Indiana University-Purdue University Indianapolis (IUPUI) / Immune response enables the body to resist foreign invasions. The Inflammatory response is an important aspect in the immune response which is articulated by elements such as cytokines, APC, T-cell and B-cell, effector cell or natural killer. Of these elements, T-cells especially T-helper cells; a sub class of T-cells plays a pivotal role in stimulating the immune response by participating in various biological reactions such as, the transcription regulatory network. Transcriptional regulatory mechanisms are mediated by a set of transcription factors (TFs), that bind to a specific region (motifs or transcription factor binding sites, TFBS), on the target gene(s) controlling the expression of genes that are involved in T-helper cell mediated immune response. Eukaryotic regulatory motifs, referred to as cis regulatory modules (CRMs) or cistrome, co-occur with the regulated gene’s transcription start site (TSS) thus, providing all the essential components for building the transcriptional regulatory networks that depends on the relevant TF-TFBS interactions. Here, we study IL-12 stimulated transcriptional regulators in STAT4 mediated T helper 1 (Th1) cell development by focusing on the identification of TFBS and CRMs using a set of Stat4 ChIP-on-chip target genes. A region containing 2000 bases of Mus musculus sequences with the Stat4 binding site, derived from the ChIP-on-chip data, has been characterized for enrichment of other motifs and, thus CRMs. Our experiments identify some potential motifs, (such as NF-κB and PPARγ/RXR) being enriched in the Stat4 binding sequences compared to neighboring background sequences. Furthermore, these predicted CRMs were observed to be associated with biologically relevant target genes in the ChIP-on-chip data set by meaningful gene ontology annotations. These analyses will enable us to comprehend the complicated transcription regulatory network and at the same time categorically analyze the IL-12 stimulated Stat4 mediated Th1 cell differentiation.

This thesis presents the design, implementation, and fabrication of an analog front-end (AFE) targeting 2x blind ADC-based receivers. The front-end consists of a combination of an anti-aliasing filter (AAF) and a 2-tap feed-forward equalizer (FFE) (AAF/FFE), the required clock generation circuitry (Ck Gen), 4 time-interleaved 4-b ADCs, and DeMUX. The contributions of this design are the AAF/FFE and the Ck Gen. The overall front-end optimizes the channel/filter characteristics for data-rates of 2-10 Gb/s. The bandwidth of the AAF is scalable with the data-rate and the analog 2-tap feed-forward equalizer (FFE) is designed without the need for noise-sensitive analog delay cells. The test-chip is implemented in 65-nm CMOS and the AAF/FFE occupies 152×86 μm2 and consumes 2.4 mW at 10 Gb/s. Measured frequency responses at data-rates of 10, 5, and 2 Gb/s confirm the scalability of the front-end bandwidth. FFE achieves 11 dB of high-frequency boost at 10 Gb/s.

This thesis presents the design, implementation, and fabrication of an analog front-end (AFE) targeting 2x blind ADC-based receivers. The front-end consists of a combination of an anti-aliasing filter (AAF) and a 2-tap feed-forward equalizer (FFE) (AAF/FFE), the required clock generation circuitry (Ck Gen), 4 time-interleaved 4-b ADCs, and DeMUX. The contributions of this design are the AAF/FFE and the Ck Gen. The overall front-end optimizes the channel/filter characteristics for data-rates of 2-10 Gb/s. The bandwidth of the AAF is scalable with the data-rate and the analog 2-tap feed-forward equalizer (FFE) is designed without the need for noise-sensitive analog delay cells. The test-chip is implemented in 65-nm CMOS and the AAF/FFE occupies 152×86 μm2 and consumes 2.4 mW at 10 Gb/s. Measured frequency responses at data-rates of 10, 5, and 2 Gb/s confirm the scalability of the front-end bandwidth. FFE achieves 11 dB of high-frequency boost at 10 Gb/s.

The transcription in the eukaryotic cells involves epigenetic regulatory mechanisms that control local and higher-order chromatin remodelling. In the skin, keratinocyte-specific genes are organized into distinct loci including Epidermal Differentiation Complex (EDC) and Keratin type I/II loci. This thesis introduces bioinformatics approaches to analyze multi-level regulatory mechanisms that control skin development and keratinocyte-specific differentiation. Firstly, integration of gene expression data with analyses of linear genome organization showed dramatic downregulation of the genes that comprise large genomic domains in the sweat glands including EDC locus, compared to ii hair follicles, suggesting substantial differences in global genome rearrangement during development of these two distinct skin appendages. Secondly, comparative analysis of the genetic programmes regulated in keratinocytes by Lhx2 transcription factor and chromatin remodeler Satb1 revealed that significant number of their target genes is clustered in the genome. Furthermore, it was shown in this study that Satb1 target genes are lineage-specific. Thirdly, analysis of the topological interactomes of Loricrin and Keratin 5 in hair follicle steam cells revealed presence of the cis- and trans-interactions and lineage specific genes (Wnt, TGF-beta/activin, Notch, etc.). Expression levels of the genes that comprise interactomes show correlation with their histone modification status. This study demonstrates the crucial role for integration of transcription factormediated and epigenetic regulatory mechanisms in establishing a proper balance of gene expression in keratinocytes during development and differentiation into distinct cell lineages and provides an integrated bioinformatics platform for further analyses of the changes in global organization of keratinocyte-specific genomic loci in normal and diseased skin.

Proper development of skeletal muscle occurs through a highly complex process where activation and repression of genes are essential. Control of this process is regulated by timely and spatial expression of specific transcription factors (TFs). Six1 and Six4 are homeodomain TFs known to be essential for skeletal muscle development in mice. Using the C2C12 cell line, a model for skeletal muscle differentiation, I used a functional genomics approach, employing siRNA specific to both these TFs, to characterize their role in skeletal myogenesis. To identify the genes that are regulated by both these TFs, gene expression profiling by microarray of cells treated with siRNA against Six1 and/or Six4 was performed. The knock-down of these TFs caused lower expression of markers of terminal differentiation genes in addition to an impairment of myoblast fusion and differentiation. Interestingly, transcript profiling of cells treated with siRNA against myogenin revealed that several of the Six1 and Six4 target genes are also regulated by myogenin. Through a combination of bioinformatic analyses it was also found that specific knock-down of Six4 causes an up-regulation of genes involved in mitosis and the cell cycle. In summary, these results show that Six1 and Six4 can both independently regulate different genes, but can also cooperate together with other TFs where they play an important role in the proper regulation of skeletal myogenesis.

Proper development of skeletal muscle occurs through a highly complex process where activation and repression of genes are essential. Control of this process is regulated by timely and spatial expression of specific transcription factors (TFs). Six1 and Six4 are homeodomain TFs known to be essential for skeletal muscle development in mice. Using the C2C12 cell line, a model for skeletal muscle differentiation, I used a functional genomics approach, employing siRNA specific to both these TFs, to characterize their role in skeletal myogenesis. To identify the genes that are regulated by both these TFs, gene expression profiling by microarray of cells treated with siRNA against Six1 and/or Six4 was performed. The knock-down of these TFs caused lower expression of markers of terminal differentiation genes in addition to an impairment of myoblast fusion and differentiation. Interestingly, transcript profiling of cells treated with siRNA against myogenin revealed that several of the Six1 and Six4 target genes are also regulated by myogenin. Through a combination of bioinformatic analyses it was also found that specific knock-down of Six4 causes an up-regulation of genes involved in mitosis and the cell cycle. In summary, these results show that Six1 and Six4 can both independently regulate different genes, but can also cooperate together with other TFs where they play an important role in the proper regulation of skeletal myogenesis.

This thesis is concerned with cutting farm machine and it consists of three parts. Theoretical part describes mechanism of chip formation, shape of the chips, temperature of the chips, technological variables describing chips, chip conveyor, chip crusher, briquetting press and coolant filter. In the next part there is proposal of design of cutting farm machine for production cell consists of six machining center. At last part there is design proposal and calculation of chip conveyor.

The transcription in the eukaryotic cells involves epigenetic regulatory mechanisms that control local and higher-order chromatin remodelling. In the skin, keratinocyte-specific genes are organized into distinct loci including Epidermal Differentiation Complex (EDC) and Keratin type I/II loci. This thesis introduces bioinformatics approaches to analyze multi-level regulatory mechanisms that control skin development and keratinocyte-specific differentiation. Firstly, integration of gene expression data with analyses of linear genome organization showed dramatic downregulation of the genes that comprise large genomic domains in the sweat glands including EDC locus, compared to ii hair follicles, suggesting substantial differences in global genome rearrangement during development of these two distinct skin appendages. Secondly, comparative analysis of the genetic programmes regulated in keratinocytes by Lhx2 transcription factor and chromatin remodeler Satb1 revealed that significant number of their target genes is clustered in the genome. Furthermore, it was shown in this study that Satb1 target genes are lineage-specific. Thirdly, analysis of the topological interactomes of Loricrin and Keratin 5 in hair follicle steam cells revealed presence of the cis- and trans-interactions and lineage specific genes (Wnt, TGF-beta/activin, Notch, etc.). Expression levels of the genes that comprise interactomes show correlation with their histone modification status. This study demonstrates the crucial role for integration of transcription factormediated and epigenetic regulatory mechanisms in establishing a proper balance of gene expression in keratinocytes during development and differentiation into distinct cell lineages and provides an integrated bioinformatics platform for further analyses of the changes in global organization of keratinocyte-specific genomic loci in normal and diseased skin.

Studies on chip ejection from tools provide important information for the design of tools and effective chip collection elements used in woodworking machines. Among the chip properties, chip motion and its distribution are of particular interest for the design process. Until now, chip velocities have only been measured by manual tracking over a high-speed image sequence, which allows only a small scope of inspection. Here, we present the use of high-speed imaging in combination with particle tracking velocimetry as a new method for the semi-automatic evaluation of the magnitudes and directions of chip velocities. The methodology was tested in groove milling of particleboard. It was found that state-of-the-art particle tracking algorithms are suitable for quantitative analysis of chip motion in high-speed images. Therefore, spatial and temporal analysis of the chip velocity along the tool circumference are feasible and are presented here. In addition to chip velocity, chip collisions with the tool or other chips can be observed. This research also shows that image evaluation of chip sizes and shapes is potentially possible. In summary, the presented work provides methods that can quantitatively describe chip motion after chip formation. The experiments indicate that with each tooth engagement, new chips are formed, which initially move into the chip space at a median velocity higher than the cutting speed. After collisions with the tool and interparticle collisions, the particles leave the chip space of the tool at lower speeds. The machining tests performed with different process settings showed differences in the analysis results of chip movement. In the future, the presented methodology offers the possibility of investigating the relationships between tooth and chip space geometries, as well as different materials and the chip ejection of tools. Thus, the presented methodology provides a basis for creating a more general understanding of chip motion from machining operations, which can lead to innovations and improvements in chip collection.

Safe, effective hydrogen fuel cells are one of the contenders for the next shift in mobile power technology. One of the solutions to the inherent risks of high pressure hydrogen fuel cells is an outer low pressure container with an inner high pressure containers containing a micro-electromechanical systems (MEMS) valve which regulating the pressure. These MEMS valves consist of several etched Si-chips stacked and bonded, which shall withstand the pressure and temperature range in the high pressure fuel cell as well as the embrittlement caused by the hydrogen gas. Hydrogen embrittlement is a phenomena where materials, mostly metals, lose their ductility due to diffusion of hydrogen atoms into their grain boundaries. A suitable method for stacking the chips is needed and thus a literature study was conducted. Several chip-to-chip bonding methods were investigated in the purpose of finding the most suitable methods tolerating temperatures between -40 to 85°C, pressure up to 1000 bar, hermetically sealing, withstanding hydrogen embrittlement and still bond with particulate contaminations caused by testing each chip. The method found to be best fitting for the purpose was anodic bonding with an alkali glass. Alternatively anodic bonding with a ceramic glass system could be considered if technique from alkali glass is adaptable.