Search results for: adipose tissue derived stromal/stem cell

Authors: Ewelina Wisnik, Zsolt Regdon, Kinga Chmielewska, Laszlo Virag, Agnieszka Robaszkiewicz

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Apart from protecting genome, PARP1 has been documented to regulate many intracellular processes inter alia gene transcription by physically interacting with chromatin bound proteins and by their ADP-ribosylation. Our recent findings indicate that expression of PARP1 decreases during the differentiation of human CD34+ hematopoietic stem cells to monocytes as a consequence of differentiation-associated cell growth arrest and formation of E2F4-RBL2-HDAC1-SWI/SNF repressive complex at the promoter of this gene. Since the RBL2 complexes repress genes in a E2F-dependent manner and are widespread in the genome in G0 arrested cells, we asked (a) if RBL2 directly contributes to defining monocyte phenotype and function by targeting gene promoters and (b) if RBL2 controls gene transcription indirectly by repressing PARP1. For identification of genes controlled by RBL2 and/or PARP1,we used primer libraries for surface receptors and TLR signaling mediators, genes were silenced by siRNA or shRNA, analysis of gene promoter occupation by selected proteins was carried out by ChIP-qPCR, while statistical analysis in GraphPad Prism 5 and STATISTICA, ChIP-Seq data were analysed in Galaxy 2.5.0.0. On the list of 28 genes regulated by RBL2, we identified only four solely repressed by RBL2-E2F4-HDAC1-BRM complex. Surprisingly, 24 out of 28 emerged genes controlled by RBL2 were co-regulated by PARP1 in six different manners. In one mode of RBL2/PARP1 co-operation, represented by MAP2K6 and MAPK3, PARP1 was found to associate with gene promoters upon RBL2 silencing, which was previously shown to restore PARP1 expression in monocytes. PARP1 effect on gene transcription was observed only in the presence of active EP300, which acetylated gene promoters and activated transcription. Further analysis revealed that PARP1 binding to MA2K6 and MAPK3 promoters enabled recruitment of EP300 in monocytes, while in proliferating cancer cell lines, which actively transcribe PARP1, this protein maintained EP300 at the promoters of MA2K6 and MAPK3. Genome-wide analysis revealed a similar distribution of PARP1 and EP300 around transcription start sites and the co-occupancy of some gene promoters by PARP1 and EP300 in cancer cells. Here, we described a new RBL2/PARP1/EP300 axis which controls gene transcription regardless of the cell type. In this model cell, cycle-dependent transcription of PARP1 regulates expression of some genes repressed by RBL2 upon cell cycle limitation. Thus, RBL2 may indirectly regulate transcription of some genes by controlling the expression of EP300-recruiting PARP1. Acknowledgement: This work was financed by Polish National Science Centre grants nr DEC-2013/11/D/NZ2/00033 and DEC-2015/19/N/NZ2/01735. L.V. is funded by the National Research, Development and Innovation Office grants GINOP-2.3.2-15-2016-00020 TUMORDNS, GINOP-2.3.2-15-2016-00048-STAYALIVE and OTKA K112336. AR is supported by Polish Ministry of Science and Higher Education 776/STYP/11/2016.

Keywords: retinoblastoma transcriptional co-repressor like 2 (RBL2), poly(ADP-ribose) polymerase 1 (PARP1), E1A binding protein p300 (EP300), monocytes

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Authors: Rukia Yosuf

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Diabetes is a national healthcare crisis related to both macrovascular and microvascular complications. We hypothesized that higher levels of physical activity are associated with lower total and visceral fat mass, lower systolic blood pressure, and increased insulin sensitivity. Participant inclusion criteria: 21-50 years old, BMI ≥ 30 kg/m2, hemoglobin A1C 5.7-6.4, fasting glucose 100-125 mg/dL, and HOMA IR ≥ 2.5. Exclusion criteria: history of diabetes, hypertension, HIV, renal disease, hearing loss, alcoholic intake over four drinks daily, use of organic nitrates or PDE5 inhibitors, and decreased cardiac function. Total physical activity was measured using accelerometers, body composition using DXA, and insulin resistance via fsIVGTT. Clinical and biochemical cardiometabolic risk factors, blood pressure and heart rate were obtained using a calibrated sphygmomanometer. Anthropometric measures, fasting glucose, insulin, lipid profile, C-reactive protein, and BMP were analyzed using standard procedures. Within our study, we found correlations between levels of physical activity in a heterogeneous group of prediabetic adults. Patients with more physical activity had a higher degree of insulin sensitivity, lower blood pressure, total visceral adipose tissue, and overall lower total mass. Total physical activity levels showed small, but significant correlations with systolic blood pressure, visceral fat, lean mass and insulin sensitivity. After normalizing for the race, age, and gender using multiple regression, these associations were no longer significant considering our small sample size. More research into prediabetes will decrease the population of diabetics overall. In the future, we could increase sample size and conduct cross sectional and longitudinal studies in various populations with prediabetes.

Keywords: diabetes, kidney disease, nephrology, prediabetes

Procedia PDF Downloads 185

Authors: Tanima Chatterjee, Maitree Bhattacharyya

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The global challenge of type 2 diabetes mellitus is a major health concern in this millennium, and researchers are continuously exploring new targets to develop a novel therapeutic strategy. Type 2 diabetes mellitus (T2DM) is often coupled with dyslipidemia increasing the risks for cardiovascular (CVD) complications. Enhanced oxidative and nitrosative stresses appear to be the major risk factors underlying insulin resistance, dyslipidemia, β-cell dysfunction, and T2DM pathogenesis. Autophagy emerges to be a promising defense mechanism against stress-mediated cell damage regulating tissue homeostasis, cellular quality control, and energy production, promoting cell survival. In this study, we have attempted to explore the pivotal role of autophagy in T2DM subjects with or without dyslipidemia in peripheral blood mononuclear cells and insulin-resistant HepG2 cells utilizing flow cytometric platform, confocal microscopy, and molecular biology techniques like western blotting, immunofluorescence, and real-time polymerase chain reaction. In the case of T2DM with dyslipidemia higher population of autophagy, positive cells were detected compared to patients with the only T2DM, which might have resulted due to higher stress. Autophagy was observed to be triggered both by oxidative and nitrosative stress revealing a novel finding of our research. LC3 puncta was observed in peripheral blood mononuclear cells and periphery of HepG2 cells in the case of the diabetic and diabetic-dyslipidemic conditions. Increased expression of ATG5, LC3B, and Beclin supports the autophagic pathway in both PBMC and insulin-resistant Hep G2 cells. Upon blocking autophagy by 3-methyl adenine (3MA), the apoptotic cell population increased significantly, as observed by caspase‐3 cleavage and reduced expression of Bcl2. Autophagy has also been evidenced to control oxidative stress-mediated up-regulation of inflammatory markers like IL-6 and TNF-α. To conclude, this study elucidates autophagy to play a protective role in the case of diabetes mellitus with dyslipidemia. In the present scenario, this study demands to have a significant impact on developing a new therapeutic strategy for diabetic dyslipidemic subjects by enhancing autophagic activity.

Keywords: autophagy, apoptosis, dyslipidemia, reactive oxygen species, reactive nitrogen species, Type 2 diabetes

Procedia PDF Downloads 127

Authors: M. Ravindiran, P. Shankar

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This paper deals with the comparison of two synthesis methods, namely, sol-gel, and combustion to prepare Fe doped ZnO nano material. Characterization results for structural, optical and magnetic properties were analyzed for the sol gel and combustion synthesis derived materials. Magnetic studies of the prepared compounds reveal that the combustion synthesis derived material has good magnetization of 50 emu/gm with a better hysteresis loop curve.

Keywords: DMS, combustion, ferromagnetic, synthesis methods

Procedia PDF Downloads 423

Authors: Thanunthon Bowornsakulwong, Charukorn Charukarn, Franck Courtes, Panit Kitsubun, Lalintip Horcharoen

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Vero cell is a continuous cell line that is widely used for the production of viral vaccines. However, due to its adherent characteristic, scaling up strategy in large-scale production remains complicated and thus limited. Consequently, suspension-like Vero cell culture processes based on microcarriers have been introduced and employed while also providing increased surface area per volume unit. However, harvesting Vero cells from microcarriers is a huge challenge due to difficulties in cells detaching, lower recovery yield, time-consuming and dissociation agent carry-over. To overcome these problems, we developed a dissociation-association platform technology for detaching and re-attaching cells during subculturing from microcarriers to microcarriers, which will be conveniently applied to seed trains strategies in large scale bioreactors. Herein, Hillex-2 was used to culture Vero cells in serum-containing media using spinner flasks as a scale-down model. The overall confluency of cells on microcarriers was observed using inverted microscope, and the sample cells were daily detached in order to obtain the kinetics data. The metabolites consumption and by-products formation were determined by Nova Biomedical BioprofileFlex.

Keywords: dissociation-reassociation, microcarrier, scale up, Vero cell

Procedia PDF Downloads 130

Authors: Xin Chen, Xinyong Li, Qidong Zhao, Dong Wang

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A new coupling system was constructed by combining photo-electrochemical cell with electro-fenton cell (PEC-EF). The electrode material in this system was derived from MnyFe₁₋yCo Prussian-Blue-Analog (PBA). Mn₀.₄Fe₀.₆Co₀.₆₇-N@C spin-coated on carbon paper behaved as the gas diffusion cathode and Mn₀.₄Fe₀.₆Co₀.₆₇O₂.₂ spin-coated on fluorine-tin oxide glass (FTO) as anode. The two separated cells could degrade Sulfamethoxazole (SMX) simultaneously and some coupling mechanisms by PEC and EF enhancing the degradation efficiency were investigated. The continuous on-site generation of H₂O₂ at cathode through an oxygen reduction reaction (ORR) was realized over rotating ring-disk electrode (RRDE). The electron transfer number (n) of the ORR with Mn₀.₄Fe₀.₆Co₀.₆₇-N@C was 2.5 in the selected potential and pH range. The photo-electrochemical properties of Mn₀.₄Fe₀.₆Co₀.₆₇O₂.₂ were systematically studied, which displayed good response towards visible light. The photoinduced electrons at anode can transfer to cathode for further use. Efficient photo-electro-catalytic performance was observed in degrading SMX. Almost 100% SMX removal was achieved in 120 min. This work not only provided a highly effective technique for antibiotic treatment but also revealed the synergic effect between PEC and EF.

Keywords: electro-fenton, photo-electrochemical, synergic effect, sulfamethoxazole

Procedia PDF Downloads 178

Authors: Subhash Reddy

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Mobile communication in today’s world means a lot to the human kind, through this many deals are made and others are broken, within seconds. That is because of our sophisticated methods of transporting the data at very high speeds and to very long distances, within no time. That is also because we kept on changing the method of serving the connections as the no of connections kept on increasing, that has led to many methods like TDMA, CDMA, and FDMA, etc. in wireless communications. And also the areas, where the connections are provided are also divided into CELLS, which are the basic blocks for cellular communications. Along with the wireless network, providing a wired network in mobile phones would serve as a very good alternative and would divert the extra traffic of a cell, so that a CELL which is providing wireless network can operate more efficiently.

Keywords: CDMA, FDMA, TDMA, CELL

Procedia PDF Downloads 483

Authors: S. A. Nirmal, J. M. Ingale, G. S. Asane, S. C. Pal, Subhash C. Mandal

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The present work was undertaken to determine the effects of Amaranthus roxburghianus Nevski. (Amaranthaceae) root alone and in combination with piperine in treating ulcerative colitis (UC) in mice. Swiss albino mice were divided into seven groups (n = 6). Standard group received prednisolone (5 mg/kg, i.p.). Treatment groups received hydroalcoholic extract of roots of A. roxburghianus (50 and 100 mg/kg, p.o.) and a combination of hydroalcoholic extract of roots of A. roxburghianus (50 and 100 mg/kg, p.o.) and piperine (5 mg/kg, p.o.). Ulcer index, colitis severity, myeloperoxidase (MPO), malondialdehyde and glutathione were estimated from blood and tissue. Column chromatography of the extract was done and purified fractions were analyzed by gas chromatography-mass spectroscopy (GC-MS). Treatment with the combination of hydroalcoholic extract of A. roxburghianus and piperine showed minimal ulceration, hemorrhage, necrosis and leucocyte infiltration by histopathological observation. Acetic acid increased MPO levels in blood and colon tissue to 355 U/mL and 385 U/mg, respectively. The combination of hydroalcoholic extract (100 mg/kg) and piperine (5 mg/kg) significantly decreased MPO in blood and tissue to 182 U/mL and 193 U/mg, respectively. Similarly, this combination significantly reduced MPO and increased glutathione levels in blood and tissue. Various phytoconstituents were detected by GC-MS. The combination of hydroalcoholic extract and piperine is effective in the treatment of UC and the effects are comparable with the standard drug prednisolone. 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, eugenol and benzene, and 1-(1,5-dimethyl-4-hexenyl)-4-methyl are reported having analgesic, anti-inflammatory, and antioxidant properties; they may play a role in the biological activity of A. roxburghianus root.

Keywords: Amaranthus roxburghianus, ulcerative colitis, anti-inflammatory, ulcerative colitis

Procedia PDF Downloads 521

Authors: S. Limpattayanate, M. Hunsom

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A comparison of activity and stability of the as-formed Pt/C, Pt-Co, and Pt-Pd/C electrocatalysts, prepared by a combined approach of impregnation and seeding, was performed. According to the activity test in a single proton exchange membrane (PEM) fuel cell, the oxygen reduction reaction (ORR) activity of the Pt-M/C electro catalyst was slightly lower than that of Pt/C. The j0.9 V and E10 mA/cm2 of the as-prepared electrocatalysts increased in the order of Pt/C>Pt-Co/C>Pt-Pd/C. However, in the medium-to-high current density region, Pt-Pd/C exhibited the best performance. With regard to their stability in a 0.5 M H2SO4 electrolyte solution, the electro chemical surface area decreased as the number of rounds of repetitive potential cycling increased due to the dissolution of the metals within the catalyst structure. For long-term measurement, Pt-Pd/C was the most stable than the other three electrocatalysts.

Keywords: ORR activity, stability, Pt-based electrocatalysts, PEM fuel cell

Procedia PDF Downloads 440

Authors: Fethi Benyettou, Abdelkader Aissat, M. A. Benammar

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In this work, we use Silvaco TCAD software for modeling and simulations of standard GaAs solar cell, InAs/GaAs and GaSb/GaAs p-i-n quantum dot solar cell. When comparing 20-layer InAs/GaAs, GaSb/GaAs quantum dots solar cells with standard GaAs solar cell, the conversion efficiency in simulation results increased from 16.48 % to 22.6% and 16.48% to 22.42% respectively. Also, the absorption range edge of photons with low energies extended from 900 nm to 1200 nm.

Keywords: SILVACO TCAD, the quantum dot, simulation, materials engineering

Procedia PDF Downloads 486

Authors: Mohamed M. Bumadian, D. James Gilmour

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Three bacterial species were isolated from the River Wye (Derbyshire, England) and identified using 16S rRNA gene sequencing as Serratia proteamaculans, Aeromonas hydrophila and Klebsiella pneumoniae. Respiration rates of the strains were measured in order to determine the metabolic activity under salt stress. The highest respiration rates of all three strains were found at 0.17 M and 0.5 M NaCl and then the respiration rate decreased with increasing concentrations of NaCl. In addition, the effect of increasing concentrations of NaCl on malate dehydrogenase activity was determined using cell-free extracts of the three strains. Malate dehydrogenase activity was stimulated at NaCl concentrations up to 0.5 M, and a small level of activity remained even at 3.5 M NaCl. The pH optimum of the malate dehydrogenase in cell-free extracts of all strains was higher than pH 7.5.

Keywords: fresh water, halotolerant pathogenic bacteria, 16S rRNA gene, cell-free extracts, respiration rates, malate dehydrogenase

Procedia PDF Downloads 457

Authors: Mohanad Alhabo, Naveed Nawaz

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The ongoing call or data session must be maintained to ensure a good quality of service. This can be accomplished by performing the handover procedure while the user is on the move. However, the dense deployment of small cells in 5G networks is a challenging issue due to the extensive number of handovers. In this paper, a neighbour cell list method is proposed to reduce the number of target small cells and hence minimizing the number of handovers. The neighbour cell list is built by omitting cells that could cause an unnecessary handover and handover failure because of short time of stay of the user in these cells. A multi-attribute decision making technique, simple additive weighting, is then applied to the optimized neighbour cell list. Multi-tier small cells network is considered in this work. The performance of the proposed method is analysed and compared with that of the existing methods. Results disclose that our method has decreased the candidate small cell list, unnecessary handovers, handover failure, and short time of stay cells compared to the competitive method.

Keywords: handover, HetNets, multi-attribute decision making, small cells

Procedia PDF Downloads 117

Authors: Sana Rezig, Yosr Ben Mlik, Mounir Jaouadi, Foued Khoffi, Slah Msahli, Bernard Durand

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Increasing interest in environmental concerns, natural fibers are once again being considered as reinforcements for polymer composites. The main objective of this study is to explore another natural resource, Typha fiber; which is renewable without production cost and available abundantly in nature. The aim of this study was to study the flexural properties of composite resin with and without reinforcing Typha leaf and stem fibers. The specimens were made by the hand-lay-up process using polyester matrix. In our work, we focused on the effect of various treatment conditions (sea water, alkali treatment and a combination of the two treatments), as a surface modifier, on the flexural properties of the Typha fibers reinforced polyester composites. Moreover, weight ratio of Typha leaf or stem fibers was investigated. Besides, both fibers from leaf and stem of Typha plant were used to evaluate the reinforcing effect. Another parameter, which is reinforcement structure, was investigated. In fact, a first composite was made with air-laid nonwoven structure of fibers. A second composite was with a mixture of fibers and resin for each kind of treatment. Results show that alkali treatment and combined process provided better mechanical properties of composites in comparison with fiber treated by sea water. The fiber weight ratio influenced the flexural properties of composites. Indeed, a maximum value of flexural strength of 69.8 and 62,32 MPa with flexural modulus of 6.16 and 6.34 GPawas observed respectively for composite reinforced with leaf and stem fibers for 12.6 % fiber weight ratio. For the different treatments carried out, the treatment using caustic soda, whether alone or after retting seawater, show the best results because it improves adhesion between the polyester matrix and the fibers of reinforcement. SEM photographs were made to ascertain the effects of the surface treatment of the fibers. By varying the structure of the fibers of Typha, the reinforcement used in bulk shows more effective results as that used in the non-woven structure. In addition, flexural strength rises with about (65.32 %) in the case of composite reinforced with a mixture of 12.6% leaf fibers and (27.45 %) in the case of a composite reinforced with a nonwoven structure of 12.6 % of leaf fibers. Thus, to better evaluate the effect of the fiber origin, the reinforcing structure, the processing performed and the reinforcement factor on the performance of composite materials, a statistical study was performed using Minitab. Thus, ANOVA was used, and the patterns of the main effects of these parameters and interaction between them were established. Statistical analysis, the fiber treatment and reinforcement structure seem to be the most significant parameters.

Keywords: flexural properties, fiber treatment, structure and weight ratio, SEM photographs, Typha leaf and stem fibers

Procedia PDF Downloads 412

Authors: Ceyda Okudu, Secil Eroglu, Khandakar A. S. M. Saadat, Sibel O. Balci

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Ring Finger Protein 2 (RNF2) is a member of polycomb repressive complex 1 (PRC1), which is one of the epigenetic regulators in the genome. When RNF2 combines with other PRC1 members, it mediates the mono-ubiquitination of Histon2A (H2A). In breast cancer, RNF2 is commonly overexpressed, and also it promotes metastasis and invasion in other aggressive tumors like melanoma, prostate, and hepatocarcinoma. The role of RNF2 in the metastasis and invasion of breast cancer has not yet been elucidated. Our aim is to observe the role of RNF2 in metastasis and invasion in this study by miRNA mediated RNF2 gene silencing in breast cancer cell lines. We selected miRNAs, targeting to RNF2 by searching online databases. miR-17-5p, miR20a-5p, and miR-106b-5p were transfected to breast cancer cell lines (MCF-7, MDA-MB-231, SK-BR-3, and ZR-75-1), and also we used normal breast epithelial cell line (hTERT-HME1) to compare RNF2 gene expression level. After 48-72 hours post-transfection, mRNAs were isolated from the cells, and gene expressions were measured by RT-qPCR after from cDNA syntheses. We observed that RNF2 was highly expressed in SK-BR-3 and MDA-MB-231 cell lines opposite to MCF-7 and ZR-75-1 cell lines. RNF2 was downregulated 5, 5 and 7 fold by miR17-5p, miR20a-5p and miR106b-5p respectively in MCF-7. However, in SK-BR-3 and ZR-75-1 cell lines, miRNAs did not affect significantly RNF2 gene expression level. miR20a-5p decreased RNF2 3 fold and miR17-5p and miR106b-5p did not affect MDA-MB-231. After gene expression analysis, we performed metastasis and invasion assay in MCF-7 cells. For metastasis, we used both wound healing assay and Transwell Cell Migration Assay, and we used Transwell Cell Invasion Assay for invasion. The data of this assay showed that miR17-5p and miR20a-5p decreased both invasion and metastasis level, but miR106b-5p has no effect. We would like to conclude that RNF2 can be targeted by miR17-5p, miR20a-5p and miR106b-5p in MCF-7 cells and also RNF2, which is one of the upregulated genes in aggressive tumor, can be decreased by using these miRNAs. In future, we would like to confirm these results at the protein level and also whether these miRNAs are direct target of RNF2 or not.

Keywords: breast cancer, epigenetic, microRNAs, RNF2

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Authors: Jehad Dumireih, Malak Dmirieh, Michael Wink

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The present work is aimed to use plant cell suspension cultures of Crataegus monogyna for biosynthesis of valuable natural products by using quercetin as an inexpensive precursor. Suspension cell cultures of C. monogyna were established by using Murashige and Skoog medium (MS) supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L kinetin. Cells were harvested from the cultures and extracted by using methanol and ethyl acetate; then the extracts were used for the identification of isoquercetin by HPLC and by mass spectrometry. The incubation of the cells with 0.24 mM quercetin for one week resulted in an 16 fold increase of isoquercetin biosynthesis; the growth rate of the cells increased by 20%. Moreover, the biosynthesis of isoquercetin was enhanced by 40% when we divided the added quercetin into three portions each one with concentration 0.12 mM supplied at 3 days intervals. In addition, we didn’t find any positive effects of adding different concentrations the precursors phenylalanine (0.2 mM) and galactose to the cell cultures. In conclusion, the efficiency of the biotransformation of quercetin into isoquercetin depended on the concentration quercetin, its incubation time and the way of its administration. The results of the present work suggest that the biotechnological methods such as cell suspension cultures could be successfully used to obtain highly valuable natural product starting from inexpensive compound.

Keywords: biosynthesis, biotransformation, Crataegus, isoquercetin

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Authors: P. S. BEDI* , Neavty Thakur, Balvınder Sıngh

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In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3, NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria eg. Bacillus subtilis, Gram negative bacteria eg. E. coli, K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant posseses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

Keywords: Girardinia heterophylla, leaves and stem extracts, Antibacterial activity, antifungal activity.

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Authors: Dimakatso B. Gumede, Nicolette N. Houreld

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Diabetic foot ulcers (DFUs) are a complication of diabetes mellitus (DM), a metabolic disease caused by insulin resistance or insufficiency, resulting in hyperglycaemia and low-grade chronic inflammation. Current therapies for treating DFUs include wound debridement, glycaemic control, and wound dressing. However, these therapies are moderately effective as there is a recurrence of these ulcers and an increased risk of lower limb amputations. Photobiomodulation (PBM), which is the application of non-invasive low-level light for wound healing at the spectrum of 660-1000 nm, has shown great promise in accelerating the healing of chronic wounds. However, its underlying mechanisms are not clearly defined. Studies have indicated that PBM induces wound healing via the activation of signaling pathways that are involved in tissue repair, such as the transforming growth factor-β (TGF-β). However, other signaling pathways, such as the WNT/β-catenin pathway, which is also critical for wound repair, have not been investigated. This study aimed to elucidate if PBM at 660 nm and a fluence of 5 J/cm² activates the WNT/β-catenin signaling pathway for wound healing in a diabetic cellular model. Human dermal fibroblasts (WS1) were continuously cultured high-glucose (26.5 mM D-glucose) environment to create a diabetic cellular model. A central scratch was created in the diabetic model to ‘wound’ the cells. The diabetic wounded (DW) cells were thereafter irradiated at 660 nm and a fluence of 5 J/cm². Cell migration, gene expression and protein assays were conducted at 24- and 48-h post-PBM. The results showed that PBM at 660 nm and a fluence of 5 J/cm² significantly increased cell migration in diabetic wounded cells at 24-h post-PBM. The expression of CTNNB1, ACTA2, COL1A1 and COL3A1 genes was also increased in DW cells post-PBM. Furthermore, there was increased cytoplasmic accumulation and nuclear localization of β-catenin at 24 h post-PBM. The findings in this study demonstrate that PBM activates the WNT/β-catenin signaling pathway by inducing the accumulation of β-catenin in diabetic wounded cells, leading to increased cell migration and expression of wound repair markers. These results thus indicate that PBM has the potential to improve wound healing in diabetic ulcers via activation of the WNT/β-catenin signaling pathway.

Keywords: wound healing, diabetic ulcers, photobiomodulation, WNT/β-catenin, signalling pathway

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Authors: Chao Wu

Abstract:

Thioredoxin reductase 2 is a mitochondrial enzyme that belongs to the cellular defense against oxidative stress. We deleted mitochondrial Txnrd2 in a KrasG12D-driven pancreatic tumor model. Despite an initial increase in precursor lesions, tumor incidence decreased significantly. We isolated cancer cell lines from these genetically engineered mice and observed an impaired proliferation and colony formation. Reactive Oxygen Species, as determined by DCF fluorescence, were increased. We detected a higher mitochondrial copy number in Txnrd2-deficient cells (KTP). However, measurement of mitochondrial bioenergetics showed no impairment of mitochondrial function and comparable O₂-consumption and extracellular acidification rates. In addition, the mitochondrial complex composition was affected in Txnrd2 deleted cell lines. To gain better insight into the role of Txnrd2, we deleted Txnrd2 in clones from parental KrasG12D cell lines using Crispr/Cas9 technology. The deletion was confirmed by western blot and activity assay. Interestingly, and in line with previous RNA expression analysis, we saw changes in EMT markers in Txnrd2 deleted cell lines and control cell lines. This might help us explain the reduced tumor incidence in KrasG12D; Txnrd2∆panc mice.

Keywords: PDAC, TXNRD2, epithelial-to-mesenchymal-transition, ROS

Procedia PDF Downloads 114

Authors: Pei-Jung Wu, Ching-Ying Huang, Chih-Chia Lin, Chun-Han Li, Chien-Yuan Wang

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The fuel cell market has considerable development potential, but the cost is still less competitive. Replacing the traditional graphite plate with a stainless steel plate as a bipolar plate can greatly reduce the weight and volume of the stack, and has more cost advantages. However, the passivation layer on the surface of stainless steel makes the contact resistance reach the ohmic level and reduces the performance of the fuel cell. Therefore, it is necessary to reduce the interfacial contact resistance through the surface treatment. In this research, the thickness, uniformity, interfacial contact resistance (ICR), and adhesion of the carbon-based layer was analyzed. On the other hand, the effect of coating properties on the performance of the fuel cell was verified through I-V tests. The results show that after coating the contact resistance is greatly reduced by three stages to the microohm level, and as the film thickness is reduced, the contact resistance is reduced from 229~118 mΩ-cm² to 135~73 mΩ-cm² at a general assembly pressure of 1 to 2 MPa., and the current density at 0.6 V increased from 485.7 mA/cm² to 575.7 mA/cm². This study verifies the importance of the uniformity and ICR of the coating on proton exchange membrane fuel cell (PEMFC), and the surface coating technology is the key to affecting the characteristics of the coating.

Keywords: contact resistance, proton exchange membrane fuel cell, PEMFC, SS bipolar plate, spray coating process

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Authors: Sushil Pradhan

Abstract:

Biotechnology contributes to sustainable development in several ways such as biofertilizer production, biopesticide production and management of environmental pollution, tissue culture and biodiversity conservation in vitro, in vivo and in situ, Insectivorous medicinal plant Drosera burmannii Vahl belongs to the Family-Droseraceae under Order-Caryophyllales, Dicotyledoneae, Angiospermeae which has 31 (thirty one) living genera and 194 species besides 7 (seven) extinct (fossil) genera. Locally it is known as “Patkanduri” in Odia. Its Hindi name is “Mukhajali” and its English name is “Sundew”. The earliest species of Drosera was first reported in 1753 by Carolous Linnaeus called Drosera indica L (Indian Sundew). The latest species of Drosera reported by Fleisch A, Robinson, AS, McPherson S, Heinrich V, Gironella E and Madulida D.A. (2011) is Drosera ultramafica from Malaysia. More than 50 % species of Drosera have been reported from Australia and next to Australia is South Africa. India harbours only 3 species such as D. indica L, Drosera burmannii Vahl and D. peltata L. From our Odisha only D. burmannii Vahl is being reported for the first time from the district of Subarnapur near Sonepur (Arjunpur Reserve Forest Area). Drosera plant is autotrophic but to supplement its Nitrogen (N2) requirement it adopts heterotrophic mode of nutrition (insectivorous/carnivorous) as well. The colour of plant in mostly red and about 20-30cm in height with beautiful pink or white pentamerous flowers. Plants grow luxuriantly during November to February in shady and moist places near small water bodies of running water stream. Medicinally it is a popular herb in the locality for the treatment of cold and cough in children in rainy season by the local Doctors (Kabiraj and Baidya). In the present field investigation an attempt has been made to understand the unique reproductive phase and life cycle of the plant thereby planning for its conservation and propagation through various techniques of tissue culture and biotechnology. More importantly besides morphological and anatomical studies, cytological investigation is being carried out to find out the number of chromosomes in the cell and its genomics as there is no such report as yet for Drosera burmannii Vahl. The ecological significance and biodiversity conservation of Drosera with special reference to energy, environmental and chemical engineering has been discussed in the research paper presentation.

Keywords: insectivorous, medicinal, drosera, biotechnology, chromosome, genome

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Authors: Jae Young Song, In-Ohk Ouh, Seyeon Park, Byeong Sul Kang, Soo Dong Cho, In-Soo Cho

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Canine parvovirus (CPV) is a major pathogen of diarrhea disease in dogs. CPV type 2 has three of antigenic variants such as 2a, 2b, and 2c. CPV constructs a small non-enveloped, icosahedral capsid that contains single-stranded DNA. It has capsids that two largely overlapping virion proteins (VP), VP1 (82 kDa), and VP2 (65 kDa). Baculoviruses are insect pathogens that regulate insect populations in nature and are being successfully used to control insect pests. The proteins produced in the baculovirus-expression system are used for instance for functional studies, vaccine preparations, or diagnostics. The vaccines produced by baculovirus-expression system showed elicitation of antibodies. The recombinant baculovirus infected SF9 cells showed broken shape. The recombinant VP2 proteins from cell pellet or supernatant were confirmed by western blotting. The result showed that the recombinant VP2 protein bands were appeared at 65 kDa molecular weight in both cell pellet and supernatant of infected SF9 cell. These results indicated that the recombinant baculovirus infected SF9 cell express the recombinant VP2 protein successfully. In addition, the expressed recombinant VP2 protein is secreted from cell to supernatant. The baculovirus expression system can be used to produce the VP2 protein of CPV 2c. In addition, the secretion property of the expression of VP2 protein may decrease the cost of production, because it can be skipped the cell breaking step. The produced VP2 protein could be used for vaccine and the agent of diagnostic tests. This study provides the foundation of the production of CPV 2c vaccine and the diagnostic agent.

Keywords: baculovirus, canine parvovirus 2c, dog, Korea

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Authors: Mustafa Al-Yaseen, Haider Mohammed Mahdi, Haider Ali Al–Zahid, Nazar S. Haddad

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Background: Early diagnosis of musculoskeletal infections is of vital importance to avoid devastating complications. There is no single laboratory marker which is sensitive and specific in diagnosing these infections accurately. White blood cell count, erythrocyte sedimentation rate, and C-reactive protein are not specific as they can also be elevated in conditions other than bacterial infections. Materials Culture and sensitivity is not a true gold standard due to its varied positivity rates. Serum Procalcitonin is one of the new laboratory markers for pyogenic infections. The objective of this study is to assess the value of PCT in the diagnosis of soft tissue, bone, and joint infections. Patients and Methods: Patients of all age groups (seventy-four patients) with a diagnosis of musculoskeletal infection are prospectively included in this study. All patients were subjected to White blood cell count, erythrocyte sedimentation rate, C-reactive protein, and serum Procalcitonin measurements. A healthy non infected outpatient group (twenty-two patients) taken as a control group and underwent the same evaluation steps as the study group. Results: The study group showed mean Procalcitonin levels of 1.3 ng/ml. Procalcitonin, at 0.5 ng/ml, was (42.6%) sensitive and (95.5%) specific in diagnosing of musculoskeletal infections with (positive predictive value of 87.5% and negative predictive value of 48.3%) and (positive likelihood ratio of 9.3 and negative likelihood ratio of 0.6). Conclusion: Serum Procalcitonin, at a cut – off of 0.5 ng/ml, is a specific but not sensitive marker in the diagnosis of musculoskeletal infections, and it can be used effectively to rule in the diagnosis of infection but not to rule out it.

Keywords: procalcitonin, infection, labratory markers, musculoskeletal

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Authors: Duygu Yüksel Deniz, Memet Vezir Kahraman, Serap Erdem Kuruca, Mediha Süleymanoğlu

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Our first aim was to synthesize Hydroxy Apatite (HAP) and then modify its surface by adding 4-Vinylbenzene boronic acid (4-VBBA). The characterization was done by FT-IR. By adding Polyvinyl alcohol (PVA) to 4- VBBA-HAP, we obtained a suitable electrospinning solution. PVA solution which was also modified by using alkoxy silanes, in order to prevent the scaffolds from being damaged by aqueous cell medium, was added. Keratin was dissolved and then added into the electrospinning solution. Keratin containing 4-VBBA- HAP/PVA composite was used to fabricate nanofiber scaffolds with the simultaneous UV-reactive electrospinning technique. The structural characterization was done by FT-IR. Thermal gravimetric analysis was also performed by using TGA. The morphological characterization was determined by SEM analyses. Our second aim was to create a scaffold where cells could grow. With this purpose, suitable nanofibers were choosen according to their SEM analysis. Keratin containing nanofibers were seeded with 3T3, ECV and SAOS cells and their cytotoxicity and cell proliferation were investigated by using MTT assay. After cell culturing process morphological characterization was determined by SEM analyses. These scaffolds were designed to be nontoxic biomaterials. Here, a comparision was made between keratin containing 3T3, ECV and SAOS seeded nanofiber scaffolds and the results were presented and discussed.

Keywords: cell culture, keratin, nanofibers, UV-reactive electrospinning

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Authors: Mashitoh Abd Rahman, Najihah Mohd Hashim, Faiqah Ramli, Syam Mohan, Noraziah Nordin, Hamed Karimian, Hapipah Mohd Ali

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Ovarian cancer is the most lethal gynecological malignancies. HME5, a prenylflavanoid has been isolated from local medicinal plant. This compound has been reported to possess a broad spectrum of biological activities including anticancer property. However, the potential of HME5 as an antiproliferative and cytotoxic agent on an ovarian cancer cells has not yet been investigated. In this present study, we examined the antiproliferative and cytotoxic effect of HME5 on Caov-3 (Human Ovarian Adenocarcinoma) cell line by using 3-[4,5-dimethylthizol-2-y]-2,5-diphenyltetrazolium bromide (MTT) assay, Acridine orange and propidium Iodide (AOPi) and cell cycle analysis study. HME5 has shown to inhibit Caov-3 in a time-dependent manner with the IC50 values of 5µg/ml, 2µg/ml and 1µg/ml after 24h, 48h and 72h treatment, respectively. Morphological study from AOPi analysis showed that HME5 induced apoptosis after 24 and 48h post-treatment. Nevertheless, HME5 exhibited cell cycle arrest at G1 phase as indicated in flow cytometry cell cycle profiling. In conclusion, HME5 inhibited proliferation of Caov-3 through induction of apoptosis and cell cycle arrest at G1 phase.

Keywords: apoptosis, prenylflavanoid, ovarian cancer, HME5

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Authors: Sana Ilyas

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Parry-Romberg syndrome is a rare condition clinically defined by slowly progressive atrophy of the skin and soft tissues. This usually effects one side of the face, although a few cases have been documented of bilateral presentation. It is more prevalent in females and usually affects the left side of the face. The syndrome can also be accompanied by neurological abnormalities. It usually occurs in the first two decades of life with a variable rate of progression. The aetiology is unknown, and the disease eventually stabilises. The treatment options usually involve surgical management. The least invasive of these options is the management of facial asymmetry, associated with Parry Romberg syndrome, through the use of tissue fillers. This paper will review the existing literature on the management of Parry Romberg syndrome with tissue filler. Aim: The aim of the study is to explore the current published literature for the management of Parry Romberg syndrome with fillers. It is to assess the development that has been made in this method of management, its benefits and limitations, and its effectiveness for the management of Parry Romberg syndrome. Methodology: There was a thorough assessment of the current literature published on this topic. PubMed database was used for search of the published literature on this method of the management. Papers were analysed and compared with one another to assess the success and limitation of the management of Parry Romberg with dermal fillers Results and Conclusion: Case reports of the use of tissue fillers discuss the varying degrees of success with the treatment. However, this procedure has it’s limitation, which are discussed in the paper in detail. However, it is still the least invasive of all the surgical options for the management of Parry Romberg Syndrome, and therefore, it is important to explore this option with patients, as they may be more comfortable with pursuingtreatment that is less invasive and can still improve their facial asymmetry

Keywords: dermal fillers, facial asymmetry, parry romberg syndrome, tissue fillers

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Authors: Maryam Zahedifard, Fadhil Lafta Faraj, Nazia Abdul Majid, Hapipah Mohd Ali, Mahmood Ameen Abdulla

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The new quinazoline Schiff bases have been synthesized through condensation reaction of 2-aminobenzhydrazide with 5-bromosalicylaldehyde and 3-methoxy-5-bromosalicylaldehyde. The compound was investigated for anticancer activity against MCF-7 human breast cancer cell line. It demonstrated a remarkable antiproliferative effect, with an IC50 value of 3.41±0.34, after 72 hours of treatment. Most apoptosis morphological features in treated MCF-7 cells were observed by AO/PI staining. The results of cell cycle analysis indicate that compounds did not induce S and M phase arrest in cell after 24 hours of treatment. Furthermore, MCF-7 cells treated with compound subjected to apoptosis death, as exhibited by perturbation of mitochondrial membrane potential and cytochrome C release as well as increase in ROS generation. We also found activation of caspases 3/7 and -9. Moreover, acute toxicity results demonstrated the nontoxic nature of the compounds in mice. Our results showed the selected compound significantly induce apoptosis in MCF-7 cells via intrinsic pathway, which might be considered as a potential candidate for further in vivo and clinical breast cancer studies.

Keywords: quinazoline Schiff base, apoptosis, MCF-7, caspase, cell cycle, acute toxicity

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Authors: Solomon Tilahun Desisa, Muktar Seid Hussen

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Hibiscus macranthus is one of family Malvaceae and genus Hibiscus plant which grows mainly in western part of Ethiopia. Hibiscus macranthus is the most adaptable and abundant plant in the nation, which are used as an ornamental plant often a hedge or fence plant, and used as a firewood after harvesting the stem together with the bark, and used also as a fiber for trying different kinds of things by forming the rope. However, Hibiscus macranthus plant fibre has not been commercially exploited and extracted properly. This study of work describes the possibility of mechanical and retting methods of Hibiscus macranthus fibre extraction and characterization. Hibiscus macranthus fibre is a bast fibre which obtained naturally from the stem or stalks of the dicotyledonous plant since it is a natural cellulose plant fiber. And the fibre characterized by studying its physical and chemical properties. The physical characteristics were investigated as follows, including the length of 100-190mm, fineness of 1.0-1.2Tex, diameter under X100 microscopic view 16-21 microns, the moisture content of 12.46% and dry tenacity of 48-57cN/Tex along with breaking extension of 0.9-1.6%. Hibiscus macranthus fiber productivity was observed that 12-18% of the stem out of which more than 65% is primary long fibers. The fiber separation methods prove to decrease of non-cellulose ingredients in the order of mechanical, water and chemical methods. The color measurement also shows the raw Hibiscus macranthus fiber has a natural golden color according to YID1925 and paler look under both retting methods than mechanical separation. Finally, it is suggested that Hibiscus macranthus fibre can be used for manufacturing of natural and organic crop and coffee packages as well as super absorbent, fine and high tenacity textile products.

Keywords: Hibiscus macranthus, bast fiber, extraction, characterization

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Authors: Federico Cevoli

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Purinergic P2X7 receptors (P2X7R) are ligand-gated non-selective cation channels involved in several physiological and pathological processes. They are particularly promising pharmacological targets as they are present in an increasing number of different cells types. P2X7R activation is triggered following elevated concentrations of extracellular ATP, similarly to those observed in tissues injury, chronic inflammation and T-cell activation, as well as in the scrambling of phospholipids leading to membrane blebbing and apoptosis. Another hallmark of P2X7 is cell permeabilization, commonly known as “macropore” formation allowing the passage of nanometer-sized molecules up to 900Da. Recently, full-length P2X7 Cryo-EM structures revealed unique functional sites, including two cytoplasmic domains - the cytoplasmic "anchor" and "ballast". To date, the molecular units/complex by which P2X7R exerts its pathophysiological functions are unknown. Using custom-made cell-penetrating HIV-1 TAT peptides, we show for the first-time potential implications of P2X7 cytoplasmic anchor in the regulation of caspase3/7 activation as well as TNFα regulation.

Keywords: P2X7R, immunology, TAT-peptide, cell death

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Authors: Amit Kumar Baghel, Sisir Kumar Nayak

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The proposed metamaterial design help to increase the radiation efficiency at 2.9 GHz by reducing the side and back lobes by making the phase difference of the waves emerging from the phase center of the horn antenna same after passing through metamaterial array. The unit cell of the metamaterial is having concentric ring structure made of copper of 0.035 mm thickness on both sides of FR4 sheet. The inner ring diameter is kept as 3 mm, and the outer ring diameters are changed according to the path and tramission phase difference of the unit cell from the phase center of the antenna in both the horizontal and vertical direction, i.e., in x- and y-axis. In this case, the ring radius varies from 3.19 mm to 6.99 mm with the respective S21 phase difference of -62.25° to -124.64°. The total phase difference can be calculated by adding the path difference of the respective unit cell in the array to the phase difference of S21. Taking one of the unit cell as the reference, the total phase difference between the reference unit cell and other cells must be integer multiple of 360°. The variation of transmission coefficient S21 with the ring radius is greater than -6 dB. The array having 5 x 5 unit cell is kept inside the pyramidal horn antenna (L X B X H = 295.451 x 384.233 x 298.66 mm3) at a distance of 36.68 mm from the waveguide throat. There is an improvement in side lobe level in E-plane by 14.6 dB when the array is used. The front to back lobe ration is increased by 1 dB by using the array. The proposed antenna with metamaterial array can be used in beam shaping for wireless power transfer applications.

Keywords: metamaterial, side lobe level, front to back ratio, beam forming

Procedia PDF Downloads 261

Authors: Marta E. Hernandez-Caballero, Veronica Borgonio-Cuadra, Antonio Miranda-Duarte, Xochitl Rojas-Toledo, Normand Garcia-Hernandez, Maura Cardenas-Garcia, Teresa Abad-Camacho

Abstract:

Breast cancer (BC) is the most common tumor in women over the world. DNA methylation is an epigenetic modification critical in CpG sites, aberrant methylation of CpG islands in promoters is a hallmark of cancer. So, gene expression can be regulated by alterations in DNA methylation. In cell lines DACT2 gene reduces the growth and migration of tumor cells by its participation in the suppression of TGFb/SMAD2/3. PHF20L1 is involved in histone acetylation therefore, it regulates transcription. Our aim was to analyze the methylation status of the DACT2 and PHF20L1 promoter regions in tumoral and healthy mammary tissue from women with BC in different progression states. The study included 77 patients from Centro Medico Nacional La Raza in Mexico City. After identifying a CpG island in DACT2 and PHF20L1 promoters, DNA methylation status was analyzed through sodium bisulfite with subsequent amplification using methylation-specific PCR. Results revealed no changes in methylation status of PHF20L1 and cancer stages (II y III) or in comparison to healthy tissues, it was demethylated. DACT2 promoter methylation was no significant between tumoral stages (II, P = 0.37; III, P = 0.17) or with healthy tissue. Previous data reported DACT2 methylated in nasopharyngeal carcinoma but in this study promoter methylation was not observed. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains, it has been suggested that can stabilize DNMT1 regulating DNA methylation, therefore, was associated with poor prognostic in BC. We found no evidence of methylation in patients and controls in PHF20L1 promoter, so its association with BC may have no direct relation with promoter methylation. More studies including other methylation sites in these genes in BC are necessary.

Keywords: bisulfite conversion, breast cancer, DACT2, DNA methylation, PHF20L1, tumoral status

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