Search results for: cell viability.

Authors: N. V. Enwuru, B. Nkeki, E. A. Adekoya, O. A. Adebesin, B. O. Ojo, R. F. Peters, V. A. Aikhomu, U. E. Mendie, O. Akinloye

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Awareness about probiotic health benefits is increasing tremendously. However, cell viability is often low due to harsh conditions exposed during processing, handling, storage, and gastrointestinal transit. Thus, encapsulation is a promising technique that increases cell viability. The study aims to encapsulate the probiotic, evaluate its viability and cytotoxic activity of its postbiotic on the Michigan Cancer Foundation (MCF)-7 breast cancer cell line. Human and animal raw milk was sampled for lactic acid bacteria. Isolated bacteria were identified using conventional and VITEK 2 systems. The identified bacteria were encapsulated using the spray-drying method. The free and encapsulated probiotic cells were exposed to simulated gastric intestinal (SGI) fluid conditions and different storage conditions for their viability. The properties of the formed probiotic granules, their disintegration time, and the weight uniformity of the microcapsules were tested. Furthermore, the postbiotic of the free cells was extracted, and its cytotoxic effect on the MCF-7 breast cancer cell line was tested through [3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide] (MTT) assay. The bacteria isolated were identified as Lactobacillus plantarum. The size of the formed probiotic granules ranges within 0.71-1.00 mm in diameter, and disintegration time ranges from 2.14 ± 0.045 to 2.91 ± 0.293 minutes, while the average weight is 502.1 mg. The viability of encapsulated cells stored at refrigerated condition (4oC) was higher than that of cells stored at room temperature (25 oC). The encapsulated probiotic cells exhibited better viability after exposure to SGI solution at different pH levels than free cells. The Postbiotic Metabolites (PM) of L. plantarum produced a cytotoxic effect that shows significant activity similar to 5FU, a standard antineoplastic agent. The inhibition concentration of 50% growth (IC50) of postbiotic metabolite was consistent with the IC50 of the positive control (Cisplatin). Lactobacillus plantarum postbiotic exhibited a cytotoxic effect on the MCF-7 breast cancer cell line and could be used as combined adjuvant therapy in breast cancer management. The microencapsulation technique protects the probiotics and maintains their viability.

Keywords: Cytotoxicity effect, encapsulation, postbiotic, probiotic.

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Authors: Kanchana Sitlaothaworn

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Yogurt capsule was made by mixing 14% w/v of reconstitution of skim milk with 2% FOS. The mixture was fermented by commercial yogurt starter comprising Lactobacillus bulgaricus and Streptococcus thermophilus. These yogurts were made as yogurt powder by freeze-dried. Yogurt powder was put into capsule then stored for 28 days at 4oc. 8ml of commercial yogurt was found to be the most suitable inoculum size in yogurt production. After freeze-dried, the viability of L. bulgaricus and S. thermophilus reduced from 109 to 107 cfu/g. The precence of sucrose cannot help to protect cell from ice crystal formation in freeze-dried process, high (20%) sucrose reduced L. bulgaricus and S. thermophilus growth during fermentation of yogurt. The addition of FOS had reduced slowly the viability of both L. bulgaricus and S. thermophilus similar to control (without FOS) during 28 days of capsule storage. The viable cell exhibited satisfactory viability level in capsule storage (6.7x106cfu/g) during 21 days at 4oC.

Keywords: Yogurt capsule, Lactobacillus bulgaricus, Streptococcus thermophilus, freeze-drying, sucrose.

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Authors: M. R. Ghalamboran, J. J. Ramsden

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The aim of this study was to investigate whether magnetite nanoparticles affect the viability of Bradyrhizobium japanicum cells residing on the surface of soybean seeds during desiccation. Different concentrations of nanoparticles suspended in liquid medium, mixed with and adhering to Bradyrhizobium japanicum, were investigated at two temperatures, using both soybean seeds and glass beads as surrogates. Statistical design was a complete randomized block (CRB) in a factorial 6×2×2×6 experimental arrangement with four replications. The most important variable was the viability of Bradyrhizobium on the surface of the seeds. The nanoparticles increased Bradyrhizobium viability and inoculated seeds stored at low temperature had greater viability when nanoparticles had been added. At the optimum nanoparticle concentration, 50% bacterium viability on the seeds was retained after 5 days at 4ºC. Possible explanations for the observed effects are proposed.

Keywords: Bradyrhizobium japanicum, magnetitenanoparticles, soybean seed, viability.

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Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

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The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: Berberine, bulls, motility, oxidative profile, spermatozoa, viability.

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Authors: S. Yodyingyong, C. Chaichana, C. Nuchsuk, S. Roytrakul, N. P. T-Thienprasert, S. Ratanapo

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Protein hydrolysates prepared from a number of medicinal plants are promising sources of various bioactive peptides. In this work, proteins from dried whole plant of Euphorbia hirta Linn. were extracted and digested with pepsin for 12h. The hydrolysates of lesser than 3 KDa were fractionated by a cut-off membrane. The peptide hydrolysate was then purified by an anion-exchange chromatography on DEAE-Sephacel™ column and reverse-phase chromatography on Sep-pak C₁₈ column, respectively. The cytotoxic effect of each peptide fraction against a gastric carcinoma cell line (KATO-III, ATCC No. HTB103) was investigated using colorimetric MTT viability assay. A human liver cell line (Chang Liver, CLS No. 300139) was used as a control normal cell line. Two purified peptide peaks, peak l and peak ll at 100µg peptides mL^-1 affected cell viability of the gastric cancer cell lines to 63.85±4.94 and 66.92±6.46%, respectively. Our result showed for the first time that the peptide fractions derived from protein hydrolysate of Euphorbia hirta Linn. have anti-gastric cancer activity, which offers a potential novel and natural anti-gastric cancer remedy.

Keywords: Cytotoxic, peptides, Euphorbia hirta Linn., gastric carcinoma.

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Authors: Sharareh Sharifi, Pargol Ghavam Mostafavi, Ali Mashinchian Moradi, Mohammad Hadi Givianrad, Hassan Niknejad

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Marine organisms such as soft coral, sponge, ascidians, and tunicate containing rich source of natural compound have been studied in last decades because of their special chemical compounds with anticancer properties. The aim of this study was to investigate anti-cancer property of ethyl acetate extracted from marine sea pen Virgularia gustaviana found from Persian Gulf coastal (Bandar Abbas). The extraction processes were carried out with ethyl acetate for five days. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were used for qualitative identification of crude extract. The viability of HeLa and MDA-Mb-231 cancer cells was investigated using MTT assay at the concentration of 25, 50, and a 100 µl/ml of ethyl acetate is extracted. The crude extract of Virgularia gustaviana demonstrated ten fractions with different Retention factor (Rf) by TLC and Retention time (Rt) evaluated by HPLC. The crude extract dose-dependently decreased cancer cell viability compared to control group. According to the results, the ethyl acetate extracted from Virgularia gustaviana inhibits the growth of cancer cells, an effect which needs to be further investigated in the future studies.

Keywords: Virgularia gustaviana, Cembrane Diterpene, anti-cancer, HeLa cancer Cell, MDA-Md-231 Cancer cell.

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Authors: Marek Halenár, Eva Tvrdá, Simona Baldovská, Ľubomír Ondruška, Peter Massányi, Adriana Kolesárová

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This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P<0.05 with respect to time 8h). At the same time, concentrations ranging between 1 and 100 µg/mL of the extract led to a significant preservation of the cell viability (P<0.05 in case of 5, 50 and 100 µg/mL; P<0.01 with respect to 1 and 10 µg/mL, time 8h). 1 and 5 µg/mL of the extract exhibited antioxidant characteristics, translated into a significant reduction of the ROS production, particularly notable at time 8h (P<0.01). The results indicate that the Viscum extract is capable of delaying the damage inflicted to the spermatozoon by the in vitro environment.

Keywords: CASA, mistletoe, mitochondrial activity, motility, reactive oxygen species, rabbits, spermatozoa, Viscum album.

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Authors: Marek Halenár, Eva Tvrdá, Tomáš Slanina, Ľubomír Ondruška, Eduard Kolesár, Peter Massányi, Adriana Kolesárová

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The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P<0.05 with respect to 0.5 mg/mL and 1 mg/mL AMG; Time 5 h) and mitochondrial activity (P< 0.05 in case of 0.5 mg/mL AMG; P< 0.01 in case of 1 mg/mL AMG; P < 0.001 with respect to 2.5 mg/mL and 5 mg/mL AMG; Time 5 h). None of the AMG doses supplemented had any significant impact of the spermatozoa viability. In conclusion, the data revealed that short-term co-incubation of spermatozoa with AMG may result in a higher preservation of the sperm structural integrity and functional activity.

Keywords: Amygdalin, CASA, mitochondrial activity, motility, rabbits, spermatozoa, viability.

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Authors: Ali A. Alshatwi, Vaiyapuri S. Periasamy, Jegan Athinarayanan

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Engineered nanoparticles’ usage rapidly increased in various applications in the last decade due to their unusual properties. However, there is an ever increasing concern to understand their toxicological effect in human health. Particularly, metal and metal oxide nanoparticles have been used in various sectors including biomedical, food and agriculture. But their impact on human health is yet to be fully understood. In this present investigation, we assessed the toxic effect of engineered nanoparticles (ENPs) including Ag, MgO and Co3O4 nanoparticles (NPs) on human mesenchymal stem cells (hMSC) adopting cell viability and cellular morphological changes as tools The results suggested that silver NPs are more toxic than MgO and Co3O4NPs. The ENPs induced cytotoxicity and nuclear morphological changes in hMSC depending on dose. The cell viability decreases with increase in concentration of ENPs. The cellular morphology studies revealed that ENPs damaged the cells. These preliminary findings have implications for the use of these nanoparticles in food industry with systematic regulations.

Keywords: Cobalt oxide, Human mesenchymal stem cells, MgO, Silver.

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Authors: Mashayekh Amir Shahriar, Akhlaghi Morteza, Rajaee Hajar, Khani Mohammad Reza, Shokri Babak

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A new and novel approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper a pin-to-hole plasma jet suitable for biological applications is investigated and characterized and the possibility and feasibility of cancer cell treatment is evaluated. The characterization includes power consumption via Lissajous method, thermal behavior of plasma using Infra-red camera as a novel method, Optical Emission Spectroscopy (OES) to determine the species that are generated. Treatment of leukemia cancer cells is also implemented and MTT assay is used to evaluate viability.

Keywords: Atmospheric Pressure Plasma Jet (APPJ), Plasma Medicine, Cancer cell treatment, leukemia.

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Authors: KyungBae Pi, KiBeom Lee, Yongil Kim, Eun-Jung Lee

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Abnormal adipocyte growth, in terms of increased cell numbers and increased cell differentiation, is considered to be a major pathological feature of obesity. Thus, the inhibition of preadipocyte mitogenesis and differentiation could help prevent and suppress obesity. The aim of this study was to assess whether extracts from Weissella koreensis 521 cells isolated from kimchi could exert anti-adipogenic effects in 3T3-L1 cells (fat cells). Differentiating 3T3-L1 cells were treated with W. koreensis 521 cell extracts (W. koreensis 521_CE), and cell viability was assessed by MTT assays. At concentrations below 0.2 mg/ml, W. koreensis 521_CE did not exert any cytotoxic effect in 3T3-L1 cells. However, treatment with W. koreensis 521_CE significantly inhibited adipocyte differentiation, as assessed by morphological analysis and Oil Red O staining of fat. W. koreensis 521_CE treatment (0.2 mg/ml) also reduced lipid accumulation by 24% in fully differentiated 3T3-L1 adipocytes. These findings collectively indicate that Weissella koreensis 521 may help prevent obesity.

Keywords: Weissella koreensis 521, 3T3-L1 cells, adipocyte differentiation, obesity.

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Authors: Ekaterina Y. Lukianova-Hleb, Dmitri O. Lapotko

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Cell processing techniques for gene and cell therapies use several separate procedures for gene transfer and cell separation or elimination, because no current technology can offer simultaneous multi-functional processing of specific cell sub-sets in heterogeneous cell systems. Using our novel on-demand nonstationary intracellular events instead of permanent materials, plasmonic nanobubbles, generated with a short laser pulse only in target cells, we achieved simultaneous multifunctional cell-specific processing with the rate up to 50 million cells per minute.

Keywords: Delivery, cell separation, graft, laser, plasmonic nanobubble, cell therapy, gold nanoparticle.

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Authors: Arash Azizi Mazreah, Mohammad Reza Sahebi, Mohammad T. Manzuri Shalmani

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To help overcome limits to the density of conventional SRAMs and leakage current of SRAM cell in nanoscaled CMOS technology, we have developed a four-transistor SRAM cell. The newly developed CMOS four-transistor SRAM cell uses one word-line and one bit-line during read/write operation. This cell retains its data with leakage current and positive feedback without refresh cycle. The new cell size is 19% smaller than a conventional six-transistor cell using same design rules. Also the leakage current of new cell is 60% smaller than a conventional sixtransistor SRAM cell. Simulation result in 65nm CMOS technology shows new cell has correct operation during read/write operation and idle mode.

Keywords: SRAM Cell, leakage current, cell area.

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Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

Abstract:

Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: Mesenchymal stem cells, Cryopreservation, Stemness, Senescence.

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Authors: M. S. Hosseini, M. Tazzoli-Shadpour, I. Amjadi, A. A. Katbab, E. Jaefargholi-Rangraz

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Bionanotechnology deals with nanoscopic interactions between nanostructured materials and biological systems. Polymer nanocomposites with optimized biological activity have attracted great attention. Nanoclay is considered as reinforcing nanofiller in manufacturing of high performance nanocomposites. In current study, organomodified-nanoclay with negatively charged silicate layers was incorporated into biomedical grade silicone rubber. Nanoparticle loading has been tailored to enhance cell behavior. Addition of nanoparticles led to improved mechanical properties of substrate with enhanced strength and stiffness while no toxic effects was observed. Results indicated improved viability and proliferation of cells by addition of nanofillers. The improved mechanical properties of the matrix result in proper cell response through adjustment and arrangement of cytoskeletal fibers. Results can be applied in tissue engineering when enhanced substrates are required for improvement of cell behavior for in vivo applications.

Keywords: Biocompatibility, Composite, Organomodified- Nanoclay, Proliferation

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Authors: J. Chojnacki, E. Dulcet, A. Grieger

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The entomopathogenic nematodes Steinernema feltiaeare are components of many biological pesticides. The biological pesticides are applicated by means a spraying machines. The influence of high pressure operating time on viability of nematodes has been experimentally investigated in order to explain if static pressure inside of the sprayers installation was able to destroy nematodes. The value of pressure was 55 MPa and its maximum operating time was 3 hours. Changes were found in viability of pressurized samples of nematodes, mixed with water.

Keywords: Entomopathogenic nematodes, biopesticides, highpressure, sprayer.

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Authors: M. Kusá, M. Matúšová, A. Javorová, K. Velí

Abstract:

This paper deals about four items assembly process of linear drive. This assembly will be realized in flexible assembly cell on Institute of Manufacturing Systems and Applied Mechanics. There is defined manufacturing cell, individual actuators created our flexible cell. Next chapter is about control type, detailed describe a sequence control type, which will be used in mentioned flexible assembly cell. All cell control is divided in individual steps instructions. There instructions illustrate table number III.

Keywords: assembly, flexible cell, sequence control

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Authors: A. Dashti, M. Eskandari, L. Farahmand, P. Parvin, A. Jafargholi

Abstract:

Breast Cancer is one of the most considerable diseases in the United States and other countries and is the second leading cause of death in women. Common breast cancer treatments would lead to adverse side effects such as loss of hair, nausea, and weakness. These complications arise because these cancer treatments damage some healthy cells while eliminating the cancer cells. In an effort to address these complications, laser radiation was utilized and tested as a targeted cancer treatment for breast cancer. In this regard, tissue engineering approaches are being employed by using an electrospun scaffold in order to facilitate the growth of breast cancer cells. Polycaprolacton (PCL) was used as a material for scaffold fabricating because of its biocompatibility, biodegradability, and supporting cell growth. The specific breast cancer cells have the ability to create a three-dimensional cell cluster due to the spontaneous accumulation of cells in the porosity of the scaffold under some specific conditions. Therefore, we are looking for a higher density of porosity and larger pore size. Fibers showed uniform diameter distribution and final scaffold had optimum characteristics with approximately 40% porosity. The images were taken by SEM and the density and the size of the porosity were determined with the Image. After scaffold preparation, it has cross-linked by glutaraldehyde. Then, it has been washed with glycine and phosphate buffer saline (PBS), in order to neutralize the residual glutaraldehyde. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor (MTT) results have represented approximately 91.13% viability of the scaffolds for cancer cells. In order to create a cluster, Michigan Cancer Foundation-7 (MCF-7, breast cancer cell line) and Michigan Cancer Foundation-10A (MCF-10A, human mammary epithelial cell line) cells were cultured on the scaffold in 24 well plate for five days. Then, we have exposed the cluster to the laser diode 808 nm radiation to investigate the effect of laser on the tumor with different power and time. Under the same conditions, cancer cells lost their viability more than the healthy ones. In conclusion, laser therapy is a viable method to destroy the target cells and has a minimum effect on the healthy tissues and cells and it can improve the other method of cancer treatments limitations.

Keywords: Breast cancer, electrospun scaffold, polycaprolacton, laser diode, cancer treatment.

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Authors: A. Rohana, A. M., Fadzilah Adibah, M. S. Muhammad Roji

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Oleic acid (C18:1) play an important role in proliferation of fat cells. In this study, the effect of oleate on cells viability in 3T3-L1 cells (fat cells) was investigated. The 3T3-L1 cells were treated with various concentrations of oleate in the presence of 23 mM glucose. Oleate was added to adipogenic media (day 0) to investigate the influence of oleate on proliferation of postconfluent preadipocytes after 24 h induction. 0.1 mM oleate promoted cell division by increasing 33.9% number of cells from basal control in postconfluent preadipocytes. However, there were no significantly different in cells viability with control cells when oleate concentrations were increased up to 0.5 mM. When added to differentiated adipocytes (day 12) for 48 h, the number of cells decreased as oleate concentrations increased. 92.7% of cells lost demonstrated apoptosis and necrosis after 48 h with 0.5 mM oleate. The fluorochrome staining was examined under fluorescence microscopy using acridine orange and ethidium bromide double staining. Furthermore, the presence of high lactate (60.6% increased from basal control) released into plasma has shown the direct cytotoxicity of 0.5 mM oleate on adipocytes.

Keywords: adipocytes, apoptosis, oleate, postconfluentpreadipocytes

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Authors: S. C. Santos, S. R. Dionísio, A. L. D. De Andrade, L. R. Roque, A. C. Da Costa, J. L. Ienczak

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In this work, two fermentations at different temperatures (25 and 30ºC), with cell recycling, were accomplished to produce ethanol, using a mix of commercial substrates, xylose (70%) and glucose (30%), as organic source for Scheffersomyces stipitis. Five consecutive fermentations of 80 g L-1 (1º, 2º and 3º recycles), 96 g L-1 (4º recycle) and 120 g L-1 (5º recycle)reduced sugars led to a final maximum ethanol concentration of 17.2 and 34.5 g L-1, at 25 and 30ºC, respectively. Glucose was the preferred substrate; moreover xylose startup degradation was initiated after a remaining glucose presence in the medium. Results showed that yeast acid treatment, performed before each cycle, provided improvements on cell viability, accompanied by ethanol productivity of 2.16 g L-1 h- 1 at 30ºC. A maximum 36% of xylose was retained in the fermentation medium and after five-cycle fermentation an ethanol yield of 0.43 g ethanol/g sugars was observed. S. stipitis fermentation capacity and tolerance showed better results at 30ºC with 83.4% of theoretical yield referenced on initial biomass.

Keywords: 5-carbon sugar, cell recycling fermenter, mixed sugars, xylose-fermenting yeast.

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Authors: Antonella Bandiera

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A new sythetic gene coding for a Human Elastin-Like Polypeptide was constructed and expressed. The recombinant product was tested as coating agent to realize a surface suitable for cell growth. Coatings showed peculiar features and different human cell lines were seeded and cultured. All cell lines tested showed to adhere and proliferate on this substrate that has been shown also to exert a specific effect on cells, depending on cell type.

Keywords: elastin, recombinant protein, coating, cell adhesion.

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Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: Bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa.

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Authors: Chih-Wei Chao, Jiashing Yu

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To mimic the natural circumstances of cell growth in an organism, we present three-dimensional (3D) scaffolds fabricated by microfluidics for cultivation. This work investigates the cellular behaviors of rat cardiomyocytes in gelatin 3D scaffolds compared to those on 2D control, such as proliferation, viability and morphology. We found that the scaffolds may induce skeletal differentiation of H9c2 cells.

Keywords: Microfluidic device, H9c2, tissue engineering, 3D scaffolds.

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Authors: Fan Gao, Lior Pachter

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The primary tool currently used to pre-process 10X chromium single-cell ATAC-seq data is Cell Ranger, which can take very long to run on standard datasets. To facilitate rapid pre-processing that enables reproducible workflows, we present a suite of tools called scATAK for pre-processing single-cell ATAC-seq data that is 15 to 18 times faster than Cell Ranger on mouse and human samples. Our tool can also calculate chromatin interaction potential matrices and generate open chromatin signal and interaction traces for cell groups. We use scATAK tool to explore the chromatin regulatory landscape of a healthy adult human brain and unveil cell-type specific features, and show that it provides a convenient and computational efficient approach for pre-processing single-cell ATAC-seq data.

Keywords: single-cell, ATAC-seq, bioinformatics, open chromatin landscape, chromatin interactome

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Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H₂O and CO₂. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: Poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblasts, rat vascular smooth muscle cells, human stem cells.

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Authors: Feng-Sheng Wang, Wu-Hsiung Wu, Kai-Cheng Hsu

Abstract:

The optimization of biological systems, which is a branch of metabolic engineering, has generated a lot of industrial and academic interest for a long time. In the last decade, metabolic engineering approaches based on mathematical optimizations have been used extensively for the analysis and manipulation of metabolic networks. In practical optimization of metabolic reaction networks, designers have to manage the nature of uncertainty resulting from qualitative characters of metabolic reactions, e.g., the possibility of enzyme effects. A deterministic approach does not give an adequate representation for metabolic reaction networks with uncertain characters. Fuzzy optimization formulations can be applied to cope with this problem. A fuzzy multi-objective optimization problem can be introduced for finding the optimal engineering interventions on metabolic network systems considering the resilience phenomenon and cell viability constraints. The accuracy of optimization results depends heavily on the development of essential kinetic models of metabolic networks. Kinetic models can quantitatively capture the experimentally observed regulation data of metabolic systems and are often used to find the optimal manipulation of external inputs. To address the issues of optimizing the regulatory structure of metabolic networks, it is necessary to consider qualitative effects, e.g., the resilience phenomena and cell viability constraints. Combining the qualitative and quantitative descriptions for metabolic networks makes it possible to design a viable strain and accurately predict the maximum possible flux rates of desired products. Considering the resilience phenomena in metabolic networks can improve the predictions of gene intervention and maximum synthesis rates in metabolic engineering. Two case studies will present in the conference to illustrate the phenomena.

Keywords: Fuzzy multi-objective optimization problem, kinetic model, metabolic engineering.

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Authors: Irina Spaka, Rita Birkenfelde, Svetlana Kozireva, Jevgenija Osmjana, Madara Upmane, ElenaKashuba, Irina Kholodnyuk Holodnuka

Abstract:

Epstein-Barr virus (EBV) is implicated in the pathogenesis of the endemic Burkitt-s lymphoma (BL). The EBVpositive BL-derived cell lines initially maintain the original tumor phenotype of EBV infection (latency I, LatI), but most of them drift toward a lymphoblast phenotype of EBV latency III (LatIII) during in vitro culturing. The aim of the present work was to characterize the B-cell subsets in EBV-positive BL cell lines and to verify whether a particular cell subset correlates with the type of EBV infection. The phenotype analysis of two EBV-negative and eleven EBV-positive (three of LatI and eight of LatIII) BL cell lines was performed by polychromatic flow cytomery, based on expression pattern of CD19, CD10, CD38, CD27, and CD5 markers. Two cell subsets, CD19+CD10+ and CD19+CD10-, were defined in LatIII BL cell lines. In both subsets, the CD27 and CD5 cell surface expression was detected in a proportion of the cells.

Keywords: B-cell subsets, Burkitt's lymphoma cell lines, EBV latency, phenotype profiles.

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Authors: R. Mahjoub, K. Maghsoudi Mehraban

Abstract:

Since the heart of the hybrid system is the fuel cell and it has vital impact on efficiency and performance of cycle, in this study, the major modeling of electrochemical reaction within the fuel cell is analyzed. Also, solid oxide fuel cell is integrated with the gas turbine and thermodynamic analysis on different elements of hybrid system is applied. Next, in predefined operational points of hybrid cycle, the simulation results are obtained. Then, different source of irreversibility in fuel cell is modeled and influence of different major parameters on different irreversibility is computed and applied. Then, the effect of important parameters such as thickness and surface of electrolyte fuel cell are simulated in fuel cell and its dependency to these parameters is explained. At the end of the paper, different impact of parameters on fuel cell with a gas turbine and current density and voltage of fuel cell are simulated.

Keywords: Electrochemical analysis, Gas turbine, Hybrid system, Irreversibility analysis.

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Authors: Saravana Kannan Thangavelu

Abstract:

Fuel and oxidant gas delivery plate, or fuel cell plate, is a key component of a Proton Exchange Membrane (PEM) fuel cell. To manufacture low-cost and high performance fuel cell plates, advanced computer modeling and finite element structure analysis are used as virtual prototyping tools for the optimization of the plates at the early design stage. The present study examines thermal stress analysis of the fuel cell plates that are produced using a patented, low-cost fuel cell plate production technique based on screen-printing. Design optimization is applied to minimize the maximum stress within the plate, subject to strain constraint with both geometry and material parameters as design variables. The study reveals the characteristics of the printed plates, and provides guidelines for the structure and material design of the fuel cell plate.

Keywords: Design optimization, FEA, PEM fuel cell, Thermal stress

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Authors: Azhar Rauf, Sareer Badshah, Shah Khusro

Abstract:

This paper analyzes different techniques of the fine grained security of relational databases for the two variables-data accessibility and inference. Data accessibility measures the amount of data available to the users after applying a security technique on a table. Inference is the proportion of information leakage after suppressing a cell containing secret data. A row containing a secret cell which is suppressed can become a security threat if an intruder generates useful information from the related visible information of the same row. This paper measures data accessibility and inference associated with row, cell, and column level security techniques. Cell level security offers greatest data accessibility as it suppresses secret data only. But on the other hand, there is a high probability of inference in cell level security. Row and column level security techniques have least data accessibility and inference. This paper introduces cell plus innocent security technique that utilizes the cell level security method but suppresses some innocent data to dodge an intruder that a suppressed cell may not necessarily contain secret data. Four variations of the technique namely cell plus innocent 1/4, cell plus innocent 2/4, cell plus innocent 3/4, and cell plus innocent 4/4 respectively have been introduced to suppress innocent data equal to 1/4, 2/4, 3/4, and 4/4 percent of the true secret data inside the database. Results show that the new technique offers better control over data accessibility and inference as compared to the state-of-theart security techniques. This paper further discusses the combination of techniques together to be used. The paper shows that cell plus innocent 1/4, 2/4, and 3/4 techniques can be used as a replacement for the cell level security.

Keywords: Fine Grained Security, Data Accessibility, Inference, Row, Cell, Column Level Security.

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