Enzymatic protein labeling and polymeric materials for cancer-therapy and anti-fouling applications

University of Minnesota Ph.D. dissertation. 2021. Major: Chemistry. Advisor: Mark Distefano. 1 computer file (PDF); vi, 193 pages.This dissertation focused on two relevant topics, cancer and antifouling. Cancer was chosen as the first target since despite the long-standing substantive efforts, it remains one of the most challenging malignant diseases. For instance, still to this date, brain cancer has a five-year survival rate of 12.8%. Two different approaches were studied to tackle cancer. In one aspect, enzymatic labeling was exploited in synthesizing protein-based therapeutics capable of cancer-targeted imaging and drug delivery. Having multiple therapeutic payloads loaded to a site-specifically conjugated hydrophilic polymer allowed highly retained cancer-binding affinity of the target protein DARPin. Therefore, the protein product exhibited high therapeutic efficacy. In another aspect, novel stimuli-responsive hydrogels were developed to understand the underlying mechanism of brain cancer metastasis. The mechanical properties of the gel could be spatiotemporally controlled with high-resolution, enabling the study of durotaxis, which is a biased directional movement of brain cancer cells. Combined with microbiological and computational studies, new insights into cellular behavior that were acquired as a result of the new hydrogel platform revealed the molecular components and their dynamics that dictate durotaxis. On a different topic, antifouling applications were explored, which is another relevant issue, especially in the marine and health care industry. Quorum quenching enzymes were site-specifically modified via protein farnesyltransferase for visualization or immobilization purposes. The modification that allows high retention of the protein activity allowed various biotechnological applications of the quorum quenching enzyme. Overall, interdisciplinary research that involves chemical biology, polymer chemistry, and organic chemistry, was utilized to study important present-day issues.Park, Keun-Young. (2021). Enzymatic protein labeling and polymeric materials for cancer-therapy and anti-fouling applications. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/224933

A Study on Thermal Modeling and Heat Load Mitigation for Satellite Electronic Components

Since most of the satellite components are using various EEE (Electrical, Electronic and Electromechanical) parts, the reliability of EEE parts acts very important in the satellite system. There are many factors that influence the reliability of EEE parts in the satellite system. Excessively dissipated heat can cause the failure of EEE parts and consequently, leading to a failure of total satellite system. In this paper, the thermal modeling using nodal network was compared with that using plate modeling to find out which one is the most suitable methodology. For a comparison, KOMPSAT- 1 SAR was modeled by two different modeling and the result was discussed. There was almost no difference in the numerical results between the two modeling methods. However, while it took much more time to perform thermal analysis using the nodal network modeling method, and the debugging was more difficult in the plate modeling method when the error is occurred. The computation time was considerably reduced by developing and implementing the input file format transfer code when using nodal network modeling method. It was found that the nodal network modeling method is suitable for the complicated components, such as SAR or transponder, because of its simple debugging ability. Excessive heat load was expected on some EEE parts of SAR such as high heat-dissipated diodes, transistors, and inductors due to increased power requirements of KOMPSAT-2 satellite system. The methods for the mitigation of heat load were studied through the design change of housing or the layout change of high power parts

Inhibition of cAMP/PKA Pathway Protects Optic Nerve Head Astrocytes against Oxidative Stress by Akt/Bax Phosphorylation-Mediated Mfn1/2 Oligomerization.

Glaucoma is characterized by a progressive optic nerve degeneration and retinal ganglion cell loss, but the underlying biological basis for the accompanying neurodegeneration is not known. Accumulating evidence indicates that structural and functional abnormalities of astrocytes within the optic nerve head (ONH) have a role in glaucomatous neurodegeneration. Here, we investigate the impact of activation of cyclic adenosine 3',5'-monophosphate (cAMP)/protein kinase A (PKA) pathway on mitochondrial dynamics of ONH astrocytes exposed to oxidative stress. ONH astrocytes showed a significant loss of astrocytic processes in the glial lamina of glaucomatous DBA/2J mice, accompanied by basement membrane thickening and collagen deposition in blood vessels and axonal degeneration. Serial block-face scanning electron microscopy data analysis demonstrated that numbers of total and branched mitochondria were significantly increased in ONH astrocytes, while mitochondrial length and volume density were significantly decreased. We found that hydrogen peroxide- (H2O2-) induced oxidative stress compromised not only mitochondrial bioenergetics by reducing the basal and maximal respiration but also balance of mitochondrial dynamics by decreasing dynamin-related protein 1 (Drp1) protein expression in rat ONH astrocytes. In contrast, elevated cAMP by dibutyryl-cAMP (dbcAMP) or isobutylmethylxanthine treatment significantly increased Drp1 protein expression in ONH astrocytes. Elevated cAMP exacerbated the impairment of mitochondrial dynamics and reduction of cell viability to oxidative stress in ONH astrocytes by decreasing optic atrophy type 1 (OPA1), and mitofusin (Mfn)1/2 protein expression. Following combined treatment with H2O2 and dbcAMP, PKA inhibition restored mitochondrial dynamics by increasing mitochondrial length and decreasing mitochondrial number, and this promoted cell viability in ONH astrocytes. Also, PKA inhibition significantly promoted Akt/Bax phosphorylation and Mfn1/2 oligomerization in ONH astrocytes. These results suggest that modulation of the cAMP/PKA signaling pathway may have therapeutic potential by activating Akt/Bax phosphorylation and promoting Mfn1/2 oligomerization in glaucomatous ONH astrocytes

Structure of shock compressed model basaltic glass: Insights from O K-edge X-ray Raman scattering and high-resolution ^(27)Al NMR spectroscopy

The detailed atomic structures of shock compressed basaltic glasses are not well understood. Here, we explore the structures of shock compressed silicate glass with a diopside–anorthite eutectic composition (Di_(64)An_(36)), a common Fe-free model basaltic composition, using oxygen K-edge X-ray Raman scattering and high- resolution ^(27)Al solid-state NMR spectroscopy and report previously unknown details of shock-induced changes in the atomic configurations. A topologically driven densification of the Di_(64)An_(36) glass is indicated by the increase in oxygen K-edge energy for the glass upon shock compression. The first experimental evidence of the increase in the fraction of highly coordinated Al in shock compressed glass is found in the ^(27)Al NMR spectra. This unambiguous evidence of shock-induced changes in Al coordination environments provides atomistic insights into shock compression in basaltic glasses and allows us to microscopically constrain the magnitude of impact events or relevant processes involving natural basalts on Earth and planetary surfaces

Generation of subspecies level-specific microbial diagnostic microarrays using genes amplified from subtractive suppression hybridization as microarray probes

The generation of microarray probes with specificity below the species level is an ongoing challenge, not least because the high-throughput detection of microorganisms would be an efficient means of identifying environmentally relevant microbes. Here, we describe how suppression subtractive hybridization (SSH) can be applied to the production of microarray probes that are useful for microbial differentiation at the subspecies level. SSH was used to initially isolate unique genomic sequences of nine Salmonella strains, and these were validated in quadruplicate by microarray analysis. The results obtained indicate that a large group of genes subtracted by SSH could serve together, as one probe, for detecting a microbial subspecies. Similarly, the whole microbial genome (not subjected to SSH) can be used as a species-specific probe. The detailed methods described herein could be used and adapted for the estimation of any cultivable bacteria from different environments

REACTION CHARACTERISTICS OF TWO WATER GAS SHIFT CATALYSTS IN A BUBBLING FLUIDIZED BED REACTOR FOR SEWGS PROCESS

Reaction characteristics of two WGS catalysts for SEWGS process were investigated in a bubbling fluidized bed reactor. The commercial low temperature WGS catalyst produced by Süd-chemie and new catalyst produced by spray-drying method were used as bed materials. Reaction temperature, steam/CO ratio, and gas velocity were considered as experimental variables. Moreover, long-term operation results of two WGS catalysts were compared as well

THE HOT GAS DESULFURIZATION IN A COMPACT TWO BEDS SYSTEM INTEGRATED WITH COAL GASIFICATION AND FISHER-TROPSCH SYSTEM

The hot gas desulfurization (HGD) technique is one of the elemental technologies of syngas purification having both thermal efficiency and very low emissions. The HGD is a novel method to efficiently remove H2S and COS in the syngas with regenerable sorbents at high temperature and high pressure condition. We propose a compact hot gas desulfurization system by which its operability is improved in stabilizing pressure balances among units. The proposed compact two beds system has two bubbling beds, solid injection nozzle, solid conveying line, and riser. The compact desulfurization system was located between coal gasifier and Fisher-Tropsch (F-T) reactor to desulfurize syngas in order not to deactivate F-T catalyst. To check feasibility of the compact desulfurization system at high pressure condition, both cold mode and hot mode tests have been performed. In the integrated system, the compact desulfurization system has removed H2S and COS in the syngas and supplied the cleaned syngas to the F-T reactor during the continuous operation at high pressure condition