Search results for: adipose stem cells
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3709

Search results for: adipose stem cells

3409 The Amount of Organic Phosphates (Like DPG) Existing in Blood is Determining Factor of Mammal’s Bulk

Authors: Ramin Amirmardfar

Abstract:

Throughout Necessary oxygen should be supplied for all cells of a mammal at any moment through blood to make it possible remain alive all cells the mammal’s body. In case a mammal’s bulk is large, there is a farther distance between cells in different tissues and mammals’ heart. Therefore red blood cells in bulky mammal’s body should be capable of conveying oxygen to farther distances. To make it practical, oxygen should be glued red blood cells tenaciously. In other words, cohesion strength of oxygen to red blood cell of bulky mammal’s blood should be much more than the same of small mammal’s blood. In mammal’s bodies, the controlling factor of amount of cohesion of oxygen to red blood cell, are organic phosphates (like DPG). The less DPG in red blood cells of a mammal, the more cohesion of oxygen to red blood cell (at the same rate). As much as oxygen is glued more tenacious to red blood cells, oxygen could been carried to farther distance and as much as oxygen could be conveyed to farther points of heart, bulk of mammal could be larger at the same rate.

Keywords: mammals size, animals size, organic phosphates, DPG, red blood cell, metabolism

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3408 Immunomodulatory Effects of Multipotent Mesenchymal Stromal Cells on T-Cell Populations at Tissue-Related Oxygen Level

Authors: A. N. Gornostaeva, P. I. Bobyleva, E. R. Andreeva, L. B. Buravkova

Abstract:

Multipotent mesenchymal stromal cells (MSCs) possess immunomodulatory properties. The effect of MSCs on the crucial cellular immunity compartment – T-cells is of a special interest. It is known that MSC tissue niche and expected milieu of their interaction with T- cells are characterized by low oxygen concentration, whereas the in vitro experiments usually are carried out at a much higher ambient oxygen (20%). We firstly evaluated immunomodulatory effects of MSCs on T-cells at tissue-related oxygen (5%) after interaction implied cell-to-cell contacts and paracrine factors only. It turned out that MSCs under reduced oxygen can effectively suppress the activation and proliferation of PHA-stimulated T-cells and can provoke decrease in the production of proinflammatory and increase in anti-inflammatory cytokines. In hypoxia some effects were amplified (inhibition of proliferation, anti-inflammatory cytokine profile shift). This impact was more evident after direct cell-to-cell interaction; lack of intercellular contacts could revoke the potentiating effect of hypoxia.

Keywords: MSCs, T-cells, activation, low oxygen, cell-to-cell interaction, immunosuppression

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3407 The Inhibitory Effect of Weissella koreensis 521 Isolated from Kimchi on 3T3-L1 Adipocyte Differentiation

Authors: Kyungbae Pi, Kibeom Lee, Yongil Kim, Eun-Jung Lee

Abstract:

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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3406 The Impact of Academic Support Practices on Two-Year College Students’ Achievement in Science, Technology, Engineering, and Math Education: An Exploration of Factors

Authors: Gisele Ragusa, Lilian Leung

Abstract:

There are essential needs for science, technology, engineering, and math (STEM) workforces nationally. This important need underscores the necessity of increasing numbers of students attending both two-year community colleges and universities, thereby enabling and supporting a larger pool of students to enter the workforce. The greatest number of students in STEM programs attend public higher education institutions, with an even larger majority beginning their academic experiences enrolled in two-year public colleges. Accordingly, this research explores the impact of experiences and academic support practices on two-year (community) college students’ academic achievement in STEM majors with a focus on supporting students who are the first in their families to attend college. This research is a result of three years of iterative trials of differing supports to improve such students’ academic success with a cross-student comparative research methodological structure involving peer-to-peer and faculty academic supports. Results of this research indicate that background experiences and a combination of peer-to-peer and faculty-led academic support practices, including supplementary instruction, peer mentoring, and study skills support, significantly improve students’ academic success in STEM majors. These results confirm the needs that first-generation students have in navigating their college careers and what can be effective in supporting them.

Keywords: higher education policy, student support, two-year colleges, STEM achievement

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3405 Perception Towards Using E-learning with Stem Students Whose Programs Require Them to Attend Practical Sections in Laboratories during Covid-19

Authors: Youssef A. Yakoub, Ramy M. Shaaban

Abstract:

Covid-19 has changed and affected the whole world dramatically in a new way that the entire world, even scientists, have not imagined before. The educational institutions around the world have been fighting since Covid-19 hit the world last December to keep the educational process unchanged for all students. E-learning was a must for almost all US universities during the pandemic. It was specifically more challenging to use eLearning instead of regular classes among students who take practical education. The aim of this study is to examine the perception of STEM students towards using eLearning instead of traditional methods during their practical study. Focus groups of STEM students studying at a western Pennsylavian, mid-size university were interviewed. Semi-structured interviews were designed to get an insight on students’ perception towards the alternative educational methods they used in the past seven months. Using convenient sampling, four students were chosen from different STEM fields: science of physics, technology, electrical engineering, and mathematics. The interview was primarily about the extent to which these students were satisfied, and their educational needs were met through distance education during the pandemic. The interviewed students were generally able to do a satisfactory performance during their virtual classes, but they were not satisfied enough with the learning methods. The main challenges they faced included the inability to have real practical experience, insufficient materials posted by the faculty, and some technical problems associated with their study. However, they reported they were satisfied with the simulation programs they had. They reported these simulations provided them with a good alternative to their traditional practical education. In conclusion, this study highlighted the challenges students face during the pandemic. It also highlighted the various learning tools students see as good alternatives to their traditional education.

Keywords: eLearning, STEM education, COVID-19 crisis, online practical training

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3404 Analysis Influence Variation Frequency on Characterization of Nano-Particles in Preteatment Bioetanol Oil Palm Stem (Elaeis guineensis JACQ) Use Sonication Method with Alkaline Peroxide Activators on Improvement of Celullose

Authors: Luristya Nur Mahfut, Nada Mawarda Rilek, Ameiga Cautsarina Putri, Mujaroh Khotimah

Abstract:

The use of bioetanol from lignocellulosic material has begone to be developed. In Indonesia the most abundant lignocellulosic material is stem of palm which contain 32.22% of cellulose. Indonesia produces approximatelly 300.375.000 tons of stem of palm each year. To produce bioetanol from lignocellulosic material, the first process is pretreatment. But, until now the method of lignocellulosic pretretament is uneffective. This is related to the particle size and the method of pretreatment of less than optimal so that led to an overhaul of the lignin insufficient, consequently increased levels of cellulose was not significant resulting in low yield of bioetanol. To solve the problem, this research was implemented by using the process of pretreatment method ultasonifikasi in order to produce higher pulp with nano-sized particles that will obtain higher of yield ethanol from stem of palm. Research methods used in this research is the RAK that is composed of one factor which is the frequency ultrasonic waves with three varians, they are 30 kHz, 40 kHz, 50 kHz, and use constant variable is concentration of NaOH. The analysis conducted in this research is the influence of the frequency of the wave to increase levels of cellulose and change size on the scale of nanometers on pretreatment process by using the PSA methods (Particle Size Analyzer), and a Cheason. For the analysis of the results, data, and best treatment using ANOVA and test BNT with confidence interval 5%. The best treatment was obtained by combination X3 (frequency of sonication 50 kHz) and lignin (19,6%) cellulose (59,49%) and hemicellulose (11,8%) with particle size 385,2nm (18,8%).

Keywords: bioethanol, pretreatment, stem of palm, cellulosa

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3403 Effects of Excess-Iron Stress on Symbiotic Nitrogen Fixation Efficiency of Yardlong-Bean Plants

Authors: Hong Li, Tingxian Li, Xudong Wang, Qinghuo Lin

Abstract:

Excess-iron (Fe) stresses involved in legume symbiotic nitrogen fixation are not understood. Our objectives were to investigate the tolerance of yardlong-bean plants to soil excess-Fe stress and antagonistic effects of organic amendments and rhizobial inoculants on plant root nodulation and stem ureide formation. The study was conducted in the tropical Hainan Island during 2012-2013. The soil was strongly acidic (pH 5.3±0.4) and highly variable in Fe concentrations(596±79 mg/kg). The treatments were arranged in a split-plot design with three blocks. The treatment effects were significant on root nodulation, stem ureide, amino acids, plant N/Fe accumulation and bean yields (P<0.05). The yardlong-bean stem allantoin, amino acids and nitrate concentrations and relative ureide % declined with high soil Fe concentrations (>300 mg/kg). It was concluded that the co-variance of excess Fe stress could inhibit legume symbiotic N fixation efficiency. Organic amendments and rhizobial inoculants could help improve crop tolerance to excess Fe stress.

Keywords: atmospheric N fixation, root nodulation, soil Fe co-variance, stem ureide, yardlong-bean plants

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3402 Hydrogel Hybridizing Temperature-Cured Dissolvable Gelatin Microspheres as Non-Anchorage Dependent Cell Carriers for Tissue Engineering Applications

Authors: Dong-An Wang

Abstract:

All kinds of microspheres have been extensively employed as carriers for drug, gene and therapeutic cell delivery. Most therapeutic cell delivery microspheres rely on a two-step methodology: fabrication of microspheres and subsequent seeding of cells onto them. In this study, we have developed a novel one-step cell encapsulation technique using a convenient and instant water-in-oil single emulsion approach to form cell-encapsulated gelatin microspheres. This technology is adopted for hyaline cartilage tissue engineering, in which autologous chondrocytes are used as therapeutic cells. Cell viability was maintained throughout and after the microsphere formation (75-100 µm diameters) process that avoids involvement of any covalent bonding reactions or exposure to any further chemicals. Further encapsulation of cell-laden microspheres in alginate gels were performed under 4°C via a prompt process. Upon the formation of alginate constructs, they were immediately relocated into CO2 incubator where the temperature was maintained at 37°C; under this temperature, the cell-laden gelatin microspheres dissolved within hours to yield similarly sized cavities and the chondrocytes were therefore suspended within the cavities inside the alginate gel bulk. Hence, the gelatin cell-laden microspheres served two roles: as cell delivery vehicles which can be removable through temperature curing, and as porogens within an alginate hydrogel construct to provide living space for cell growth and tissue development as well as better permeability for mutual diffusions. These cell-laden microspheres, namely “temperature-cured dissolvable gelatin microsphere based cell carriers” (tDGMCs), were further encapsulated in a chondrocyte-laden alginate scaffold system and analyzed by WST-1, gene expression analyses, biochemical assays, histology and immunochemistry stains. The positive results consistently demonstrated the promise of tDGMC technology in delivering these non-anchorage dependent cells (chondrocytes). It can be further conveniently translated into delivery of other non-anchorage dependent cell species, including stem cells, progenitors or iPS cells, for regeneration of tissues in internal organs, such as engineered hepatogenesis or pancreatic regeneration.

Keywords: biomaterials, tissue engineering, microsphere, hydrogel, porogen, anchorage dependence

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3401 Biological Activity of Mesenchymal Stem Cells in the Surface of Implants

Authors: Saimir Heta, Ilma Robo, Dhimiter Papakozma, Eduart Kapaj, Vera Ostreni

Abstract:

Introduction: The biocompatible materials applied to the implant surfaces are the target of recent literature studies. Methodologies: Modification of implant surfaces in different ways such as application of additional ions, surface microstructure change, surface or laser ultrasound alteration, or application of various substances such as recombinant proteins are among the most affected by articles published in the literature. The study is of review type with the main aim of finding the different ways that the mesenchymal cell reaction to these materials is, according to the literature, in the same percentage positive to the osteointegration process. Results: It is emphasized in the literature that implant success as a key evaluation key has more to implement implant treatment protocol ranging from dental health amenity and subsequent of the choice of implant type depending on the alveolar shape of the ridge level. Conclusions: Osteointegration is a procedure that should initially be physiologically independent of the type of implant pile material. With this physiological process, it can not "boast" for implant success or implantation depending on the brand of the selected implant, as the breadth of synthetic or natural materials that promote osteointegration is relatively large.

Keywords: mesenchymal cells, implants, review, biocompatible materials

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3400 [Keynote Speech]: Guiding Teachers to Make Lessons Relevant, Appealing, and Personal (RAP) for Academically-Low-Achieving Students in STEM Subjects

Authors: Nazir Amir

Abstract:

Teaching approaches to present science and mathematics content amongst academically-low-achieving students may need to be different than approaches that are adopted for the more academically-inclined students, primarily due to the different learning needs and learning styles of these students. In crafting out lessons to motivate and engage these students, teachers need to consider the backgrounds of these students and have a good understanding of their interests so that lessons can be presented in ways that appeal to them, and made relevant not just to the world around them, but also to their personal experiences. This presentation highlights how the author worked with a Professional Learning Community (PLC) of teachers in crafting out fun and feasible classroom teaching approaches to present science and mathematics content in ways that are made Relevant, Appealing, and Personal (RAP) to groups of academically-low-achieving students in Singapore. Feedback from the students and observations from their work suggest that they were engaged through the RAP-modes of instruction, and were able to appreciate the role of science and mathematics through a variety of low-cost design-based STEM (Science, Technology, Engineering, and Mathematics) activities. Such results imply that teachers teaching academically-low-achieving students, and those in under-resourced communities, could consider infusing RAP-infused instructions into their lessons in getting students develop positive attitudes towards STEM subjects.

Keywords: STEM Education, STEAM Education, Curriculum Instruction, Academically At-Risk Students, Singapore

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3399 Seed Priming Winter Wheat (Triticum aestivum L.) for Germination and Emergence

Authors: Pakize Ozlem Kurt Polat, Gizem Metin, Koksal Yagdi

Abstract:

In order to evaluate the effect of the different sources of salt on germination and early growth of five wheat cultivars (Katea, Bezostaja, Koksal-2000, Golia, Pehlivan) an experiment was conducted at the seed laboratory of the Uludag University, Agricultural Faculty, Department of Field Crops in Bursa/Turkey. Seeds were applied in five different resources media (KCl % 2, KCl %4, KNO₃ %0,5, KH₂PO₄ %0,5, PEG %10) and distilled water as the control). The seed was fully immersed in priming media at a temperature of 24ᵒC for durations of 12 and 24hours. Six different agronomic characters (seed germination, stem length, stem weight, radicle length, fresh weight, dry weight) were measured in 7th days and 14th days. Maximum seed germination percentage of seven days are Pehlivan was observed when the seeds were applied by KH₂PO₄ and Katea by distilled water as a control. The most stem length and stem weight were obtained for seeds were applied by KH₂PO₄ %0,5 with Katea and Bezostja immersed in priming media at 12h intervals beginning 7d after planting. Seeds were applied KH₂PO₄ %0,5 media produced maximum radicle length by Koksal and dry weight by Katea. The freshest weight obtains in Katea by KNO₃ %0,5 immersed in priming media at 24h. The most germination percent, dry weight, stem length of fourteen days was observed in Katea which subjected to KH₂PO₄ %0,5 solution. The most radicle length was observed Katea and Koksal in media of KH₂PO₄ %0,5. The most stem length was obtained for seeds were applied by KH₂PO₄ %0,5 and KNO₃ with Katea and Bezostaja. When the applied chemicals and all days examined KH₂PO₄ %0,5 treatment in fourteen days and immersed for the duration of 24 hours had better effects than other medias, seven days treatments and 12hours immersed. As a result of this research, the best response of media for the wheat germination can be said that the KH₂PO₄ %0,5 immersed in priming media at 24h intervals beginning 14 days after planting.

Keywords: germination, priming, seedling growth, wheat

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3398 Single Cell Rna Sequencing Operating from Benchside to Bedside: An Interesting Entry into Translational Genomics

Authors: Leo Nnamdi Ozurumba-Dwight

Abstract:

Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.

Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq

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3397 SEM Detection of Folate Receptor in a Murine Breast Cancer Model Using Secondary Antibody-Conjugated, Gold-Coated Magnetite Nanoparticles

Authors: Yasser A. Ahmed, Juleen M Dickson, Evan S. Krystofiak, Julie A. Oliver

Abstract:

Cancer cells urgently need folate to support their rapid division. Folate receptors (FR) are over-expressed on a wide range of tumor cells, including breast cancer cells. FR are distributed over the entire surface of cancer cells, but are polarized to the apical surface of normal cells. Targeting of cancer cells using specific surface molecules such as folate receptors may be one of the strategies used to kill cancer cells without hurting the neighing normal cells. The aim of the current study was to try a method of SEM detecting FR in a murine breast cancer cell model (4T1 cells) using secondary antibody conjugated to gold or gold-coated magnetite nanoparticles. 4T1 cells were suspended in RPMI medium witth FR antibody and incubated with secondary antibody for fluorescence microscopy. The cells were cultured on 30mm Thermanox coverslips for 18 hours, labeled with FR antibody then incubated with secondary antibody conjugated to gold or gold-coated magnetite nanoparticles and processed to scanning electron microscopy (SEM) analysis. The fluorescence microscopy study showed strong punctate FR expression on 4T1 cell membrane. With SEM, the labeling with gold or gold-coated magnetite conjugates showed a similar pattern. Specific labeling occurred in nanoparticle clusters, which are clearly visualized in backscattered electron images. The 4T1 tumor cell model may be useful for the development of FR-targeted tumor therapy using gold-coated magnetite nano-particles.

Keywords: cancer cell, nanoparticles, cell culture, SEM

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3396 Quantitative Analysis of (+)-Catechin and (-)-Epicatechin in Pentace burmanica Stem Bark by HPLC

Authors: Thidarat Duangyod, Chanida Palanuvej, Nijsiri Ruangrungsi

Abstract:

Pentace burmanica Kurz., belonging to the Malvaceae family, is commonly used for anti-diarrhea in Thai traditional medicine. A method for quantification of (+)-catechin and (-)-epicatechin in P. burmanica stem bark from 12 different Thailand markets by reverse-phase high performance liquid chromatography (HPLC) was investigated and validated. The analysis was performed by a Shimadzu DGU-20A3 HPLC equipped with a Shimadzu SPD-M20A photo diode array detector. The separation was accomplished with an Inersil ODS-3 column (5 µm x 4.6 x 250 mm) using 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B) as mobile phase at the flow rate of 1 ml/min. The isocratic was set at 20% B for 15 min and the column temperature was maintained at 40 ºC. The detection was at the wavelength of 280 nm. Both (+)-catechin and (-)-epicatechin existed in the ethanolic extract of P. burmanica stem bark. The content of (-)-epicatechin was found as 59.74 ± 1.69 µg/mg of crude extract. In contrast, the quantitation of (+)-catechin content was omitted because of its small amount. The method was linear over a range of 5-200 µg/ml with good coefficients (r2 > 0.99) for (+)-catechin and (-)-epicatechin. Limit of detection values were found to be 4.80 µg/ml for (+)-catechin and 5.14 µg/ml for (-)-epicatechin. Limit of quantitation of (+)-catechin and (-)-epicatechin were of 14.54 µg/ml and 15.57 µg/ml respectively. Good repeatability and intermediate precision (%RSD < 3) were found in this study. The average recoveries of both (+)-catechin and (-)-epicatechin were obtained with good recovery in the range of 91.11 – 97.02% and 88.53 – 93.78%, respectively, with the %RSD less than 2. The peak purity indices of catechins were more than 0.99. The results suggested that HPLC method proved to be precise and accurate and the method can be conveniently used for (+)-catechin and (-)-epicatechin determination in ethanolic extract of P. burmanica stem bark. Moreover, the stem bark of P. burmanica was found to be a rich source of (-)-epicatechin.

Keywords: pentace burmanica, (+)-catechin, (-)-epicatechin, high performance liquid chromatography

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3395 Effect of Dietary Supplementation of Allium Hookeri Root and Processed Sulfur on the Growth Performance of Guinea Pigs

Authors: Nayeon, Lee, Won-Young, Cho, Hyun Joo, Jang, Chi-Ho, Lee

Abstract:

This study investigated the effects of the dietary supplementation of the Allium hookeri root, and processed sulfur, on the growth performance of guinea pigs. The guinea pigs were fed a control diet (CON), as well as the control diet including 1% freeze-dried Allium hookeri root (AH), or 0.1% processed sulfur (S), or including both the freeze-dried Allium hookeri root and the processed sulfur (AHS). The weight of perirenal adipose tissue (PAT) and the epididymal adipose tissue (EAT) in the AH were significantly lower than CON (p < 0.05). The serum cholesterols levels of the AH and the AHS were significantly lower than the S (p < 0.05). While the total saturated fatty acid content in the serum of the AH and AHS groups showed a tendency to decrease, the total monounsaturated fatty acid increased. The results of this study suggested that dietary consumption of Allium hookeri root may help to decrease fat accumulation, lower serum cholesterol levels, and control serum free fatty acid contents in the guinea pigs.

Keywords: Allium hookeri, dietary supplementation, growth performance, processed sulfur, Guinea pig

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3394 STEAM and Project-Based Learning: Equipping Young Women with 21st Century Skills

Authors: Sonia Saddiqui, Maya Marcus

Abstract:

UTS STEAMpunk Girls is an educational program for young women (aged 12-16), to empower them to be more informed and active members of the 21st century workforce. With the number of STEM graduates on the decline, especially among young women, an additional aim of the program is to trial a STEAM (Science, Technology, Engineering, Arts/Humanities/Social Sciences, Mathematics), inter-disciplinary approach to improving STEM engagement. In-line with UNESCO’s recent focus on promoting ‘transversal competencies’ in future graduates, the program utilised co-design, project-based learning, entrepreneurial processes, and inter-disciplinary learning. The program consists of two phases. Taking a participatory design approach, the first phase (co-design workshops) provided valuable insight into student perspectives around engaging young women in STEM and inter-disciplinary thinking. The workshops positioned 26 young women from three schools as subject matter experts (SMEs), providing a platform for them to share their opinions, experiences and findings around the STEAM disciplines. The second (pilot) phase put the co-design phase findings into practice, with 64 students from four schools working in groups to articulate problems with real-world implications, and utilising design-thinking to solve them. The pilot phase utilised project-based learning to engage young women in entrepreneurial and STEAM frameworks and processes. Scalable program design and educational resources were trialed to determine appropriate mechanisms for engaging young women in STEM and in STEAM thinking. Across both phases, data was collected via longitudinal surveys to obtain pre-program, baseline attitudinal information, and compare that against post-program responses. Preliminary findings revealed students’ improved understanding of the STEM disciplines, industries and professions, improved awareness of STEAM as a concept, and improved understanding regarding inter-disciplinary and design thinking. Program outcomes will be of interest to high-school educators in both STEM and the Arts, Humanities and Social Sciences fields, and will hopefully inform future programmatic approaches to introducing inter-disciplinary STEAM learning in STEM curriculum.

Keywords: co-design, STEM, STEAM, project-based learning, inter-disciplinary

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3393 Natural Honey and Effect on the Activity of the Cells

Authors: Abujnah Dukali

Abstract:

Natural honey was assessed in cell culture system for its anticancer activity. Human leukemic cell line HL 60 was treated with honey and cultured for 5 days and cytotoxicity was calculated by MTT assay. Honey showed cytotoxicity with CC50 value of 174.20 µg/ml. Radical modulation activities was assessed by lipid peroxidation assay using egg lecithin. Honey showed antioxidant activity with EC50 value of 159.73 µg/ml. In addition, treatment with HL60 cells also resulted in nuclear DNA fragmentation, as seen in agarose gel electrophoresis. This is a hallmark of cells undergoing apoptosis. Confirmation of apoptosis was performed by staining the cells with Annexin V and FACS analysis. Apoptosis is an active, genetically regulated disassembly of the cell form within. Disassembly creates changes in the phospholipid content of the cytoplasmic membrane outer leaflet. Phosphatidylserine (PS) is translocated from the inner to the outer surface of the cell for phagocytic cell recognition. The human anticoagulant, annexin V, is a Ca2+-dependent phospholipid protein with a high affinity for PS. Annexin V labeled with fluorescein can identify apoptotic cells in the population It is a confirmatory test for apoptosis. Annexin V-positive cells were defined as apoptotic cells. Since honey shows both antioxidant activity and cytotoxicity at almost the same concentration, it can prevent the free radical induced cancer as prophylactic agent and kill the cancer cells by apoptotic process as a chemotherapeutic agent. Everyday intake of honey can prevent the cancer induction.

Keywords: anticancer, cells, DNA, honey

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3392 U11 Functionalised Luminescent Gold Nanoclusters for Pancreatic Tumor Cells Labelling

Authors: Regina M. Chiechio, Rémi Leguevél, Helene Solhi, Marie Madeleine Gueguen, Stephanie Dutertre, Xavier, Jean-Pierre Bazureau, Olivier Mignen, Pascale Even-Hernandez, Paolo Musumeci, Maria Jose Lo Faro, Valerie Marchi

Abstract:

Thanks to their ultra-small size, high electron density, and low toxicity, gold nanoclusters (Au NCs) have unique photoelectrochemical and luminescence properties that make them very interesting for diagnosis bio-imaging and theranostics. These applications require control of their delivery and interaction with cells; for this reason, the surface chemistry of Au NCs is essential to determine their interaction with the targeted biological objects. Here we demonstrate their ability as markers of pancreatic tumor cells. By functionalizing the surface of the NCs with a recognition peptite (U11), the nanostructures are able to preferentially bind to pancreatic cancer cells via a receptor (uPAR) overexpressed by these cells. Furthermore, the NCs can mark even the nucleus without the need of fixing the cells. These nanostructures can therefore be used as a non-toxic, multivalent luminescent platform, capable of selectively recognizing tumor cells for bioimaging, drug delivery, and radiosensitization.

Keywords: gold nanoclusters, luminescence, biomarkers, pancreatic cancer, biomedical applications, bioimaging, fluorescent probes, drug delivery

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3391 Molecular Pathogenesis of NASH through the Dysregulation of Metabolic Organ Network in the NASH-HCC Model Mouse Treated with Streptozotocin-High Fat Diet

Authors: Bui Phuong Linh, Yuki Sakakibara, Ryuto Tanaka, Elizabeth H. Pigney, Taishi Hashiguchi

Abstract:

NASH is an increasingly prevalent chronic liver disease that can progress to hepatocellular carcinoma and now is attracting interest worldwide. The STAM™ model is a clinically-correlated murine NASH model which shows the same pathological progression as NASH patients and has been widely used for pharmacological and basic research. The multiple parallel hits hypothesis suggests abnormalities in adipocytokines, intestinal microflora, and endotoxins are intertwined and could contribute to the development of NASH. In fact, NASH patients often exhibit gut dysbiosis and dysfunction in adipose tissue and metabolism. However, the analysis of the STAM™ model has only focused on the liver. To clarify whether the STAM™ model can also mimic multiple pathways of NASH progression, we analyzed the organ crosstalk interactions between the liver and the gut and the phenotype of adipose tissue in the STAM™ model. NASH was induced in male mice by a single subcutaneous injection of 200 µg streptozotocin 2 days after birth and feeding with high-fat diet after 4 weeks of age. The mice were sacrificed at NASH stage. Colon samples were snap-frozen in liquid nitrogen and stored at -80˚C for tight junction-related protein analysis. Adipose tissue was prepared into paraffin blocks for HE staining. Blood adiponectin was analyzed to confirm changes in the adipocytokine profile. Tight junction-related proteins in the intestine showed that expression of ZO-1 decreased with the progression of the disease. Increased expression of endotoxin in the blood and decreased expression of Adiponectin were also observed. HE staining revealed hypertrophy of adipocytes. Decreased expression of ZO-1 in the intestine of STAM™ mice suggests the occurrence of leaky gut, and abnormalities in adipocytokine secretion were also observed. Together with the liver, phenotypes in these organs are highly similar to human NASH patients and might be involved in the pathogenesis of NASH.

Keywords: Non-alcoholic steatohepatitis, hepatocellular carcinoma, fibrosis, organ crosstalk, leaky gut

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3390 Profiling, Antibacterial and Antioxidant Activity of Acacia decurrens (Willd) an Invasive South Africa Tree

Authors: Joe Modise, Bamidel Joseph Okoli, Nas Molefe, Imelda Ledwaba

Abstract:

The present study describes the chemical profile and antioxidant potential of the stem bark of Acacia decurrens. The methanol fraction of A. decurrens stem bark gave the highest yield (20 %), while the hexane fraction had the lowest yield (0.2 %). The GC-MS spectra of the hexane, chloroform and ethyl acetate fractions confirm the presence of fifty two major compounds and the ICP-OES analysis of the stem bark was found to contain Co(0.41), Zn(1.75), Mn(3.69), Ca(8.67), Ni(10.54), Mg(12.98), Cr(24.38), K(47.88), Fe(154.62) ppm; which is an indication of hyper-accumulation capacity. The UV-Visible spectra of showed four absorption maxima for hexane fraction at 665 (0.028), 410 (0.116), 335 (0.278) and 250 (0.007) nm, three for chloroform fraction at 665 (0.028), 335 (0.278) and 250 (0.007) nm , three for ethyl acetate fraction at 665 (0.070), 390 (0.648) and 345 (0.663) nm and three for methanol fraction at 385 (0.508), 310 (0.886) and 295 (0.899) nm respectively. Quantitative phytochemical screening indicated that the alkaloid (0.6-3.3) % and saponins (5.1-8.6) % contents of the various fractions were significantly lower than the tannin (30.9-55.8) mg TAE/g, steroid(13.92-41.2) %, phenol (40.6-65.5) mgGAE/g and flavonoids (210.2 -284.9) mg RUE/g contents. The antioxidant activity of the fractions was analysed by different methods and revealed good to moderate antioxidant potential with different IC50 values viz. (42.2-49.6) mg/mL for ABTS and (37.8-75.0) μg/ml for DPPH respectively, compared to standard antioxidants. Based on obtained results, the A.decurrens stem bark fractions can be a source of safe, sustainable natural antioxidant drug and can be exploited as a source of controlled green-heavy metal cleaner.

Keywords: Acacia decurrens, antioxidant, DPPH, ABTS, hyperaccumulation, Menstruum, ICP-OES, GC-MS, UV/visible

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3389 Comparison of Phenolic and Urushiol Contents of Different Parts of Rhus verniciflua and Their Antimicrobial Activity

Authors: Jae Young Jang, Jong Hoon Ahn, Jae-Woong Lim, So Young Kang, Mi Kyeong Lee

Abstract:

Rhus verniciflua is commonly known as a lacquer tree in Korea. Stem barks of R. verniciflua have been used as an immunostimulator in traditional medicine. It contains phenolic compounds and is known for diverse biological activities such as antioxidant and antimicrobial activity. However, it also causes allergic dermatitis due to urushiols derivatives. For the development of active natural resources with less toxicity, the content of phenolic compounds and urushiols of different parts of R. verniciflua such as stem barks, lignum and leaves were quantitated by colorimetric assay and HPLC analysis. The urushiols content were the highest in stem barks, and followed by leaves. The lignum contained trace amount of urushiols. The phenolic contents, however, were the most abundant in lignum, and followed by leaves and stem barks. These results clear showed that the content of urushiols and phenolic differs depending on the parts of R. verniciflua. Antimicrobial activity of different parts of R. verniciflua against fish pathogenic bacteria was also investigated using Edwardsiella tarda. Lignum of R. verniciflua was the most effective in antimicrobial activity against E. tarda and phenolic constituents are suggested to be active constituents for activity. Taken together, phenolic compounds are responsible for antimicrobial activity of R. verniciflua. The lignum of R. verniciflua contains high content of phenolic compounds with less urushiols, which suggests efficient antimicrobial activity with less toxicity. Therefore, lignum of R. verniciflua are suggested as good sources for antimicrobial activity against fish bacterial diseases.

Keywords: different parts, phenolic compounds, Rhus verniciflua, urushiols

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3388 In vivo Protective Effects of Ginger Extract on Cyclophosphamide Induced Chromosomal Aberrations in Bone Marrow Cells of Swiss Mice

Authors: K. Yadamma, K. Rudrama Devi

Abstract:

The protective effect of Ginger Extract against cyclophosphamide induced cytotoxicity was evaluated in in vivo animal model using analysis of chromosomal aberrations in somatic cells of mice. Three doses of Ginger Extract (150mg/kg, 200mg/kg, and 250mg/kg body weight) were selected for modulation and given to animals after priming. The animals were sacrificed 24, 48, 72 hrs after the treatment and slides were prepared for the incidence of chromosomal aberrations in bone marrow cells of mice. When animals were treated with cyclophosphamide 50mg/kg, showed cytogenetic damage in somatic cells. However, a significant decrease was observed in the percentage of chromosomal aberrations when animals were primed with various doses of Ginger Extract. The present results clearly indicate the protective nature of Ginger Extract against cyclophosphamide induced genetic damage in mouse bone marrow cells.

Keywords: ginger extract, protection, bone marrow cells, swiss albino mice

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3387 Learning Instructional Managements between the Problem-Based Learning and Stem Education Methods for Enhancing Students Learning Achievements and their Science Attitudes toward Physics the 12th Grade Level

Authors: Achirawatt Tungsombatsanti, Toansakul Santiboon, Kamon Ponkham

Abstract:

Strategies of the STEM education was aimed to prepare of an interdisciplinary and applied approach for the instructional of science, technology, engineering, and mathematics in an integrated students for enhancing engagement of their science skills to the Problem-Based Learning (PBL) method in Borabu School with a sample consists of 80 students in 2 classes at the 12th grade level of their learning achievements on electromagnetic issue. Research administrations were to separate on two different instructional model groups, the 40-experimental group was designed with the STEM instructional experimenting preparation and induction in a 40-student class and the controlling group using the PBL was designed to students identify what they already know, what they need to know, and how and where to access new information that may lead to the resolution of the problem in other class. The learning environment perceptions were obtained using the 35-item Physics Laboratory Environment Inventory (PLEI). Students’ creating attitude skills’ sustainable development toward physics were assessed with the Test Of Physics-Related Attitude (TOPRA) The term scaling was applied to the attempts to measure the attitude objectively with the TOPRA was used to assess students’ perceptions of their science attitude toward physics. Comparisons between pretest and posttest techniques were assessed students’ learning achievements on each their outcomes from each instructional model, differently. The results of these findings revealed that the efficiency of the PLB and the STEM based on criteria indicate that are higher than the standard level of the 80/80. Statistically, significant of students’ learning achievements to their later outcomes on the controlling and experimental physics class groups with the PLB and the STEM instructional designs were differentiated between groups at the .05 level, evidently. Comparisons between the averages mean scores of students’ responses to their instructional activities in the STEM education method are higher than the average mean scores of the PLB model. Associations between students’ perceptions of their physics classes to their attitudes toward physics, the predictive efficiency R2 values indicate that 77%, and 83% of the variances in students’ attitudes for the PLEI and the TOPRA in physics environment classes were attributable to their perceptions of their physics PLB and the STEM instructional design classes, consequently. An important of these findings was contributed to student understanding of scientific concepts, attitudes, and skills as evidence with STEM instructional ought to higher responding than PBL educational teaching. Statistically significant between students’ learning achievements were differentiated of pre and post assessments which overall on two instructional models.

Keywords: learning instructional managements, problem-based learning, STEM education, method, enhancement, students learning achievements, science attitude, physics classes

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3386 Erythrophagocytic Role of Mast Cells in vitro and in vivo during Oxidative Stress

Authors: Priyanka Sharma, Niti Puri

Abstract:

Anemia develops when blood lacks enough healthy erythrocytes. Past studies indicated that anemia, inflammatory process, and oxidative stress are interconnected. Erythrocytes are continuously exposed to reactive oxygen species (ROS) during circulation, due to normal aerobic cellular metabolism and also pathology of inflammatory diseases. Systemic mastocytosis and genetic depletion of mast cells have been shown to affect anaemia. In the present study, we attempted to reveal whether mast cells have a direct role in clearance or erythrophagocytosis of normal or oxidatively damaged erythrocytes. Murine erythrocytes were treated with tert-butyl hydroperoxidase (t-BHP), an agent that induces oxidative damage and mimics in vivo oxidative stress. Normal and oxidatively damaged erythrocytes were labeled with carboxyfluorescein succinimidyl ester (CFSE) to track erythrophagocytosis. We show, for the first time, direct erythrophagocytosis of oxidatively damaged erythrocytes in vitro by RBL-2H3 mast cells as well as in vivo by murine peritoneal mast cells. Also, activated mast cells, as may be present in inflammatory conditions, showed a significant increase in the uptake of oxidatively damaged erythrocytes than resting mast cells. This suggests the involvement of mast cells in erythrocyte clearance during oxidative stress or inflammatory disorders. Partial inhibition of phagocytosis by various inhibitors indicated that this process may be controlled by several pathways. Hence, our study provides important evidence for involvement of mast cells in severe anemia due to inflammation and oxidative stress and might be helpful to circumvent the adverse anemic disorders.

Keywords: mast cells, anemia, erythrophagocytosis, oxidatively damaged erythrocytes

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3385 An Exploration of Science, Technology, Engineering, Arts, and Mathematics Competition from the Perspective of Arts

Authors: Qiao Mao

Abstract:

There is a growing number of studies concerning STEM (Science, Technology, Engineering, and Mathematics) and STEAM (Science, Technology, Engineering, Arts, and Mathematics). However, the research is little on STEAM competitions from Arts' perspective. This study takes the annual PowerTech STEAM competition in Taiwan as an example. In this activity, students are asked to make wooden bionic mechanical beasts on the spot and participate in a model and speed competition. This study aims to explore how Arts influences STEM after it involves in the making of mechanical beasts. A case study method is adopted. Through expert sampling, five prize winners in the PowerTech Youth Science and Technology Creation Competition and their supervisors are taken as the research subjects. Relevant data which are collected, sorted out, analyzed and interpreted afterwards, derive from observations, interview and document analyses, etc. The results of the study show that in the PowerTech Youth Science and Technology Creation Competition, when Arts involves in STEM, (1) it has an impact on the athletic performance, balance, stability and symmetry of mechanical beasts; (2) students become more interested and more creative in making STEAM mechanical beasts, which can promote students' learning of STEM; (3) students encounter more difficulties and problems when making STEAM mechanical beasts, and need to have more systematic thinking and design thinking to solve problems.

Keywords: PowerTech, STEAM contest, mechanical beast, arts' role

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3384 Practical Evaluation of High-Efficiency Si-based Tandem Solar Cells

Authors: Sue-Yi Chen, Wei-Chun Hsu, Jon-Yiew Gan

Abstract:

Si-based double-junction tandem solar cells have become a popular research topic because of the advantages of low manufacturing cost and high energy conversion efficiency. However, there is no set of calculations to select the appropriate top cell materials. Therefore, this paper will propose a simple but practical selection method. First of all, we calculate the S-Q limit and explain the reasons for developing tandem solar cells. Secondly, we calculate the theoretical energy conversion efficiency of the double-junction tandem solar cells while combining the commercial monocrystalline Si and materials' practical efficiency to consider the actual situation. Finally, we conservatively conclude that if considering 75% performance of the theoretical energy conversion efficiency of the top cell, the suitable bandgap energy range will fall between 1.38eV to 2.5eV. Besides, we also briefly describe some improvements of several proper materials, CZTS, CdSe, Cu2O, ZnTe, and CdS, hoping that future research can select and manufacture high-efficiency Si-based tandem solar cells based on this paper successfully. Most importantly, our calculation method is not limited to silicon solely. If other materials’ performances match or surpass silicon's ability in the future, researchers can also apply this set of deduction processes.

Keywords: high-efficiency solar cells, material selection, Si-based double-junction solar cells, Tandem solar cells, photovoltaics.

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3383 Development of a Myocardial Patch with 3D Hydrogel Electrical Stimulation System

Authors: Yung-Gi Chen, Pei-Leun Kang, Yu-Hsin Lin, Shwu-Jen Chang

Abstract:

Myocardial tissue has limited self-repair ability due to its loss of differentiation characteristic for most mature cardiomyocytes. Therefore, the effective use of stem cell technology in regenerative medicine is an important development to alleviate the current difficulties in cardiac disease treatment. The main purpose of this project was to develop a 3-D hydrogel electrical stimulating system for promoting the differentiation of stem cells into myocardial cells, and the patch will be used to repair damaged myocardial tissue. This project was focused on the preparation of the electrical stimulation system with carbon/CaCl₂ electrodes covered with carbon nanotube-hydrogel. In this study, we utilized screen imprinting techniques and used Poly(lactic-co-glycolic acid)(PLGA) membranes as printing substrates to fabricate a carbon/CaCl₂ interdigitated electrode that covered with alginate/carbon nanotube hydrogels. The single-walled carbon nanotube was added in the hydrogel to enhance the mechanical strength and conductivity of hydrogel. In this study, we used PLGA (85:15) as electrode preparing substrate. The CaCl₂/ EtOH solution (80% w/v) was mixed into carbon paste to prepare various concentration calcium-containing carbon paste (2.5%, 5%, 7.5%, 10% v/v). Different concentrations of alginate (1%, 1.5%, 2% v/v) and SWCNT(Diameter < 2nm, length between 5-15μm) (1, 1.5, 3 mg/ml) are gently immobilized on the electrode by cross-linking with calcium chloride. The three-dimensional hydrogel electrode was tested for its redox efficiency by cyclic voltammetry to determine the optimal parameters for the hydrogel electrode preparation. From the result of the final electrodes, it indicated that the electrode was not easy to maintain the pattern of the interdigitated electrode when the concentration of calcium of chloride was more than 10%. According to the gel rate test and cyclic voltammetry experiment results showed the SWCNT could increase the electron conduction of hydrogel electrodes significantly. So far the 3D electrode system has been completed, 2% alginate mixed with 3mg SWCNT is the optimal condition to construct the most complete structure for the hydrogel preparation.

Keywords: myocardial tissue engineering, screen printing technology, poly (lactic-co-glycolic acid), alginate, single walled carbon nanotube

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3382 Assessing Storage of Stability and Mercury Reduction of Freeze-Dried Pseudomonas putida within Different Types of Lyoprotectant

Authors: A. A. M. Azoddein, Y. Nuratri, A. B. Bustary, F. A. M. Azli, S. C. Sayuti

Abstract:

Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage in 4oC without vacuum. Polyethylene glycol (PEG) pre-treated freeze dry cells and broth pre-treated freeze dry cells after freeze-dry recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introduce freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 56.78% and 17.91%. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks were 26.35% and 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been growth in agar. Result from this study may be beneficial and useful as initial reference before commercialize freeze-dried P. putida.

Keywords: Pseudomonas putida, freeze-dry, PEG, tween80/Sucrose, mercury, cell viability

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3381 The Development of Student Core Competencies through the STEM Education Opportunities in Classroom

Authors: Z. Dedovets, M. Rodionov

Abstract:

The goal of the modern education system is to prepare students to be able to adapt to ever-changing life situations. They must be able to acquire required knowledge independently; apply such knowledge in practice to solve various problems by using modern technologies; think critically and creatively; competently use information; be communicative, work in a team; and develop their own moral values, intellect and cultural awareness. As a result, the status of education significantly increases; new requirements to its quality have been formed. In recent years, the competency-based approach in education has become of significant interest. This approach is a strengthening of applied and practical characteristics of a school education and leads to the forming of the key students’ competencies which define their success in future life. In this article, the authors’ attention focuses on a range of key competencies, educational, informational and communicative and on the possibility to develop such competencies via STEM education. This research shows the change in students’ attitude towards scientific disciplines such as mathematics, general science, technology and engineering as a result of STEM education. Two-staged analyzes questionnaires completed by students of forms II to IV in the republic of Trinidad and Tobago allowed the authors to categorize students between two levels that represent students’ attitude to various disciplines. The significance of differences between selected levels was confirmed with the use of Pearsons’ chi-squared test. In summary, the analysis of obtained data makes it possible to conclude that STEM education has a great potential for development of core students’ competencies and encourages the development of positive student attitude towards the above mentioned above scientific disciplines.

Keywords: STEM, science, technology, engineering, mathematics, students’ competency, Pearson's chi-squared test

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3380 An Increase in Glucose Uptake per se is Insufficient to Induce Oxidative Stress and Vascular Endothelial Cell Dysfunction

Authors: Heba Khader, Victor Solodushko, Brian Fouty

Abstract:

Hyperglycemia is a hallmark of uncontrolled diabetes and causes vascular endothelial dysfunction. An increase in glucose uptake and metabolism by vascular endothelial cells is the presumed trigger for this hyperglycemia-induced dysfunction. Glucose uptake into vascular endothelial cells is mediated largely by Glut-1. Glut-1 is an equilibrative glucose transporter with a Km value of 2 mM. At physiologic glucose concentrations, Glut-1 is almost saturated and, therefore, increasing glucose concentration does not increase glucose uptake unless Glut-1 is upregulated. However, hyperglycemia downregulates Glut-1 and decreases rather than increases glucose uptake in vascular endothelial cells. This apparent discrepancy necessitates further study on the effect of increasing glucose uptake on the oxidative state and function of vascular endothelial cells. To test this, a Tet-on system was generated to conditionally regulate Glut-1 expression in endothelial cells by the addition and removal of doxycycline. Glut-1 overexpression was confirmed by Western blot and radiolabeled glucose uptake measurements. Upregulation of Glut-1 resulted in a 4-fold increase in glucose uptake into endothelial cells as determined by 3H deoxy-D-glucose uptake. Increased glucose uptake through Glut-1 did not induce an oxidative stress nor did it cause endothelial dysfunction in rat pulmonary microvascular endothelial cells determined by monolayer resistance, cell proliferation or advanced glycation end product formation. Increased glucose uptake through Glut-1did not lead to an increase in glucose metabolism, due in part to inhibition of hexokinase in Glut-1 overexpressing cells. In summary, this study demonstrates that increasing glucose uptake and intracellular glucose by overexpression of Glut-1 does not alter the oxidative state of rat pulmonary microvascular endothelial cells or cause endothelial cell dysfunction. These results conflict with the current paradigm that hyperglycemia leads to oxidative stress and endothelial dysfunction in vascular endothelial cells through an increase in glucose uptake.

Keywords: endothelial cells, glucose uptake, Glut1, hyperglycemia

Procedia PDF Downloads 328