Thesis (Ph. D.)--University of Hawaii at Manoa, 1991. / Includes bibliographical references (leaves 167-173) / Microfiche. / xxiv, 212 leaves, bound 29 cm

Thesis (Ph. D.)--Ohio State University, 2008. / Title from first page of PDF file. Includes bibliographical references (p. 161-172).

No description available.

No sistema imune de artrópodes, em contraste com o dos vertebrados, não ocorre uma resposta a antígenos por meio da produção de imunoglobulinas específicas contra agentes infecciosos. Portanto, a imunidade adaptativa está ausente, sendo que o sistema imune nesses animais baseia-se somente numa resposta inata. Os mecanismos presentes na resposta imune inata dos artrópodes incluem: (i) o sistema de coagulação; (ii) a cascata da profenoloxidase; (iii) a liberação de moléculas que possuem ação direta contra microorganismos dentre elas, os peptídeos antimicrobianos (PAMs). Estas moléculas podem ser constitutivas, como observado em aracnídeos, ou terem sua expressão induzida após desafio imune, como observado em alguns insetos. PAMs são moléculas anfipáticas, geralmente catiônicas e compostas por 9 a 100 resíduos. O estudo desses peptídeos, além de possibilitar a descoberta de novas moléculas que tenham modo de ação alternativo aos antibióticos convencionais, permite uma compreensão mais ampla do sistema imunológico de diferentes grupos de animais, bem como a origem dos seus mecanismos na história evolutiva. Neste trabalho foi utilizado o opilião Acutisoma longipes como modelo experimental para a caracterização de PAMs presentes na sua hemolinfa, tendo em vista que não há registros de estudos dessa natureza que utilizaram representantes da ordem Opiliones. Primeiramente, foi demonstrada a ocorrência de diversas frações, obtidas da purificação da hemolinfa, com atividade anti-M. luteus, consistidas por peptídeos que aparentemente são constitutivos. Uma das frações mostrou-se pura e apresentou um peptídeo de 2,1 kDa cuja estrutura primária, composta por 18 resíduos, foi completamente elucidada (SGYLPGKEYVYKYKGKVF) por sequenciamento de novo e sequenciamento do N-terminal. Este peptídeo linear foi nomeado longipina. O peptídeo sintético apresentou atividade antimicrobiana contra as bactérias Escherichia coli e Micrococcus luteus, e as leveduras Candida albicans e C. tropicalis, além de não apresentar atividade hemolítica na máxima concentração testada (100 mM). Foi demonstrado que a longipina liga-se preferencialmente a vesículas unilamelares grandes (LUVs), constituídas de fosfolipídios aniônicos (POPG), que mimetizam as cargas da superfície das células de microorganismos. A ligação do peptídeo a LUVs compostas, na razão molar de 1:1, por POPG e POPC (zwiteriônico) provoca o extravasamento de marcadores (carboxifluoresceína) aprisionados nos seus interiores. Esse peptídeo encontra-se majoritariamente desestruturado em solução ou na presença de sistema mimético composto somente por POPC. Porém, na sua ligação com POPG:POPC 1:1, adota estruturas b e estrutura relacionada à presente em agregados intermoleculares de fibras amiloides. / The arthropods immune system, in contrast to the vertebrates one, lacks a response to antigens through the production of specific immunoglobulins to fight against infectious agents. Therefore, the adaptive immunity is absent, and their immune system is only supported by an innate response. The mechanisms present in the arthropods innate immune response include: (i) the clotting system; (ii) the prophenoloxidase cascade; (ii) the release of molecules that can directly act against microorganisms among them, the antimicrobial peptides (AMPs). These molecules can be constitutive, as observed in arachnids, or have their expression induced upon imune challenge, as observed in some insects. AMPs are amphipathic molecules, usually cationic and formed by 9 to 100 residues. The study of these peptides, besides allowing the discovery of new molecules that exert their mode of action alternatively from the conventional antibiotics, permits a wider understanding of the immunological system from different groups of animals, as well as the origin of their mechanisms in the evolutionary history. The harvestman Acutisoma longipes was used in this work as na experimental model for the characterization of AMPs from its haemolymph, taking into account that there are no records of using representatives of the Opiliones order in such kind of study. Firstly, it was demonstrated the occurrence of several fractions, obtained from the haemolymph purification, that presented anti-M. luteus activity, consisted by peptides that are apparently constitutive. One of these fractions was pure and presented a 2.1 kDa peptide which had its primary structure, composed by 18 residues, completely elucidated by de novo and N-terminal sequencing (SGYLPGKEYVYKYKGKVF). This linear peptide was named longipin. The synthetic peptide presented antimicrobial activity against Escherichia coli and Micrococcus luteus bacteria and Candida albicans and C. tropicalis yeasts, in addition to the absence of hemolytic activity at the highest concentration used (100mM). It was shown that longipin preferentially binds to large unilamellar vesicles (LUVs), constituted by anionic phospholipds (POPG), that mimics the membrane surface of microbe cells. The peptide biding to LUVs composed, in 1:1 molar ratio, by POPG and POPC (zwitterionic) causes dye leakage (carboxyfluorescein) from their inside. This peptide in solution or in the presence of POPC solely containing mimic system is mostly unstructured. However, upon its binding to POPG:POPC 1:1, it adopts b and intermolecular aggregates amyloid-like fibrils structures.

Low noise, directional microphones are critical for hearing aid applications. This thesis is focused on further development of a biomimetic micromachined directional microphone based on the ear structure of the Ormia Ochracea, a parasitic fly able to locate sound sources in the audio frequency range with high accuracy. The development efforts have been on implementing a version of the microphone for a behind the ear (BTE) package while improving the overall optical efficiency and noise level, demonstrating pulsed laser operation for reduced power consumption, and electrostatic control of the microphone diaphragm position for stable operation over a long time. The new packaging method for the microphone addressed the need for tighter placement tolerances along with a redesigned diaphragm and integration of a microscale optical lens array to improve the optical efficiency of the device. The completed packages were characterized for sensitivity improvement and optical efficiency. The overall optical efficiency was significantly increased from less than 1% to the photo diode array collecting 50% of the emitted optical power from the Vertical Cavity Surface Emitting Laser (VCSEL). This, coupled with the new diaphragm design, improved the acoustic performance of the microphones. Consequently, the noise levels recorded on the devices were about 31 dBA SPL, more than 15dB better than conventional directional microphones with nearly 10 times larger port spacing. Since the application for this technology is hearing aids, the power consumed by the working device needs to be at an acceptable level. The majority of the power used by the microphone is from continuously operating the VCSEL with 2mW optical output power. To reduce this power requirement, it was suggested to pulse the VCSEL at high enough frequency with low duty cycle so that the acoustic signals can be recovered from its samples. In this study, it was found that the VCSEL can be pulsed with little to no degradation in signal to noise ratio as long as the thermal mechanical noise dominated the noise spectrum. The results also indicated that a pulse train with a duty cycle of around 20% can be used without a major loss of performance in the device, meaning the device can effectively run at 1/5 of its original power under pulsed operation mode. Finally, a control technique to overcome some inherent problems of the microphone was demonstrated. Since the optical sensitivity of the microphone depends on the gap between the diaphragm grating and the integrated mirror, it is important to keep that bias gap constant during long term operation against environmental variations and charging effects. Using a simple electrostatic bias controller scheme, the sensitivity variation of the microphone was improved by a factor of 7.68 with bias control. Overall, this thesis has addressed several important aspects of a micromachined biomimetic microphone and further demonstrated its feasibility for hearing aid applications.

No sistema imune de artrópodes, em contraste com o dos vertebrados, não ocorre uma resposta a antígenos por meio da produção de imunoglobulinas específicas contra agentes infecciosos. Portanto, a imunidade adaptativa está ausente, sendo que o sistema imune nesses animais baseia-se somente numa resposta inata. Os mecanismos presentes na resposta imune inata dos artrópodes incluem: (i) o sistema de coagulação; (ii) a cascata da profenoloxidase; (iii) a liberação de moléculas que possuem ação direta contra microorganismos dentre elas, os peptídeos antimicrobianos (PAMs). Estas moléculas podem ser constitutivas, como observado em aracnídeos, ou terem sua expressão induzida após desafio imune, como observado em alguns insetos. PAMs são moléculas anfipáticas, geralmente catiônicas e compostas por 9 a 100 resíduos. O estudo desses peptídeos, além de possibilitar a descoberta de novas moléculas que tenham modo de ação alternativo aos antibióticos convencionais, permite uma compreensão mais ampla do sistema imunológico de diferentes grupos de animais, bem como a origem dos seus mecanismos na história evolutiva. Neste trabalho foi utilizado o opilião Acutisoma longipes como modelo experimental para a caracterização de PAMs presentes na sua hemolinfa, tendo em vista que não há registros de estudos dessa natureza que utilizaram representantes da ordem Opiliones. Primeiramente, foi demonstrada a ocorrência de diversas frações, obtidas da purificação da hemolinfa, com atividade anti-M. luteus, consistidas por peptídeos que aparentemente são constitutivos. Uma das frações mostrou-se pura e apresentou um peptídeo de 2,1 kDa cuja estrutura primária, composta por 18 resíduos, foi completamente elucidada (SGYLPGKEYVYKYKGKVF) por sequenciamento de novo e sequenciamento do N-terminal. Este peptídeo linear foi nomeado longipina. O peptídeo sintético apresentou atividade antimicrobiana contra as bactérias Escherichia coli e Micrococcus luteus, e as leveduras Candida albicans e C. tropicalis, além de não apresentar atividade hemolítica na máxima concentração testada (100 mM). Foi demonstrado que a longipina liga-se preferencialmente a vesículas unilamelares grandes (LUVs), constituídas de fosfolipídios aniônicos (POPG), que mimetizam as cargas da superfície das células de microorganismos. A ligação do peptídeo a LUVs compostas, na razão molar de 1:1, por POPG e POPC (zwiteriônico) provoca o extravasamento de marcadores (carboxifluoresceína) aprisionados nos seus interiores. Esse peptídeo encontra-se majoritariamente desestruturado em solução ou na presença de sistema mimético composto somente por POPC. Porém, na sua ligação com POPG:POPC 1:1, adota estruturas b e estrutura relacionada à presente em agregados intermoleculares de fibras amiloides. / The arthropods immune system, in contrast to the vertebrates one, lacks a response to antigens through the production of specific immunoglobulins to fight against infectious agents. Therefore, the adaptive immunity is absent, and their immune system is only supported by an innate response. The mechanisms present in the arthropods innate immune response include: (i) the clotting system; (ii) the prophenoloxidase cascade; (ii) the release of molecules that can directly act against microorganisms among them, the antimicrobial peptides (AMPs). These molecules can be constitutive, as observed in arachnids, or have their expression induced upon imune challenge, as observed in some insects. AMPs are amphipathic molecules, usually cationic and formed by 9 to 100 residues. The study of these peptides, besides allowing the discovery of new molecules that exert their mode of action alternatively from the conventional antibiotics, permits a wider understanding of the immunological system from different groups of animals, as well as the origin of their mechanisms in the evolutionary history. The harvestman Acutisoma longipes was used in this work as na experimental model for the characterization of AMPs from its haemolymph, taking into account that there are no records of using representatives of the Opiliones order in such kind of study. Firstly, it was demonstrated the occurrence of several fractions, obtained from the haemolymph purification, that presented anti-M. luteus activity, consisted by peptides that are apparently constitutive. One of these fractions was pure and presented a 2.1 kDa peptide which had its primary structure, composed by 18 residues, completely elucidated by de novo and N-terminal sequencing (SGYLPGKEYVYKYKGKVF). This linear peptide was named longipin. The synthetic peptide presented antimicrobial activity against Escherichia coli and Micrococcus luteus bacteria and Candida albicans and C. tropicalis yeasts, in addition to the absence of hemolytic activity at the highest concentration used (100mM). It was shown that longipin preferentially binds to large unilamellar vesicles (LUVs), constituted by anionic phospholipds (POPG), that mimics the membrane surface of microbe cells. The peptide biding to LUVs composed, in 1:1 molar ratio, by POPG and POPC (zwitterionic) causes dye leakage (carboxyfluorescein) from their inside. This peptide in solution or in the presence of POPC solely containing mimic system is mostly unstructured. However, upon its binding to POPG:POPC 1:1, it adopts b and intermolecular aggregates amyloid-like fibrils structures.