Search results for: packed red blood cells
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 5277

Search results for: packed red blood cells

4467 Systolic Blood Pressure and Its Determinants: Study in a Population Attending Pharmacies in a Portuguese Coastal City

Authors: M. J. Reis Lima, J. Oliveira, M. Brito, C. Lemos, A. Mascarenhas, E. Teixeira Lemos

Abstract:

Hypertension is a common condition causing cardio and cerebrovascular complications. Portugal has one of the highest mortality rates from stroke and a high prevalence of hypertension. Systolic blood pressure (SBP) is an important risk factor for cardiovascular events (myocardial infarction and stroke) and premature mortality, particularly in the elderly population. The present study aims to estimate the prevalence of hypertension in a Portuguese population living in a coastal city and to identify some of its determinants (namely gender, age, the body mass index and physical activity frequency). A total of 91 adults who attended three pharmacies of a coastal city in the center of Portugal, between May and August of 2013 were evaluated. Attendants who reported to have diabetes or taking antihypertensive drugs in the 2 previous weeks were excluded from the study. Sociodemographic factors, BMI, habits of exercise and BP were assessed. Hypertension was defined as blood pressure ≥140/90 mmHg. The majority of the studied population was constituted by women (75.8%), with a mean age of 54.2±1.6 years old, married or living in civil union and that had completed secondary school or had higher education (40%). They presented a mean BMI of 26.2±4.76 Kg/m2. and were sedentary. The mean BP was 127.0±17.77mmHg- 74.69 ± 9.53. In this population, we found 4.3% of people with hypertension and 16.1% with normal high blood pressure. Men exhibit a tendency to present higher systolic blood pressure values than women. Of all the factors considered, SBP values also tended to be higher with age and higher BMI values. Despite the fact that the mean values of SBP did not present values higher than 140 mmHg we must be concerned because the studied population is undiagnosed for hypertension. Our study even with some limitations might be a prelude to the upcoming research about the underlying factors responsible for the occurrence of SBP.

Keywords: hypertension, age, exercise, obesity and gender

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4466 Investigating the Role of Circular RNA GATAD2A on H1N1 Replication

Authors: Tianqi Yu, Yingnan Ding, Yina Zhang, Yulan Liu, Yahui Li, Jing Lei, Jiyong Zhou, Suquan Song, Boli Hu

Abstract:

Circular RNAs (circRNAs) play critical roles in various diseases. However, whether and how circular RNA regulates influenza A virus (IAV) infection is unknown. Here, we studied the role of circular RNA GATA Zinc Finger Domain Containing 2A (circ-GATAD2A) in the replication of IAV H1N1 in A549 cells. Circ-GATAD2A was formed upon H1N1 infection. Knockdown of circ-GATAD2A in A549 cells enhanced autophagy and inhibited H1N1 replication. By contrast, overexpression of circ-GATAD2A impaired autophagy and promoted H1N1 replication. Similarly, knockout of vacuolar protein sorting 34 (VPS34) blocked autophagy and increased H1N1 replication. However, the expression of circ-GATAD2A could not further enhance H1N1 replication in VPS34 knockout cells. Collectively, these data indicated that circ-GATAD2A promotes the replication of H1N1 by inhibiting autophagy.

Keywords: autophagy, circ-GATAD2A, H1N1, replication

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4465 Colonization of Embrionic Gonads of Nile Tilapia by Giant Gourami Testicular Germ Cells

Authors: Irma Andriani, Ita Djuwita, Komar Sumantadinata, Alimuddin

Abstract:

The recent study has been conducted to develop testicular germ cell transplantation as a tool for preservation and propagation of male germ-plasm from endangered fish species, as well as to produce surrogate broodstock of commercially valuable fish. Giant gourami testis had been used as a model for donor and Nile tilapia larvae as recipient. We developed testicular cell xenotransplantation by optimizing the timing of intraperitoneal cell transplantation to recipient larvae aged 1, 3, 5 and 7 days post hatching (dph). Freshly isolated testis of giant gourami weighing 600–800 g were minced in dissociation medium and then incubated for 3 hours in room temperature to collect monodisperce cell suspension. Donor cells labeled with PKH 26 were transplanted into the peritoneal cavity of Nile tilapia larvae using glass micropipettes. Parameters observed were survival rate of Nile tilapia larvae at 24 hours post transplantation (pt) and colonization efficiency of donor cells at 2 and 3 months pt. The incorporated donor cells were observed under fluorescent microscope. The result showed that the lowest survival rate at 24 hours pt was 1 dph larvae (82.74±6.76%) and the highest survival rate were 3 and 5 dph larvae (95.00±5.00% and 95.00±2.50%, respectively). The highest colonization efficiency was on 3 dph larvae (61.1±34.71%) and the lowest colonization efficiency was on 7 dph larvae (19.43±17.33%). In conclusion, 3 dph Nile tilapia larvae was the best recipient for giant gourami testicular germ cells xenotransplantation.

Keywords: xenotransplantation, testicular germ cell, giant gourami, Nile tilapia, colonization efficiency

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4464 Experimental Study of Boost Converter Based PV Energy System

Authors: T. Abdelkrim, K. Ben Seddik, B. Bezza, K. Benamrane, Aeh. Benkhelifa

Abstract:

This paper proposes an implementation of boost converter for a resistive load using photovoltaic energy as a source. The model of photovoltaic cell and operating principle of boost converter are presented. A PIC micro controller is used in the close loop control to generate pulses for controlling the converter circuit. To performance evaluation of boost converter, a variation of output voltage of PV panel is done by shading one and two cells.

Keywords: boost converter, microcontroller, photovoltaic power generation, shading cells

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4463 The Possible Antioxidant, Hypoglycemic Effect and Antimicrobial Potential of Mangifera Indicia Leaves Aqueous Extract in Albino Rats

Authors: Sahar B. Ahmed, M. Mostafa Said, Mona I. Mohamed

Abstract:

Streptozotocin (STZ) caused a significant increase in blood glucose and malondialdehyde (MDA) levels in serum accompanied by a significant decrease in blood reduced glutathione (GSH) and superoxide dismutase (SOD) activities. Also, ALT, AST, albumin and urea were markedly affected by STZ injection. The oral administration of Mango leaves extract (MLE) one hour before STZ injection was significantly improved the blood glucose level, ALT, AST activities, albumin and urea that associated with the regulation of MDA, GSH and SOD levels. The antimicrobial activity of MLE showed a significant inhibitory activity against multidrug resistant gram positive and gram negative bacteria isolated from patients in Egyptian hospitals especially Salmonella typhi and typhimurium. In conclusion, results revealed the antioxidant, hypoglycemic effect and antimicrobial potentials of MLE under investigation. Further studies will be needed to investigate the prolonged period of MLE administration and its possible side effects.

Keywords: aqueous extract of mango leaves, STZ, antioxidant, hypoglycemic effect, antimicrobial potentials.

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4462 Effect of Varying Scaffold Architecture and Porosity of Calcium Alkali Orthophosphate Based-Scaffolds for Bone Tissue Engineering

Authors: D. Adel, F. Giacomini, R. Gildenhaar, G. Berger, C. Gomes, U. Linow, M. Hardt, B. Peleskae, J. Günster, A. Houshmand, M. Stiller, A. Rack, K. Ghaffar, A. Gamal, M. El Mofty, C. Knabe

Abstract:

The goal of this study was to develop 3D scaffolds from a silica containing calcium alkali orthophosphate utilizing two different fabrication processes, first a replica technique namely the Schwartzwalder Somers method (SSM), and second 3D printing, i.e. Rapid prototyping (RP). First, the mechanical and physical properties of the scaffolds (porosity, compressive strength, and solubility) was assessed and second their potential to facilitate homogenous colonization with osteogenic cells and extracellular bone matrix formation throughout the porous scaffold architecture. To this end murine and rat calavarie osteoblastic cells were dynamically seeded on both scaffold types under perfusion with concentrations of 3 million cells. The amount of cells and extracellular matrix as well as osteogenic marker expression was evaluated using hard tissue histology, immunohistochemistry, and histomorphometric analysis. Total porosities of both scaffolds were 86.9 % and 50% for SSM and RP respectively, Compressive strength values were 0.46 ± 0.2 MPa for SSM and 6.6± 0.8 MPa for RP. Regarding the cellular behavior, RP scaffolds displayed a higher cell and matrix percentage of 24.45%. Immunoscoring yielded strong osteocalcin expression of cells and matrix in RP scaffolds and a moderate expression in SSM scaffolds. 3D printed RP scaffolds displayed superior mechanical and biological properties compared to SSM. 3D printed scaffolds represent excellent candidates for bone tissue engineering.

Keywords: calcium alkali orthophosphate, extracellular matrix mineralization, osteoblast differentiation, rapid prototyping, scaffold

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4461 Effectiveness of Homoeopathic Medicine Conium Maculatum 200 C for Management of Pyuria

Authors: Amir Ashraf

Abstract:

Homoeopathy is an alternative system of medicine discovered by German physician Samuel Hahnemann in 1796. It has been used by several people for various health conditions globally for more than last 200 years. In India, homoeopathy is considered as a major system of alternative medicine. Homoeopathy is found effective in various medical conditions including Pyuria. Pyuria is the condition in which pus cells are found in urine. Homoeopathy is very useful for reducing pus cells, and homeopathically potentized Conium Mac (Hemlock) is an important remedy commonly used for reducing pyuria. Aim: To reduce the amount pus cells found in urine using Conium Mac 200C. Methods: Design. Small N Design. Samples: Purposive Sampling with 5 cases diagnosed as pyuria. Tools: Personal Data Schedule and ICD-10 Criteria for Pyuria. Techniques: Potentized homoeopathic medicine, Conium Mac 200th potency is used. Statistical Analysis: The statistical analyses were done using non-parametric tests. Results: There is significant pre/post difference has been identified. Conclusion: Homoeopathic potency, Conium Mac 200 C is effective in reducing the increased level of pus cells found in urine samples.

Keywords: homoeopathy, alternative medicine, Pyuria, Conim Mac, small N design, non-parametric tests, homeopathic physician, Ashirvad Hospital, Kannur

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4460 Fabrication of Graphene Oxide Based Planar Hetero-Junction Perovskite Solar Cells

Authors: Khursheed Ahmad, Shaikh M. Mobin

Abstract:

In this work, we have developed a highly stable planar heterojunction perovskite solar cells (PSCs) with a architecture (ITO/GO/PEDOT:PSS/MAPbI3/PCBM/Carbon tape). The PSCs was fabricated under air using GO/PEDOT:PSS as hole transport layer while the carbon tape used as a back contact to complete the device. The fabricated PSCs device exhibited good stability and performance in terms of power conversion efficiency of 5.2%. The PSCs devices were exposed to ambient condition for 4 days which shows excellent stability confirmed by XRD analysis. We believed that the stability of the planar heterojunction perovskite solar cell may be due the presence of GO which inhibits the direct contact between PEDOT:PSS and MAPbI3.

Keywords: graphene oxide, perovskite solar cells, hole transport layer, PEDOT:PSS

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4459 Temporal Changes of Heterogeneous Subpopulations of Human Adipose-Derived Stromal/Stem Cells in vitro

Authors: Qiuyue Peng, Vladimir Zachar

Abstract:

The application of adipose-derived stromal/stem cells (ASCs) in regenerative medicine is gaining more awareness due to their advanced translational potential and abundant source preparations. However, ASC-based translation has been confounded by high subpopulation heterogeneity, causing ambiguity about its precise therapeutic value. Some phenotypes defined by a unique combination of positive and negative surface markers have been found beneficial to the required roles. Therefore, the immunophenotypic repertoires of cultured ASCs and temporal changes of distinct subsets were investigated in this study. ASCs from three donors undergoing cosmetic liposuction were cultured in standard culturing methods, and the co-expression patterns based on the combination of selected markers at passages 1, 4, and 8 were analyzed by multi-chromatic flow cytometry. The results showed that the level of heterogeneity of subpopulations of ASCs became lower by in vitro expansion. After a few passages, most of the CD166⁺/CD274⁺/CD271⁺ based subpopulations converged to CD166 single positive cells. Meanwhile, these CD29⁺CD201⁺ double-positive cells, in combination with CD36/Stro-1 expression or without, feathered only the major epitopes and maintained prevailing throughout the whole process. This study suggested that, upon in vitro expansion, the phenotype repertoire of ASCs redistributed and stabilized in a way that cells co-expressing exclusively the strong markers remained dominant. These preliminary findings provide a general overview of the distribution of heterogeneous subsets residents within human ASCs during expansion in vitro. It is a critical step to fully characterize ASCs before clinical application, although the biological effects of heterogeneous subpopulations still need to be clarified.

Keywords: adipose-derived stromal/stem cells, heterogeneity, immunophenotype, subpopulations

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4458 Effect of Blood Sugar Levels on Short Term and Working Memory Status in Type 2 Diabetics

Authors: Mythri G., Manjunath ML, Girish Babu M., Shireen Swaliha Quadri

Abstract:

Background: The increase in diabetes among the elderly is of concern because in addition to the wide range of traditional diabetes complications, evidence has been growing that diabetes is associated with increased risk of cognitive decline. Aims and Objectives: To find out if there is any association between blood sugar levels and short-term and working memory status in patients of type 2 diabetes. Materials and Methods: The study was carried out in 200 individuals aged between 40-65 years consisting of 100 diagnosed cases of Type 2 Diabetes Mellitus and 100 non-diabetics from OPD of Mc Gann Hospital, Shivamogga. Rye’s Auditory Verbal Learning Test, Verbal Fluency Test and Visual Reproduction Test, Working Digit Span Test and Validation Span Test were used to assess short-term and working memory. Fasting and Post Prandial blood sugar levels were estimated. Statistical analysis was done using SPSS 21. Results: Memory test scores of type 2 diabetics were significantly reduced (p < 0.001) when compared to the memory scores of age and gender matched non-diabetics. Fasting blood sugar levels were found to have a negative correlation with memory scores for all 5 tests: AVLT (r=-0.837), VFT (r=-0.888), VRT(r=-0.787), WDST (r=-0.795) and VST (r=-0.943). Post- Prandial blood sugar levels were found to have a negative correlation with memory scores for all 5 tests: AVLT (r=-0.922), VFT (r=-0.848), VRT(r=-0.707),WDST (r=-0.729) and VST (r=-0.880) Memory scores in all 5 tests were found to be negatively correlated with the FBS and PPBS levels in diabetic patients (p < 0.001). Conclusion: The decreased memory status in diabetic patients may be due to many factors like hyperglycemia, vascular disease, insulin resistance, amyloid deposition and also some of the factor combine to produce additive effects like, type of diabetes, co-morbidities, age of onset, duration of the disease and type of therapy. These observed effects of blood sugar levels of diabetics on memory status are of potential clinical importance because even mild cognitive impairment could interfere with todays’ activities.

Keywords: diabetes, cognition, diabetes, HRV, respiratory medicine

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4457 Peptidoglycan Vaccine-On-Chip against a Lipopolysaccharide-Induced Experimental Sepsis Model

Authors: Katerina Bakela, Ioanna Zerva, Irene Athanassakis

Abstract:

Lipopolysaccharide (LPS) is commonly used in murine sepsis models, which are largely associated with immunosuppression (incretion of MDSCs cells and Tregs, imbalance of inflammatory/anti-inflammatory cytokines) and collapse of the immune system. After adapting the LPS treatment to the needs of locally bred BALB/c mice, the present study explored the protective role of Micrococcus luteus peptidoglycan (PG) pre-activated vaccine-on chip in endotoxemia. The established protocol consisted of five daily intraperitoneal injections of 0.2mg/g LPS. Such protocol allowed longer survival, necessary in the prospect of the therapeutic treatment application. The so-called vaccine-on-chip consists of a 3-dimensional laser micro-texture Si-scaffold loaded with BALB/c mouse macrophages and activated in vitro with 1μg/ml PG, which exert its action upon subcutaneous implantation. The LPS treatment significantly decreased CD4+, CD8+, CD3z+, and CD19+ cells, while increasing myeloid-derived suppressor cells (MDSCs), CD25+, and Foxp3+ cells. These results were accompanied by increased arginase-1 activity in spleen cell lysates and production of IL-6, TNF-a, and IL-18 while acquiring severe sepsis phenotype as defined by the murine sepsis scoring. The in vivo application of PG pre-activated vaccine-on chip significantly decreased the percent of CD11b+, Gr1+, CD25+, Foxp3+ cells, and arginase-1 activity in the spleen of LPS-treated animals, while decreasing IL-6 and TNF-a in the serum, allowing survival to all animals tested and rescuing the severity of sepsis phenotype. In conclusion, these results reveal a promising mode of action of PG pre-activated vaccine-on chip in LPS endotoxemia, strengthening; thus, the use of treatment is septic patients.

Keywords: myeloid-derived suppressor cells, peptidoglycan, sepsis, Si-scaffolds

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4456 Investigation of Wood Chips as Internal Carbon Source Supporting Denitrification Process in Domestic Wastewater Treatment

Authors: Ruth Lorivi, Jianzheng Li, John J. Ambuchi, Kaiwen Deng

Abstract:

Nitrogen removal from wastewater is accomplished by nitrification and denitrification processes. Successful denitrification requires carbon, therefore, if placed after biochemical oxygen demand (BOD) and nitrification process, a carbon source has to be re-introduced into the water. To avoid adding a carbon source, denitrification is usually placed before BOD and nitrification processes. This process however involves recycling the nitrified effluent. In this study wood chips were used as internal carbon source which enabled placement of denitrification after BOD and nitrification process without effluent recycling. To investigate the efficiency of a wood packed aerobic-anaerobic baffled reactor on carbon and nutrients removal from domestic wastewater, a three compartment baffled reactor was presented. Each of the three compartments was packed with 329 g wood chips 1x1cm acting as an internal carbon source for denitrification. The proposed mode of operation was aerobic-anoxic-anaerobic (OAA) with no effluent recycling. The operating temperature, hydraulic retention time (HRT), dissolved oxygen (DO) and pH were 24 ± 2 , 24 h, less than 4 mg/L and 7 ± 1 respectively. The removal efficiencies of chemical oxygen demand (COD), ammonia nitrogen (NH4+-N) and total nitrogen (TN) attained was 99, 87 and 83% respectively. TN removal rate was limited by nitrification as 97% of ammonia converted into nitrate and nitrite was denitrified. These results show that application of wood chips in wastewater treatment processes is an efficient internal carbon source. 

Keywords: aerobic-anaerobic baffled reactor, denitrification, nitrification, wood chip

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4455 Comparison of Bismuth-Based Nanoparticles as Radiosensitization Agents for Radiotherapy

Authors: Merfat Algethami, Anton Blencowe, Bryce Feltis, Stephen Best, Moshi Geso

Abstract:

Nano-materials with high atomic number atoms have been demonstrated to enhance the effective radiation dose and thus potentially could improve therapeutic efficacy in radiotherapy. The optimal nanoparticulate agents require high X-ray absorption coefficients, low toxicity, and should be cost effective. The focus of our research is the development of a nanoparticle therapeutic agent that can be used in radiotherapy to provide optimal enhancement of the radiation effects on the target. In this study, we used bismuth (Bi) nanoparticles coated with starch and bismuth sulphide nanoparticles (Bi2S3) coated with polyvinylpyrrolidone (PVP). These NPs are of low toxicity and are one of the least expensive heavy metal-based nanoparticles. The aims of this study were to synthesise Bi2S3 and Bi NPs, and examine their cytotoxicity to human lung adenocarcinoma epithelial cells (A549). The dose enhancing effects of NPs on A549 cells were examined at both KV and MV energies. The preliminary results revealed that bismuth based nanoparticles show increased radio-sensitisation of cells, displaying dose enhancement with KV X-ray energies and to a lesser degree for the MV energies. We also observed that Bi NPs generated a greater dose enhancement effect than Bi2S3 NPs in irradiated A549 cells. The maximum Dose Enhancement Factor (DEF) was obtained at lower energy KV range when cells treated with Bi NPs (1.5) compared to the DEF of 1.2 when cells treated with Bi2S3NPs. Less radiation dose enhancement was observed when using high energy MV beam with higher DEF value of Bi NPs treatment (1.26) as compared to 1.06 DEF value with Bi2S3 NPs. The greater dose enhancement was achieved at KV energy range, due the effect of the photoelectric effect which is the dominant process of interaction of X-ray. The cytotoxic effect of Bi NPs on enhancing the X-ray dose was higher due to the higher amount of elemental Bismuth present in Bi NPs compared to Bi2S3 NPs. The results suggest that Bismuth based NPs can be considered as valuable dose enhancing agents when used in clinical applications.

Keywords: A549 lung cancer cells, Bi2S3 nanoparticles, dose enhancement effect, radio-sensitising agents

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4454 Potential Activities of Human Endogenous Retroviral kDNA in Melanoma Pathogenesis and HIV-1 Infection

Authors: Jianli Dong, Fangling Xu, Gengming Huang

Abstract:

Human endogenous retroviral elements (HERVs) comprise approximately 8% of the human genome. They are thought to be germline-integrated genetic remnants of retroviral infections. Although HERV sequences are highly defective, some, especially the K type (HERV-K), have been shown to be expressed and may have biological activities in the pathogenesis of cancer, chronic inflammation and autoimmune diseases. We found that HERV-K GAG and ENV proteins were strongly expressed in pleomorphic melanoma cells. We also detected a critical role of HERV-K ENV in mediating intercellular fusion and colony formation of melanoma cells. Interestingly, we found that levels of HERV-K GAG and ENV expression correlated with the activation of ERK and loss of p16INK4A in melanoma cells, and inhibition of MEK or CDK4, especially in combination, reduced HERV-K expression in melanoma cells. We also performed a reverse transcription-polymerase chain reaction (RT-PCR) assay using DNase I digestion to remove “contaminating” HERV-K genomic DNA and examined HERV-K RNA expression in plasma samples from HIV-1 infected individuals. We found a covariation between HERV-K RNA expression and CD4 cell counts in HIV-1 positive samples. Although a causal link between HERV-K activation and melanoma development, and between HERV-K activation, HIV-1 infection and CD4 cell count have yet to be determined, existing data support the further research efforts in HERV-K.

Keywords: CD4 cell, HERV-K, HIV-1, melanoma

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4453 Effect of the Polymer Modification on the Cytocompatibility of Human and Rat Cells

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

Abstract:

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 osteoblast, rat vascular smooth muscle cells, human stem cells

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4452 Cognitive Model of Analogy Based on Operation of the Brain Cells: Glial, Axons and Neurons

Authors: Ozgu Hafizoglu

Abstract:

Analogy is an essential tool of human cognition that enables connecting diffuse and diverse systems with attributional, deep structural, casual relations that are essential to learning, to innovation in artificial worlds, and to discovery in science. Cognitive Model of Analogy (CMA) leads and creates information pattern transfer within and between domains and disciplines in science. This paper demonstrates the Cognitive Model of Analogy (CMA) as an evolutionary approach to scientific research. The model puts forward the challenges of deep uncertainty about the future, emphasizing the need for flexibility of the system in order to enable reasoning methodology to adapt to changing conditions. In this paper, the model of analogical reasoning is created based on brain cells, their fractal, and operational forms within the system itself. Visualization techniques are used to show correspondences. Distinct phases of the problem-solving processes are divided thusly: encoding, mapping, inference, and response. The system is revealed relevant to brain activation considering each of these phases with an emphasis on achieving a better visualization of the brain cells: glial cells, axons, axon terminals, and neurons, relative to matching conditions of analogical reasoning and relational information. It’s found that encoding, mapping, inference, and response processes in four-term analogical reasoning are corresponding with the fractal and operational forms of brain cells: glial, axons, and neurons.

Keywords: analogy, analogical reasoning, cognitive model, brain and glials

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4451 Experimental Measurements for the Effect of Dilution Procedure in Blood Esterases as Animals Biomarker for Exposure to Organophosphate Compounds

Authors: Kasim Sakran Abass

Abstract:

This main aim of this study was to confirm and extend our current knowledge about the effects of dilutions on esterases activities in the blood for birds with respect to protecting the enzyme from organophosphate inhibition. There were significantly higher esterases activities in dilution 1:10 in all blood samples from quail, duck, and chick compared to other dilutions (1:5, 1:15, 1:20, and 1:25). Furthermore, our results also pointed to the importance of estimating different dilutions effects prior to using in birds as biomarker tools of environmental exposure. Concentration–inhibition curves were determined for the inhibitor in the presence of dilutions 1:5, 1:10 plus 1:15 (to stimulate carboxylesterase). Point estimates (concentrations calculated to produce 20, 50, and 80% inhibition) were compared across conditions and served as a measure of esterase-mediated detoxification. Among the thiol esters (dilution 1:5) was observed to have the highest specificity constant (kcat/Km), and the Km and kcat values were 176 μM and 16,765 s−1, respectively for S-phenyl thioacetate ester, while detected in (dilution 1:15) the lowest specificity constant (kcat/Km), and the Km and kcat values were 943 μM and 1154 s−1, respectively for acetylthiocholine iodide ester.

Keywords: esterase, animal, dilution, pesticides

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4450 Blood Analysis of Diarrheal Calves Using Portable Blood Analyzer: Analysis of Calves by Age

Authors: Kwangman Park, Jinhee Kang, Suhee Kim, Dohyeon Yu, Kyoungseong Choi, Jinho Park

Abstract:

Statement of the Problem: Diarrhea is a major cause of death in young calves. This causes great economic damage to the livestock industry. These diarrhea cause dehydration, decrease blood flow, lower the pH and degrade enzyme function. In the past, serum screening was not possible in the field. However, now with the spread of portable serum testing devices, it is now possible to conduct tests directly on field. Thus, accurate serological changes can be identified and used in the field of large animals. Methodology and Theoretical Orientation: The test groups were calves from 1 to 44 days old. The status of the feces was divided into four grade to determine the severity of diarrhea (grade 0,1,2,3). Grade 0, 1 is considered to have no diarrhea. Grade 2, 3 is considered to diarrhea positive group. One or more viruses were detected in this group. Diarrhea negasitive group consisted of 57 calves (Asan=30, Samrye=27). Diarrhea positive group consisted of 34 calves (Kimje=27, Geochang=7). The feces of all calves were analyzed by PCR Test. Blood sample was measured using an automatic blood analyzer(i-STAT, Abbott inc. Illinois, US). Calves were divided into 3 groups according to age. Group 1 is 1 to 14 days old. Group 2 is 15 to 28 days old. Group 3 is more than 28 days old. Findings: Diarrhea caused an increase in HCT due to dehydration. The difference from normal was highest in 15 to 28 days old (p < 0.01). At all ages, bicarbonate decreased compared to normal, and therefore pH decreased. Similar to HCT, the largest difference was observed between 15 and 28 days (p < 0.01). The pCO₂ decreases to compensate for the decrease in pH. Conclusion and Significance: At all ages, HCT increases, and bicarbonate, pH, and pCO₂ decrease in diarrhea calves. The calf from 15 days to 28 days shows the most difference from normal. Over 28 days of age, weight gain and homeostasis ability increase, diarrhea is seen in the stool, there are fewer hematologic changes than groups below 28 days of age.

Keywords: calves, diarrhea, hematological changes, i-STAT

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4449 Effects of IMUNO-2865® as Immune Supplement for the Aquaculture Industry

Authors: Ivan Zupan, Tomislav Saric, Suzana Tkalcic

Abstract:

IMUNO-2865® is a commercially available, β–glucan based, natural hemicellulose compound with proven immunostimulative properties in people, domestic and some aquatic animals. During the experimental feeding trial with IMUNO-2865® in juvenile wild-caught chub under laboratory conditions, supplementation resulted in overall higher growth performance for all experimental groups regardless of the concentration of the added compound. The maximum, 5% concentration of the supplement, resulted in highest weight gain and calculated specific growth rate. In sea bream, as economically most important species in the Mediterranean aquaculture, significant increases in numbers of monocytes and heterophils were observed in the group supplemented with 2.5 % of IMUNO-2865® in the feed. An overall increase of erythrocytes was noted by the end of the experiment, although with variable distribution among groups. Blood Ca++ levels, total proteins, and total NH₃ were significantly higher after 60 days of feeding in all treatment groups compared to the control and remained elevated in the treated group following the secession of supplementation. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum paraoxonase PON1 (U/L) showed similar trends. All these parameters are playing a significant role in either oxygen supplementation of tissues, or anabolic and catabolic processes that on molecular levels contribute to the overall health and immune-building capacity of cells and tissues. The complete lack of mortality in sea bream and presented increases in cellular, biochemical and oxidative stress parameters in the blood suggest that the IMUNO-2865® represents a safe dietary supplement for in aquaculture, with an overall positive and potentially immunostimulative effect on farmed fish.

Keywords: IMUNO-2865®, β–glucans, Mediterranean aquaculture, fish imunnostimulans

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4448 Lipoic Acid Accelerates Wound Healing by Diminishing Pro-Inflammatory Markers and Chemokine Expression in Rheumatoid Arthritis Mouse Model

Authors: Khairy M. A. Zoheir

Abstract:

One of the most severe complications of Rheumatoid arthritis is delayed recovery. lipoic acid possesses antioxidant, hypoglycemic, and anti-inflammatory activity. In the present study, the effects of lipoic acid was investigated on the key mediators of Rheumatoid arthritis, namely, CD4+CD25+ T cell subsets, GITR expressing cells, CD4+CD25+Foxp3+ regulatory T (Treg) cells, T-helper-17 (Th17) cells, and pro-inflammatory cytokines Interleukin-1β (IL-1β), Interleukin-6 (IL-6) and Tumor Necrosis Factor- α (TNF-α)] through flow-cytometry and qPCR analyses. Lipoic acid treated mice showed a significant decrease in the Rheumatoid arthritis, the frequency of GITR-expressing cells, and Th1 cytokines (IL-17A, TNF-αand Interferon- γ (IFN-γ) compared with positive and negative controlled mice. Lipoic acid treatment also down regulated the mRNA expression of the inflammatory mediators compared with the Rheumatoid arthritis mouse model and untreated mice. The number of Tregs also found to be significantly upregulated in lipoic acid treated mice. Our results were confirmed by the histopathological examination. This study showed the beneficial role of lipoic acid in promoting a well-balanced tool for therapy Rheumatoid arthritis.

Keywords: lipoic acid, chemokines, inflammatory, rheumatoid arthritis

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4447 Study of Mechanical Properties of Large Scale Flexible Silicon Solar Modules on the Various Substrates

Authors: M. Maleczek, Leszek Bogdan, Kazimierz Drabczyk, Agnieszka Iwan

Abstract:

Crystalline silicon (Si) solar cells are the main product in the market among the various photovoltaic technologies concerning such advantages as: material richness, high carrier mobilities, broad spectral absorption range and established technology. However, photovoltaic technology on the stiff substrates are heavier, more fragile and less cost-effective than devices on the flexible substrates to be applied in special applications. The main goal of our work was to incorporate silicon solar cells into various fabric, without any change of the electrical and mechanical parameters of devices. This work is realized for the GEKON project (No. GEKON2/O4/268473/23/2016) sponsored by The National Centre for Research and Development and The National Fund for Environmental Protection and Water Management. In our work, the polyamide or polyester fabrics were used as a flexible substrate in the created devices. Applied fabrics differ in tensile and tear strength. All investigated polyamide fabrics are resistant to weathering and UV, while polyester ones is resistant to ozone, water and ageing. The examined fabrics are tight at 100 cm water per 2 hours. In our work, commercial silicon solar cells with the size 156 × 156 mm were cut into nine parts (called single solar cells) by diamond saw and laser. Gap and edge after cutting of solar cells were checked by transmission electron microscope (TEM) to study morphology and quality of the prepared single solar cells. Modules with the size of 160 × 70 cm (containing about 80 single solar cells) were created and investigated by electrical and mechanical methods. Weight of constructed module is about 1.9 kg. Three types of solar cell architectures such as: -fabric/EVA/Si solar cell/EVA/film for lamination, -backsheet PET/EVA/Si solar cell/EVA/film for lamination, -fabric/EVA/Si solar cell/EVA/tempered glass, were investigated taking into consideration type of fabric and lamination process together with the size of solar cells. In investigated devices EVA, it is ethylene-vinyl acetate, while PET - polyethylene terephthalate. Depend on the lamination process and compatibility of textile with solar cell an efficiency of investigated flexible silicon solar cells was in the range of 9.44-16.64 %. Multi folding and unfolding of flexible module has no impact on its efficiency as was detected by Instron equipment. Power (P) of constructed solar module is 30 W, while voltage about 36 V. Finally, solar panel contains five modules with the polyamide fabric and tempered glass will be produced commercially for different applications (dual use).

Keywords: flexible devices, mechanical properties, silicon solar cells, textiles

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4446 Research on the Role of Platelet Derived Growth Factor Receptor Beta in Promoting Dedifferentiation and Pulmonary Metastasis of Osteosarcoma Under Hypoxic Microenvironment

Authors: Enjie Xu, Zhen Huang, Kunpeng Zhu, Jianping Hu, Xiaolong Ma, Yongjie Wang, Jiazhuang Zhu, Chunlin Zhang

Abstract:

Abstract: Hypoxia and dedifferentiation of osteosarcoma (OS) cells leads to poor prognosis. We plan to identify the role of hypoxia on dedifferentiation and the associated signaling pathways. We performed a sphere formation assay and determined spheroid cells as dedifferentiated cells by detecting stem cell-like markers. RNAi assay was used to explore the expression relationship between hypoxia inducible factor 1 subunit alpha (HIF1A) and platelet derived growth factor receptor beta (PDGFRB). We obtained PDGFRB knockdown and overexpression cells through lentiviral infection experiments and the effects of PDGFRB on cytoskeleton rearrangement and cell adhesion were explored by immunocytochemistry. Wound-healing experiments, transwell assays, and animal trials were employed to investigate the effect of PDGFRB on OS metastasis. Dedifferentiated OS cells were found to exhibit high expression of HIF1A and PDGFRB, and HIF1A promoted the expression of PDGFRB, subsequently activated ras homolog family member A (RhoA), and increased the phosphorylation of myosin light chain (MLC). PDGFRB also enhanced the phosphorylation of focal adhesion kinase (FAK). The OS cell morphology and vinculin distribution were altered by PDGFRB. PDGFRB also promoted cell dedifferentiation and had a significant impact on the metastasis of OS cells both in vitro and in vivo. Our results demonstrated that HIF1A up-regulated PDGFRB under hypoxic conditions, and PDGFRB regulated the actin cytoskeleton by activating RhoA and subsequently phosphorylating MLC, thereby promoting OS dedifferentiation and pulmonary metastasis.

Keywords: osteosarcoma, dedifferentiation, metastasis, cytoskeleton rearrangement, PDGFRB, hypoxia

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4445 A 3d Intestine-On-Chip Model Allows Colonization with Commensal Bacteria to Study Host-Microbiota Interaction

Authors: Michelle Maurer, Antonia Last, Mark S. Gresnigt, Bernhard Hube, Alexander S. Mosig

Abstract:

The intestinal epithelium forms an essential barrier to prevent translocation of microorganisms, toxins or other potentially harmful molecules into the bloodstream. In particular, dendritic cells of the intestinal epithelium orchestrate an adapted response of immune tolerance to commensals and immune defense against invading pathogens. Systemic inflammation is typically associated with a dysregulation of this adapted immune response and is accompanied by a disruption of the epithelial and endothelial gut barrier which enables dissemination of pathogens within the human body. To understand the pathophysiological mechanisms underlying the inflammation-associated gut barrier breakdown, it is crucial to elucidate the complex interplay of the host and the intestinal microbiome. A microfluidically perfused three-dimensional intestine-on-chip model was established to emulate these processes in the presence of immune cells, commensal bacteria, and facultative pathogens. Multi-organ tissue flow (MOTiF) biochips made from polystyrene were used for microfluidic perfusion of the intestinal tissue model. The biochips are composed of two chambers separated by a microporous membrane. Each chamber is connected to inlet and outlet channels allowing independent perfusion of the individual channels and application of microfluidic shear stress. Human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages and intestinal epithelial cells (Caco-2) were assembled on the biochip membrane. Following 7 – 14 days of growth in the presence of physiological flow conditions, the epithelium was colonized with the commensal bacterium Lactobacillus rhamnosus, while the endothelium was perfused with peripheral blood mononuclear cells (PBMCs). Additionally, L. rhamnosus was co-cultivated with the opportunistic fungal pathogen Candida albicans. Within one week of perfusion, the epithelial cells formed self-organized and well-polarized villus- and crypt-like structures that resemble essential morphological characteristics of the human intestine. Dendritic cells were differentiated in the epithelial tissue that specifically responds to bacterial lipopolysaccharide (LPS) challenge. LPS is well-tolerated at the luminal epithelial side of the intestinal model without signs of tissue damage or induction of an inflammatory response, even in the presence of circulating PBMC at the endothelial lining. In contrast, LPS stimulation at the endothelial side of the intestinal model triggered the release of pro-inflammatory cytokines such as TNF, IL-1β, IL-6, and IL-8 via activation of macrophages residing in the endothelium. Perfusion of the endothelium with PBMCs led to an enhanced cytokine release. L. rhamnosus colonization of the model was tolerated in the immune competent tissue model and was demonstrated to reduce damage induced by C. albicans infection. A microfluidic intestine-on-chip model was developed to mimic a systemic infection with a dysregulated immune response under physiological conditions. The model facilitates the colonization of commensal bacteria and co-cultivation with facultative pathogenic microorganisms. Both, commensal bacteria alone and facultative pathogens controlled by commensals, are tolerated by the host and contribute to cell signaling. The human intestine-on-chip model represents a promising tool to mimic microphysiological conditions of the human intestine and paves the way for more detailed in vitro studies of host-microbiota interactions under physiologically relevant conditions.

Keywords: host-microbiota interaction, immune tolerance, microfluidics, organ-on-chip

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4444 Stem Cell Fate Decision Depending on TiO2 Nanotubular Geometry

Authors: Jung Park, Anca Mazare, Klaus Von Der Mark, Patrik Schmuki

Abstract:

In clinical application of TiO2 implants on tooth and hip replacement, migration, adhesion and differentiation of neighboring mesenchymal stem cells onto implant surfaces are critical steps for successful bone regeneration. In a recent decade, accumulated attention has been paid on nanoscale electrochemical surface modifications on TiO2 layer for improving bone-TiO2 surface integration. We generated, on titanium surfaces, self-assembled layers of vertically oriented TiO2 nanotubes with defined diameters between 15 and 100 nm and here we show that mesenchymal stem cells finely sense TiO2 nanotubular geometry and quickly decide their cell fate either to differentiation into osteoblasts or to programmed cell death (apoptosis) on TiO2 nanotube layers. These cell fate decisions are critically dependent on nanotube size differences (15-100nm in diameters) of TiO2 nanotubes sensing by integrin clustering. We further demonstrate that nanoscale topography-sensing is feasible not only in mesenchymal stem cells but rather seems as generalized nanoscale microenvironment-cell interaction mechanism in several cell types composing bone tissue network including osteoblasts, osteoclast, endothelial cells and hematopoietic stem cells. Additionally we discuss the synergistic effect of simultaneous stimulation by nanotube-bound growth factor and nanoscale topographic cues on enhanced bone regeneration.

Keywords: TiO2 nanotube, stem cell fate decision, nano-scale microenvironment, bone regeneration

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4443 The Nature of the Complicated Fabric Textures: How to Represent in Primary Visual Cortex

Authors: J. L. Liu, L. Wang, B. Zhu, J. Zhou, W. D. Gao

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Fabric textures are very common in our daily life. However, we never explore the representation of fabric textures from neuroscience view. Theoretical studies suggest that primary visual cortex (V1) uses a sparse code to efficiently represent natural images. However, how the simple cells in V1 encode the artificial textures is still a mystery. So, here we will take fabric texture as stimulus to study the response of independent component analysis that is established to model the receptive field of simple cells in V1. Experimental results based on 140 classical fabric images indicate that the receptive fields of simple cells have obvious selectivity in orientation, frequency, and phase when drifting gratings are used to determine their tuning properties. Additionally, the distribution of optimal orientation and frequency shows that the patch size selected from each original fabric image has a significant effect on the frequency selectivity.

Keywords: fabric texture, receptive filed, simple cell, spare coding

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4442 Investigation and Estimation of State of Health of Battery Pack in Battery Electric Vehicles-Online Battery Characterization

Authors: Ali Mashayekh, Mahdiye Khorasani, Thomas Weyh

Abstract:

The tendency to use the Battery-Electric vehicle (BEV) for the low and medium driving range or even high driving range has been growing more and more. As a result, higher safety, reliability, and durability of the battery pack as a component of electric vehicles, which has a great share of cost and weight of the final product, are the topics to be considered and investigated. Battery aging can be considered as the predominant factor regarding the reliability and durability of BEV. To better understand the aging process, offline battery characterization has been widely used, which is time-consuming and needs very expensive infrastructures. This paper presents the substitute method for the conventional battery characterization methods, which is based on battery Modular Multilevel Management (BM3). According to this Topology, the battery cells can be drained and charged concerning their capacity, which allows varying battery pack structures. Due to the integration of the power electronics, the output voltage of the battery pack is no longer fixed but can be dynamically adjusted in small steps. In other words, each cell can have three different states, namely series, parallel, and bypass in connection with the neighbor cells. With the help of MATLAB/Simulink and by using the BM3 modules, the battery string model is created. This model allows us to switch two cells with the different SoC as parallel, which results in the internal balancing of the cells. But if the parallel switching lasts just for a couple of ms, we can have a perturbation pulse which can stimulate the cells out of the relaxation phase. With the help of modeling the voltage response pulse of the battery, it would be possible to characterize the cell. The Online EIS method, which is discussed in this paper, can be a robust substitute for the conventional battery characterization methods.

Keywords: battery characterization, SoH estimation, RLS, BEV

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4441 Histone Deacetylases Inhibitor - Valproic Acid Sensitizes Human Melanoma Cells for alkylating agent and PARP inhibitor

Authors: Małgorzata Drzewiecka, Tomasz Śliwiński, Maciej Radek

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The inhibition of histone deacetyles (HDACs) holds promise as a potential anti-cancer therapy because histone and non-histone protein acetylation is frequently disrupted in cancer, leading to cancer initiation and progression. Additionally, histone deacetylase inhibitors (HDACi) such as class I HDAC inhibitor - valproic acid (VPA) have been shown to enhance the effectiveness of DNA-damaging factors, such as cisplatin or radiation. In this study, we found that, using of VPA in combination with talazoparib (BMN-637 – PARP1 inhibitor – PARPi) and/or Dacarabazine (DTIC - alkylating agent) resulted in increased DNA double strand break (DSB) and reduced survival (while not affecting primary melanocytes )and proliferation of melanoma cells. Furthermore, pharmacologic inhibition of class I HDACs sensitizes melanoma cells to apoptosis following exposure to DTIC and BMN-637. In addition, inhibition of HDAC caused sensitization of melanoma cells to dacarbazine and BMN-637 in melanoma xenografts in vivo. At the mRNA and protein level histone deacetylase inhibitor downregulated RAD51 and FANCD2. This study provides that combining HDACi, alkylating agent and PARPi could potentially enhance the treatment of melanoma, which is known for being one of the most aggressive malignant tumors. The findings presented here point to a scenario in which HDAC via enhancing the HR-dependent repair of DSBs created during the processing of DNA lesions, are essential nodes in the resistance of malignant melanoma cells to methylating agent-based therapies.

Keywords: melanoma, hdac, parp inhibitor, valproic acid

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4440 PDMS-Free Microfluidic Chips Fabrication and Utilisation for Pulsed Electric Fields Applications

Authors: Arunas Stirke, Neringa Bakute, Gatis Mozolevskis

Abstract:

A technology of microfluidics is an emerging tool in the field of biology, medicine and chemistry. Microfluidic device is also known as ‘lab-on-a-chip’ technology [1]. In moving from macro- to microscale, there is unprecedented control over spatial and temporal gradients and patterns that cannot be captured in conventional Petri dishes and well plates [2]. However, there is not a single standard microfluidic chip designated for all purposes – every different field of studies needs a specific microchip with certain geometries, inlet/outlet, channel depth and other parameters to precisely regulate the required function. Since our group is studying an effect of pulsed electric field (PEF) to the cells, we have manufactured a microfluidic chip designated for high-throughput electroporation of cells. In our microchip, a cell culture chamber is divided into two parallel channels by a membrane, meanwhile electrodes for electroporation are attached to the wall of the channels. Both microchannels have their own inlet and outlet, enabling injection of transfection material separately. Our perspective is to perform electroporation of mammalian cells in two different ways: (1) plasmid and cells are injected in the same microchannel and (2) injected into separate microchannels. Moreover, oxygen and pH sensors are integrated on order to analyse cell viability parameters after PEF treatment.

Keywords: microfluidics, chip, fabrication, electroporation

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4439 Culture of Human Mesenchymal Stem Cells Culture in Xeno-Free Serum-Free Culture Conditions on Laminin-521

Authors: Halima Albalushi, Mohadese Boroojerdi, Murtadha Alkhabori

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Introduction: Maintenance of stem cell properties during culture necessitates the recreation of the natural cell niche. Studies reported the promising outcome of mesenchymal stem cells (MSC) properties maintenance after using extracellular matrix such as CELLstart™, which is the recommended coating material for stem cells cultured in serum-free and xeno-free conditions. Laminin-521 is known as a crucial adhesion protein, which is found in natural stem cell niche, and plays an important role in facilitating the maintenance of self-renewal, pluripotency, standard morphology, and karyotype of human pluripotent stem cells (PSCs). The aim of this study is to investigate the effects of Laminin-521 on human umbilical cord-derived mesenchymal stem cells (UC-MSC) characteristics as a step toward clinical application. Methods: Human MSC were isolated from the umbilical cord via the explant method. Umbilical cord-derived-MSC were cultured in serum-free and xeno-free conditions in the presence of Laminin-521 for six passages. Cultured cells were evaluated by morphology and expansion index for each passage. Phenotypic characterization of UC-MSCs cultured on Laminin-521 was evaluated by assessment of cell surface markers. Results: Umbilical cord derived-MSCs formed small colonies and expanded as a homogeneous monolayer when cultured on Laminin-521. Umbilical cord derived-MSCs reached confluence after 4 days in culture. No statistically significant difference was detected in all passages when comparing the expansion index of UC-MSCs cultured on LN-521 and CELLstart™. Phenotypic characterization of UC-MSCs cultured on LN-521 using flow cytometry revealed positive expression of CD73, CD90, CD105 and negative expression of CD34, CD45, CD19, CD14 and HLA-DR.Conclusion: Laminin-521 is comparable to CELLstart™ in supporting UC-MSCs expansion and maintaining their characteristics during culture in xeno-free and serum-free culture conditions.

Keywords: mesenchymal stem cells, culture, laminin-521, xeno-free serum-free

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4438 Role of Endonuclease G in Exogenous DNA Stability in HeLa Cells

Authors: Vanja Misic, Mohamed El-Mogy, Yousef Haj-Ahmad

Abstract:

Endonuclease G (EndoG) is a well conserved mitochondrio-nuclear nuclease with dual lethal and vital roles in the cell. The aim of our study was to examine whether EndoG exerts its nuclease activity on exogenous DNA substrates such as plasmid DNA (pDNA), considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus, targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. These findings make it unlikely that targeting of EndoG is a viable strategy for improving the duration and level of transgene expression from non-viral DNA vectors in gene therapy efforts.

Keywords: EndoG, silencing, exogenous DNA stability, HeLa cells

Procedia PDF Downloads 447