New discoveries in the field of metabolism by applying single-cell and spatial omics

Single-cell multi-Omics(SCM-Omics)and spatial multi-Omics(SM-Omics)technologies provide state-of-the-art methods for exploring the composition and function of cell types in tissues/organs.Since its emergence in 2009,single-cell RNA sequencing(scRNA-seq)has yielded many groundbreaking new discoveries.The combination of this method with the emergence and development of SM-Omics techniques has been a pioneering strategy in neuroscience,developmental biology,and cancer research,especially for assessing tumor heterogeneity and T-cell infiltration.In recent years,the application of these methods in the study of metabolic diseases has also increased.The emerging SCM-Omics and SM-Omics approaches allow the molecular and spatial analysis of cells to explore regulatory states and determine cell fate,and thus provide promising tools for unraveling heterogeneous metabolic processes and making them amenable to intervention.Here,we review the evolution of SCM-Omics and SM-Omics technologies,and describe the progress in the application of SCM-Omics and SM-Omics in metabolism-related diseases,including obesity,diabetes,nonalcoholic fatty liver disease(NAFLD)and cardiovascular disease(CVD).We also conclude that the application of SCM-Omics and SM-Omics approaches can help resolve the molecular mechanisms underlying the pathogenesis of metabolic diseases in the body and facilitate therapeutic measures for metabolism-related diseases.This review concludes with an overview of the current status of this emerging field and the outlook for its future.

Phytochemical composition and toxicity assessment of Ammi majus L.

Objective:To assess the acute and subacute toxicity as well as the phytochemical composition of two extracts and three fractions of Ammi majus L.Methods:The aqueous extracts were prepared separately by maceration for 48 h and by infusion for 1 h,while the fractions were prepared by the Soxhlet extractor,successively employing cyclohexane,ethyl acetate,and ethanol.The acute toxicity study was carried out in accordance with the OECD N°423 guideline at a single dose(2000 mg/kg)in mice for 14 days.The subacute toxicity study was performed by a daily oral administration of 250 mg/kg 2 for 10 d and 100 mg/kg doses for 28 d.Phytochemical screening was performed using staining and precipitation reactions,while the chemical characterization of some analytes was detected by HPLC-MS/MS analysis.Results:In the acute toxicity study,no signs of toxicity such as convulsion,salivation,diarrhea,sleep and coma were observed during 30 minutes and 14 days,so the lethal dose was higher than 2000 mg/kg for each extract and fraction.The subacute toxicity results showed that at a dose of 250 mg/kg,61.10%of the animals died and the rest developed morbidity.On the other hand,at a dose of 100 mg/kg,all the animals were still alive after 28 days,with no morbidity and the biochemical parameters were normal with no abnormalities in the liver,kidneys and pancreas.Phytochemical screening indicated the presence of flavonoids,tannins,coumarins,and free quinones and the absence of alkaloids and anthocyanins.Conclusions:The extracts and fractions of Ammi majus L.are not toxic in the short and long term with a varied chemical composition.Toxicological tests on animals other than rodents and in the long term(more than 28 days)are needed to further confirm the safety of Ammi majus extracts.

Customized scaffolds for large bone defects using 3D‑printed modular blocks from 2D‑medical images

Additive manufacturing(AM)has revolutionized the design and manufacturing of patient-specific,three-dimensional(3D),complex porous structures known as scaffolds for tissue engineering applications.The use of advanced image acquisition techniques,image processing,and computer-aided design methods has enabled the precise design and additive manufacturing of anatomically correct and patient-specific implants and scaffolds.However,these sophisticated techniques can be timeconsuming,labor-intensive,and expensive.Moreover,the necessary imaging and manufacturing equipment may not be readily available when urgent treatment is needed for trauma patients.In this study,a novel design and AM methods are proposed for the development of modular and customizable scaffold blocks that can be adapted to fit the bone defect area of a patient.These modular scaffold blocks can be combined to quickly form any patient-specific scaffold directly from two-dimensional(2D)medical images when the surgeon lacks access to a 3D printer or cannot wait for lengthy 3D imaging,modeling,and 3D printing during surgery.The proposed method begins with developing a bone surface-modeling algorithm that reconstructs a model of the patient’s bone from 2D medical image measurements without the need for expensive 3D medical imaging or segmentation.This algorithm can generate both patient-specific and average bone models.Additionally,a biomimetic continuous path planning method is developed for the additive manufacturing of scaffolds,allowing porous scaffold blocks with the desired biomechanical properties to be manufactured directly from 2D data or images.The algorithms are implemented,and the designed scaffold blocks are 3D printed using an extrusion-based AM process.Guidelines and instructions are also provided to assist surgeons in assembling scaffold blocks for the self-repair of patient-specific large bone defects.

Large-Scale Membrane Bioreactors for Industrial Wastewater Treatment in China:Technical and Economic Features,Driving Forces,and Perspectives

Membrane bioreactors(MBRs)have been and will continue playing an important role in industrial wastewater treatment and reuse in China.The sustainable development of MBR technology in its mature-application stage requires reciprocal interactions between engineering and research participants.Thus,in this study,a total of 182 large-scale MBR projects treating industrial wastewater(with individual treatment capacities5000 m3d1)commissioned and under construction from 2003 to 2019 were analyzed comprehensively.Fast growth of the cumulative treatment capacity was observed,with extension to diverse industries,and the super large-scale was enhanced recently.The treatment processes,pollutant removal efficiencies,and actual operational parameters were summarized regarding the particularity of industrial wastewater compared to municipal wastewater.Economic features including the total investment costs of the projects,their total footprint,and their operational energy consumption were analyzed as well.A vigorous MBR market has formed in China with the fast development of membrane elements and engineering suppliers,continuously increasing official oriented projects,and responsive and innovative business modes.MBR technology has been mostly applied in specific economic zones and water-deficient areas,but its widespread use all over China is foreseeable considering the vast future market for industrial wastewater treatment and recycling.The policy–economy and market–technology driving forces revealed that MBR is consistent with the national development demand.According to the survey and analysis,prospective development in both engineering and research aspects of MBR is proposed to maintain its competitive edge.

Effect of Helicobacter pylori infection on p53 expression of gastric mucosa and adenocarcinoma with microsatellite instability

AIM: To investigate the relationship between Helicobacter pylori(H pylori) infection, microsatellite instability and the expressions of the p53 in gastritis, intestinal metaplasia and gastric adenocarcinoma and to elucidate the mechanism of gastric carcinogenesis relating to H pylori infection.METHODS: One hundred and eight endoscopic biopsies and gastric adenocarcinoma were available for the study including 33 cases of normal, 45 cases of gastritis, 30 cases of intestinal metaplasia, and 46 cases of gastric adenocarcinoma. Peripheral blood samples of these patients were also collected. H pyloriinfection and p53 expressions were detected by means of streptavidin-peroxidase (SP)immunohistochemical method. Microsatellite loci were studied by PCR-SSCP-CE using the markers BAT-26,D17S261, D3S1283, D2S123, and D3S1611. MSI was defined as the peak shift in the DNA of the gastric tissue compared with that of the peripheral blood samples. Based on the number of mutated MSI markers, specimens were charac-terized as high MSI (MSI-H) if they manifested instability at two or more markers, low MSI (MSI-L) if unstable at only one marker, and microsatellite stable (MSS) if they showed no instability at any marker.RESULTS: H pyloriinfection was detected in the samples of gastritis, intestinal metaplasia, and gastric adenocarcinoma and the infection frequencies were 84.4%, 76.7%, and 65.2%, respectively, whereas no H pylori infection was detected in the samples of normal control. There was a significant difference in the infection rates between gastritis and carcinoma samples (P = 0.035). No MSI was detected in gastritis samples, one MSI-H and two MSI-L were detected among the 30 intestinal metaplasia samples, and 12 MSI-H and 3 MSI-L were detected in the 46 gastric carcinomas.In those gastric carcinomas, the MSI-H frequency in H pyloripositive group was significantly higher than that in H pylorinegative group. No p53 expression was detected in the normal and gastritis samples from dyspeptic patients. P53-positive immunohistochemical staining was detected in 13.3% of intestinal metaplasia samples and in 43.5% of gastric carcinoma samples. The levels of p53 in H pyloripositive samples were higher than those in the negative group when the carcinoma samples were subdivided into H pylori-positive and -negative groups (P = 0.013). Eight samples were detected with positive p53 expression out of the 11 MSI-H carcinomas with H pylorl infection and no p53 expression could be seen in the H pylori-negative samples.CONCLUSION: H pylori affect the p53 pattern in gastric mucosa when MMR system fails to work. Mutations of the p53 gene seem to be an early event in gastric carcinogenesis.

Detection of Promoter Hypermethylation of GSTP1 and CDH1 Genes and the Relationship of Histopathological Parameters of the Breast

Background: Breast cancer is the most common cancer in women. Histopathology plays an important part in determining the treatment strategy for women with breast cancer. GSTP1 plays an important role in protecting cells from cytotoxic and carcinogenic agents and it is expressed in normal tissues at variable levels in different cell types. CDH1 plays a critical role for establishment and maintenance of polarity and differentiation of epithelium during the development period. Also, it plays an important role in signal transduction, differentiation, gene expression, cellmotility and inflammations. Methods: In this study the promoter methylation levels of GSTP1 and CDH1 gene which are associated with breast cancer were investigated by technique of Methylation Sensitive High Resolution Melting Analysis (MS-HRM). We analysed primary tumor core biopsies from 80 high-risk primary breast cancer patients (tumors ≥ 2 cm and/or lymphatic metastase and/or distant metastases and/or under 40 years). Also the patients’ histopathologic types were associated with the methylation levels. Results: In our study the promoter hypermethylation status was observed at different rates;GSTP1 and CDH1 hypermethylation frequencies were 82% and 95% respectively. The promoter hypermethylation levels of the genes were found to be significant with lymph node positivity, ER positivity and HER2/neu negativity. Conclusion: Our study is important as being the first study that analyzes association between histopathologic type and GSTP1 and CDH1 gene promotor methylation status in Turkish population.

Separation of methane from different gas mixtures using modified silicon carbide nanosheet: Micro and macro scale numerical studies

This research discusses the separation of methane gas from three different gas mixtures,CH4/H2 S,CH4/N2 and CH4/CO2,using a modified silicon carbide nanosheet(Si CNS)membrane using both molecular dynamics(MD)and computational fluid dynamics(CFD)methods.The research examines the effects of different structures of the Si CNSs on the separation of these gas mixtures.Various parameters including the potential of the mean force,separation factor,permeation rate,selectivity and diffusivity are discussed in detail.Our MD simulations showed that the separation of CH4/H2 S,and CH4/CO2 mixtures was successful,while simulation demonstrated a poor result for the CH4/N2 mixture.The effect of temperature on the diffusivity of gas is also discussed,and a correlation is introduced for diffusivity as a function of temperature.The evaluated value for diffusivity is then used in the CFD method to investigate the permeation rate of gas mixtures.

Hydrophobic coating of surfaces by plasma polymerization in an RF plasma reactor with an outer planar electrode:synthesis,characterization and biocompatibility

This paper presents the plasma polymerization of poly（hexafluorobutyl acrylate）（PHFBA） thin films on different substrates in an RF plasma reactor with an outer planar electrode. This reactor configuration allows？large area uniformity and fast processing times. Deposition rates of up to60 nm min-1were observed. The influence？of plasma power and substrate temperature on the？deposition rate, structure and wettability of the as-deposited films was？investigated. It was observed that better hydrophobicity was obtained at high plasma power and in low temperature conditions. PHFBA thin films deposited on electrospun poly（acrylonitrile） fiber mats under such conditions resulted in superhydrophobic surfaces with？contact angle values greater than 150°.In？vitro cell studies using human epithelial cells demonstrated the non-toxic nature of the？plasma-polymerized PHFBA films.

Exploring a natural MDR reversal agent: potential of medicinal food supplement Nerium oleander leaf distillate

Objective:To investigate the molecular effects ofNerium oleanderleaf distillate on paclitaxel and vincristine resistant(MCF-7/Pac and MCF-7/Vinc)cells and sensitive(MCF-7/S)cell lines.Methods:Nerium oleander(N.oleander)leaf extract was obtained by hydrodistillation method.The toxicological effects ofN.oleanderdistillate,previously suggested as medicinal food supplement,on drug resistant cells were evaluated by XTT tests.MDR modulation potential of the plant material was evaluated by flow cytometry and fluorescent microscopy.Paclitaxel and vincristine were applied to the sublines in combination with N.oleanderdistillate.Results:Fractional inhibitory indices show thatN.oleanderdistillate did not increase the antiproliferative effects of anticancer drugs.N.oleandertreatment in to MCF-7/Pac and MCF-7/Vinc did not inhibit P-gp activity and MDR1 gene expression level.Conclusions:As a result it may be suggested that althoughN.oleanderdistillate has some medicinal effects as food supplement it may not be suitable as an MDR modulator for drug resistant breast cancer cells.

Effects of chelating agents on the sol-gel synthesis of nano-zirconia:Comparison of the Pechini and sugar-based methods

This study focused on the comparison of the Pechini and sugar-based combustion synthesis methods to produce nano-zirconia. Zirconium hydroxide was utilized as metal precursor and citric acid, sucrose, and fructose were used as chelating agents, followed by calcination at 500, 600, and 700°C in air, respectively. Characterization was conducted by thermal analysis, specific surface area measurement, Fourier transform infrared spectroscopy, X-ray diffraction, and scanning and transmission electron microscopy. When sucrose and citric acid were used as chelating agents during synthesis, mixtures of monoclinic and tetragonal phases were formed after calcination at 600 and 700°C. In the fructose samples, the tetragonal structure was the unique characterized phase. The tetragonal parameters in the fructose samples were determined using the diffraction data and the lattice parameter ratio was proven to increase with the temperature increase. Compared with the citrate and sucrose samples, the largest specific surface area(27 m2·g-1) and smallest particle size(39.1 nm) were obtained for the fructose sample after calcination at 700°C. The study revealed the formation of single-phase stabilized tetragonal zirconia using fructose as chelating agent after calcination at 500°C, and the presence and formation mechanism of stabilized tetragonal phase were also discussed on the basis of the X-ray and electron diffraction studies.

Organic Material and Trace Elements of Bituminous Rocks in the Ozank(?)y Field,Ankara,Turkey

Bituminous rocks in the Ozankoey （Ankara） field are different from those of the Paleocene- Eocene Mengen and Giineytepe （Bolu） regions in metal enrichment levels. Organic carbon （Corg） content of organic material-rich rocks in the Ozankoey （Ankara） field is 3.66-40.72% wt averaging 14.34%. The dominant organic materials are algae/amorphous accompanied by minor amount of herbaceous material （The dominant kerogen type is Type-I with a limited amount of Type-Ⅱ kerogen.）. The bituminous rocks in the Ozankoey field are enriched in heavy metals such as Ni, Mn, As and Cr. In comparison with the average enrichment values of dements, Ni, Mn, As and Cr in bituminous shales of the Ozankoey field are as about 4.38, 14.93, 10.90 and 5.58 times as average values. The average concentrations of these heavy metals are also as high as 215× 10^-6, 828 × 10^-6, 58.54 × 10^-6, and 148 × 10^-6 respectively. In addition, sorption properties of day and organic materials are also important for metal enrichments in the bituminous shales.

Mathematical modeling and numerical computation of the effective interfacial conditions for Stokes flow on an arbitrarily rough solid surface

The present work is concerned with a two-dimensional(2D)Stokes flow through a channel bounded by two parallel solid walls.The distance between the walls may be arbitrary,and the surface of one of the walls can be arbitrarily rough.The main objective of this work consists in homogenizing the heterogeneous interface between the rough wall and fluid so as to obtain an equivalent smooth slippery fluid/solid interface characterized by an effective slip length.To solve the corresponding problem,two efficient numerical approaches are elaborated on the basis of the method of fundamental solution(MFS)and the boundary element methods(BEMs).They are applied to different cases where the fluid/solid interface is periodically or randomly rough.The results obtained by the proposed two methods are compared with those given by the finite element method and some relevant ones reported in the literature.This comparison shows that the two proposed methods are particularly efficient and accurate.

The synthesis of boronic-imine structured compounds and identification of their anticancer, antimicrobial and antioxidant activities

Boronic acid compounds with different substituted groups were handled to synthesize various ligands encoded as B1, B2, B3, B4, BS, B6, B7 and BS. B5 and B7 were tested for the cytotoxic activity against the prostate cancer cells and it was found that the cell viability of cancer cells was decreased while most of the healthy cells could still be viable. 5 μM solutions of B5 and B7 decreased the cell viability to 33% and 44%; whereas healthy cells were 71% and 95%, respectively, after treatment. Antimicrobial properties were explored against the bacterial and fungal microorganisms with B1, B5 and BZ The inhibition zones were evaluated for all boronic structures, and the growth inhibition zones were determined in a range of 7-13 mm diameter for different microorganism species. Staphylococcus aureus was the common micro- organism that three boronic compounds with imine ligands showed the activity. Antioxidant features of B2, B3, B4, B5, B6, B7 and B8 were investigated by different processes such as Beta-carotene bleaching （BCB）, 2,2-diphenyl picryl hydrazyl （DPPH）, 2,2″-azino-bis（ 3-ethylbenzothiazoline-6-sulphonic acid） （ABTS） and CUPric reducing antioxidant capacity （CLIPRAC） methods. Significant antioxidant activity was achieved by the phenyl boronic based ligands and these compounds demonstrated as much activity as standards （α-Toc and BHT）. In addition, all structures were applied properly without any decomposition during the experiments. They were rather stable both in aqueous media and solid state.

Acute myeloid leukemia cells inhibit the differentiation and maturation of dendritic cells and induce the generation of regulatory T cells

Objective:To investigate the effects of soluble factors secreted by acute myeloid leukemia(AML) cells on the phenotypical and functional properties of DCs derived from normal mononuclear cells. Methods:Mononuclear cells were cul-tured with interleukin-4(IL-4) and granulocyte-macrophage colony-stimulating factor(GM-CSF),in the presence or absence of 24 h culture supernatants from fresh primary AML cells,to generate immature DCs. The maturation of DCs was induced by cytokines IL-1beta,IL-6,tumor necrosis factor-alpha(TNF-alpha),and prostaglandin-2(PGE-2). The phenotypic alterations of DCs and DCs-primed CD4+ T cells were evaluated using flow cytometry. Precursor frequency(PF) was calculated to monitor the allostimulatory effects of DCs on CD4+ and CD8+ T cells. Results:AML cell supernatant-treated DCs showed significantly lower expression of co-stimulatory molecules CD80 and CD86,and reduced response to cytokines IL-1beta,IL-6,TNF-alpha,and PGE-2. The allostimulatory effects of AML cell supernatant-treated DCs on CD4+ and CD8+ T cells were significantly lower than those of normal mature DCs [PF:(1.8 ± 0.5)% vs.(5.2 ± 1.6)% for CD4+ T cells,(2.1 ± 0.6)% vs.(6.5 ± 2.0)% for CD8+ T cells,P < 0.01]. These AML supernatant-induced DCs could also induce allogeneic CD4+ T cells to differentiate into CD4+CD25high T cells,which had immunophenotyping characteristics of regulatory T cells,i.e. they expressed Foxp3 but not active maker CD69. Conclusion:This study demonstrates that soluble factors secreted by AML cells can inhibit development and functions of DCs. In addition,AML supernatant-induced DCs can induce the generation of CD4+CD25high T cells from CD4+ T cells,which may be a mechanism of increased prevalence of CD4+CD25high regulatory T cells and immune dysfunction in AML patients.

Fundamental Open Questions on Engineering of “Super” Hydrogen Sorption in Graphite Nanofibers: Relevance for Clean Energy Applications

Herein, some fundamental open questions on engineering of “super” hydrogen sorption (storage) in carbonaceous nanomaterials are considered, namely: 1) on thermodynamic stability and related characteristics of some hydrogenated graphene layers nanostructures: relevance to the hydrogen storage problem;2) determination of thermodynamic characteristics of graphene hydrides;3) a treatment and interpretation of some recent STM, STS, HREELS/LEED, PES, ARPS and Raman spectroscopy data on hydrogensorbtion with epitaxial graphenes;4) on the physics of intercalation of hydrogen into surface graphene-like nanoblisters in pyrolytic graphite and epitaxial graphenes;5) on the physics of the elastic and plastic deformation of graphene walls in hydrogenated graphite nanofibers;6) on the physics of engineering of “super” hydrogen sorption (storage) in carbonaceous nanomaterials, in the light of analysis of the Rodriguez-Baker extraordinary data and some others. These fundamental open questions may be solved within several years.

Investigation of DNAmethylation of TWIST Gene in Breast Cancer and Its Relationship to Histopathological Features

BACKGROUND: Promoter hypermethylation and global hypomethylation in the human genome are hallmarks of most cancers. OBJECTIVE: The aim of this study is to assess the methylation profile patterns of TWIST gene and to investigate the relationship of methylation with pathological features. METHODS: Romoter CpGisland methylation of TWIST which can be related in breast cancer was performed by methylation-sensitive high resolution melting (MS-HRM) analysis. Formalin-fixed, paraffin-embedded (FFPE) tissue samples of 80 patients with a diagnosis of primary breast cancer from Eskisehir Osmangazi University medical faculty of Oncology Clinic were included. RESULTS: In our study, the promoter hypermethylation frequency of TWIST gene was 25.0%. With these results, when the prognostic factors of the patients were analyzed, tumor stage and age were found to be meaningless with the hypermethylation of TWIST gene, but found to be significant with lymph node positivity, ER positivity, PR negativity, and HER2/NEU negativity. CONCLUSION: Our study is important as being the first study that analyzes association between histopathologic type and TWIST gene promotor methylation status in Turkish population.

Investigation of Methylation Profiles of TP53, Caspase 9, Caspase 8, Caspase 3 Genes Treated with DNA Methyl Transferase Inhibitor (DNMTi) Zebularine (ZEB) and Caffeic Acid Phenethyl Ester (CAPE) on MCF-7 and MDA-MB-231 Breast Cancer Cell Lines

Purpose: MCF-7 (ER+, WTP53) and MDA-MB-231 (ER Met, Mutant P53) Caffeic Acid Phenethyl Ester (CAPE) and DNA Methyl Transferase Inhibitor (DNMTi) in breast cancer cell lines of Zebularine (ZEB) single and combined application of TP53, caspase-9, caspase 8 and caspase-3 genes as a result of the use of single and combined drug methylation profiles are aimed to be evaluated by specific PCR method. Material-Metods: In the MCF-7 and MDA-MB-231 breast cancer cell lines, MTT test and survival analysis were performed as a result of single and combined application of CAPE and Zebularine and Methylation Specific PCR was performed to examine the methylation of caspase-3, caspase-8, caspase-9 and TP53 genes. Results: According to the results of 24-hour drug administration, the IC50 for the MCF-7 cell line was determined as 200 μM, for CAPE 40 μM and for the combined values of 50 μM ZEB + 5 μM CAPE. The effects of caspase-3, caspase-8, caspase-9 and TP53 genes on the methylation level of ZEB, CAPE and ZEB + CAPE drug combination were determined by using bisulfite modified DNAs in MCF-7 and MDA-MB-231 cell lines. Discussion: In the MCF-7 cell line, the 120 μM ZEB viability rate was 51%, and the viability of 80 μM ZEB MDA-MB-231 breast cancer cells decreased by 59.7%. After 20 μM CAPE, viability in MCF-7 cells decreased by 31% in 120 μM CAPE and MDA-MB-231 cells decreased by 41%. The viability with 40 μM CAPE decreased by 19% in MDA-MB-231 cells. It was found that 20 μM CAPE concentration was associated with TP53 methylation in MCF-7 cell lines. The 80 μM ZEB concentration was found to be closely related to the unmethylated status of the TP53 gene. These results obtained with 50 μM ZEB + 5 μM CAPE application were found to be related to the methylated-unmetylated status of the TP53 gene in half (50%). For the caspase-9 gene of MDA-MB-231 cells, 80 μM ZEB concentration was found to be associated with unmetylated status. The effective use of drugs with low concentrations of the drug dose provides a more appropriate approach in terms of treatment.

Investigation of the Effects of Zebularine on Caspase-3 and Caspase-9 Involved in Anticancer and Apoptotic Mechanisms in SKBR3 Breast Cancer Cell Line

The aim of this study is to investigate the anti-proliferative effects of Zebularine on caspase-3 and 9 genes by methylation and protein expression on SKBR3 cells. The SKBR3 cells were treated with Zebularine at different concentrations (0 - 140 μM) for 24 - 96 hours and the effects on the cell viability were shown. The effects on the cell migration and cell transformation were also demonstrated at the specified dose (IC50 = 40 μM). At the same time, the HRM method and western-blot analysis were used to understand the effects in the apoptotic mechanism. According to the obtained results, it was observed that Zebularine significantly decreased the cell proliferation, cell migration and the cell growth (p < 0.001). As a result of the methylation analyzes performed on SKBR-3 cells at the applied dose, it was observed that Zebularine caused a decrease in both caspase-3 and 9 methylation rates at the 72nd hour. However, both reductions in the methylation levels didn’t cause a significant change (p > 0.05). At the same time, it was detected by the western-blot analyses that there was a time dependent increase in caspase-3 genes while a decrease was detected in the caspase-9 gene in progress of time. The obtained results showed that the methylation changes occurring in the caspase genes weren’t related to the protein levels. According to these results, supportive results are obtained showing that the Zebularine can be used in chemotherapy and that this study is unique since it is the first study in the literature intending to investigate the effects of Zebularine on the SKBR3 cells.

Reinforcement of Polypropylene Composite system via Fillers and Compatibilizers

The purpose of this work is to compare the mechanical properties of the different PP/ Filler composites. We considered the effect of treatment of the filler on the adhesion between the filler and the matrix. Modification of polypropylene (PP) with different coated and uncoated fillers is investigated in this paper. Six types of different fillers are used for preparation of filled PP composites. The composite samples were homogenized in a twin screw extruder. The compatibility of PP and fillers focused in this study and modified by using coupling agent. The obtained values of composites were cor- related with mechanical properties.

The Evaluation of the Effects of Temozolomide on MGMT Gene Expression in MCF-7 and SKBR3 Human Breast Cancer Cell Lines

Background and Aim: In this study, it was aimed to examine the cytotoxic effect of temozolomide (TMZ) treatment, on MCF-7 and SKBR3 cell lines, to study the methylation levels of MGMT gene expression and gene promoter region. Methods: The MTT test was performed to determine the effective dose of TMZ. The time-dependent cell survival test was performed after the IC50 value was found. Western blotting was performed to determine MGMT gene expression levels. High Resolution Melting (HRM) technique was used to determine the methylation levels of MGMT gene promoter region. Results: TMZ has been shown to have a high cytotoxic effect on SKBR3 cell line and low cytotoxicity on MCF-7. When MGMT expression levels before and after TMZ treatment were observed by western blotting, the gene expression levels of TMZ treatment were shown to decrease in both cell lines. It was observed that MGMT gene promoter region was hypermethylated in two cell lines, and that the application of TMZ further increased the methylation levels in the promoter region. Conclusions: It was seen that TMZ could be used as a single agent in SKBR-3 cell line. With this study on breast cancer, it is expected that temozolomide treatment will lead future in vitro and in vivo studies for breast cancer.