Search results for: STEM (science
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3288

Search results for: STEM (science

2958 Study of the Combinatorial Impact of Substrate Properties on Mesenchymal Stem Cell Migration Using Microfluidics

Authors: Nishanth Venugopal Menon, Chuah Yon Jin, Samantha Phey, Wu Yingnan, Zhang Ying, Vincent Chan, Kang Yuejun

Abstract:

Cell Migration is a vital phenomenon that the cells undergo in various physiological processes like wound healing, disease progression, embryogenesis, etc. Cell migration depends primarily on the chemical and physical cues available in the cellular environment. The chemical cue involves the chemokines secreted and gradients generated in the environment while physical cues indicate the impact of matrix properties like nanotopography and stiffness on the cells. Mesenchymal Stem Cells (MSCs) have been shown to have a role wound healing in vivo and its migration to the site of the wound has been shown to have a therapeutic effect. In the field of stem cell based tissue regeneration of bones and cartilage, one approach has been to introduce scaffold laden with MSCs into the site of injury to enable tissue regeneration. In this work, we have studied the combinatorial impact of the substrate physical properties on MSC migration. A microfluidic in vitro model was created to perform the migration studies. The microfluidic model used is a three compartment device consisting of two cell seeding compartments and one migration compartment. Four different PDMS substrates with varying substrate roughness, stiffness and hydrophobicity were created. Its surface roughness and stiffness was measured using Atomic Force Microscopy (AFM) while its hydrphobicity was measured from the water contact angle using an optical tensiometer. These PDMS substrates are sealed to the microfluidic chip following which the MSCs are seeded and the cell migration is studied over the period of a week. Cell migration was quantified using fluorescence imaging of the cytoskeleton (F-actin) to find out the area covered by the cells inside the migration compartment. The impact of adhesion proteins on cell migration was also quantified using a real-time polymerase chain reaction (qRT PCR). These results suggested that the optimal substrate for cell migration would be one with an intermediate level of roughness, stiffness and hydrophobicity. A higher or lower value of these properties affected cell migration negatively. These observations have helped us in understanding that different substrate properties need to be considered in tandem, especially while designing scaffolds for tissue regeneration as cell migration is normally impacted by the combinatorial impact of the matrix. These observations may lead us to scaffold optimization in future tissue regeneration applications.

Keywords: cell migration, microfluidics, in vitro model, stem cell migration, scaffold, substrate properties

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2957 The Future Shrouded by the Ghost of History: Palestinian Science Fiction "The Key"

Authors: KongQue

Abstract:

Palestinian novelist Anwar Hamed’s work "The Key" is a science fiction that profoundly metaphors the Palestinian issue. In the novel, Anwar Hamed uses the "key", a popular symbol of Palestinian resistance culture, to symbolize the history of "Nakba" and the Palestinians' right to return, and uses the ghost lingering outside the door of the Israelis as a metaphor Palestinians and the Palestinian issue, for being covered, hidden, and turned a blind eye. Through the literary genre of science fiction, Anwar Hamed continues the literary tradition of Palestinian writers recreating the historical trauma of the Palestinians, explores the causes of the inability to communicate between Palestine and Israel, and attempts to reawaken concern over the plight of Palestinian refugees.

Keywords: palestine, key, science fiction, arabic literature

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2956 Inverted Diameter-Limit Thinning: A Promising Alternative for Mixed Populus tremuloides Stands Management

Authors: Ablo Paul Igor Hounzandji, Benoit Lafleur, Annie DesRochers

Abstract:

Introduction: Populus tremuloides [Michx] regenerates rapidly and abundantly by root suckering after harvest, creating stands with interconnected stems. Pre-commercial thinning can be used to concentrate growth on fewer stems to reach merchantability faster than un-thinned stands. However, conventional thinning methods are typically designed to reach even spacing between residual stems (1,100 stem ha⁻¹, evenly distributed), which can lead to treated stands consisting of weaker/smaller stems compared to the original stands. Considering the nature of P. tremuloides's regeneration, with large underground biomass of interconnected roots, aiming to keep the most vigorous and largest stems, regardless of their spatial distribution, inverted diameter-limit thinning could be more beneficial to post-thinning stand productivity because it would reduce the imbalance between roots and leaf area caused by thinning. Aims: This study aimed to compare stand and stem productivity of P. tremuloides stands thinned with a conventional thinning treatment (CT; 1,100 stem ha⁻¹, evenly distributed), two levels of inverted diameter-limit thinning (DL1 and DL2, keeping the largest 1100 or 2200 stems ha⁻¹, respectively, regardless of their spatial distribution) and a control unthinned treatment. Because DL treatments can create substantial or frequent gaps in the thinned stands, we also aimed to evaluate the potential of this treatment to recreate mixed conifer-broadleaf stands by fill-planting Picea glauca seedlings. Methods: Three replicate 21 year-old sucker-regenerated aspen stands were thinned in 2010 according to four treatments: CT, DL1, DL2, and un-thinned control. Picea glauca seedlings were underplanted in gaps created by the DL1 and DL2 treatments. Stand productivity per hectare, stem quality (diameter and height, volume stem⁻¹) and survival and height growth of fill-planted P. glauca seedlings were measured 8 year post-treatments. Results: Productivity, volume, diameter, and height were better in the treated stands (CT, DL1, and DL2) than in the un-thinned control. Productivity of CT and DL1 stands was similar 4.8 m³ ha⁻¹ year⁻¹. At the tree level, diameter and height of the trees in the DL1 treatment were 5% greater than those in the CT treatment. The average volume of trees in the DL1 treatment was 11% higher than the CT treatment. Survival after 8 years of fill planted P. glauca seedlings was 2% greater in the DL1 than in the DL2 treatment. DL1 treatment also produced taller seedlings (+20 cm). Discussion: Results showed that DL treatments were effective in producing post-thinned stands with larger stems without affecting stand productivity. In addition, we showed that these treatments were suitable to introduce slower growing conifer seedlings such as Picea glauca in order to re-create or maintain mixed stands despite the aggressive nature of P. tremuloides sucker regeneration.

Keywords: Aspen, inverted diameter-limit, mixed forest, populus tremuloides, silviculture, thinning

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2955 The Need for the Utilization of Instructional Materials on the Teaching and Learning of Agricultural Science Education in Developing Countries

Authors: Ogoh Andrew Enokela

Abstract:

This paper dwelt on the need for the utilization of instructional materials with highlights on the type of instructional materials, selection, uses and their importance on the learning and teaching of Agricultural Science Education in developing countries. It further discussed the concept of improvisation with some recommendation in terms of availability, utilization on the teaching and learning of Agricultural Science Education.

Keywords: instructional materials, agricultural science education, improvisation, teaching and learning

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2954 Wave Powered Airlift PUMP for Primarily Artificial Upwelling

Authors: Bruno Cossu, Elio Carlo

Abstract:

The invention (patent pending) relates to the field of devices aimed to harness wave energy (WEC) especially for artificial upwelling, forced downwelling, production of compressed air. In its basic form, the pump consists of a hydro-pneumatic machine, driven by wave energy, characterised by the fact that it has no moving mechanical parts, and is made up of only two structural components: an hollow body, which is open at the bottom to the sea and partially immersed in sea water, and a tube, both joined together to form a single body. The shape of the hollow body is like a mushroom whose cap and stem are hollow; the stem is open at both ends and the lower part of its surface is crossed by holes; the tube is external and coaxial to the stem and is joined to it so as to form a single body. This shape of the hollow body and the type of connection to the tube allows the pump to operate simultaneously as an air compressor (OWC) on the cap side, and as an airlift on the stem side. The pump can be implemented in four versions, each of which provides different variants and methods of implementation: 1) firstly, for the artificial upwelling of cold, deep ocean water; 2) secondly, for the lifting and transfer of these waters to the place of use (above all, fish farming plants), even if kilometres away; 3) thirdly, for the forced downwelling of surface sea water; 4) fourthly, for the forced downwelling of surface water, its oxygenation, and the simultaneous production of compressed air. The transfer of the deep water or the downwelling of the raised surface water (as for pump versions indicated in points 2 and 3 above), is obtained by making the water raised by the airlift flow into the upper inlet of another pipe, internal or adjoined to the airlift; the downwelling of raised surface water, oxygenation, and the simultaneous production of compressed air (as for the pump version indicated in point 4), is obtained by installing a venturi tube on the upper end of the pipe, whose restricted section is connected to the external atmosphere, so that it also operates like a hydraulic air compressor (trompe). Furthermore, by combining one or more pumps for the upwelling of cold, deep water, with one or more pumps for the downwelling of the warm surface water, the system can be used in an Ocean Thermal Energy Conversion plant to supply the cold and the warm water required for the operation of the same, thus allowing to use, without increased costs, in addition to the mechanical energy of the waves, for the purposes indicated in points 1 to 4, the thermal one of the marine water treated in the process.

Keywords: air lifted upwelling, fish farming plant, hydraulic air compressor, wave energy converter

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2953 Induction of Different Types of Callus and Somatic Embryogenesis in Various Explants of Taraxacum Kok-Saghyz Rodin

Authors: Kairat Uteulin, Azhar Iskakova, Serik Mukhambetzhanov, Bayan Yesbolayeva, Gabit Bari, Aslan Zheksenbai, Kabyl Zhambakin, Chingis Dzhabykbayev, Vladimir Piven, Izbasar Rakhimbaiev

Abstract:

To explore the potential for in vitro rapid regeneration of Russian dandelion (Taraxacum kok-saghyz Rodin), different concentrations of 6-Benzylaminopurine (BAP), 2,4-Dichlorophenoxyacetic acid (2.4-D) and BAP combined with Indole-3-acetic acid (IAA) were evaluated for their effects on the induction of somatic embryos from leaf, seed stem and root explants. Different explants were cultured on MS medium supplemented with various concentrations (0, 0.5, 1, 1.5, 2, 2.5 and 3 mg/l) of each kind of hormone. Callus induction percentage, fresh weight, color and texture of the callus were assessed after 14 and 28 days of culture. The optimum medium for the proliferation of embryogenic calli from leaf and root explants was MS supplemented with 2.5 mg/L BAP and 0.5 mg/L 2.4-D. Concentrations of 2.5 mg/L BAP and 1.5 mg/L IAA also had a remarkable effect on root and stem explants. The best concentration to produce callus from stem explants was 0.5 mg/L BAP and 1 mg/L IAA. Results of mean comparison showed that BAP and 2.4-D were more effective on different explants than BAP and IAA. Results of the double staining method proved that somatic embryogenesis occurred in the most concentrations of BAP and 2.4-D. Under microscopic observations, the different developmental stages of the embryos (globular, heart, torpedo and cotyledonary) were revealed together in callus cells, indicating that the most tested hormone combinations were effective for somatic embryogenesis formation in this species. Seed explants formed torpedo and cotyledonary stages faster than leaf and root explants in the most combinations. Most calli from seed explants were cream colored and friable, while calli were compact and light green from leaf and root explants. Some combinations gave direct regeneration and (3 mg/L BAP and 2 mg/L IAA) in seed explants and (0.5 mg/L BAP and 2.5 mg/L IAA) in leaf explants had the highest number of shoots with average of 21 and 27 shoots per callus. The developed protocol established the production of different callus types from seed, leaf, and root explants and plant regeneration through somatic embryogenesis.

Keywords: taraxacum kok-saghyz Rodin, callus, somatic embryogenesis

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2952 Attitudes of Secondary School Students towards Science and Technical Education in Yauri Metropolis Kebbi State, Nigeria

Authors: Ibrahim Alhassan Libata

Abstract:

This study was carried out to assess attitude of secondary school students towards science and technical education in Yauri metropolis, Kebbi State, Nigeria. The population of the study was 200. Proportionate random sampling method was used in selecting 132 as sample size. Science and technical education is the most powerful forces for change in the world today, and students who hope to have a hand in shaping a better future must participate for their advancements. Four Null hypotheses were generated to guide the conduct of the study, questionnaire was the only instrument used in the study; the instrument was subjected to test-retest reliability. The reliability index of the instrument was 0.69. Overall scores of the Students were analyzed and a mean score was determined, the mean score of students was 85. There were no significant differences between the attitudes of male and female students towards science and technical education. The results also revealed that there was significant difference between the attitude of boding and day school students towards science and technical education, personality constraints of students is one factor militating against the participation of students in science and technical education, socio-economic status of the parents over the years have been the dominant factor of student’s inadequate representation in the field of science and technical education. Based on the findings of this study, the researcher recommended that teachers should motivate students, which they can do through their teaching styles and by showing them the relevance of the learning topics to their everyday lives. Government and the school management should create the learning environment that helps motivate students not only to come to classes but also want to learn and enjoy learning science and technical education, establishment of more Science and Technical Colleges education, more Public enlightenment campaigns to motivate parents and the entire community to support their children in studying science and technical education.

Keywords: attitude, students, science, Yauri

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2951 Development of a Natural Anti-cancer Formulation Which Can Target Triple Negative Breast Cancer Stem Cells

Authors: Samashi Munaweera

Abstract:

Cancer stem cells (CSC) are responsible for the initiation, extensive proliferation and metastasis of cancer. CSCs, including breast cancer stem cells (bCSCs) have a capacity to generate chemo and radiotherapy resistance heterogeneous population of cells. Over-expressed ABCB1 has been reported as a main reason for drug resistance of CSCs via activating drug efflux pumps by creating pores in the cell membrane. The overall efficiency of chemotherapeutic agents might be enhanced by blocking the ABCB protein efflux pump in the CSC membrane. There is an urgent need to search for persuasive natural drugs which can target CSCs. Anti-cancer properties of Hylocereus undatus on cancer CSCs have not yet been studied. In the present study, the anti-cancer effects of the peel and flesh of H. undatus fruit on bCSCs were evaluated with the aim of developing a marketable anti-cancer nutraceutical formulation. The flesh and peel of H. undatus were freeze-dried and sequentially extracted into four different solvents (hexane, chloroform, ethyl acetate and ethanol). All extracts (eight extracts) were dried under reduced pressure, and different concentrations (12.5-400 µg/mL) were treated on bCSCs isolated from a triple-negative chemo-resistant breast cancer phenotype (MDA-MB-231 cells). Anti-proliferative effects of all extracts and paclitaxel (positive control) were determined by a colorimetric assay (WST-1 based). Since peel-chloroform (IC50= 54.8 µg/mL) and flesh-ethyl acetate (IC50= 150.5 µg/mL) extras exerted a potent anti-proliferative effect at 72 h post-incubation, a combinatorial formulation (CF) was developed with the most active peel-chloroform extract and 20 µg/mL of verapamil (a known ABCB1 drug efflux pump blocker) first time in the world. Anti-proliferative effects and pro-apoptotic effects of CF were confirmed by estimating activated caspase3 and caspase7 levels and apoptotic morphological features in the CF-treated bCSCs compared to untreated and only verapamil (20 µg/mL) treated bCSCs, and CF treated normal mammary epithelial cells (MCF-10A). The antiproliferative effects of CF (16.4 µg/mL) are greater than paclitaxel (19.2 µg/mL) and three folds greater than peel-chloroform extract (IC50= 54.8 µg/mL) on bCSCs while exerting less effects on normal cells (> 400 µg/mL). Collectively, CF can be considered as a potential initiative of a nutraceutical formulation that can target CSCs.

Keywords: breast cancer stem cells (bCSCs), Hylocereus undatus, combinatorial formulation (CF), ABCB 1 protein, verapamil

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2950 Analysis of the Potential of Biomass Residues for Energy Production and Applications in New Materials

Authors: Sibele A. F. Leite, Bernno S. Leite, José Vicente H. D´Angelo, Ana Teresa P. Dell’Isola, Julio CéSar Souza

Abstract:

The generation of bioenergy is one of the oldest and simplest biomass applications and is one of the safest options for minimizing emissions of greenhouse gasses and replace the use of fossil fuels. In addition, the increasing development of technologies for energy biomass conversion parallel to the advancement of research in biotechnology and engineering has enabled new opportunities for exploitation of biomass. Agricultural residues offer great potential for energy use, and Brazil is in a prominent position in the production and export of agricultural products such as banana and rice. Despite the economic importance of the growth prospects of these activities and the increasing of the agricultural waste, they are rarely explored for energy and production of new materials. Brazil products almost 10.5 million tons/year of rice husk and 26.8 million tons/year of banana stem. Thereby, the aim of this study was to analysis the potential of biomass residues for energy production and applications in new materials. Rice husk (specify the type) and banana stem (specify the type) were characterized by physicochemical analyses using the following parameters: organic carbon, nitrogen (NTK), proximate analyses, FT-IR spectroscopy, thermogravimetric analyses (TG), calorific values and silica content. Rice husk and banana stem presented attractive superior calorific (from 11.5 to 13.7MJ/kg), and they may be compared to vegetal coal (21.25 MJ/kg). These results are due to the high organic matter content. According to the proximate analysis, biomass has high carbon content (fixed and volatile) and low moisture and ash content. In addition, data obtained by Walkley–Black method point out that most of the carbon present in the rice husk (50.5 wt%) and in banana stalk (35.5 wt%) should be understood as organic carbon (readily oxidizable). Organic matter was also detected by Kjeldahl method which gives the values of nitrogen (especially on the organic form) for both residues: 3.8 and 4.7 g/kg of rice husk and banana stem respectively. TG and DSC analyses support the previous results, as they can provide information about the thermal stability of the samples allowing a correlation between thermal behavior and chemical composition. According to the thermogravimetric curves, there were two main stages of mass-losses. The first and smaller one occurred below 100 °C, which was suitable for water losses and the second event occurred between 200 and 500 °C which indicates decomposition of the organic matter. At this broad peak, the main loss was between 250-350 °C, and it is because of sugar decomposition (components readily oxidizable). Above 350 °C, mass loss of the biomass may be associated with lignin decomposition. Spectroscopic characterization just provided qualitative information about the organic matter, but spectra have shown absorption bands around 1030 cm-1 which may be identified as species containing silicon. This result is expected for the rice husk and deserves further investigation to the stalk of banana, as it can bring a different perspective for this biomass residue.

Keywords: rice husk, banana stem, bioenergy, renewable feedstock

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2949 The in Vitro and in Vivo Antifungal Activity of Terminalia Mantaly on Aspergillus Species Using Drosophila melanogaster (UAS-Diptericin) As a Model

Authors: Ponchang Apollos Wuyep, Alice Njolke Mafe, Longchi Satkat Zacheaus, Dogun Ojochogu, Dabot Ayuba Yakubu

Abstract:

Fungi causes huge losses when infections occur both in plants and animals. Synthetic Antifungal drugs are mostly very expensive and highly cytotoxic when taken. This study was aimed at determining the in vitro and in vivo antifungal activities of the leaves and stem extracts of Terminalia mantaly (Umbrella tree)H. Perrier on Aspergillus species in a bid to identify potential sources of cheap starting materials for the synthesis of new drugs to address the growing antimicrobial resistance. T. mantaly leave and stem powdered plant was extracted by fractionation using the method of solvent partition co-efficient in their graded form in the order n-hexane, Ethyl acetate, methanol and distilled water and phytochemical screening of each fraction revealed the presence of alkaloids, saponins, Tannins, flavonoids, carbohydrates, steroids, anthraquinones, cardiac glycosides and terpenoids in varying degrees. The Agar well diffusion technique was used to screen for antifungal activity of the fractions on clinical isolates of Aspergillus species (Aspergillus flavus and Aspergillus fumigatus). Minimum inhibitory concentration (MIC50) of the most active extracts was determined by the broth dilution method. The fractions test indicated a high antifungal activity with zones of inhibition ranging from 6 to 26 mm and 8 to 30mm (leave fractions) and 10mm to 34mm and 14mm to36mm (stem fractions) on A. flavus and A. fumigatus respectively. All the fractions indicated antifungal activity in a dose response relationship at concentrations of 62.5mg/ml, 125mg/ml, 250mg/ml and 500mg/ml. Better antifungal efficacy was shown by the Ethyl acetate, Hexane and Methanol fractions in the in vitro as the most potent fraction with MIC ranging from 62.5 to 125mg/ml. There was no statistically significant difference (P>0.05) in the potency of the Eight fractions from leave and stem (Hexane, Ethyl acetate, methanol and distilled water, antifungal (fluconazole), which served as positive control and 10% DMSO(Dimethyl Sulfoxide)which served as negative control. In the in vivo investigations, the ingestion technique was used for the infectious studies Female Drosophilla melanogaster(UAS-Diptericin)normal flies(positive control),infected and not treated flies (negative control) and infected flies with A. fumigatus and placed on normal diet, diet containing fractions(MSM and HSM each at concentrations of 10mg/ml 20mg/ml, 30mg/ml, 40mg/ml, 50mg/ml, 60mg/ml, 70mg/ml, 80mg/ml, 90mg/ml and 100mg/ml), diet containing control drugs(fluconazole as positive control)and infected flies on normal diet(negative control), the flies were observed for fifteen(15) days. Then the total mortality of flies was recorded each day. The results of the study reveals that the flies were susceptible to infection with A. fumigatus and responded to treatment with more effectiveness at 50mg/ml, 60mg/ml and 70mg/ml for both the Methanol and Hexane stem fractions. Therefore, the Methanol and Hexane stem fractions of T. mantaly contain therapeutically useful compounds, justifying the traditional use of this plant for the treatment of fungal infections.

Keywords: Terminalia mantaly, Aspergillus fumigatus, cytotoxic, Drosophila melanogaster, antifungal

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2948 Analyzing Information Management in Science and Technology Institute Libraries in India

Authors: P. M. Naushad Ali

Abstract:

India’s strength in basic research is recognized internationally. Science and Technology research in India has been performed by six distinct bodies or organizations such as Cooperative Research Associations, Autonomous Research Council, Institute under Ministries, Industrial R&D Establishment, Universities, Private Institutions. All most all these institutions are having a well-established library/information center to cater the information needs of their users like scientists and technologists. Information Management (IM) comprises disciplines concerned with the study and the effective and efficient management of information and resources, products and services as well as the understanding of the involved technologies and the people engaged in this activity. It is also observed that the libraries and information centers in India are also using modern technologies for the management of various activities and services to serve their users in a better way. Science and Technology libraries in the country are usually better equipped because the investment in Science and Technology in the country are much larger than those in other fields. Thus, most of the Science and Technology libraries are equipped with modern IT-based tools for handling and management of library services. In spite of these facts Science and Technology libraries are having all the characteristics of a model organization where computer application is found most successful, however, the adoption of this IT based management tool is not uniform in these libraries. The present study will help to know about the level use of IT-based management tools for the information management of Science and Technology libraries in India. The questionnaire, interview, observation and document review techniques have been used in data collection. Finally, the author discusses findings of the study and put forward some suggestions to improve the quality of Science and Technology institute library services in India.

Keywords: information management, science and technology libraries, India, IT-based tools

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2947 Improved Mechanical Properties and Osteogenesis in Electrospun Poly L-Lactic Ultrafine Nanofiber Scaffolds Incorporated with Graphene Oxide

Authors: Weili Shao, Qian Wang, Jianxin He

Abstract:

Recently, the applications of graphene oxide in fabricating scaffolds for bone tissue engineering have been received extensive concern. In this work, poly l-lactic/graphene oxide composite nanofibers were successfully fabricated by electrospinning. The morphology structure, porosity and mechanical properties of the composite nanofibers were characterized using different techniques. And mouse mesenchymal stem cells were cultured on the composite nanofiber scaffolds to assess their suitability for bone tissue engineering. The results indicated that the composite nanofiber scaffolds had finer fiber diameter and higher porosity as compared with pure poly l-lactic nanofibers. Furthermore, incorporation of graphene oxide into the poly l-lactic nanofibers increased protein adsorptivity, boosted the Young’s modulus and tensile strength by nearly 4.2-fold and 3.5-fold, respectively, and significantly enhanced adhesion, proliferation, and osteogenesis in mouse mesenchymal stem cells. The results indicate that composite nanofibers could be excellent and versatile scaffolds for bone tissue engineering.

Keywords: poly l-lactic, graphene oxide, osteogenesis, bone tissue engineering

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2946 Science and Technology as Contemporary Epistemological Conditions of Literature

Authors: Lin Zou

Abstract:

This paper explores how the development of science and technology in the recent decades has created new conditions for literature and aesthetics. These are epistemological conditions that not only offer empirical understandings of the human mentality, behavior, emotions, and humanity in general, but reshape how value and the ontological questions are understood and linked with humanity. This paper will discuss the implications of these epistemological conditions for the depiction and interpretation of human subjectivity in literature. The paper will first seek to present the argument that science and technology have created new conditions for literature and aesthetics. It then outlines the implications of these new conditions for literature and aesthetics. The main methodologies used are close reading and case studies.

Keywords: epistemological conditions, literature and aesthetics, science and technology, subjectivity

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2945 Mesenchymal Stem Cells on Fibrin Assemblies with Growth Factors

Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

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Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

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2944 Wharton's Jelly-Derived Mesenchymal Stem Cells Modulate Heart Rate Variability and Improve Baroreflex Sensitivity in Septic Rats

Authors: Cóndor C. José, Rodrigues E. Camila, Noronha L. Irene, Dos Santos Fernando, Irigoyen M. Claudia, Andrade Lúcia

Abstract:

Sepsis induces alterations in hemodynamics and autonomic nervous system (ASN). The autonomic activity can be calculated by measuring heart rate variability (HRV) that represents the complex interplay between ASN and cardiac pacemaker cells. Wharton’s jelly mesenchymal stem cells (WJ-MSCs) are known to express genes and secreted factors involved in neuroprotective and immunological effects, also to improve the survival in experimental septic animals. We hypothesized, that WJ-MSCs present an important role in the autonomic activity and in the hemodynamic effects in a cecal ligation and puncture (CLP) model of sepsis. Methods: We used flow cytometry to evaluate WJ-MSCs phenotypes. We divided Wistar rats into groups: sham (shamoperated); CLP; and CLP+MSC (106 WJ-MSCs, i.p., 6 h after CLP). At 24 h post-CLP, we recorded the systolic arterial pressure (SAP) and heart rate (HR) over 20 min. The spectral analysis of HR and SAP; also the spontaneous baroreflex sensitivity (measure by bradycardic and tachycardic responses) were evaluated after recording. The one-way ANOVA and the post hoc Student– Newman– Keuls tests (P< 0.05) were used to data comparison Results: WJ-MSCs were negative for CD3, CD34, CD45 and HLA-DR, whereas they were positive for CD73, CD90 and CD105. The CLP group showed a reduction in variance of overall variability and in high-frequency power of HR (heart parasympathetic activity); furthermore, there is a low-frequency reduction of SAP (blood vessels sympathetic activity). The treatment with WJ-MSCs improved the autonomic activity by increasing the high and lowfrequency power; and restore the baroreflex sensitive. Conclusions: WJ-MSCs attenuate the impairment of autonomic control of the heart and vessels and might therefore play a protective role in sepsis. (Supported by FAPESP).

Keywords: baroreflex response, heart rate variability, sepsis, wharton’s jelly-derived mesenchymal stem cells

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2943 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

Abstract:

Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

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2942 The Need for Interdisciplinary Approach in Studying Archaeology: An Evolving Cultural Science

Authors: Inalegwu Stephany Akipu

Abstract:

Archaeology being the study of mans past using the materials he left behind has been argued to be classified under sciences while some scholars are of the opinion that it does not deserve the status of being referred to as ‘science’. However divergent the opinions of scholars may be on the classification of Archaeology as a science or in the humanities, the discipline has no doubt, greatly aided in shaping the history of man’s past. Through the different stages that the discipline has transgressed, it has encountered some challenges. This paper therefore, attempts to highlight the need for the inclusion of branches of other disciplines when using Archaeology in reconstructing man’s history. The objective of course, is to add to the existing body of knowledge but specifically to expose the incomparable importance of archaeology as a discipline and to place it on such a high scale that it will not be regulated to the background as is done in some Nigerian Universities. The paper attempts a clarification of some conceptual terms and discusses the developmental stages of archaeology. It further describes the present state of the discipline and concludes with the disciplines that need to be imbibed in the use of Archaeology which is an evolving cultural science to obtain the aforementioned interdisciplinary approach.

Keywords: archaeology, cultural, evolution, interdisciplinary, science

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2941 The Impact of Science Teachers' Epistemological Beliefs and Metacognition on Their Use of Inquiry Based Teaching Approaches

Authors: Irfan Ahmed Rind

Abstract:

Science education has recently become the top priority of government of Pakistan. Number of schemes has been initiated for the improvement of science teaching and learning at primary and secondary levels of education, most importantly training in-service science teachers on inquiry based teaching and learning to empower students and encourage creativity, critical thinking, and innovation among them. Therefore, this approach has been promoted in the recent continuous professional development trainings for the in-service teachers. However, the follow ups on trained science teachers and educators suggest that these teachers fail to implement the inquiry based teaching and learning in their classes. In addition, these trainings also fail to bring any significant change in students’ science content knowledge and understanding as per the annual national level surveys conducted by government and independent agencies. Research suggests that science has been taught using scientific positivism, which supports objectivity based on experiments and mathematics. In contrary, the inquiry based teaching and learning are based on constructivism, which conflicts with the positivist epistemology of science teachers. It was, therefore, assumed that science teachers struggle to implement the inquiry based teaching approach as it conflicts with their basic epistemological beliefs. With this assumption, this research aimed to (i) understand how science teachers conceptualize the nature of science, and how this influence their understanding of learning, learners, their own roles as teachers and their teaching strategies, (ii) identify the conflict of science teachers’ epistemological beliefs with the inquiry based teaching approach, and (iii) find the ways in which science teachers epistemological beliefs may be developed from positivism to constructivism, so that they may effectively use the inquiry based teaching approach in teaching science. Using qualitative case study approach, thirty six secondary and higher secondary science teachers (21 male and 15 female) were selected. Data was collected using interviewed, participatory observations (sixty lessons were observed), and twenty interviews from students for verifications of teachers’ responses. The findings suggest that most of the science teacher were positivist in defining the nature of science. Most of them limit themselves to one fix answer that is provided in the books and that there is only one 'right' way to teach science. There is no room for students’ or teachers’ own opinion or bias when it comes to scientific concepts. Inquiry based teaching seems 'no right' to them. They find it difficult to allow students to think out of the box. However, some interesting exercises were found to be very effective in bringing the change in teachers’ epistemological beliefs. These will be discussed in detail in the paper. The findings have major implications for the teachers, educators, and policymakers.

Keywords: science teachers, epistemology, metacognition, inquiry based teaching

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2940 Gamification Beyond Competition: the Case of DPG Lab Collaborative Learning Program for High-School Girls by GameLab KBTU and UNICEF in Kazakhstan

Authors: Nazym Zhumabayeva, Aleksandr Mezin, Alexandra Knysheva

Abstract:

Women's underrepresentation in STEM is critical, worsened by ineffective engagement in educational practices. UNICEF Kazakhstan and GameLab KBTU's collaborative initiatives aim to enhance female STEM participation by fostering an inclusive environment. Learning from LEVEL UP's 2023 program, which featured a hackathon, the 2024 strategy pivots towards non-competitive gamification. Although the data from last year's project showed higher than average student engagement, observations and in-depth interviews with participants showed that the format was stressful for the girls, making them focus on points rather than on other values. This study presents a gamified educational system, DPG Lab, aimed at incentivizing young women's participation in STEM through the development of digital public goods (DPGs). By prioritizing collaborative gamification elements, the project seeks to create an inclusive learning environment that increases engagement and interest in STEM among young women. The DPG Lab aims to find a solution to minimize competition and support collaboration. The project is designed to motivate female participants towards the development of digital solutions through an introduction to the concept of DPGs. It consists of a short online course, a simulation videogame, and a real-time online quest with an offline finale at the KBTU campus. The online course offers short video lectures on open-source development and DPG standards. The game facilitates the practical application of theoretical knowledge, enriching the learning experience. Learners can also participate in a quest that encourages participants to develop DPG ideas in teams by choosing missions throughout the quest path. At the offline quest finale, the participants will meet in person to exchange experiences and accomplishments without engaging in comparative assessments: the quest ensures that each team’s trajectory is distinct by design. This marks a shift from competitive hackathons to a collaborative format, recognizing the unique contributions and achievements of each participant. The pilot batch of students is scheduled to commence in April 2024, with the finale anticipated in June. It is projected that this group will comprise 50 female high-school students from various regions across Kazakhstan. Expected outcomes include increased engagement and interest in STEM fields among young female participants, positive emotional and psychological impact through an emphasis on collaborative learning environments, and improved understanding and skills in DPG development. GameLab KBTU intends to undertake a hypothesis evaluation, employing a methodology similar to that utilized in the preceding LEVEL UP project. This approach will encompass the compilation of quantitative metrics (conversion funnels, test results, and surveys) and qualitative data from in-depth interviews and observational studies. For comparative analysis, a select group of participants from the previous year's project will be recruited to engage in the DPG Lab. By developing and implementing a gamified framework that emphasizes inclusion, engagement, and collaboration, the study seeks to provide practical knowledge about effective gamification strategies for promoting gender diversity in STEM. The expected outcomes of this initiative can contribute to the broader discussion on gamification in education and gender equality in STEM by offering a replicable and scalable model for similar interventions around the world.

Keywords: collaborative learning, competitive learning, digital public goods, educational gamification, emerging regions, STEM, underprivileged groups

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2939 The Context of Teaching and Learning Primary Science to Gifted Students: An Analysis of Australian Curriculum and New South Wales Science Syllabus

Authors: Rashedul Islam

Abstract:

A firmly-validated aim of teaching science is to support student enthusiasm for science learning with an outspread interest in scientific issues in future life. This is in keeping with the recent development in Gifted and Talented Education statement which instructs that gifted students have a renewed interest and natural aptitude in science. Yet, the practice of science teaching leaves many students with the feeling that science is difficult and compared to other school subjects, students interest in science is declining at the final years of the primary school. As a curriculum guides the teaching-learning activities in school, where significant consequences may result from the context of the curricula and syllabi, are a major feature of certain educational jurisdictions in NSW, Australia. The purpose of this study was an exploration of the curriculum sets the context to identify how science education is practiced through primary schools in Sydney, Australia. This phenomenon was explored through document review from two publicly available documents namely: the NSW Science Syllabus K-6, and Australian Curriculum: Foundation - 10 Science. To analyse the data, this qualitative study applied themed content analysis at three different levels, i.e., first cycle coding, second cycle coding- pattern codes, and thematic analysis. Preliminary analysis revealed the phenomenon of teaching-learning practices drawn from eight themes under three phenomena aligned with teachers’ practices and gifted student’s learning characteristics based on Gagné’s Differentiated Model of Gifted and Talent (DMGT). From the results, it appears that, overall, the two documents are relatively well-placed in terms of identifying the context of teaching and learning primary science to gifted students. However, educators need to make themselves aware of the ways in which the curriculum needs to be adapted to meet gifted students learning needs in science. It explores the important phenomena of teaching-learning context to provide gifted students with optimal educational practices including inquiry-based learning, problem-solving, open-ended tasks, creativity in science, higher order thinking, integration, and challenges. The significance of such a study lies in its potential to schools and further research in the field of gifted education.

Keywords: teaching primary science, gifted student learning, curriculum context, science syllabi, Australia

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2938 Evaluation of the Self-Efficacy and Learning Experiences of Final year Students of Computer Science of Southwest Nigerian Universities

Authors: Olabamiji J. Onifade, Peter O. Ajayi, Paul O. Jegede

Abstract:

This study aimed at investigating the preparedness of the undergraduate final year students of Computer Science as the next entrants into the workplace. It assessed their self-efficacy in computational tasks and examined the relationship between their self-efficacy and their learning experiences in Southwest Nigerian universities. The study employed a descriptive survey research design. The population of the study comprises all the final year students of Computer Science. A purposive sampling technique was adopted in selecting a representative sample of interest from the final year students of Computer Science. The Students’ Computational Task Self-Efficacy Questionnaire (SCTSEQ) was used to collect data. Mean, standard deviation, frequency, percentages, and linear regression were used for data analysis. The result obtained revealed that the final year students of Computer Science were averagely confident in performing computational tasks, and there is a significant relationship between the learning experiences of the students and their self-efficacy. The study recommends that the curriculum be improved upon to accommodate industry experts as lecturers in some of the courses, make provision for more practical sessions, and the learning experiences of the student be considered an important component in the undergraduate Computer Science curriculum development process.

Keywords: computer science, learning experiences, self-efficacy, students

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2937 Active Learning Based on Science Experiments to Improve Scientific Literacy

Authors: Kunihiro Kamataki

Abstract:

In this study, active learning based on simple science experiments was developed in a university class of the freshman, in order to improve their scientific literacy. Through the active learning based on simple experiments of generation of cloud in a plastic bottle, students increased the interest in the global atmospheric problem and were able to discuss and find solutions about this problem positively from various viewpoints of the science technology, the politics, the economy, the diplomacy and the relations among nations. The results of their questionnaires and free descriptions of this class indicate that they improve the scientific literacy and motivations of other classroom lectures to acquire knowledge. It is thus suggested that the science experiment is strong tool to improve their intellectual curiosity rapidly and the connections that link the impression of science experiment and their interest of the social problem is very important to enhance their learning effect in this education.

Keywords: active learning, scientific literacy, simple scientific experiment, university education

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2936 The Effect of Nanoscience and Nanotechnology Education on Preservice Science Teachers' Awareness of Nanoscience and Nanotechnology

Authors: Tuba Senel Zor, Oktay Aslan

Abstract:

With current trends in nanoscience and nanotechnology (NST), scientists have paid much attention to education and nanoliteracy in parallel with the developments on these fields. To understand the advances in NST research requires a population with a high degree of science literacy. All citizens should soon need nanoliteracy in order to navigate some of the important science-based issues faced to their everyday lives. While the fields of NST are advancing rapidly and raising their societal significance, general public’s awareness of these fields has remained at a low level. Moreover, students enrolled different education levels and teachers don’t have awareness at expected level. This problem may be stemmed from inadequate education and training. To remove the inadequacy, teachers have greatest duties and responsibilities. Especially science teachers at all levels need to be made aware of these developments and adequately prepared so that they are able to teach about these advances in a developmentally appropriate manner. If the teachers develop understanding and awareness of NST, they can also discuss the topic with their students. Therefore, the awareness and conceptual understandings of both the teachers who will teach science to students and the students who will be introduced about NST should be increased, and the necessary training should be provided. The aim of this study was to examine the effect of NST education on preservice science teachers’ awareness of NST. The study was designed in one group pre-test post-test quasi-experimental pattern. The study was conducted with 32 preservice science teachers attending the Elementary Science Education Program at a large Turkish university in central Anatolia. NST education was given during five weeks as two hours per week. Nanoscience and Nanotechnology Awareness Questionnaire was used as data collected tool and was implemented for pre-test and post-test. The collected data were analyzed using Statistical package for the Social Science (SPSS). The results of data analysis showed that there was a significant difference (z=6.25, p< .05) on NST awareness of preservice science teachers after implemented NST education. The results of the study indicate that NST education has an important effect for improving awareness of preservice science teachers on NST.

Keywords: awareness level, nanoliteracy, nanoscience and nanotechnology education, preservice science teachers

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2935 Nature of Science in Physics Textbooks – Example of Quebec Province

Authors: Brahim El Fadil

Abstract:

The nature of science as a solution (NOS) to life problems is well established in school activities the world over. However, this study reveals the lack of representation of the NOS in science textbooks used in Quebec Province. A content analysis method was adopted to analyze the NOS in relation to optics knowledge and teaching-learning activities in Grade 9 science and technology textbooks and Grade 11 physics textbooks. The selected textbooks were approved and authorized by the Provincial Ministry of Education. Our analysis points out that most of these editions provided a poor representation of NOS. None of them indicates that scientific knowledge is subject to change, even though the history of optics reveals evolutionary and revolutionary changes. Moreover, the analysis shows that textbooks place little emphasis on the discussion of scientific laws and theories. Few of them argue that scientific inquiries are required to gain a deep understanding of scientific concepts. Moreover, they rarely present empirical evidence to support their arguments.

Keywords: nature of science, history of optics, geometrical theory of optics, wave theory of optics

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2934 Nanotechnology: A New Revolution to Increase Agricultural Production

Authors: Reshu Chaudhary, R. S. Sengar

Abstract:

To increase the agricultural production Indian farmer needs to aware of the latest technology i.e. precision farming to maximize the crop yield and minimize the input (fertilizer, pesticide etc.) through monitoring the environmental factors. Biotechnology and information technology have provided lots of opportunities for the development of agriculture. But, still we have to do much more for increasing our agricultural production in order to achieve the target growth of agriculture to secure food, to eliminate poverty and improve living style, to enhance agricultural exports and national income and to improve quality of agricultural products. Nanotechnology can be a great element to satisfy these requirements and to boost the multi-dimensional development of agriculture in order to fulfill the dream of Indian farmers. Nanotechnology is the most rapidly growing area of science and technology with its application in physical science, chemical science, life science, material science and earth science. Nanotechnology is a part of any nation’s future. Research in nanotechnology has extremely high potential to benefit society through application in agricultural sciences. Nanotechnology has greater potential to bring revolution in the agricultural sector.

Keywords: agriculture, biotechnology, crop yield, nanotechnology

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2933 Satisfaction in Supreme Financial Disbursement in the Faculty of Science and Technology, Suan Sunandha Rajabhat University

Authors: Adisai Thovicha, Jiranan Pattaphong

Abstract:

The objective of this research is to study the satisfaction of the disbursement of the Faculty of Science and Technology, Suan Sunandha Rajabhat University. The sample of this study consisted of 98 participants who are faculty members and staff of the Faculty of Science and Technology. Sample was drawn by systematic random sampling technique. Questionnaire was used to collect data. Analysis involves frequency, percentage, mean and standard deviation. It was found that: (1) Most of the 98 faculty members and staff are female, aged between 31-40 years and they have been working at the university for 1-5 years. (2) The satisfaction level of the disbursement of the Faculty of Science and Technology, Suan Sunandha Rajabhat University is high. When each aspect is considered, the satisfaction level of faculty members and staff of the Faculty of Science and Technology is high in service providing staff, process and facilitation.

Keywords: satisfaction of disbursement, petition financing, faculty members, staff

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2932 Attitudes of the Staff in the Faculty of Science and Technology towards the E-Office Documentary System of the Faculty of Science and Technology, Suan Sunandha Rajabhat University

Authors: Narinee Sophatsathit, Yuwadee Nitutorn

Abstract:

The purpose of this study was to investigate the attitudes of the staff in the Faculty of Science and Technology towards the e-office documentary system of the Faculty of Science and Technology, Suan Sunandha Rajabhat University. The subjects of this study included 98 staffs of the Faculty of Science and Technology, Suan Sunandha Rajabhat University. Questionnaires were used to ask the attitude of the staffs towards the implementation of e-office system. The results showed that most of the users of e-office are female with the age between 31-40 years old, master degree of education and in the academic positions. They have working experiences between 1-5 years and reported the time of using e-office between 8:30-12:30 during the weekday with the frequency of 3-5 times/day. Most of them reported their opinions on the e-office at high level (x=3.84) and problems and obstacles in using e-office at high level, as well (x=3.63).

Keywords: attitudes, e-office, staff, documentary system

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2931 Discussion on Big Data and One of Its Early Training Application

Authors: Fulya Gokalp Yavuz, Mark Daniel Ward

Abstract:

This study focuses on a contemporary and inevitable topic of Data Science and its exemplary application for early career building: Big Data and Leaving Learning Community (LLC). ‘Academia’ and ‘Industry’ have a common sense on the importance of Big Data. However, both of them are in a threat of missing the training on this interdisciplinary area. Some traditional teaching doctrines are far away being effective on Data Science. Practitioners needs some intuition and real-life examples how to apply new methods to data in size of terabytes. We simply explain the scope of Data Science training and exemplified its early stage application with LLC, which is a National Science Foundation (NSF) founded project under the supervision of Prof. Ward since 2014. Essentially, we aim to give some intuition for professors, researchers and practitioners to combine data science tools for comprehensive real-life examples with the guides of mentees’ feedback. As a result of discussing mentoring methods and computational challenges of Big Data, we intend to underline its potential with some more realization.

Keywords: Big Data, computation, mentoring, training

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2930 Reconstruction of Alveolar Bone Defects Using Bone Morphogenetic Protein 2 Mediated Rabbit Dental Pulp Stem Cells Seeded on Nano-Hydroxyapatite/Collagen/Poly(L-Lactide)

Authors: Ling-Ling E., Hong-Chen Liu, Dong-Sheng Wang, Fang Su, Xia Wu, Zhan-Ping Shi, Yan Lv, Jia-Zhu Wang

Abstract:

Objective: The objective of the present study is to evaluate the capacity of a tissue-engineered bone complex of recombinant human bone morphogenetic protein 2 (rhBMP-2) mediated dental pulp stem cells (DPSCs) and nano-hydroxyapatite/collagen/poly(L-lactide)(nHAC/PLA) to reconstruct critical-size alveolar bone defects in New Zealand rabbit. Methods: Autologous DPSCs were isolated from rabbit dental pulp tissue and expanded ex vivo to enrich DPSCs numbers, and then their attachment and differentiation capability were evaluated when cultured on the culture plate or nHAC/PLA. The alveolar bone defects were treated with nHAC/PLA, nHAC/PLA+rhBMP-2, nHAC/PLA+DPSCs, nHAC/PLA+DPSCs+rhBMP-2, and autogenous bone (AB) obtained from iliac bone or were left untreated as a control. X-ray and a polychrome sequential fluorescent labeling were performed post-operatively and the animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis. Results: Our results showed that DPSCs expressed STRO-1 and vementin, and favoured osteogenesis and adipogenesis in conditioned media. DPSCs attached and spread well, and retained their osteogenic phenotypes on nHAC/PLA. The rhBMP-2 could significantly increase protein content, alkaline phosphatase (ALP) activity/protein, osteocalcin (OCN) content, and mineral formation of DPSCs cultured on nHAC/PLA. The X-ray graph, the fluorescent, histological observation and histomorphometric analysis showed that the nHAC/PLA+DPSCs+rhBMP-2 tissue-engineered bone complex had an earlier mineralization and more bone formation inside the scaffold than nHAC/PLA, nHAC/PLA+rhBMP-2 and nHAC/PLA+DPSCs, or even autologous bone. Implanted DPSCs contribution to new bone were detected through transfected eGFP genes. Conclutions: Our findings indicated that stem cells existed in adult rabbit dental pulp tissue. The rhBMP-2 promoted osteogenic capability of DPSCs as a potential cell source for periodontal bone regeneration. The nHAC/PLA could serve as a good scaffold for autologous DPSCs seeding, proliferation and differentiation. The tissue-engineered bone complex with nHAC/PLA, rhBMP-2, and autologous DPSCs might be a better alternative to autologous bone for the clinical reconstruction of periodontal bone defects.

Keywords: nano-hydroxyapatite/collagen/poly (L-lactide), dental pulp stem cell, recombinant human bone morphogenetic protein, bone tissue engineering, alveolar bone

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2929 Oxidative Damage to Lipids, Proteins, and DNA during Differentiation of Mesenchymal Stem Cells Derived from Umbilical Cord into Biologically Active Hepatocytes

Authors: Abdolamir Allameh, Shahnaz Esmaeili, Mina Allameh, Safoura Khajeniazi

Abstract:

Stem cells with therapeutic applications can be isolated from human placenta/umblical cord blood (UCB) as well as the cord tissue (UC). Stem cells in culture are vulnerable to oxidative stress, particularly when subjected to differentiation process. The aim of this study was to examine the chnages in the rate of oxidation that occurs to cellular macromolecules during hepatic differentiation of mononuclear cells (MSCs). In addition, the impact of the hepatic differentiation process of MSC on cellular and biological activity of the cells will be undertaken. For this purpose, first mononuclear cells (MNCs) were isolated from human UCB which was obtained from a healthy full-term infant. The cells were cultured at a density of 3×10⁵ cells/cm² in DMEM- low-glucose culture media supplemented with 20% FBS, 2 mM L-glutamine, 100 μg/ml streptomycin and 100 U/ml penicillin. Cell cultures were then incubated at 37°C in a humidified 5% CO₂ incubator. After removing non-adherent cells by replacing culture medium, fibroblast-like adherent cells were resuspended in 0.25% trypsin-EDTA and plated in 25 cm² flasks (1×10⁴/ml). Characterization of the MSCs was routinely done by observing their morphology and growth curve. MSCs were subjected to a 2-step hepatocyte differentiation protocol in presence of hepatocyte growth factor (HGF), dexamethazone (DEX) and oncostatin M (OSM). The hepatocyte-like cells derived from MSCs were checked every week for 3 weeks for changes in lipid peroxidation, protein carbonyl formation and DNA oxidation i.e., 8-hydroxy-2'-deoxyguanosine (8-OH-dG) assay. During the 3-week differentiation process of MSCs to hepatocyte-like cells we found that expression liver-specific markers such as albumin, was associated with increased levels of lipid peroxidation and protein carbonyl formation. Whereas, undifferentiated MSCs has relatively low levels of lipid peroxidation products. There was a significant increase ( p < 0.05) in lipid peroxidation products in hepatocytes on days 7, 14, and 21 of differentiation. Likewise, the level of protein carbonyls in the cells was elevated during the differentiation. The level of protein carbonyls measured in hepatocyte-like cells obtained 3 weeks after differentiation induction was estimated to be ~6 fold higher compared to cells recovered on day 7 of differentiation. On the contrary, there was a small but significant decrease in DNA damage marker (8-OH-dG) in hepatocytes recovered 3 weeks after differentiation onset. The level of 8-OHdG which was in consistent with formation of reactive oxygen species (ROS). In conclusion, this data suggest that despite the elevation in oxidation of lipid and protein molecules during hepatocyte development, the cells were normal in terms of DNA integrity, morphology, and biologically activity.

Keywords: adult stem cells, DNA integrity, free radicals, hepatic differentiation

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