Salvia genus is a widely cultivated genus and used in medicine for various purposes as having antimicrobial, antioxidant, anticarcinogen and anti-inflammatory features. In this study the aim was to investigate phenolic composition of Salvia absconditiflora and understand the possible effects of those constituents in cancer related drug metabolizing enzymes. Salvia absconditiflora showed 80,43 % Radical Scavenging Activity against DPPH radical. Total flavonoid content was found as one third of total phenolic content. Presence of important phenolic acids and flavonoids such as caffeic acid, luteolin, coumaric acid are validated with LC-MS/MS analysis. Cytotoxicity of Salvia absconditiflora treatment on MCF-7 and MDA-MB-231 breast cancer cell lines were investigated through XTT and TBE assays both dose and time dependent manner. Cell proliferation was inhibited 50 % by different IC50 values calculated in different assays and different time intervals. This suggests that two breast cancer cell lines response in a different way to cytotoxic treatments. Cancer related drug metabolizing enzyme gene modulations were investigated with qRT-PCR. CYP1A1 and CYP1B1 were upregulated in MCF-7 but down-regulated in MDA-MB-231 cells in response to Salvia absconditiflora treatment.

Salvia genus is a widely cultivated genus and used in medicine for various purposes as having antimicrobial, antioxidant, anticarcinogen and anti-inflammatory features. In this study the aim was to investigate phenolic composition of Salvia absconditiflora and understand the possible effects of those constituents in cancer related drug metabolizing enzymes. Salvia absconditiflora showed 80,43 % Radical Scavenging Activity against DPPH radical. Total flavonoid content was found as one third of total phenolic content. Presence of important phenolic acids and flavonoids such as caffeic acid, luteolin, coumaric acid are validated with LC-MS/MS analysis. Cytotoxicity of Salvia absconditiflora treatment on MCF-7 and MDA-MB-231 breast cancer cell lines were investigated through XTT and TBE assays both dose and time dependent manner. Cell proliferation was inhibited 50 % by different IC50 values calculated in different assays and different time intervals. This suggests that two breast cancer cell lines response in a different way to cytotoxic treatments. Cancer related drug metabolizing enzyme gene modulations were investigated with qRT-PCR. CYP1A1 and CYP1B1 were up-regulated in MCF-7 but down-regulated in MDA-MB-231 cells in response to Salvia absconditiflora treatment.

The objective of this study was to investigate the exposure assessment and health risk of particulate matter emitted from steel industry for high-risk inhabitants living in metro Kaohsiung. A steel plant and an electric arc plant were selected for conducting the sampling of particulate matter (PM). The physical and chemical properties of PM sampled at the working place and at the fence of selected plants were also analyzed to establish the fingerprints of PM in the steel industry. In order to accomplish this study, both personal sampling and environmental sampling were conducted in steel plants. Personal exposure sampling was undertaken at working places, while the conduction of environmental sampling might affected by various uncertain environmental factors (such as meteorological condition and other emission sources), but it could characterize the overall environmental situation and help exposure assessment. Moreover, The exposure of employers (including manufacturing workers and supporting staffs) could be further assessed based on the concentration and duration of PM exposure. Specific metal concentration was obtained from working environment, searching for exposure parameter (such as exposure frequency, exposure duration, body weight, average time, and etc.) and toxicity database (e.g. cancer slope factor and reference dose). The data could be used for assessing both cancer risk and non-cancer risk of specific heavy metal. Sampling data obtained from working places showed that the highest PM concentration were observed during the electric arc process, especially the charging and discharging procedures. The concentration of PM1.0, PM10, and TSP at the working places were 53.3~195.6 £gg/m3 ,365.7~550.0 £gg/m3, and 1085.5~2755.0 £gg/m3, respectively. Measured at the outdoor environments of working places, the highest PM10 and TSP concentration, ranging from 365.7~550.0 £gg/m3 and 1085.5 ~2755.0 £gg/m3, were observed at the sinter plants of a steel manufacturer. Sampling data obtained at the fence of steel plants (a steel plant and electric arc plant) indicated that the concentration of PM10 and TSP exceeded the ambient air quality standards. Modification of manufacturing process and improvement of PM collection system are highly required to reduce the concentration level as well as the emission of PM. Results of fingerprint of PM (PM2.5, PM2.5-10, TSP) obtained from working places showed that iron, calcium, sulfate, nitrate, calcium ion, and elemental carbon were the major chemical content of PM at the melting plant, the sinter plant, and the storage field. While, iron, calcium, sulfate, nitrate, calcium ion, and organic carbon were the major chemical content of PM at electric arc plant. Results of personal exposure concentration and exposure dose of PM showed that the highest exposure level of both manufacturing workers and supporting staffs were observed at the storage field, while the lowest exposure level was found at the sinter plant. Therefore, enforcement of wearing maskers and/or shelters at the storage field is also highly recommended to prevent manufacturing workers from the exposure of high-level PM at working places. Moreover, the cancer risk of manufacturing workers exposured to hexavalent chromium (Cr6+) at the melting plant, the sinter plant, the storage field, and the electric arc plant exceeded acceptable cancer risk standard (10-5~10-6), while the hazard index of manganese (Mn) was much higher than other metals.Meanwhile, exposuring to Cr6+, Ni, and Mn at the electric arc plant also exceeded acceptable standards (Cr6+ and Ni¡G10-5~10-6 , Mn¡G1). Several control strategies, including pollution reduction measures such as the usage of clean fuel, process modification, the improvement of collection system, the enhancement of manufacture management, the conduction of environmental monitoring and exposure assessment, floor cleanup and truck entrance management, are recommended to improve the contamination of PM at working places and surrounding environments for metallurgic industries. Moreover, enforcement of wearing maskers and/or shelters at high-risk environments is also highly recommended to prevent manufacturing workers from the exposure of high-level PM at working places. Keywords: steel industry, particulate matter sampling, physical and chemical characteristics, exposure assessment, health risk

In this study, collagen based scaffolds were prepared for cornea tissue engineering. Three different cell carriers (rat tail collagen foam, insoluble collagen foam and patterned collagen film) were produced using two different collagen sources. Scaffolds were designed to mimic the unique topographical features of the corneal stroma. A novel crosslinking method was developed to achieve constant foam thickness. All scaffolds were tested with the primary cells of the native corneal stroma, human keratocytes. Although both foams promoted cell growth and penetration, rat tail foams were found to be superior for keratocyte proliferation. Their degradation rates were high enough but did not compromise their structural integrity during testing. Transparency studies with the foams revealed a progressive improvement. Collagen films degraded significantly over a one month period / however, the presence of cells increased the tensile strength of the films over a 21 day period to close to that of the native cornea and compensated for the loss of strength due to degradation. The micropatterned films proved to have higher transparency than the unpatterned scaffolds. In this study, it was possible to prepare collagen based micropatterned scaffolds using a silicon wafer and then a silicone template, successively, starting from original designs. The resultant collagen films were able to control cell growth through contact guidance, restricted cells and secreted-ECM within the pattern grooves, resulting in a higher transparency in comparison to unpatterned films. Thus, the tissue engineered constructs revealed a significant potential for use as total artificial corneal substitutes.

Interactions of cholesterol reducing agent simvastatin with charged model membranes were investigated. Effects of cholestrol reducing agent simvastatin on the phase transition behaviour and physical properties of the anionic dipalmitoyl phosphatidylglycerol (DPPG) multilamellar liposome were studied as a function of temperature and simvastatin concentration. Moreover the effect of acyl chain length on the simvastatin model membrane interactions was monitored using dipalmitoyl phosphatidylglycerol (DPPG) and dimyristoyl phosphatidylglycerol (DMPG) lipids. All experiments were carried out by two non-invasive techniques namely Fourier Transform Infrared (FTIR) Spectroscopy and Differential Scanning Calorimetry (DSC). The observations made in the this study clearly showed that simvastatin interacts with the lipids of multilamellar liposomes and induces some variations in the structure of membranes. These effects are seen in the thermotropic phase transition profile of the membranes, on membrane order, acyl chain flexibility, lipid head group structures and membrane fluidity. The analysis of the C-H stretching region of FTIR spectra showed that, as simvastatin concentration increased, the phase transition curve broadened, pretransition temperature diminished, membrane order and membrane fluidity increased for anionic DPPG membrane. Moreover analysis of the C=O stretching and PO2 - stretching bands showed that simvastatin caused dehydration effect by decreasing of hydrogen bonding capacity in the glycerol backbone and also around the lipid head groups. DSC studies showed that as the simvastatin concentration increased, DSC curves broadened. In addition, simvastatin-induced lateral phase separation was observed in the DSC thermograms. In the second part of the study, the effect of acyl chain length on the simvastatin - membrane interactions was investigated for DPPG and DMPG lipid membranes. All parameters used in the FTIR studies are compared for DMPG and DPPG membranes. Similar results were observed for both membranes, except for the CH2 antisymmetric stretching band frequency at gel phase. Results showed that there are no significant effect of acyl chain length on simvastin - membrane interactions.

Amifostine is the only approved radioprotective agent by the Food and Drug Administration for reducing the damaging effects of radiation on healthy tissues. In this study, the effects of ionizing radiation on rat liver microsomal membrane and brain tissue and the protecting effects of amifostine on these systems were investigated at molecular level. Sprague-Dawley rats, which were administered amifostine or not, were whole-body irradiated and liver microsomal membranes and different regions of the brain of these rats were analyzed using FTIR spectroscopy, FTIR microspectroscopy and synchrotron FTIR microspectroscopy. The first part of this study revealed that ionizing radiation caused a decrease in the total lipid content and CH2 groups of lipids, an increase in the carbonyl esters, olefinic=CH and CH3 groups of lipids in the white matter and grey matter regions of the brain, which could be interpreted as a result of lipid peroxidation. In addition, radiation altered the protein structure of the brain. Amifostine caused significant protective effect against all the radiation induced damages in the brain. In the second part of the study, FTIR results showed that radiation induced a decrease in the lipid/protein ratio and a degradation of lipids into smaller fragments that contain less CH2 and more carbonyl esters, olefinic=CH and CH3 groups in microsomal membranes. In addition, radiation caused an alteration in the secondary structure of proteins, an increase in lipid order and a decrease in the membrane dynamics. Amifostine prevented all the radiation induced compositional, structural and functional damages in the liver microsomal membranes.

Rheum species are medicinally important plants due to the presence of anthracene derivatives and in this study antioxidative, cytotoxic, apoptotic and chemopreventive characteristics of R. ribes extracts were evaluated. R. ribes shoot and root dry powder samples were prepared and extracted with ethyl acetate, ethanol and water. The extracts were revealed to be a potential scavenger of DPPH radicals and the chemical composition of the extracts was quantified by colorimetric determination of total phenol (GAE) and flavonoid (CAE) contents. HL&ndash / 60 cells were cultured in the presence of various concentrations of extracts up to 72 hr. R. ribes inhibited the surviv al of HL-60 cells in a concentration- and time-dependent manner, shown by trypan blue and XTT. R. ribes caused HL-60 cells apoptosis via formation of phosphatidylserine externalization, as evidenced by flow cytometry. Exposure of HL-60 cells to higher concentrations of extracts for 72 h resulted in a shift of 87% of the cell population from normal to the early/late apoptotic stage. The R. ribes induced apoptosis may be partially attributed to the activation of caspase-3 and up-regulation of caspase-3 expression was detected in western blot. The significant release of cytochrome c from the mitochondria into the cytosol was observed. The mRNA expression ratio of Bax/Bcl-2 was increased. The apoptosis was also demonstrated by DNA ladder and TUNEL. Chemopreventive effects of R.ribes were investigated at the gene level of CYP1B1 and CYP1A1, and GST enzyme activity against cDNB and concluded that R.ribes modulated activities of these enzymes generally at a time dependent level. T h ese findings suggest that Rheum ribes exhibits potential antioxidant and anticancer properties by inducing caspase-dependent cell death in HL-60 cells.

Quantum dots (QDs) are increasingly attracting attention in recent years due to their potential in biological imaging and drug delivery applications. Despite their significant advantages over organic dyes and fluorescent proteins, cytotoxicity is still a major problem in live-cell QD labeling. In this work, in-vitro characterization of a novel CdTe/2MPA quantum dot capped with CdS-DMSA was conducted on human cervical cancer (HeLa) and mouse fibroblast (NIH/3T3) cell lines. Biocompatibility of this novel particle was evaluated in comparison to a commercial quantum dot (Qdot 565) and various QDs with CdTe core. Cytotoxicity of quantum dots was investigated using XTT and proliferation assays. Cellular internalization and localization of particles were studied using confocal laser scanning microscopy. For quantitative determination of internalization and intracellular QD stability, we also performed uptake and cadmium release assays. Optimal cell imaging concentration with CdTe-CdS/2MPA-DMSA was determined as 10-50 ug/mL in HeLa cells. Localization of the internalized QD particles was observed in the perinuclear region of the cells. XTT and proliferation assays provided identical viability results for the tested QDs. CdS-DMSA capping increased cytocompatibility of CdTe/2MPA by 15% in NIH/3T3 cells. Biocompatibility of this capped particle was further increased by 3-folds with pegylation. For pegylated CdTe-CdS/2MPA-DMSA and commercial Qdot 565, we have not observed QD-related cytotoxicity on NIH/3T3 cells following 24-hr QD exposure at 50 ug/mL. Our in-vitro characterization studies indicate that CdTe-CdS/2MPA-DMSA is a promising live-cell imaging probe which can be effectively excited in the visible range of the electromagnetic spectrum.

Drug delivery systems are designed to achieve low, local doses at the target site. Delivery systems can provide the drug in a continuous manner or in response to environmental stimuli such as temperature, pH or UV. This study aimed to develop photosensitive liposomes that achieve UV-responsive release of their content. The main mechanism was to incorporate a light sensitive molecule into the liposomal bilayer then achieve destabilization of the membrane by exposure to UV. This would result in an on demand release of the bioactive content. Suprofen, a nonstereoidal anti-inflammatory drug, also a light sensitive molecule, was selected to achieve the destabilization in this study. Lipid vesicles were prepared with different ratios of phosphatidyl choline, cholesterol and Suprofen (PC:CHOL:SPF) and characterized in terms of encapsulation efficiency, release rate and responsiveness to UV. Preliminary studies were carried out with calcein (CAL), a fluorescent dye, due to the ease of detection and the in vitro studies were carried out with the cancer drug Cisplatin.

Colorectal carcinoma (CRC) is one of the most common cancers and is the leading cause of cancer deaths in much of the developed world. Owing to the high incidence of drug resistance and potential toxic effects of chemotherapy drugs, much research is currently underway to design better strategies for smart drug delivery systems. Cyclooxygenase-2 (COX-2) pathway is associated with poor prognosis in colon carcinomas. The selective COX-2 inhibitor drug Celecoxib (CLX) has been shown to posses COX-2 independent anti-carcinogenic effects in addition to inhibition of prostaglandins synthesis. The aim of the presented thesis was to develop a liposomal delivery system for CLX and to evaluate functional effects in CRC cell lines. Starting with multilamellar vesicles capable of CLX encapsulation and retention, nano sized liposomes were prepared and characterized in vitro. The optimum composition was determined as 10:1 DSPC: Cholesterol molar ratio and Polyethylene glycol (PEG) grafting at 2% of phospholipids. The extent of cellular association of PEGylated liposome formulation was analyzed quantitatively and cellular localization was analyzed qualitatively. We detected that CLX loaded PEGylated liposomes inhibited proliferation and cellular motility of cancer cells in a 2D model system. Our results showed that, Epidermal Growth Factor Receptor (EGFR) targeted CLX loaded immunoliposomes were extremely cytotoxic in cancer cells with high EGFR expression but not in cells devoid of EGFR expression. This delivery system may pioneer studies that may potentially circumvent the harmful systemic side effects of cancer preventive and chemotherapy drugs as well as allow the use of targeted combinatorial therapies.