marcadores de adiposidade.

Dynamic Compartmentalization of Persistent UPEC in the Superficial Bladder Epithelium

Urinary tract infections (UTIs) are typically caused by bacteria that colonize different regions of the urinary tract, mainly the bladder and the kidney. Approximately 25% of women that suffer from UTIs experience a recurrent infection within 6 months of the initial bout, making UTIs a serious economic burden resulting in more than 10 million hospital visits and $3.5 billion in healthcare costs in the United States alone. Type-1 fimbriated Uropathogenic E. coli (UPEC) is the major causative agent of UTIs, accounting for almost 90 % of bacterial UTIs. The unique ability of UPEC to bind and invade the superficial bladder epithelium allows the bacteria to persist inside epithelial niches and survive antibiotic treatment. Persistent, intracellular UPEC are retained in the bladder epithelium for long periods, making them a source of recurrent UTIs. Hence, the ability of UPEC to persist in the bladder is a matter of major health and economic concern, making studies exploring the underlying mechanism of UPEC persistence highly relevant.

In my thesis, I will describe how intracellular Uropathogenic E.coli (UPEC) evade host defense mechanisms in the superficial bladder epithelium. I will also describe some of the unique traits of persistent UPEC and explore strategies to induce their clearance from the bladder. I have discovered that the UPEC virulence factor Alpha-hemolysin (HlyA) plays a key role in the survival and persistence of UPEC in the superficial bladder epithelium. In-vitro and in-vivo studies comparing intracellular survival of wild type (WT) and hemolysin deficient UPEC suggested that HlyA is vital for UPEC persistence in the superficial bladder epithelium. Further in-vitro studies revealed that hemolysin helped UPEC persist intracellularly by evading the bacterial expulsion actions of the bladder cells and remarkably, this virulence factor also helped bacteria avoid t degradation in lysosomes.

To elucidate the mechanistic basis for how hemolysin promotes UPEC persistence in the urothelium, we initially focused on how hemolysin facilitates the evasion of UPEC expulsion from bladder cells. We found that upon entry, UPEC were encased in “exocytic vesicles” but as a result of HlyA expression these bacteria escaped these vesicles and entered the cytosol. Consequently, these bacteria were able to avoid expulsion by the cellular export machinery.

Since bacteria found in the cytosol of host cells are typically recognized by the cellular autophagy pathway and transported to the lysosomes where they are degraded, we explored why this was not the case here. We observed that although cytosolic HlyA expressing UPEC were recognized and encased by the autophagy system and transported to lysosomes, the bacteria appeared to avoid degradation in these normally degradative compartments. A closer examination of the bacteria containing lysosomes revealed that they lacked V-ATPase. V-ATPase is a well-known proton pump essential for the acidification of mammalian intracellular degradative compartments, allowing for the proper functioning of degradative proteases. The absence of V-ATPase appeared to be due to hemolysin mediated alteration of the bladder cell F-actin network. From these studies, it is clear that UPEC hemolysin facilitates UPEC persistence in the superficial bladder epithelium by helping bacteria avoid expulsion by the exocytic machinery of the cell and at the same time enabling the bacteria avoid degradation when the bacteria are shuttled into the lysosomes.

Interestingly even though UPEC appear to avoid elimination from the bladder cell their ability to multiple in bladder cells seem limited.. Indeed, our in-vitro and in-vivo experiments reveal that UPEC survive in superficial bladder epithelium for extended periods of time without a significantly change in CFU numbers. Indeed, we observed these bacteria appeared quiescent in nature. This observation was supported by the observation that UPEC genetically unable to enter a quiescence phase exhibited limited ability to persist in bladder cells in vitro and in vivo, in the mouse bladder.

The studies elucidated in this thesis reveal how UPEC toxin, Alpha-hemolysin plays a significant role in promoting UPEC persistence via the modulation of the vesicular compartmentalization of UPEC at two different stages of the infection in the superficial bladder epithelium. These results highlight the importance of UPEC Alpha-hemolysin as an essential determinant of UPEC persistence in the urinary bladder.

Discerning the sub-phonemic identity of pre-nasal /æ/ in British Columbia English

There are two features of /æ/ in British Columbia (BC) English that are widely attested in the literature: it is undergoing retraction and lowering and it is sensitive to the influence of certain following consonants. The present study aims to utilize both features to evaluate the phonological status of /æ/ before nasal consonants in BC English by examining the progression of sound change and the phonemic organization of /æ/ in different environments. Specifically, production and perception results are taken together to evaluate the phonetic position of pre-nasal /æ/ relative to other environments. These results are interpreted within a modular feedforward architecture of phonology to establish the phonological (allophonic) and phonetic (shallowphonic) rules that govern the internal relationships between the subphonemic elements of /æ/ in BC English. Further, the findings of this study provide evidence for the allophone being the target of sound change, rather than the phoneme.

In Vitro modelling of RSV infection and cytopathogenesis in well-differentiated human primary airway epithelial cells (WD-PAECs).

The choice of model used to study human respiratory syncytial virus (RSV) infection is extremely important. RSV is a human pathogen that is exquisitely adapted to infection of human hosts. Rodent models, such as mice and cotton rats, are semi-permissive to RSV infection and do not faithfully reproduce hallmarks of RSV disease in humans. Furthermore, immortalized airway-derived cell lines, such as HEp-2, BEAS-2B, and A549 cells, are poorly representative of the complexity of the respiratory epithelium. The development of a well-differentiated primary pediatric airway epithelial cell models (WD-PAECs) allows us to simulate several hallmarks of RSV infection of infant airways. They therefore represent important additions to RSV pathogenesis modeling in human-relevant tissues. The following protocols describe how to culture and differentiate both bronchial and nasal primary pediatric airway epithelial cells and how to use these cultures to study RSV cytopathogenesis.

Slow Oscillations in the Mouse Hippocampus Entrained by Nasal Respiration

Different types of network oscillations occur in different behavioral, cognitive, or vigilance states. The rodent hippocampus expresses prominentoscillations atfrequencies between 4 and 12Hz,which are superimposed by phase-coupledoscillations (30 –100Hz).These patterns entrain multineuronal activity over large distances and have been implicated in sensory information processing and memory formation. Here we report a new type of oscillation at near- frequencies (2– 4 Hz) in the hippocampus of urethane-anesthetized mice. The rhythm is highly coherent with nasal respiration and with rhythmic field potentials in the olfactory bulb: hence, we called it hippocampal respiration-induced oscillations. Despite the similarity in frequency range, several features distinguish this pattern from locally generatedoscillations: hippocampal respiration-induced oscillations have a unique laminar amplitude profile, are resistant to atropine, couple differentlytooscillations, and are abolished when nasal airflow is bypassed bytracheotomy. Hippocampal neurons are entrained by both the respiration-induced rhythm and concurrent oscillations, suggesting a direct interaction between endogenous activity in the hippocampus and nasal respiratory inputs. Our results demonstrate that nasal respiration strongly modulates hippocampal network activity in mice, providing a long-range synchronizing signal between olfactory and hippocampal networks.

Regenerative Cues of Myeloid Cells on Pancreatic Epithelium

Thesis (Ph.D.)--University of Washington, 2016-07

Dacriocistorrinostomia endoscópica nasal: resultados do Hospital Professor Doutor Fernando Fonseca

Objectivo: analisar os resultados da experiência de um hospital distrital na realização de dacriocistorrinostomias (DCR) endoscópicas e determinar o papel actual desta técnica para tratamento de obstruções da via nasolacrimal. Materiais e Métodos: Reviram-se os processos clínicos de 67 dacriocistorrinostomias endoscópicas consecutivas realizadas no Hospital Fernando Fonseca entre Janeiro de 1997 e Dezembro de 2012. Analisou-se a clínica apresentada pelos doentes, o nível da estenose detectado na dacriocistografia, as complicações intra e pós-operatórias e os resultados funcionais e anatómicos pós-operatórios. Resultados: Dos 51 doentes, 12 eram do sexo masculino e 34 do sexo feminino, com média de idades de 57 anos. A epífora foi a queixa predominante. Com a excepção de dois casos, todas as cirurgias foram executadas com instrumentos de dissecção motorizados. O sucesso da DCR endoscópica foi de 78,1%. Conclusões: A DCR endoscópica é uma técnica simples, segura e eficaz para o tratamento das obstruções distais da via nasolacrimal.

New approach to study the real exposure to fungi in cork industry: nasal swabs mycobiota investigation coupled with screening on fungal resistance to azoles

The permanent contact with cork may lead to constant exposure to fungi, raising awareness as a potential occupational hazard in the cork industry.The presence of fungi belonging to the Penicillium glabrum complex has been associated with the development of respiratory diseases such as suberosis, one of the most prevalent diseases among workers from cork industries, besides occupational asthma. Azoles are used as pesticides but also the first line therapy in the treatment of Aspergillus infections; azole-resistance as been described as to have also an environmental source and is considered an emerging public health problem.The aim of this work was to characterize fungal distribution and to evaluate the presence of azole-resistant Aspergillus isolates in nose swab samples from the cork industry workers.

Slow Oscillations in the Mouse Hippocampus Entrained by Nasal Respiration

Different types of network oscillations occur in different behavioral, cognitive, or vigilance states. The rodent hippocampus expresses prominentoscillations atfrequencies between 4 and 12Hz,which are superimposed by phase-coupledoscillations (30 –100Hz).These patterns entrain multineuronal activity over large distances and have been implicated in sensory information processing and memory formation. Here we report a new type of oscillation at near- frequencies (2– 4 Hz) in the hippocampus of urethane-anesthetized mice. The rhythm is highly coherent with nasal respiration and with rhythmic field potentials in the olfactory bulb: hence, we called it hippocampal respiration-induced oscillations. Despite the similarity in frequency range, several features distinguish this pattern from locally generatedoscillations: hippocampal respiration-induced oscillations have a unique laminar amplitude profile, are resistant to atropine, couple differentlytooscillations, and are abolished when nasal airflow is bypassed bytracheotomy. Hippocampal neurons are entrained by both the respiration-induced rhythm and concurrent oscillations, suggesting a direct interaction between endogenous activity in the hippocampus and nasal respiratory inputs. Our results demonstrate that nasal respiration strongly modulates hippocampal network activity in mice, providing a long-range synchronizing signal between olfactory and hippocampal networks.

Nuclear alterations in nasal mucosa epithelial cells of students exposed to formaldehyde

Introduction: Formaldehyde is a compound with a wide range and is commonly used in anatomy and pathology laboratories. At room temperature is quickly volatilized to a pungent and suffocating gas and its inhalation has been correlated to nuclear alterations in different tissues. We aimed to investigate whether exposure to this compound was correlated with the appearance of cytotoxic and genotoxic features in the nasal epithelial cells of students enrolled in a human anatomy course. Material and Methods: This prospective study collected periodically nasal cells from mucosa of 17 volunteers from two different undergraduate programs with different workloads of practical lessons in an anatomy laboratory, 30 and 90 hours per semester. Cells were staining according to Feulgen method and nuclear morphology was analyzed to detect possible damage. Dunn's post hoc test was used in the statistical analysis. Pearson's correlation was performed for gender, age and questionnaire responses. Results: Epithelial cells showed indicators of cytotoxicity and mutagenicity. Students with a more extensive workload in anatomy laboratory displayed a more severe profile with an increase in karyorrhexis (p < 0.05) over time. The micronucleus analysis showed difference between first and second collection (p < 0.01), although it was not maintained over the time. Students with a less extensive workload display no differences in most of cytological features. Despite karyorrhexis was present in a greater number of cells, for this group no significant difference was observed between any range. The same was observed to karyolysis and micronucleus (p > 0.05). Conclusion: Individuals exposed for short periods of time to formaldehyde are subject to the toxic action of this gas. Karyorrhexis was the most frequently observed cytotoxic feature and micronucleus showed an increase between the first time point. The patterns observed between the student's groups suggest a negative effect due to exposure time.

Determinación de la Flora Bacteriana Nasal y Cloacal de la Tortuga “golfina” Lepidochelys olivacea, Especie Anidante en el Área Natural Protegida Complejo Los Cóbanos. Sonsonate, El Salvador

Durante su ciclo de vida las tortugas marinas están expuestas a muchos peligros, cada año mueren al ser capturadas accidentalmente en las redes de pesca, víctimas de la pérdida de hábitat de áreas de alimentación y anidación, debido al desarrollo costero e incremento del turismo, lo anterior ha ocasionado que se les considere como especies en peligro o amenazadas de extinción. La mayoría de las enfermedades de las tortugas marinas aún no han sido descritas; por lo que es necesario conocer las bacterias que se encuentran presente en el organismo y los posibles vectores de enfermedades. El objetivo del presente estudio fue determinar la composición de la flora bacteriana en el área nasal y cloacal de las tortugas marinas Lepidochelys olivacea durante su época de anidación entre los meses de junio a octubre del año 2012, se colectaron muestras bacteriológicas provenientes de 20 hembras anidantes mediante recorridos nocturnos en tres playas El Faro, El Cocal y Salinitas pertenecientes al Área Natural Protegida Complejos Los Cóbanos Departamento de Sonsonate, El Salvador. Las muestras se colectaron a través del uso de hisopos estériles que se introdujeron en la cloaca durante el desove y en uno de los conductos nasales. Estas fueron trasladadas en al laboratorio de Biología de la Universidad de El Salvador en menos de 24 horas para su posterior análisis. En el laboratorio las muestras nasales fueron cultivadas en agar sangre, MacConkey y agar manitol sal, las cloacales fueron cultivadas en agar sangre, MacConkey, TCBS y XLD, a todas las colonias aisladas se les realizaron pruebas bioquímicas rutinarias IMVIC. Además se utilizó API 20E.

Conidiobolomicosis nasal en una oveja (Ovis aries) en Uruguay

Resumen La conidiobolomicosis es una enfermedad causada por Conidiobolus spp. que afecta a varias especies de animales, incluyendo ovinos, llamas, ciervos, caballos, perros y humanos. En ovinos se caracteriza por lesiones granulomatosas en la cavidad nasal. Aunque la enfermedad es endémica en distintas regiones de Brasil, aún no ha sido reportada en Uruguay. El objetivo de este trabajo es describir un caso de conidiobolomicosis nasal en una oveja en Uruguay y proveer una breve revisión bibliográfica. El caso ocurrió en marzo del 2015 en una oveja de 19 meses de edad en un establecimiento ubicado en el departamento de Colonia. Los principales signos clínicos fueron debilidad, secreción nasal, obnubilación, torneo y exoftalmos unilateral. Los principales hallazgos patológicos fueron rinosinusitis granulomatosa necrotizante localmente extensiva, afectando la región posterior de la cavidad nasal, con desplazamiento del tabique nasal y osteítis necrotizante/osteolítica de los huesos faciales, con extensión al tejido subcutáneo facial y espacio retro-ocular derechos. Microscópicamente se constató la presencia de hifas intralesionales, identificadas por inmunohistoquímica como Conidiobolus spp., rodeadas por material de Splendore-Hoeppli. Se concluye que la conidiobolomicosis ocurre esporádicamente en Uruguay y que debe ser considerada dentro de los diagnósticos diferenciales de rinitis en ovinos en este País