Malignant mesothelioma (MM) is an aggressive and highly chemo-resistant tumour of the mesothelium. Asbestos is indicated as the environmental factor most commonly associated with mesothelioma. The chemo-resistance is possibly due to impaired apoptotic mechanisms since it is known most chemotherapeutic drugs act via apoptosis. Murine MM cell lines that have been derived from tumours induced by inoculation of crocidolite asbestos into mice provide a suitable model, since both phenotypic and biological properties are closely similar to the human disease. Four murine MM cell lines were used in this study, namely ABl, AB12, AC29 and AC34. The aim of this study was to determine susceptibility of those MM cell lines to induction of apoptosis and the expression of key molecules in those cells. Many apoptosis-related genes are known, expression of some of these genes in four murine MM cell lines were investigated in this study. Gene expression was determined using conventional reverse transcription PCR (RT-PCR) and quantitated using real-time RT-PCR with SYBR-Green I detection on a Rotor-Gene 2000 (Corbett Research, N.S.W., Australia). Gene expression data was normalised against the most stable housekeeping genes as determined by the geNonn software. Susceptibility of the four murine MM cell lines to apoptosis induction was determined using cisplatin or TNF-a or IFN-y at different concentrations and at different times in the case of cisplatin. Apoptosis was assessed by a DNA laddering assay. Conventional RT-PCR results showed that all four murine MM cell lines expressed DR5, Bax, Bcl-xL, FLIP-L, c-Myc and caspase-3. Fas mRNA was detected in all cell lines except AC29. Neither FasL nor Bcl-2 was expressed in the four murine MM cell lines. / Quantitation of gene expression showed that there were significant differences in Fas, DRS, Bax, Bcl-xL, c-Myc, FLIP-L and caspase-3 mRNA levels across the cell lines (P<0.05). Absence of Fas receptor in AC29 may play a role in the immunogenicity of this cell line. DNA laddering results indicated that cisplatin induced apoptosis in a dose- and time- dependent manner in those MM cell lines. Susceptibility as reflected by the minimum apoptosis-inducing dose varied among the cell lines from 1 pdml (AB12) to 10 & n l (AC34). Susceptibility to TNF-a or IFN-y also varied among the cell lines where AB12 was sensitive while AC29 was resistant to those cytokines. The AC29 and AB1 cell lines were used to examine cisplatin or TNF-a induced genes related to apoptosis, expression of Fas, caspase-3, Bax and Bcl-xL mRNA was determined using real-time RT-PCR after 6 and 24 hours induction with cisplatin or TNF-a. The results demonstrated that caspase-3 and Bax were up-regulated whereas Bcl-xL was down-regulated in AC29 after 6 hours treatment with cisplatin. Although only Bcl-xL was down-regulated in ABl after 6 hours treatment with cisplatin, the down regulation was more pronounced than in AC29. TNF-a induction of gene expression showed that Bcl-xl decreased and Fas was up-regulated significantly in AB1 after 6 hours whereas only Bcl-xL was down-regulated in AC29 after 6 hours and continued to decrease until 24 hours. The differences in gene expression changes were noticed not only between cell lines but also between two inducer agents. There were several significant results of this study. Firstly, gene expression in murine MM was seen to parallel those that have previously been described for the human disease. / Therefore murine MM is likely to be a useful in in vivo model for future studies targeting apoptotic molecules. Secondly, a number of genes were not previously examined in MM were characterized in the murine model. Finally, differences in basal and induced gene expression between cell lines and inducing agents were characterized which should be followed in further studies at the protein level (eg caspase-3 activity).

This thesis presents the design and implementation of 140 GHz to 170 GHz transceivers in SiGe HBT technologies and a 95 GHz receiver in 65 nm CMOS technology. Optimization and modeling of all passive components and transistor biasing at peak-fT and peak-fMAX current densities are employed to obtain higher frequency operation of circuit blocks compared to state of the art. These circuit blocks include static and dynamic frequency dividers, voltage-controlled oscillators, and tuned mm-wave amplifiers. Design procedures for a 100 GHz static divider, a 136 GHz dynamic divider, as well as low-power divider topologies are presented. A methodology for the design of quadrature voltage-controlled oscillators in CMOS and SiGe technologies is described, together with a technique for reduced-power LO-path design. Tuned 5-stage 140 GHz, 160 GHz, and 170 GHz amplifiers with more than 15 dB in SiGe HBT technology are reported. Using these circuit building blocks, a 95 GHz receiver in 65 nm CMOS technology with 12.5 dB gain, 7 dB noise figure, and 206mW. Also, several 165 GHz transceivers are implemented in SiGe HBT technology. The 165 GHz transceivers that include an oscillator, a divider, RX, LO, and TX amplifiers, and a mixer, were designed with and without on-chip antennas. They have -3 dB conversion gain, all achieved at RF, and -3.5 dBm output power. Following that, a 140 GHz fully-integrated transceiver and a 140 GHz transceiver array with on-chip patch antennas were designed in a 0.13micron SiGe BiCMOS technology, demonstrating the highest integration levels at this frequency in silicon to date. These transceivers feature 136-145 GHz voltage-controlled oscillators, 20 dB receive-amplifiers and mixers,transmit-amplifiers with amplitude-shift keying modulation, and variable-gain IF amplifiers. At 140 GHz the transceivers have up to 32 dB conversion gain and -8 dBm output power. Wireless data transmission at 4 Gb/s was demonstrated over 1.15m with off-chip horn antennas, and over 2 cm with the on chip antennas. With on-chip antennas, the transceiver could detect a Doppler shift of as little as 25 Hz. Both transceivers were also operational in frequency-modulated continuous-wave radar mode.

This thesis presents the design and implementation of 140 GHz to 170 GHz transceivers in SiGe HBT technologies and a 95 GHz receiver in 65 nm CMOS technology. Optimization and modeling of all passive components and transistor biasing at peak-fT and peak-fMAX current densities are employed to obtain higher frequency operation of circuit blocks compared to state of the art. These circuit blocks include static and dynamic frequency dividers, voltage-controlled oscillators, and tuned mm-wave amplifiers. Design procedures for a 100 GHz static divider, a 136 GHz dynamic divider, as well as low-power divider topologies are presented. A methodology for the design of quadrature voltage-controlled oscillators in CMOS and SiGe technologies is described, together with a technique for reduced-power LO-path design. Tuned 5-stage 140 GHz, 160 GHz, and 170 GHz amplifiers with more than 15 dB in SiGe HBT technology are reported. Using these circuit building blocks, a 95 GHz receiver in 65 nm CMOS technology with 12.5 dB gain, 7 dB noise figure, and 206mW. Also, several 165 GHz transceivers are implemented in SiGe HBT technology. The 165 GHz transceivers that include an oscillator, a divider, RX, LO, and TX amplifiers, and a mixer, were designed with and without on-chip antennas. They have -3 dB conversion gain, all achieved at RF, and -3.5 dBm output power. Following that, a 140 GHz fully-integrated transceiver and a 140 GHz transceiver array with on-chip patch antennas were designed in a 0.13micron SiGe BiCMOS technology, demonstrating the highest integration levels at this frequency in silicon to date. These transceivers feature 136-145 GHz voltage-controlled oscillators, 20 dB receive-amplifiers and mixers,transmit-amplifiers with amplitude-shift keying modulation, and variable-gain IF amplifiers. At 140 GHz the transceivers have up to 32 dB conversion gain and -8 dBm output power. Wireless data transmission at 4 Gb/s was demonstrated over 1.15m with off-chip horn antennas, and over 2 cm with the on chip antennas. With on-chip antennas, the transceiver could detect a Doppler shift of as little as 25 Hz. Both transceivers were also operational in frequency-modulated continuous-wave radar mode.

The demand for high data rate transfers in short range areas have been increasing significantly. Millimeter wave communication systems can fulfill the requirements for such applications due to the availability of wide bandwidths at these frequencies. Particularly, 60 GHz frequency band is more appropriate among other mm-wave bands because of the oxygen energy absorption resonance at this frequency. Millimeter wave antennas are one of the desired components in short range wireless communications. High gain and broadband antennas are required for this purpose. In this study, a fully planar 60 GHz antenna is introduced. Tapered dielectric rod antenna is chosen to achieve high radiation efficiency. The antenna is designed on two common substrates with high permittivity: alumina (Al2O3) and high resistive Silicon. Both substrates are very low loss and many designs for front-end components are developed on these substrates due to their high permittivity. In other words, the proposed antenna can be integrated with the front-end platform in the same substrate. In addition, the antenna feeding is the CPW line which makes it a convenient solution for integration of the antenna with RF front-ends such as MMICs or MEMS circuits in this range of frequency.

The demand for high data rate transfers in short range areas have been increasing significantly. Millimeter wave communication systems can fulfill the requirements for such applications due to the availability of wide bandwidths at these frequencies. Particularly, 60 GHz frequency band is more appropriate among other mm-wave bands because of the oxygen energy absorption resonance at this frequency. Millimeter wave antennas are one of the desired components in short range wireless communications. High gain and broadband antennas are required for this purpose. In this study, a fully planar 60 GHz antenna is introduced. Tapered dielectric rod antenna is chosen to achieve high radiation efficiency. The antenna is designed on two common substrates with high permittivity: alumina (Al2O3) and high resistive Silicon. Both substrates are very low loss and many designs for front-end components are developed on these substrates due to their high permittivity. In other words, the proposed antenna can be integrated with the front-end platform in the same substrate. In addition, the antenna feeding is the CPW line which makes it a convenient solution for integration of the antenna with RF front-ends such as MMICs or MEMS circuits in this range of frequency.

Orientador: Fausto Berzin / Dissertação (mestrado) - Universidade Estadula de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-07-24T19:49:56Z (GMT). No. of bitstreams: 1 Fornasari_CarlosAlberto_M.pdf: 1832465 bytes, checksum: 93ee85cbc557a85a6cb4c3005870e44e (MD5) Previous issue date: 1999 / Resumo: Considerando a importância das ações dos músculos (MM.) rombóides maior e menor na manutenção, fixação e estabilização da escápula a proposta deste estudo foi analisá-los eletromiograficamente durante os movimentos dinâmicos e livres de adução do úmero de 150° a 0°, abdução horizontal de 90° a 0° e abdução horizontal de 0° a 50°, em decúbito ventral, combinando-os com as variações de posições do úmero: neutra, rotação medial e rotação lateral. Os registros eletromiográficos foram coletados em oito homens, saudáveis, destros, com idade entre 22 e 25 anos (média 22,8 '+ ou -' 1,0), a partir de um eletromiógrafo multicanal Nicolet Viking, com a utilização do programa MVA (Maximum Voluntary Activity), por meio de utilização de eletrodos de fios Karma Allong, 25 mícron. Os resultados estão apresentados em RMS (Root Mean Square) em 'mu 'V. A análise estatística dos resultados foi realizada por meio dos teste de FRI DMAN e WlLCOXON, em função da não distribuição normal dos dados, com nível de % de significância ( 'alfa '=0,05). Os músculos apresentaram sinal eletromiográfico em todos os movimentos estudados, em todas as variações de posição do úmero. Em todos os movimentos estudados foi observado maior sinal eletromiográfico para o M. rombóide maior do que para o M. rombóide menor (p<0,05). Considerando o M. rombóide maior, este apresentou maior sinal eletromiográfico no movimento de abdução horizontal 0° a 50° e menor sinal eletromiográfico no movimento de adução 150° a 0°, independente da posição do úmero (p<0,05). Quando foi analisado com relação às varias posições em que o úmero foi colocado, o maior sinal eletromiográfico ocorreu na posição de rotação lateral no movimento de adução 150° a 0° para o M. rombóide maior (p<0,05). Nos demais movimentos e variações de posição do úmero os resultados não foram estatisticamente significantes tanto para o M. rombóide maior como para oM. rombóide menor (p>0,05). Na análise particular do M. rombóide menor, este apresentou maior sinal eletromiográfico no movimento de abdução horizontal de 0° a 50°, que foi maior que o sinal eletromiográfico para o movimento de abdução horizontal de 90° a 0°, que por sua vez obteve umsinal eletromiográfico maior que o movimento de adução de 150° a 0° (p<0,05). Os resultados indicam que há um recrutamento diferenciado das unidades motoras entre os músculos quando se variam os movimentos e as posições. Os resultados apontam ainda, para necessidade de novos estudos comparativos, com métodos similares para investigar as relações funcionais entre estes músculos / Abstract: The purpose of the present study was to analyze electromyographically the MM. rhomboideus major and minor in shoulder movements, considering both the importance in the maintenance, fixation and stability of the scapula to the movements of shoulder joint, and the absence of additional studies of these movements. The records of eight healthy man aged 22 to 25 were collected (Nicolet Vicking electromyograph) with fine wire electrodes. The statical analyses of the results was carried out using tests FRIDMAN and WILCOXON (p<0,05). The muscles presented activities for all the movements studied. The M. rhomboideus major presented higher activity than the M. rhomboideus minor. Concerning the movements, the M. rhomboideus major it was verified that the activity for the adduction movement of 150°-0° was lower than the horizontal abduction of 0°-50°. The M. rhomboideus minor presented higher activity in horizontal abduction of 0°50°, for the adduction movemenl of 150° - 0° and horizontal- abduction of 90° -0° movement, with no significant results for the humerus positioning. The results indicate that there is a differential recruitment of the motor units between the muscles, when the moviments and situations vary. Our results indicate the necessity for additional comparative studies to investidate the functional retalions between these muscles / Mestrado / Biologia e Patologia Buco-Dental / Mestre em Odontologia

Iron containing metalloenzymes are an extremely important class of biocatalysts conserved throughout evolution because of their vital role in the biochemistry of life. Here we discuss a specific class of these enzymes that use molecular oxygen binding to enable there activity. We also attempt to describe synthetic analogues whose chemistry is based on that seen in those natural systems. This dissertation will highlight how computational research can illuminate specific aspects of the reaction mechanisms that these systems catalyse, which in many cases are unable to be understood purely experimentally. We report on two combined QM/MM and density functional theory (DFT) projects, which describe the AlkB demethylation enzyme and the SyrB2 halogenase; both highlight the strengths and weaknesses of each method. Our DFT work on an i-propyl-bis(imino)pyridine, an equatorial tridentate ligand, developed by one of the papers’ co-authors (Badiei, Siegler et al. 2011) exampifies superoxo chemistry based on the dioxygenases. Our other projects focus on monooxygenase catalysed chemistry one based on the biomimic [FeIV(O)(TPA)Cl]+ reports on a halogenase mimic that shows exciting chemoselectivity in halogenation vs. hydroxylation. I also report on publications examining two other biomimetic ligands. A imido-bridged diiron-oxo phtalocyanine complex capable of hydroxylating methane and a nonheme iron system which gives us a good deal of insight into the effects of secondary coordination sphere chemistry [FeII(N4Py2Ph)(NCCH3)](BF4)2. My computational studies have given insight into the chemical properties of metal-oxo oxidants and their reactivity patterns with substrate and have been utilized to explain experimentally observed data.

Cytochrome P450 (P450) enzymes are found in all kingdoms of life, catalysing a wide range of biosynthetic and metabolic processes. They are, in fact, of particular interest in a variety of applications such as the design of agents for the inhibition of a particular P450 to combat pathogens or the engineering of enzymes to produce a particular activity. Bacterial P450BM3 is of particular interest as it is a self-sufficient multi-domain protein with high reaction rates and a primary structure and function similar to mammalian isoforms. It is an attractive enzyme to study due to its potential for engineering catalysts with fast reaction rates which selectively produce molecules of high value.In order to study this enzyme in detail and characterise intermediate species and reactions, the first step was to design a general hybrid quantum mechanical /molecular mechanics (QM/MM) computational method for their investigation. Two QM/MM approaches were developed and tested against existing experimental and theoretical data and were then applied to subsequent investigations.The dissociation of water from the water-bound resting state was scrutinised to determine the nature of the spin conversion that occurs during this transformation. A displacement of merely 0.5 Å from the starting state was found to trigger spin crossing, with no requirement for the presence of a substrate or large conformational changes in the enzyme.A detailed investigation of the sulfoxidation reaction was undertaken to establish the nature of the oxidant species. Both reactions involving Compound 0 (Cpd0) and Compound I (CpdI) confirmed a concerted pathway proceeding via a single-state reactivity mechanism. As the reaction involving Cpd0 was found to be unrealistically high, the reaction proceeds preferentially via the quartet state of CpdI. This QM/MM study revealed that the preferred spin-state and the transition state structure for sulfoxidation are influenced by the protein environment. P450cam and P450BM3 were found to have CpdI species with different Fe-S distances and spin density distributions, and the latter having a larger reaction barrier for sulfoxidation.A novel P450 species, the doubly-reduced pentacoordinated system, was characterised using gas-phase and QM/MM methods. It was discovered to have a heme radical coupled to two unpaired electrons on the iron centre, making it the only P450 species to have similar characteristics to CpdI. Calculated spectroscopic parameters may assist experimentalists in the identification of the elusive CpdI.

Recently, there is great interest in achieving higher-level integration, higher data rates, and reduced overall costs. At millimeter-wave (mm-wave) bands, the wavelength is small enough to realize an antenna-on-chip (AoC), which is an ideal solution for high compactness and lower costs. However, the main drawback of AoC is the low resistivity (10 Ω-cm) Si substrate used in the standard CMOS technology, which absorbs most radio-frequency (RF) power that was supposed to be radiated by the on-chip antenna. Moreover, due to the high relative permittivity (11.9) and relatively large electrical thickness of the Si, higher order surface wave modes get excited, which further degrade the antenna radiation performance. In order to alleviate the above-mentioned issues with the low gain of AoC, a combination of an artificial magnetic conductor (AMC) surface, a high dielectric constant superstrate, and a Fresnel lens is presented in this work. The AMC is realized in standard CMOS technology along with the AoC, whereas the superstrate and lens are part of a smart packaging solution. The AMC surface can change wave propagation characteristics at the operating frequency to achieve in-phase reflection, resulting in gain enhancement by reducing the loss in the substrate. The high dielectric constant superstrate behaves as an impedance transformer between the Si substrate and air, thus enhancing the coupling to air. Finally, the Fresnel lens enhances the gain by focusing the electromagnetic (EM) radiation beam at the boresight. For AoC realization, a standard 0.18 μm CMOS process was utilized. A coplanar waveguide (CPW) fed monopole on-chip antenna at 71 GHz, along with the corresponding driving circuit, was designed and fabricated. The AMC enhances the gain by 3 dB. Since the chip needs to be packaged anyways, in this work, we optimize the package to provide further gain enhancement. This smart package, comprising a superstrate and a Fresnel lens, provides a gain enhancement of 16 dB. The overall combination of the optimized AMC surface, superstrate layer, and lens package can provide a gain enhancement of around 19 dB. Furthermore, the package has been realized through additive manufacturing techniques that ensure lower costs for the overall system.

No description available.