A key component of neural circuit formation is the elaboration of complex dendritic arbors, the pattern of which constrains inputs to the neuron and thus, the information it processes. As such, many neurodevelopmental disorders such as autism and Down, Rett, and Fragile X Syndromes are associated with reduced forebrain dendrite arborization. Identifying molecules involved in regulating dendrite arborization and neural circuitry formation therefore, is a start to understanding these disorders. Nearly 70 cadherin superfamily adhesion molecules are encoded by the Pcdha, Pcdhb, and Pcdhg gene clusters. These so-called clustered protocadherins (Pcdhs) are broadly expressed throughout the nervous system, with lower levels found in a few non-neuronal tissues. Each neuron expresses a limited repertoire of clustered Pcdh genes, a complicated process controlled by differential methylation and promoter choice. The clustered Pcdh proteins interact homophilically in trans as cis-multimers, which has the potential to generate a combinatorially explosive number of distinct adhesive interfaces that may give neurons unique molecular identities important for circuit formation. Functional studies of animals in which clustered Pcdhs have been deleted or disrupted demonstrate that these proteins play critical roles in neuronal survival, axon and dendrite arborization, and synaptogenesis. Additionally, they have been implicated in the progression of several cancers, suggesting that basic studies of their function and signaling pathways will have important future clinical applications. Recent work has shown that γ-Pcdhs can regulate the Wnt signaling pathway, a common tumorigenic pathway which also play roles in neurodevelopment, but the molecular mechanisms remain unknown. I determined that γ-Pcdhs differentially regulate Wnt signaling: the C3 isoform uniquely inhibits the pathway while 13 other isoforms upregulate Wnt signaling. Focusing on γ-Pcdh-C3, I show that the variable cytoplasmic domain (VCD) is critical for Wnt signaling inhibition. γ-Pcdh-C3, but not other isoforms, physically interacts with Axin1, a key component of the canonical Wnt pathway. The C3 VCD competes with Dishevelled for binding to the DIX domain of Axin1, which stabilizes Axin1 at the membrane and leads to reduced phosphorylation of Wnt co-receptor Lrp6. I also present evidence that the Wnt pathway can be modulated up (by γ-Pcdh-A1) or down (by γ-Pcdh-C3) in the cerebral cortex in vivo, using conditional transgenic alleles. Studies have implicated γ-Pcdhs as a whole, in many neurodevelopmental processes but little is known if distinct roles exists for individual isoforms. By using a specific C3-isoform knockout mouse line engineered in collaboration with Dr. Robert Burgess of The Jackson Laboratory, I was able to uncover a unique role for the C3-isoform in the regulation of dendrite arborization. Mice without γ-Pcdh-C3 exhibit significantly reduced dendrite complexity in cortical neurons. This phenotype was recapitulated in cultured cortical neurons in vitro, which can be rescued by reintroducing the C3-isoform. The ability of γ-Pcdh-C3 to promote dendrite arborization cell-autonomously was abrogated when Axin1 was depleted with an shRNA, indicating that this process by which γ-Pcdh-C3 regulates dendrite arborization is mediated by its interaction with Axin1, which I had previously demonstrated. Together, these data suggest that γ-Pcdh-C3 has unique roles distinct from other γ-Pcdhs, in the regulation of Wnt signaling and dendrite arborization, both of which are mediated by interaction with Axin1.

Cell attachment is vital for the integration of biomaterials in the body. Surface modification using cell adhesive peptides, such as Arginine-Glycine-Aspartic (RGD), has showed promise for enhancing cell adhesion. Cell adhesion on glass and polyethylene glycol (PEGylated) surfaces modified with active RGD and Proline- Histidine-Serine-Arginine-Asparagine (PHSRN) peptides as well as inactive RDG and HRPSN control peptides was investigated in serum free conditions using three cell lines; NIH3T3 fibroblasts, MC3T3 pre-osteoblasts and C2C12 pre-myoblasts. Peptide attachment to glass surfaces was confirmed by x-ray photoelectron spectroscopy and contact angle measurements. Cell attachment and spreading was equivalent on all peptide and fibronectin coated glass surfaces and was significantly higher than on unmodified glass after 3 hours. Cell attachment to the peptide modified glass was reduced in the presence of soluble RGD and RDG peptides, indicating that cell attachment to these surfaces may be integrin mediated, but not specific for RGD. Inhibition of protein synthesis with cycloheximide revealed that endogenous protein synthesis did not influence the specificity of cell attachment to the peptide modified glass surfaces in all cell types within a 3 hour period. However, cycloheximide treatment inhibited cell spreading on the peptide modified glass surfaces, suggesting that proteins synthesis was required for spreading. Long term adhesion studies, within a 24 hour period, showed that all cell lines were able to remain attached to the peptide modified glass surfaces, while C2C12 and MC3T3 cells were also able to form focal adhesions during this period. Cell attachment to peptide modified PEGylated surfaces over a 3 hour period showed that NIH3T3 and C2C12 cells experienced significantly higher levels of cell attachment on the RGD modified surface compared to the other peptides. MC3T3 cells attached to all the peptide modified PEGylated surfaces to the same extent, suggesting that cell attachment to peptide modified PEGlyated surfaces, can be cell type dependent. In conclusion all the peptides were able to promote cell adhesion on glass surfaces in the absence of a PEG linker. In the presence of a PEG linker cellular response to the peptide surfaces was both peptide and cell type dependent.

Thesis (Ph. D.)--University of Chicago, Dept. of Biochemistry and Molecular Biology, June 1999. / Includes bibliographical references. Also available on the Internet.

We theoretically study the adhesion and membrane-mediated interaction of cylindrical colloids to a flat fluid membrane. There are two ways to approach this problem. The first way, based on energy, requires finding the equilibrium shape of the membrane given the placement of the particle(s). In order to do so, we need to know how the energy of the surface depends on its shape (i.e. the surface Hamiltonian), as well as how the adhered colloid deforms the membrane. The second way to approach this class of problems is “geometrical”, where forces between the membrane-adhered particles are related directly to the geometry of the deformed membrane via the surface stress tensor. The surface Hamiltonian allows finding the stress at any point on the membrane in terms of local geometry. The force acting on the colloid can then be found by integrating this surface stress tensor along any contour enclosing the colloid. In this thesis, using the approach based on free energy calculations, we look into the problem of cylindrical colloids adhering to a membrane with fixed constant adhesion energy between the membrane and the colloids. Angle-arclength parameterization is used in order to treat the problem beyond small gradient approximation. We present three different cases here: single cylinder adhering on a membrane, two cylinders adhering on the same side of the membrane, and two cylinders adhering on different sides of the membrane. For the single cylinder case we present a structural phase diagram to separate no wrapping, partial wrapping and closure states and we compare it to the phase diagram obtained for a related system of spherical colloids. For two cylinders adhered on the same side of the membrane we obtain repulsive interaction and transition from shallow to deep wrapping as the cylinders move apart from each other. We also look into a phase where two cylinders are vertically stacked and discuss its energetics. For two cylinders adhering to the opposite sides of the membrane, attractive interaction is obtained in accordance with previous results and we further show that in that case two cylinders are generally in contact and a first-order transition from shallow to full wrapping is possible. In the last section, we put a framework for the class of problems where the particle is between the membrane and the supporting interface, where adhesion is assumed between the interface and the membrane.

We theoretically study the adhesion and membrane-mediated interaction of cylindrical colloids to a flat fluid membrane. There are two ways to approach this problem. The first way, based on energy, requires finding the equilibrium shape of the membrane given the placement of the particle(s). In order to do so, we need to know how the energy of the surface depends on its shape (i.e. the surface Hamiltonian), as well as how the adhered colloid deforms the membrane. The second way to approach this class of problems is “geometrical”, where forces between the membrane-adhered particles are related directly to the geometry of the deformed membrane via the surface stress tensor. The surface Hamiltonian allows finding the stress at any point on the membrane in terms of local geometry. The force acting on the colloid can then be found by integrating this surface stress tensor along any contour enclosing the colloid. In this thesis, using the approach based on free energy calculations, we look into the problem of cylindrical colloids adhering to a membrane with fixed constant adhesion energy between the membrane and the colloids. Angle-arclength parameterization is used in order to treat the problem beyond small gradient approximation. We present three different cases here: single cylinder adhering on a membrane, two cylinders adhering on the same side of the membrane, and two cylinders adhering on different sides of the membrane. For the single cylinder case we present a structural phase diagram to separate no wrapping, partial wrapping and closure states and we compare it to the phase diagram obtained for a related system of spherical colloids. For two cylinders adhered on the same side of the membrane we obtain repulsive interaction and transition from shallow to deep wrapping as the cylinders move apart from each other. We also look into a phase where two cylinders are vertically stacked and discuss its energetics. For two cylinders adhering to the opposite sides of the membrane, attractive interaction is obtained in accordance with previous results and we further show that in that case two cylinders are generally in contact and a first-order transition from shallow to full wrapping is possible. In the last section, we put a framework for the class of problems where the particle is between the membrane and the supporting interface, where adhesion is assumed between the interface and the membrane.

In this research, a systematic and multi-disciplinary study was conducted to understand the fundamental science of moisture-induced degradation of interfacial adhesion. The research is comprised of both experimental and modeling components of analysis and consists of four primary components. First, the moisture transport behavior within underfill adhesives is experimentally characterized and incorporated into a finite element model to depict the moisture ingress and interfacial moisture concentration for each respective level of moisture preconditioning. Second, the effect of moisture on the variation of the underfill elastic modulus is demonstrated and the physical mechanisms for the change identified. Third, the aggregate effect of moisture on the interfacial fracture toughness of underfill to both copper and FR-4 board substrates is determined. This includes the primary effect of moisture being physically present at the interface and the secondary effect of moisture changing the elastic modulus of the adhesive when absorbed. Last, the recovery of both the elastic modulus and interfacial fracture toughness from moisture preconditioning is assessed with reversible and irreversible components identified. Using adsorption theory in conjunction with fracture mechanics, an analytical model is developed that predicts the loss in interfacial fracture toughness as a function of moisture content. The model incorporates key parameters relevant to the problem of moisture in epoxy joints identified from the experimental portion of this research, including the interfacial hydrophobicity, epoxy nanopore density, saturation concentration, and density of water. This research results in a comprehensive understanding of the primary mechanisms responsible for the interfacial degradation due to the presence of moisture. The experimental results obtained through this research provide definitive data for the electronics industry to use in their product design, failure analysis, and reliability modeling. The predictive model developed in this research provides a useful tool for developing new adhesives, innovative surface treatment methods, and effective protection methodologies for enhancing interfacial adhesion.

In this study, it was nanoimprint focused on the anti-adhesion technique between the grating structure silicon molds below 200nm half-pitch and polymer materials (H-PDMS). The nano-groove structure molds with different depths and widths were made by FIB. During the process of molding by soft-lithography, an anti-adhesion layer needed being plated between the silicon and PDMS mold, which was in order to get completely formed H-PDMS soft mold and prevent defective mold caused by the adhesion problem on the surface. There were three kinds of method of plating anti-adhesion layer which were the liquid immersion, vapor deposition, and fluorine doped DLC film. The PFOTCS was used as mold releasing agent in the methods of liquid immersion and vapor deposition, and the contact angle was measured to realize the ability of anti-adhesion. In the method of fluorine doped DLC film, in addition to measuring the anti-adhesion ability for each sample through contact angle with water, the AFM was also applied to measure the degree of adhesion on the surface for each film. And the contact angles with water between each film were also compared. The methods of plating anti-adhesion film with lower degree of adhesion on the surface could be acquired and discussed by means of the above-mentioned ways to fabricate the molds with good formability

To reduce electronics packaging lead time and potentially to reduce manufacturing cost, an innovative packaging process targeting rapid package prototyping (RPP) has been developed. The developed RPP process, which is based on a data-driven chip-first approach, provides electrical functionality as well as form factors for micro-systems packages. The key component of the RPP process is the nano-particle silver (NPS) interconnect. However, NPS has not yet been adequately proven for use in electronics packaging applications. Moreover, its adhesion to electronics packaging materials such as polyimide, benzocyclobutene (BCB), copper, and aluminum is found to be weak. Thus, improving the adhesion strength of NPS will be a key issue for reliable package prototypes with NPS interconnects. In this research, the adhesion of NPS to substrate materials is found to be attributed to particle adhesion and more specifically, van der Waals forces. An adhesion model based on the van der Waals force is suggested in order to predict NPS adhesion strength to packaging materials. A new adhesion test method that is based on a die shear test and a button shear test is developed to validate the NPS adhesion prediction model. The newly developed adhesion test method is generic in nature and can be extended to other thin films' adhesion tests. The NPS adhesion model provides a general and explicit relation between NPS tensile bond strength and adhesion factors such as substrate hardness, adhesion distance, van der Waals constant, and particle diameter. The NPS adhesion model is verified as a first order adhesion model using experimental data from seventeen packaging materials. Substrate hardness is identified as a primary factor in NPS adhesion. Adhesion distance and van der Waals constant are also significant in organic and inorganic materials. Diffusion or other interfacial reaction between NPS and metal substrates such as copper and silver seems to exist. Finally, guidelines to improve the adhesion strength of NPS are suggested based on the adhesion model and on external adhesion factors such as Silane coupling agents and plasma treatment.

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Current understanding of molecular-level interactions is inadequate to explain the initial moments of bacterial adhesion. Such information is required to develop appropriate models for bacteria-surface interactions and predictions of cell transport in subsurface environments. Bacterial adhesion is influenced by bacterial surfaces, substratum physical-chemical characteristics, and solution chemistry. Extracellular polymeric substances (EPS), surface proteins, and lipopolysaccharides (LPS) mediate cell adhesion and conditioning film formation via direct bonding to a substrate. The goal of this dissertation is to probe molecular-scale interactions of cell surface macromolecules at mineral surfaces under environmentally-relevant conditions. Four primary investigations are presented in this dissertation. The first study uses in situ attenuated total reflectance (ATR) Fourier transform infrared (FTIR) spectroscopy to reveal that prior to Mn-oxidation via Pseudomonas putida GB-1, cell adhesion to ZnSe is favorable. Subsequent Mn-oxidation results in increased extracellular proteins expression. Conversely, planktonic cell adhesion is inhibited for Mn-oxide coated cells via blocking of surface proteins. The second investigation reveals the formation of inner-sphere complexes between bacteria surface phosphoryl groups and nanohematite (α-Fe₂O₃). Spectra of bacteria (P. aeruginosa PAO1, Shewanella oneidensis MR-1, and Bacillus subtilis) on α- Fe₂O₃ contain peaks indicative of P-OFe inner-sphere bonding. Spectra collected for oxide-adsorbed model P-containing compounds give spectral signatures similar to those P-OFe bonding interactions observed for whole cell and EPS. The behavior of P. aeruginosa serotype 10 LPS in aqueous solutions was investigated in the third study. Ionic strength, pH, and electrolyte composition were varied during collection of ATR-FTIR and dynamic light scattering (DLS) data. Results reveal stable aggregate Na-LPS aggregates, whereas binding of Ca²⁺ to phosphate groups in the lipid A region leads to aggregate reorientation and increased interaction with ZnSe (hydrophobic). DLS data demonstrate decreasing hydrodynamic radius of LPS aggregates with increasing I and decreasing pH. In the fourth investigation, ATR-FTIR was used to probe the solid-solution interface of LPS on surfaces of ZnSe, Ge, α-Fe₂O₃, and α-Al₂O₃ in solutions of varying ionic composition and pH. Na-LPS aggregates remain stable and spectra are biased towards solution phase LPS. Ca-LPS aggregates are disrupted, leading to enhanced interaction with surfaces via hydrophobic (lipid A- ZnSe) and electrostatic (O-antigenhydrophilic surfaces) interactions.