The genus Salvia L. is significantly important with regard to both its worldwide distribution and usage areas including food, medical and perfumary industries. In this current study, it is targeted to address the chromosome numbers and karyomorphology of the ten species and one variety of the genus Salvia. All of the eleven taxa examined in this study are economically significant and nine of these are endemic to Turkey. To define the chromosome numbers and karyomorphology of these eleven taxa somatic chromosomes of the each were examined. Mitotic metaphase chromosomes were obtained from root meristems of germinating seeds, which were pre-treated in &amp / #945 / -bromonaphtalene at 4&ordm / C for 16 h, then fixed in Carnoy solution (3 parts of ethanol: 1 parts of glacial acetic acid) at 4&ordm / C for 24h and stored in 70 % ethanol. Fixed root tips were stained in 2 % aceto-orcein and squashed in a drop of 45 % acetic acid. Long arm, short arm, total length of the each chromosomes were measured / relative length, arm ratio, centromeric index of the each chromosome were calculated. Karyogram and haploid idiograms were drawn by computer-aided analysis programme (Bs200pro). A cluster analysis of the karyotype data was carried out to examine karyotype similarity among taxa. Somatic chromosome numbers have been counted as 2n=2x=14 for the endemic taxa S. divaricata Montbret &amp / Aucher, S. euphratica Montbret &amp / Aucher ex Bentham (var. leiocalycina (Rech. fil.) Hedge) and S. recognita Fisch. &amp / Mey. / 2n=2x=14-1B for Salvia rosifolia Sm. / 2n=20 for S. longipedicellata Hedge, S. vermifolia Hedge &amp / Hub.-Mor. and S. yosgadensis Freyn &amp / Bornm. / 2n=2x=22 for S. aethiopis L., S. cilicica Boiss. &amp / Kotschy, S. hypargeia Fisch. &amp / Mey. and 2n=2x=32 for S. napifolia Jacq. respectively. In general, the chromosomes are short with median and submedian centromeres. The current study is essential for being the first report about chromosome numbers and karyomorphology of the six endemic taxa, namely S. divaricata, S. euphratica var. leiocalycina, S. longipedicellata, S. rosifolia, S. vermifolia and S. yosgadensis. Moreover, in spite of the chromosome numbers of S. aethiopis, S. cilicica, S. hypargeia and S. recognita are known, this research is the first study for their karyomorphologies.

Emodin (1,3,8-trihydroxy-6-methylanthraquinone) is a phytoestrogenic component of Rheum plant extracts which has been used for medical treatment since ancient times. It has been shown to have anti-inflammatory and anti-cancer effects. In our research, we aimed to study the biological effects of emodin on MCF-7 and MDA-231 cell lines. Cytotoxicity assays showed that emodin treatment for 48hours caused a concentration dependent decrease in viable cell numbers of both cell lines. As determined by cell counting with tryphan blue, IC50 values were 8.40 and 12.17 &micro / g/ml for MCF-7 and MDA-231 cells, respectively. Apoptotic effects of emodin was investigated by measuring the changes in apoptotic and antiapoptotic gene expressions by qRT-PCR. In MCF-7 cells, Bax expression increased with increasing emodin concentrations, while Bcl-2 expression was downregulated. Bax/Bcl-2 ratio was calculated as 9.2 fold at 10&micro / g emodin/ml treatment for 48 hours, indicating stimulation of apoptosis. However, Bax/Bcl-2 ratio was found 1.6 fold for MDA-231 cells. These results were in accordance with the results obtained from microarray analysis of related gene expressions. MCF-7 cells were more apoptotic in comparison to MDA-231 cells. DNA fragmentation was observed in MCF-7 cells by TUNEL method. GST enzyme activity that was measured using CDNB as substrate, was increased 100% with respect to control up to 5&micro / g emodin/ml treatment of MCF-7 cells for 48 hours. However, effect of emodin on GST activity in MDA-231 cells was found insignificant. According to microarray analysis results, emodin affected the gene expressions of GST isozymes in MCF-7 cells much more than in MDA-231 cells.

Meniscus is a wedge shaped structure, with a convex base attached to a flat tibial surface, and with a concave femoral surface, on which femur and tibia articulate. It has several functions including joint lubrication, shock absorption, load transmission and joint stability. Various methods were tried to treat meniscal tears but each has its own drawbacks. Tissue engineering seems to be a promising solution that avoids all the problems associated with the other approaches. In this study, a three dimensional (3D) collagen-based structure was prepared by tissue engineering to mimic the natural human meniscus. Three different foams prepared under different conditions were combined and nano/microfibrous layers were placed in between them. To mimic the properties of the natural tissue, the top layer was composed of collagen-chondroitin sulfate-hyaluronic acid (Coll-CS-HA) prepared by freezing at -20&ordm / C followed by lyophilization. The middle and bottom layers were made with just collagen after freezing at -20&ordm / C and -80&ordm / C, respectively and lyophilization. Aligned nano/microfibers were prepared using collagen-poly(L-lactic-co-glycolic acid (Coll-PLGA). Various crosslinking procedures such as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS), genipin (GP), glutaraldehyde (GLU) either alone or in combination with dehydrothermal treatment (DHT) were used and based on both compressive and tensile properties, the best crosslinker was chosen to be DHT+EDC/NHS. Mechanical properties (compressive, tensile and shear) of the dry foams and the final 3D construct were evaluated. The highest mechanical properties were obtained with the 3D construct. Then all these foams and the 3D construct were seeded with human fibrochondrocytes to study the cell behavior such as attachment, proliferation, and extracellular matrix (ECM) and glucosaminoglycan (GAG) production. Furthermore, the influence of cell seeding on the compressive properties of wet individual foams and the 3D construct was observed. As expected, the highest cell proliferation and compressive properties were obtained with the 3D construct. Finally, the 3D constructs, seeded with fibrochondrocytes, were implanted in New Zealand rabbits after meniscectomy. The initial microscopical examination show that the 3D construct has a significant potential as a meniscus substitute.

ABSTRACT INVESTIGATON OF CHEMOPREVENTIVE PROPERTIES OF URTICA DIOICA L., IN MCF-7 AND MDA231 BREAST CANCER CELL LINES. G&uuml / ler, Elif Ph.D., Biological Sciences Department Supervisor : Prof. Dr. Mesude

This study investigated the 9th grade students&#039 / achievement regarding diffusion and osmosis in relation to reasoning ability, prior knowledge and gender. A total of 397 ninth grade students participated in the study. The Test of logical thinking (TOLT) and the Diffusion and Osmosis Diagnostic Test (DODT) were administered to determine students&#039 / reasoning ability and achievement in diffusion and osmosis, respectively. DODT results showed that the range of correct answers for the first tier was 41 % to 91%. When both tiers were combined, the correct responses were reduced to a range of 21% to 61%. This result reveals that students have enough content knowledge but they don&rsquo / t know the underlying reason of their choice in diffusion and osmosis concepts. Pearson Product Moment correlations showed a statistically significant positive correlation between achievement and students&#039 / prior knowledge &amp / reasoning ability. MRC Analysis was conducted to determine the contribution of prior knowledge, reasoning ability and gender to the achievement. Prior knowledge and reasoning ability, but not gender, made a statistically significant contribution to the variation on achievement. Prior knowledge and reasoning ability together predicted 37 % of the variation on achievement. Stepwise multiple regression analysis was computed to determine the variables were best predicting students&rsquo / achievement. While prior knowledge explains 33 % of the variation in achievement, reasoning ability explains only 4 % of the variation in achievement. Results indicate that prior knowledge is a better predictor than reasoning ability in students&rsquo / achievement.

Nerve injury is a serious clinical problem that has a direct impact on the quality of life. Nerve tissue engineering (NTE) is one of the most promising methods in human health care to restore the function of damaged neural tissues. The current state of the art involves the construction of a tissue engineered, nano or micropatterned 3-D nerve tube that has fibers or channels in the inside. The scope of this study is to construct a 3-D, biodegradable nerve tube which consists of an aligned, electrospun mat seeded with stem cells that is wrapped in a porous micropatterned film which contains support cells. In two separate approaches human mesenchymal stem cells (MSCs) and mouse neural stem cells (NSCs) were used. In the design with the MSCs, the micropatterned exterior part of the nerve tube contained undifferentiated MSCs as support cells and this was wrapped around the fibers seeded with MSCs which were induced to neural differentiation. In the other case, NSCs differentiated into astrocytes were used as support cells seeded on the micropatterned film and the mat was loaded with undifferentiated NSCs. Differentiation into neural cells and astrocytes were shown with immunocytochemistry and RT-PCR. The neuron-like MSCs and NSCs were shown to express neural marker &amp / #946 / -Tubulin III whereas astrocytes expressed glial fibrillary acidic protein (GFAP), an astrocyte marker. RT-PCR showed that early neural markers, nestin and Nurr 1, were expressed at passage 4 by undifferentiated MSCs and by MSCs induced to neural differentiation, while these markers were not expressed in undifferentiated MSCs at passages 2 and 3. The cells aligned along the axis of the micropattern of the film and along the axis of the fiber on the fibrous mat. This behavior was also maintained after construct formation. MTS and confocal microscopy revealed that the cells were viable and homogeneously distributed over the two parts of the scaffold. This indicates that the construct has a potential to be tested in vivo for nerve tissue engineering purposes.

ABSTRACT INTERACTION BETWEEN MICRO AND NANO PATTERNED POLYMERIC SURFACES AND DIFFERENT CELL TYPES &Ouml / z&ccedil / elik, Hayriye Ph.D., Department of Biology Supervisor: Prof. Dr. Vasif Hasirci Co-Supervisor: Dr. Celestino Padeste August 2012, 139 pages Micro and nanopatterned surfaces are powerful experimental platforms for investigating the mechanisms of cell adhesion, cell orientation, differentiation and they enable significant contributions to the fields of basic cell and stem cell biology, and tissue engineering. In this study, interaction between micro and nanopatterned polymeric surfaces and different cell types was investigated. Three types of micropillars were produced by photolithography (Type 1-3), while nanometer sized pillars were produced in the form of an array by electron beam lithography (EBL). Replica of silicon masters were made of polydimethylsiloxane (PDMS). Polymeric [P(L-D,L)LA and a P(L-D,L)LA:PLGA blend] replica were prepared by solvent casting of these on the PDMS template and used in in vitro studies. The final substrates were characterized by various microscopic methods such as light microscopy, atomic force microscopy (AFM) and scanning electron microscopy (SEM). In order to investigate deformation of the nucleus in response to the physical restrictions imposed by micropillars, Type 1 and Type 2 pillars were used. These substrates were covered with pillars with different interpillar distances. While Type 1 is covered with symmetrically (in X-Y directions) distributed pillars, Type 2 pillars were distributed asymmetrically and the inter-pillar distances were increased. Nuclei deformation of five cell v types, two cancer cell lines (MCF7 and Saos-2), one healthy bone cell (hFOB1.19), one stem cell (bone marrow origined mesemchymal stem cells, BMSCs) and one standard biomaterial test cell type, (L929) fibroblasts was examined by using fluorescence microscopy and SEM. The nuclei of Saos-2 and MCF7 cells were found to be deformed most drastically. Nucleus deformation and intactness of nuclear membrane was examined by Anti- Lamin A staining. The interaction of the cells with micropillars was visualized by labelling focal adhesion complexes (FAC). Wettabilities of patterned and smooth surfaces were determined. As the patterns become denser (closer micropillars, Type 1) the hydrophobicity increased. Similar to water droplets, the cells were mostly spread at the top of the Type 1 pillars. The number of cells spread on the substrate surface was much higher on Type 2 patterned films. In order to support these qualitative findings, nucleus deformation was quantified by image analysis. Frequency of nucleus deformation was determined as the ratio of deformed to the total number of nuclei (%). In order to quantify the intensity of nuclei deformation, their circularity was evaluated. In addition to nucleus deformation, alterations in the ratio of cell area-to-nucleus area in response to micropillars were determined by image analysis. The results indicated that cancerous cells were more deformable. The qualitative microscopic evaluation and the data obtained by quantification of the nucleus and cellular deformation were in good agreement. In addition, the findings were consistent with expectations which suggest that cancerous cells are &ldquo / softer&rdquo / . In the second part of the research the force applied by the cells on arrays of micropillars with high aspect ratios (Type 3 substrates) during tugging at the pillars was investigated. Micropillars were produced using P(L-D,L)LA as well as a 60:40 blend of P(L-D,L)LA with PLGA. The blend is a material with lower stiffness than P(L-D,L)LA. The mechanical properties of the two materials were determined by tensile testing of solvent cast films. Deformation of Type 3 micropillars by the cellular tugging force of Saos-2 and L929 was studied by fluorescence and SEM microscopy, both on stiff and softer substrates. Displacements of the centers nodes of the pillars were evaluated from SEM micrographs. On the stiff surface, the two cell types bent the pillars to the same extent. On the other softer substrate (blends), however, the maximum displacements observed with Saos-2 cells were higher than the ones caused on the stiffer substrate or the ones caused by L929 cells. It is reported that stiffness of the substrate can determine stem cell lineage commitment. In order to examine the effects of change of substrate stiffness on osteogenic differentiation of BMSCs, osteopontin (OPN) expression was determined microscopically. It was found that osteogenic differentiation is enhanced when BMSCs are cultured on P(L-D,L)LA Type 3 pillars. vi In the last part of research, arrays of nanopillars whose interpillar distances systematically varied to form different fields were examined in terms of adhesion and alignment in order to determine the differential adhesion of BMSCs and Saos-2 cells. The difference in their adhesion preference on nanopillar arrays was quantified by image analysis. It was observed that BMSCs and Saos-2 cells behaved in an opposite manner with respect to each other on the fields with the highest density of nanopillars. The BMSCs avoided the most densely nanopillar covered fields and occupied the pattern free regions. The Saos-2, on the other hand, occupied the most densely nanopillar covered fields and left the pattern free regions almost unpopulated. It was also found that both BMSCs and Saos-2 cells aligned in the direction of the shorter distance between the pillars. Both BMSCs and Saos-2 cells started to align on the pillars if the distance in any direction was &gt / 1.5 &mu / m. To better understand the effects of chemical and physical cues, protein coating and material stiffness were tested as two additional parameters. After fibronectin coating, the surfaces of P(L-D,L)LA films with the highly dense pillar covered fields, which were avoided when uncoated, were highly populated by the BMSC. Similarly, decreasing the stiffness of a surface which was normally avoided by the BMSCs made it more acceptable for the cells to attach.

Bone marrow mesenchymal stem cells (BM-MSCs) are pluripotent cells that can differentiate into a variety of non-hematopoietic tissues. They also maintain healthy heamatopoiesis by providing supportive cellular microenvironment into BM. In this thesis, MSCs were characterized in terms of their morphological, immunophenotypical and differentiation properties. Then, they were examined by attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy together with hierarchical clustering, and FTIR microspectroscopy. In the first part of this study, global structural and compositional changes in BM-MSCs during beta thallasemia major (