Search results for: protein functional module
Commenced in January 2007
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Edition: International
Paper Count: 5574

Search results for: protein functional module

3264 Neuro-Epigenetic Changes on Diabetes Induced-Synaptic Fidelity in Brain

Authors: Valencia Fernandes, Dharmendra Kumar Khatri, Shashi Bala Singh

Abstract:

Background and Aim: Epigenetics are the inaudible signatures of several pathological processes in the brain. This study understands the influence of DNA methylation, a major epigenetic modification, in the prefrontal cortex and hippocampus of the diabetic brain and its notable effect on the cellular chaperones and synaptic proteins. Method: Chronic high fat diet and STZ-induced diabetic mice were studied for cognitive dysfunction, and global DNA methylation, as well as DNA methyltransferase (DNMT) activity, were assessed. Further, the cellular chaperones and synaptic proteins were examined using DNMT inhibitor, 5-aza-2′-deoxycytidine (5-aza-dC)-via intracerebroventricular injection. Moreover, % methylation of these synaptic proteins were also studied so as to correlate its epigenetic involvement. Computationally, its interaction with the DNMT enzyme were also studied using bioinformatic tools. Histological studies for morphological alterations and neuronal degeneration were also studied. Neurogenesis, a characteristic marker for new learning and memory formation, was also assessed via the BrdU staining. Finally, the most important behavioral studies, including the Morris water maze, Y maze, passive avoidance, and Novel object recognition test, were performed to study its cognitive functions. Results: Altered global DNA methylation and increased levels of DNMTs within the nucleus were confirmed in the cortex and hippocampus of the diseased mice, suggesting hypermethylation at a genetic level. Treatment with AzadC, a global DNA demethylating agent, ameliorated the protein and gene expression of the cellular chaperones and synaptic fidelity. Furthermore, the methylation analysis profile showed hypermethylation of the hsf1 protein, a master regulator for chaperones and thus, confirmed the epigenetic involvement in the diseased brain. Morphological improvements and decreased neurodegeneration, along with enhanced neurogenesis in the treatment group, suggest that epigenetic modulations do participate in learning and memory. This is supported by the improved behavioral test battery seen in the treatment group. Conclusion: DNA methylation could possibly accord in dysregulating the memory-associated proteins at chronic stages in type 2 diabetes. This could suggest a substantial contribution to the underlying pathophysiology of several metabolic syndromes like insulin resistance, obesity and also participate in transitioning this damage centrally, such as cognitive dysfunction.

Keywords: epigenetics, cognition, chaperones, DNA methylation

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3263 Prevalence of Breast Cancer Molecular Subtypes at a Tertiary Cancer Institute

Authors: Nahush Modak, Meena Pangarkar, Anand Pathak, Ankita Tamhane

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Background: Breast cancer is the prominent cause of cancer and mortality among women. This study was done to show the statistical analysis of a cohort of over 250 patients detected with breast cancer diagnosed by oncologists using Immunohistochemistry (IHC). IHC was performed by using ER; PR; HER2; Ki-67 antibodies. Materials and methods: Formalin fixed Paraffin embedded tissue samples were obtained by surgical manner and standard protocol was followed for fixation, grossing, tissue processing, embedding, cutting and IHC. The Ventana Benchmark XT machine was used for automated IHC of the samples. Antibodies used were supplied by F. Hoffmann-La Roche Ltd. Statistical analysis was performed by using SPSS for windows. Statistical tests performed were chi-squared test and Correlation tests with p<.01. The raw data was collected and provided by National Cancer Insitute, Jamtha, India. Result: Luminal B was the most prevailing molecular subtype of Breast cancer at our institute. Chi squared test of homogeneity was performed to find equality in distribution and Luminal B was the most prevalent molecular subtype. The worse prognostic indicator for breast cancer depends upon expression of Ki-67 and her2 protein in cancerous cells. Our study was done at p <.01 and significant dependence was observed. There exists no dependence of age on molecular subtype of breast cancer. Similarly, age is an independent variable while considering Ki-67 expression. Chi square test performed on Human epidermal growth factor receptor 2 (HER2) statuses of patients and strong dependence was observed in percentage of Ki-67 expression and Her2 (+/-) character which shows that, value of Ki depends upon Her2 expression in cancerous cells (p<.01). Surprisingly, dependence was observed in case of Ki-67 and Pr, at p <.01. This shows that Progesterone receptor proteins (PR) are over-expressed when there is an elevation in expression of Ki-67 protein. Conclusion: We conclude from that Luminal B is the most prevalent molecular subtype at National Cancer Institute, Jamtha, India. There was found no significant correlation between age and Ki-67 expression in any molecular subtype. And no dependence or correlation exists between patients’ age and molecular subtype. We also found that, when the diagnosis is Luminal A, out of the cohort of 257 patients, no patient shows >14% Ki-67 value. Statistically, extremely significant values were observed for dependence of PR+Her2- and PR-Her2+ scores on Ki-67 expression. (p<.01). Her2 is an important prognostic factor in breast cancer. Chi squared test for Her2 and Ki-67 shows that the expression of Ki depends upon Her2 statuses. Moreover, Ki-67 cannot be used as a standalone prognostic factor for determining breast cancer.

Keywords: breast cancer molecular subtypes , correlation, immunohistochemistry, Ki-67 and HR, statistical analysis

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3262 Simple and Scalable Thermal-Assisted Bar-Coating Process for Perovskite Solar Cell Fabrication in Open Atmosphere

Authors: Gizachew Belay Adugna

Abstract:

Perovskite solar cells (PSCs) shows rapid development as an emerging photovoltaic material; however, the fast device degradation due to the organic nature, mainly hole transporting material (HTM) and lack of robust and reliable upscaling process for photovoltaic module hindered its commercialization. Herein, HTM molecules with/without fluorine-substituted cyclopenta[2,1-b;3,4-b’]dithiophene derivatives (HYC-oF, HYC-mF, and HYC-H) were developed for PSCs application. The fluorinated HTM molecules exhibited better hole mobility and overall charge extraction in the devices mainly due to strong molecular interaction and packing in the film. Thus, the highest power conversion efficiency (PCE) of 19.64% with improved long stability was achieved for PSCs based on HYC-oF HTM. Moreover, the fluorinated HYC-oF demonstrated excellent film processability in a larger-area substrate (10 cm×10 cm) prepared sequentially with the absorption perovskite underlayer via a scalable bar coating process in ambient air and owned a higher PCE of 18.49% compared to the conventional spiro-OMeTAD (17.51%). The result demonstrates a facile development of HTM towards stable and efficient PSCs for future industrial-scale PV modules.

Keywords: perovskite solar cells, upscaling film coating, power conversion efficiency, solution processing

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3261 Simple Ways to Enhance the Security of Web Services

Authors: Majid Azarniush, Soroush Mokallaei

Abstract:

Although robust security software, including anti-viruses, anti spy wares, anti-spam and firewalls, are amalgamated with new technologies such as Safe Zone, Hybrid Cloud, Sand Box etc., and it can be said that they have managed to prepare highest level of security against viruses, spy wares and other malwares in 2012, but in fact hackers' attacks to websites are increasingly becoming more and more complicated. Because of security matters and developments, it can be said that it was expected to happen so. Here in this work, we try to point out to some functional and vital notes to enhance security on the web enabling the user to browse safely in no limit web world and to use virtual space securely.

Keywords: firewalls, security, web services, software

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3260 Quality and Shelf life of UHT Milk Produced in Tripoli, Libya

Authors: Faozia A. S. Abuhtana, Yahia S. Abujnah, Said O. Gnann

Abstract:

Ultra High Temperature (UHT) processed milk is widely distributed and preferred in numerous countries all over the world due its relatively high quality and long shelf life. Because of the notable high consumption rate of UHT in Libya in addition to negligible studies related to such product on the local level, this study was designed to assess the shelf life of locally produced as well as imported reconstituted sterilized whole milk samples marketed in Tripoli, Libya . Four locally produced vs. three imported brands were used in this study. All samples were stored at room temperature (25± 2C ) for 8 month long period, and subjected to physical, chemical, microbiological and sensory tests. These tests included : measurement of pH, specific gravity, percent acidity, and determination of fat, protein and melamine content. Microbiological tests included total aerobic count, total psychotropic bacteria, total spore forming bacteria and total coliform counts. Results indicated no detection of microbial growth of any type during the study period, in addition to no detection of melamine in all samples. On the other hand, a gradual decline in pH accompanied with gradual increase in % acidity of both locally produced and imported samples was observed. Such changes in both pH and % acidity reached their lowest and highest values respectively during the 24th week of storage. For instance pH values were (6.40, 6.55, 6.55, 6.15) and (6.30, 6.50, 6.20) for local and imported brands respectively. On the other hand, % acidity reached (0.185, 0181, 0170, 0183) and (0180, 0.180, 0.171) at the 24th week for local and imported brands respectively. Similar pattern of decline was also observed in specific gravity, fat and protein content in some local and imported samples especially at later stages of the study. In both cases, some of the recorded pH values, % acidity, sp. gravity and fat content were in violation of the accepted limits set by Libyan standard no. 356 for sterilized milk. Such changes in pH, % acidity and other UHT sterilized milk constituents during storage were coincided with a gradual decrease in the degree of acceptance of the stored milk samples of both types as shown by sensory scores recorded by the panelists. In either case degree of acceptance was significantly low at late stages of storage and most milk samples became relatively unacceptable after the 18th and 20th week for both untrained and trained panelists respectively.

Keywords: UHT milk, shelf life, quality, gravity, bacteria

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3259 Quality of Donut Supplemented with Hom Nin Rice Flour

Authors: Supatchalee Sirichokworrakit, Pannin Intasen, Chansuda Angkawut

Abstract:

Hom Nin rice (Oryza Sativa L.) was processed into flour and used to substitute wheat flour in donuts. The donuts were prepared with 0, 20, 40, 60, and 80% Hom Nin rice flour (HNF). The donuts were subjected to proximate, texture, color and sensory evaluations. The results of the study revealed that the ash, moisture, crude fiber contents increased while crude fat and protein contents decreased as the level of HNF increased. The hardness and chewiness of donut increased as the HNF increased but the cohesiveness, springiness, and specific volume decreased. Color of donut (L*, a*, and b* values) decreased with the addition of HNF. Overall acceptability for the 20-40% HNF additions did not differ significantly from the score of the 100% wheat flour.

Keywords: Hom Nin rice, donut, texture evaluation, sensory evaluation

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3258 The Design of Intelligent Passenger Organization System for Metro Stations Based on Anylogic

Authors: Cheng Zeng, Xia Luo

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Passenger organization has always been an essential part of China's metro operation and management. Facing the massive passenger flow, stations need to improve their intelligence and automation degree by an appropriate integrated system. Based on the existing integrated supervisory control system (ISCS) and simulation software (Anylogic), this paper designs an intelligent passenger organization system (IPOS) for metro stations. Its primary function includes passenger information acquisition, data processing and computing, visualization management, decision recommendations, and decision response based on interlocking equipment. For this purpose, the logical structure and intelligent algorithms employed are particularly devised. Besides, the structure diagram of information acquisition and application module, the application of Anylogic, the case library's function process are all given by this research. Based on the secondary development of Anylogic and existing technologies like video recognition, the IPOS is supposed to improve the response speed and address capacity in the face of emergent passenger flow of metro stations.

Keywords: anylogic software, decision-making support system, intellectualization, ISCS, passenger organization

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3257 Optimal Design Solution in "The Small Module" Within the Possibilities of Ecology, Environmental Science/Engineering, and Economics

Authors: Hassan Wajid

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We will commend accommodating an environmentally friendly architectural proposal that is extremely common/usual but whose features will make it a sustainable space. In this experiment, the natural and artificial built space is being proposed in such a way that deals with Environmental, Ecological, and Economic Criteria under different climatic conditions. Moreover, the criteria against ecology-environment-economics reflect in the different modules which are being experimented with and analyzed by multiple research groups. The ecological, environmental, and economic services are provided used as units of production side by side, resulting in local job creation and saving resources, for instance, conservation of rainwater, soil formation or protection, less energy consumption to achieve Net Zero, and a stable climate as a whole. The synthesized results from the collected data suggest several aspects to consider when designing buildings for beginning the design process under the supervision of instructors/directors who are responsible for developing curricula and sustainable goals. Hence, the results of the research and the suggestions will benefit the sustainable design through multiple results, heat analysis of different small modules, and comparisons. As a result, it is depleted as the resources are either consumed or the pollution contaminates the resources.

Keywords: optimization, ecology, environment, sustainable solution

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3256 Designing Information Systems in Education as Prerequisite for Successful Management Results

Authors: Vladimir Simovic, Matija Varga, Tonco Marusic

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This research paper shows matrix technology models and examples of information systems in education (in the Republic of Croatia and in the Germany) in support of business, education (when learning and teaching) and e-learning. Here we researched and described the aims and objectives of the main process in education and technology, with main matrix classes of data. In this paper, we have example of matrix technology with detailed description of processes related to specific data classes in the processes of education and an example module that is support for the process: ‘Filling in the directory and the diary of work’ and ‘evaluation’. Also, on the lower level of the processes, we researched and described all activities which take place within the lower process in education. We researched and described the characteristics and functioning of modules: ‘Fill the directory and the diary of work’ and ‘evaluation’. For the analysis of the affinity between the aforementioned processes and/or sub-process we used our application model created in Visual Basic, which was based on the algorithm for analyzing the affinity between the observed processes and/or sub-processes.

Keywords: designing, education management, information systems, matrix technology, process affinity

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3255 Assessment of the Effect of Ethanolic Leaf Extract of Annona squamosa L. on Den Induced Hepatocellular Carcinoma in Experimental Animals

Authors: Vanitha Varadharaj, Vijalakshmi Krishnamurthy

Abstract:

Annona squamosa Linn, commonly known as Sugar apple, belonging to the family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The alkaloid and flavonoids present in Annona squamosa leaf has proved to have antioxidant activity. The present work has been planned to investigate the effect of ethanolic leaf extract of Annona squamosa leaf on Den Induced wistar albino rats. The study was carried out to analyze the biochemical Parmeters like Total Proteins, Bilirubin, Enzymatic and Non –Enzymatic enzymes, Marker enzymes and Tumor markers in serum and also the histopathological studies in liver is carried out in control and DEN induced rats. Supplementation of ELAS (Ethanolic Leaf Extract Of Annona squamosa) reduced the liver weight and also reduced the tumour incidence. Chemoprevention group showed near normal values of bilirubin when compared with the control rats. Total protein was decreased in the cancer bearing group and on treatment with the extract the levels of protein were restored. Both in pre and post treatment group, the activities of enzymatic antioxidants such as superoxide dismutase, catalase, and Glutathione peroxidase were increased but in pre treated animals it was more effective than post treated animals. The non- enzymatic antioxidants such as vitamin C and vitamin E were brought back to normal level significantly in post and pre treated animals. Activities of marker enzymes such as SGOT, SGPT, ALP, γ GT were significantly elevated in the serum of cancer animals and the values returned to normal after treatment with the extract suggesting the hepato protective effect of the extract. Lipid peroxide was found to be elevated in the cancer induced group. This condition was brought back to the normal in the pre and post treated animals with ELAS. Histological examination also confirmed the anti- carcinogenic potential of ELAS, Cancer induced groups had a triple fold increase in their AFP values when compared to other groups. DEN treatment increased the level of AFP expression while ELAS partially counteracted the effect of it. So the scientific validation obtained from this study may pave way to many budding scientists to find new drugs from Annona squamosa for various ailments.

Keywords: annona squamosa, biochemical parmeters, cancer, leaf extract

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3254 Mechanical Properties of Biological Tissues

Authors: Young June Yoon

Abstract:

We will present four different topics in estimating the mechanical properties of biological tissues. First we elucidate the viscoelastic behavior of collagen molecules whose diameter is a couple of nanometers. By using the molecular dynamics simulation, we observed the viscoelastic behavior in different pulling velocity. Second, the protein layer, so called ‘sheath’ in enamel microstructure reduces the stress concentration in enamel minerals. We examined the result by using the finite element methods. Third, the anisotropic elastic constants of dentin are estimated by micromechanical analysis and estimated results are close to the experimentally measured data. Last, new formulation between the fabric tensor and the wave velocity is established for calcaneus by employing the poroelasticity. This formulation can be simply used for future experiments.

Keywords: tissues, mechanics, mechanical properties, wave propagation

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3253 Control of Helminthosporiosis in Oryza sativa Varieties Treated with 24-Epibrassinolide

Authors: Kuate Tueguem William Norbert, Ngoh Dooh Jules Patrice, Kone Sangou Abdou Nourou, Mboussi Serge Bertrand, Chewachang Godwill Mih, Essome Sale Charles, Djuissi Tohoto Doriane, Ambang Zachee

Abstract:

The objectives of this study were to evaluate the effects of foliar application of 24-epibrassinolide (EBR) on the development of rice helminthosporiosis caused by Bipolaris oryzae and its influence on the improvement of growth parameters and induction of the synthesis of defense substances in the rice plants. The experimental asset up involved a multifactorial split-plot with two varieties (NERICA 3 and local variety KAMKOU) and five treatments (T0: control, T1: EBR, T2: BANKO PLUS (fungicide), T3: NPK (chemical fertilizer), T4: mixture: NPK + BANKO PLUS + EBR) with three repetitions. Agro-morphological and epidemiological parameters, as well as substances for plant resistance, were evaluated over two growing seasons. The application of the EBR induced significant growth of the rice plants for the 2015 and 2016 growing seasons on the two varieties tested compared to the T0 treatment. At 74 days after sowing (DAS), NERICA 3 showed plant heights of 58.9 ± 5.4; 83.1 ± 10.4; 86.01 ± 9.4; 69.4 ± 11.1 and 87.12 ± 7.4 cm at T0; T1; T2; T3, and T4, respectively. Plant height for the variety KAMKOU varied from 87,12 ± 8,1; 88.1 ± 8.1 and 92.02 ± 6.3 cm in T1, T2, and T3 to 74.1 ± 8.6 and 74.21 ± 11.4 cm in T0 and T3. In accordance with the low rate of expansion of helminthosporiosis in experimental plots, EBR (T1) significantly reduced the development of the disease with severities of 0.0; 1.29, and 2.04%, respectively at 78; 92, and 111 DAS on the variety NERICA 3 compared with1; 3.15 and 3.79% in the control T0. The reduction of disease development/severity as a result of the application of EBR is due to the induction of acquired resistance of rice varieties through increased phenol (13.73 eqAG/mg/PMF) and total protein (117.89 eqBSA/mg/PMF) in the T1 treatment against 5.37 eqAG/mg/PMF and 104.97 eqBSA/mg/PMF in T0 for the NERICA 3 variety. Similarly, on the KAMKOU variety, 148.53 eqBSA/mg/PMF were protein and 6.10 eqAG/mg/PMF of phenol in T1. In summary, the results show the significant effect of EBR on plant growth, yield, synthesis of secondary metabolites and defense proteins, and disease resistance. The EBR significantly reduced losses of rice grains by causing an average gain of about 1.55 t/ha compared to the control and 1.00 t/ha compared to the NPK-based treatment for the two varieties studied. Further, the enzymatic activities of PPOs, POXs, and PR2s were higher in leaves from treated EBR-based plants. These results show that 24-epibrassinolide can be used in the control of helminthosporiosis of rice to reduce disease and increase yields.

Keywords: Oryza sativa, 24-epibrassinolide, helminthosporiosis, secondary metabolites, PR proteins, acquired resistance

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3252 Seal and Heal Miracle Ointment: Effects of Cryopreserved and Lyophilized Amniotic Membrane on Experimentally Induced Diabetic Balb/C Mice

Authors: Elizalde D. Bana

Abstract:

Healing restores continuity and form through cell replication; hence, conserving structural integrity. In response to the worldwide pressing problem of chronic wounds in the healthcare delivery system, the researcher aims to provide effective intervention to preserve the structural integrity of the person. The wound healing effects of cryopreserved and lyophilized amniotic membrane (AM) of a term fetus embedded into two (2) concentrations (1.5 % and 1.0 %) of absorption-based ointment has been evaluated in vivo using the excision wound healing model 1x1 cm size. The total protein concentration in full term fetus was determined by the Biuret and Bradford methods, which are based on UV-visible spectroscopy. The percentages of protein presence in 9.5 mg (Mass total sample) of Amniotic membrane ranges between 14.77 – 14.46 % in Bradford method, while slightly lower to 13.78 – 13.80 % concentration in Biuret method, respectively. Bradford method evidently showed higher sensitivity for proteins than Biuret test. Overall, the amniotic membrane is composed principally of proteins in which a copious amount of literature substantially proved its healing abilities. After which, an area of 1 cm by 1 cm skin tissue was excised to its full thickness from the dorsolateral aspect of the isogenic mice and was applied twice a day with the ointment formulation having two (2) concentrations for the diabetic group and non-diabetic group. The wounds of each animal were left undressed and its area was measured every other day by a standard measurement formula from day 2,4,6,8,10,12 and 14. By the 14th day, the ointment containing 1.5 % of AM in absorption-based ointment applied to non-diabetic and diabetic group showed 100 % healing. The wound areas in the animals treated with the standard antibiotic, Mupirocin Ointment (Brand X) showed a 100% healing by the 14th day but with traces of scars, indicating that AM prepared from cryopreservation and lyophilization, at that given concentration, had a better wound healing property than the standard antibiotic. Four (4) multivariate tests were used which showed a significant interaction between days and treatments, meaning that the ointments prepared in two differing concentrations and induced in different groups of the mice had a significant effect on the percent of contraction over time. Furthermore, the evaluations of its effectiveness to wound healing were all significant although in differing degrees. It is observed that the higher the concentrations of amniotic membrane, the more effective are the results.

Keywords: wounds, healing, amniotic membrane ointments, biomedical, stem cell

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3251 Experimental and Numerical Analysis of Built-In Thermoelectric Generator Modules with Elliptical Pin-Fin Heat Sink

Authors: J. Y Jang, C. Y. Tseng

Abstract:

A three-dimensional numerical model of thermoelectric generator (TEG) modules attached to a large chimney plate is proposed and solved numerically using a control volume based finite difference formulation. The TEG module consists of a thermoelectric generator, an elliptical pin-fin heat sink, and a cold plate for water cooling. In the chimney, the temperature of flue gases is 450-650K. Therefore, the effects of convection and radiation heat transfer are considered. Although the TEG hot-side temperature and thus the electric power output can be increased by inserting an elliptical pin-fin heat sink into the chimney tunnel to increase the heat transfer area, the pin fin heat sink would cause extra pumping power at the same time. The main purpose of this study is to analyze the effects of geometrical parameters on the electric power output and chimney pressure drop characteristics. In addition, the effects of different operating conditions, including various inlet velocities (Vin = 1, 3, 5 m/s) and inlet temperatures (Tgas = 450, 550, 650K) are discussed in detail. The predicted numerical data for the power vs. current (P-I) curve are in good agreement (within 11%) with the experimental data.

Keywords: thermoelectric generator, waste heat recovery, pin-fin heat sink, experimental and numerical analysis

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3250 Pressure Angle and Profile Shift Factor Effects on the Natural Frequency of Spur Tooth Design

Authors: Ali Raad Hassan

Abstract:

In this paper, an (irregular) case relating to base circle, root circle, and pressure angle has been discussed and a computer programme has been developed to simulate and plot spur gear tooth profile, including involute and trochoid curves based on the formulation of rack cutter using different values of pressure angle and profile shift factor and it gave the values of all important geometric parameters. The results showed the flexibility of this approach and versatility of the programme to draw many different cases of spur gear teeth of any module, pressure angle, profile shift factor, number of teeth and rack cutter tip radius. The procedure developed can be extended to produce finite element models of heretofore intractable geometrical forms, to exploring fabrication of nonstandard tooth forms also. Finite elements model of these irregular cases have been built using above programme, and modal analysis has been done using ANSYS software, and natural frequencies of these selected cases have been obtained and discussed.

Keywords: involute, trochoid, pressure angle, profile shift factor, natural frequency

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3249 Conformational Switch of hRAGE upon Self-Association

Authors: Ikhlas Ahmed, Jamillah Zamoon

Abstract:

The human receptor for advanced glycation end product is a plasma membrane receptor with an intrinsically disordered region. The protein consists of three extracellular domains, a single membrane spanning transmembrane domain, and a cytosolic domain which is intrinsically disordered and responsible for signaling. The disordered nature of the cytosolic domain allows it to be dynamic in solution. This receptor self-associates to higher forms. The association is triggered by ligand, metal or by the extracellular domain. Fluorescence spectroscopy technique is used to test the self-association of the different concentrations of the cytosolic domain. This work has concluded that the cytosolic domain of this receptor also self-associates. Moreover, the self-association does not require ligand or metal.

Keywords: fluorescence spectroscopy, hRAGE, IDP, Self-association

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3248 Experimental and Numerical Investigation of Micro-Welding Process and Applications in Digital Manufacturing

Authors: Khaled Al-Badani, Andrew Norbury, Essam Elmshawet, Glynn Rotwell, Ian Jenkinson , James Ren

Abstract:

Micro welding procedures are widely used for joining materials, developing duplex components or functional surfaces, through various methods such as Micro Discharge Welding or Spot Welding process, which can be found in the engineering, aerospace, automotive, biochemical, biomedical and numerous other industries. The relationship between the material properties, structure and processing is very important to improve the structural integrity and the final performance of the welded joints. This includes controlling the shape and the size of the welding nugget, state of the heat affected zone, residual stress, etc. Nowadays, modern high volume productions require the welding of much versatile shapes/sizes and material systems that are suitable for various applications. Hence, an improved understanding of the micro welding process and the digital tools, which are based on computational numerical modelling linking key welding parameters, dimensional attributes and functional performance of the weldment, would directly benefit the industry in developing products that meet current and future market demands. This paper will introduce recent work on developing an integrated experimental and numerical modelling code for micro welding techniques. This includes similar and dissimilar materials for both ferrous and non-ferrous metals, at different scales. The paper will also produce a comparative study, concerning the differences between the micro discharge welding process and the spot welding technique, in regards to the size effect of the welding zone and the changes in the material structure. Numerical modelling method for the micro welding processes and its effects on the material properties, during melting and cooling progression at different scales, will also be presented. Finally, the applications of the integrated numerical modelling and the material development for the digital manufacturing of welding, is discussed with references to typical application cases such as sensors (thermocouples), energy (heat exchanger) and automotive structures (duplex steel structures).

Keywords: computer modelling, droplet formation, material distortion, materials forming, welding

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3247 Exploring Students’ Voices in Lecturers’ Teaching and Learning Developmental Trajectory

Authors: Khashane Stephen Malatji, Makwalete Johanna Malatji

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Student evaluation of teaching (SET) is the common way of assessing teaching quality at universities and tracing the professional growth of lecturers. The aim of this study was to investigate the role played by student evaluation in the teaching and learning agenda at one South African University. The researchers used a qualitative approach and a case study research design. With regards to data collection, document analysis was used. Evaluation reports were reviewed to monitor the growth of lecturers who were evaluated during the academic years 2020 and 2021 in one faculty. The results of the study reveal that student evaluation remains the most relevant tool to inform the teaching agenda at a university. Lecturers who were evaluated were found to grow academically. All lecturers evaluated during 2020 have shown great improvement when evaluated repeatedly during 2021. Therefore, it can be concluded that student evaluation helps to improve the pedagogical and professional proficiency of lecturers. The study therefore, recommends that lecturers conduct an evaluation for each module they teach every semester or annually in case of year modules. The study also recommends that lecturers attend to all areas that draw negative comments from students in order to improve.

Keywords: students’ voices, teaching agenda, evaluation, feedback, responses

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3246 Modular Probe for Basic Monitoring of Water and Air Quality

Authors: Andrés Calvillo Téllez, Marianne Martínez Zanzarric, José Cruz Núñez Pérez

Abstract:

A modular system that performs basic monitoring of both water and air quality is presented. Monitoring is essential for environmental, aquaculture, and agricultural disciplines, where this type of instrumentation is necessary for data collection. The system uses low-cost components, which allows readings close to those with high-cost probes. The probe collects readings such as the coordinates of the geographical position, as well as the time it records the target parameters of the monitored. The modules or subsystems that make up the probe are the global positioning (GPS), which shows the altitude, latitude, and longitude data of the point where the reading will be recorded, a real-time clock stage, the date marking the time, the module SD memory continuously stores data, data acquisition system, central processing unit, and energy. The system acquires parameters to measure water quality, conductivity, pressure, and temperature, and for air, three types of ammonia, dioxide, and carbon monoxide gases were censored. The information obtained allowed us to identify the schedule of modification of the parameters and the identification of the ideal conditions for the growth of microorganisms in the water.

Keywords: calibration, conductivity, datalogger, monitoring, real time clock, water quality

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3245 Analysis on Thermococcus achaeans with Frequent Pattern Mining

Authors: Jeongyeob Hong, Myeonghoon Park, Taeson Yoon

Abstract:

After the advent of Achaeans which utilize different metabolism pathway and contain conspicuously different cellular structure, they have been recognized as possible materials for developing quality of human beings. Among diverse Achaeans, in this paper, we compared 16s RNA Sequences of four different species of Thermococcus: Achaeans genus specialized in sulfur-dealing metabolism. Four Species, Barophilus, Kodakarensis, Hydrothermalis, and Onnurineus, live near the hydrothermal vent that emits extreme amount of sulfur and heat. By comparing ribosomal sequences of aforementioned four species, we found similarities in their sequences and expressed protein, enabling us to expect that certain ribosomal sequence or proteins are vital for their survival. Apriori algorithms and Decision Tree were used. for comparison.

Keywords: Achaeans, Thermococcus, apriori algorithm, decision tree

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3244 Determination of Activation Energy for Thermal Decomposition of Selected Soft Tissues Components

Authors: M. Ekiert, T. Uhl, A. Mlyniec

Abstract:

Tendons are the biological soft tissue structures composed of collagen, proteoglycan, glycoproteins, water and cells of extracellular matrix (ECM). Tendons, which primary function is to transfer force generated by the muscles to the bones causing joints movement, are exposed to many micro and macro damages. In fact, tendons and ligaments trauma are one of the most numerous injuries of human musculoskeletal system, causing for many people (particularly for athletes and physically active people), recurring disorders, chronic pain or even inability of movement. The number of tendons reconstruction and transplantation procedures is increasing every year. Therefore, studies on soft tissues storage conditions (influencing i.e. tissue aging) seem to be an extremely important issue. In this study, an atomic-scale investigation on the kinetics of decomposition of two selected tendon components – collagen type I (which forms a 60-85% of a tendon dry mass) and elastin protein (which combine with ECM creates elastic fibers of connective tissues) is presented. A molecular model of collagen and elastin was developed based on crystal structure of triple-helical collagen-like 1QSU peptide and P15502 human elastin protein, respectively. Each model employed 4 linear strands collagen/elastin strands per unit cell, distributed in 2x2 matrix arrangement, placed in simulation box filled with water molecules. A decomposition phenomena was simulated with molecular dynamics (MD) method using ReaxFF force field and periodic boundary conditions. A set of NVT-MD runs was performed for 1000K temperature range in order to obtained temperature-depended rate of production of decomposition by-products. Based on calculated reaction rates activation energies and pre-exponential factors, required to formulate Arrhenius equations describing kinetics of decomposition of tested soft tissue components, were calculated. Moreover, by adjusting a model developed for collagen, system scalability and correct implementation of the periodic boundary conditions were evaluated. An obtained results provide a deeper insight into decomposition of selected tendon components. A developed methodology may also be easily transferred to other connective tissue elements and therefore might be used for further studies on soft tissues aging.

Keywords: decomposition, molecular dynamics, soft tissue, tendons

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3243 Angiomotin Regulates Integrin Beta 1-Mediated Endothelial Cell Migration and Angiogenesis

Authors: Yuanyuan Zhang, Yujuan Zheng, Giuseppina Barutello, Sumako Kameishi, Kungchun Chiu, Katharina Hennig, Martial Balland, Federica Cavallo, Lars Holmgren

Abstract:

Angiogenesis describes that new blood vessels migrate from pre-existing ones to form 3D lumenized structure and remodeling. During directional migration toward the gradient of pro-angiogenic factors, the endothelial cells, especially the tip cells need filopodia to sense the environment and exert the pulling force. Of particular interest are the integrin proteins, which play an essential role in focal adhesion in the connection between migrating cells and extracellular matrix (ECM). Understanding how these biomechanical complexes orchestrate intrinsic and extrinsic forces is important for our understanding of the underlying mechanisms driving angiogenesis. We have previously identified Angiomotin (Amot), a member of Amot scaffold protein family, as a promoter for endothelial cell migration in vitro and zebrafish models. Hence, we established inducible endothelial-specific Amot knock-out mice to study normal retinal angiogenesis as well as tumor angiogenesis. We found that the migration ratio of the blood vessel network to the edge was significantly decreased in Amotec- retinas at postnatal day 6 (P6). While almost all the Amot defect tip cells lost migration advantages at P7. In consistence with the dramatic morphology defect of tip cells, there was a non-autonomous defect in astrocytes, as well as the disorganized fibronectin expression pattern correspondingly in migration front. Furthermore, the growth of transplanted LLC tumor was inhibited in Amot knockout mice due to fewer vasculature involved. By using MMTV-PyMT transgenic mouse model, there was a significantly longer period before tumors arised when Amot was specifically knocked out in blood vessels. In vitro evidence showed that Amot binded to beta-actin, Integrin beta 1 (ITGB1), Fibronectin, FAK, Vinculin, major focal adhesion molecules, and ITGB1 and stress fibers were distinctly induced by Amot transfection. Via traction force microscopy, the total energy (force indicater) was found significantly decreased in Amot knockdown cells. Taken together, we propose that Amot is a novel partner of the ITGB1/Fibronectin protein complex at focal adhesion and required for exerting force transition between endothelial cell and extracellular matrix.

Keywords: angiogenesis, angiomotin, endothelial cell migration, focal adhesion, integrin beta 1

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3242 Evaluation of Dry Matter Yield of Panicum maximum Intercropped with Pigeonpea and Sesbania Sesban

Authors: Misheck Musokwa, Paramu Mafongoya, Simon Lorentz

Abstract:

Seasonal shortages of fodder during the dry season is a major constraint to smallholder livestock farmers in South Africa. To mitigate the shortage of fodder, legume trees can be intercropped with pastures which can diversify the sources of feed and increase the amount of protein for grazing animals. The objective was to evaluate dry matter yield of Panicum maximum and land productivity under different fodder production systems during 2016/17-2017/18 seasons at Empangeni (28.6391° S and 31.9400° E). A randomized complete block design, replicated three times was used, the treatments were sole Panicum maximum, Panicum maximum + Sesbania sesban, Panicum maximum + pigeonpea, sole Sesbania sesban, Sole pigeonpea. Three months S.sesbania seedlings were transplanted whilst pigeonpea was direct seeded at spacing of 1m x 1m. P. maximum seeds were drilled at a respective rate of 7.5 kg/ha having an inter-row spacing of 0.25 m apart. In between rows of trees P. maximum seeds were drilled. The dry matter yield harvesting times were separated by six months’ timeframe. A 0.25 m² quadrant randomly placed on 3 points on the plot was used as sampling area during harvesting P. maximum. There was significant difference P < 0.05 across 3 harvests and total dry matter. P. maximum had higher dry matter yield as compared to both intercrops at first harvest and total. The second and third harvest had no significant difference with pigeonpea intercrop. The results was in this order for all 3 harvest: P. maximum (541.2c, 1209.3b and 1557b) kg ha¹ ≥ P. maximum + pigeonpea (157.2b, 926.7b and 1129b) kg ha¹ > P. maximum + S. sesban (36.3a, 282a and 555a) kg ha¹. Total accumulation of dry matter yield of P. maximum (3307c kg ha¹) > P. maximum + pigeonpea (2212 kg ha¹) ≥ P. maximum + S. sesban (874 kg ha¹). There was a significant difference (P< 0.05) on seed yield for trees. Pigeonpea (1240.3 kg ha¹) ≥ Pigeonpea + P. maximum (862.7 kg ha¹) > S.sesbania (391.9 kg ha¹) ≥ S.sesbania + P. maximum. The Land Equivalent Ratio (LER) was in the following order P. maximum + pigeonpea (1.37) > P. maximum + S. sesban (0.84) > Pigeonpea (0.59) ≥ S. Sesbania (0.57) > P. maximum (0.26). Results indicates that it is beneficial to have P. maximum intercropped with pigeonpea because of higher land productivity. Planting grass with pigeonpea was more beneficial than S. sesban with grass or sole cropping in terms of saving the shortage of arable land. P. maximum + pigeonpea saves a substantial (37%) land which can be subsequently be used for other crop production. Pigeonpea is recommended as an intercrop with P. maximum due to its higher LER and combined production of livestock feed, human food, and firewood. Panicum grass is low in crude protein though high in carbohydrates, there is a need for intercropping it with legume trees. A farmer who buys concentrates can reduce costs by combining P. maximum with pigeonpea this will provide a balanced diet at low cost.

Keywords: fodder, livestock, productivity, smallholder farmers

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3241 Identification of Odorant Receptors through the Antennal Transcriptome of the Grapevine Pest, Lobesia botrana (Lepidoptera: Tortricidae)

Authors: Ricardo Godoy, Herbert Venthur, Hector Jimenez, Andres Quiroz, Ana Mutis

Abstract:

In agriculture, grape production has great economic importance at global level, considering that in 2013 it reached 7.4 million hectares (ha) covered by plantations of this fruit worldwide. Chile is the number one exporter in the world with 800,000 tons. However, these values have been threatened by the attack of the grapevine moth, Lobesia botrana (Denis & Schiffermuller) (Lepidoptera: Tortricidae), since its detection in 2008. Nowadays, the use of semiochemicals, in particular the major component of the sex pheromone, (E,Z)-7.9-dodecadienil acetate, are part of mating disruption methods to control L. botrana. How insect pests can recognize these molecules, is being part of huge efforts to deorphanize their olfactory mechanism at molecular level. Thus, an interesting group of proteins has been identified in the antennae of insects, where odorant-binding proteins (OBPs) are known by transporting molecules to odorant receptors (ORs) and a co-receptor (ORCO) causing a behavioral change in the insect. Other proteins such as chemosensory proteins (CSPs), ionotropic receptors (IRs), odorant degrading enzymes (ODEs) and sensory neuron membrane proteins (SNMPs) seem to be involved, but few studies have been performed so far. The above has led to an increasing interest in insect communication at a molecular level, which has contributed to both a better understanding of the olfaction process and the design of new pest management strategies. To date, it has been reported that the ORs can detect one or a small group of odorants in a specific way. Therefore, the objective of this study is the identification of genes that encode these ORs using the antennal transcriptome of L. botrana. Total RNA was extracted for females and males of L. botrana, and the antennal transcriptome sequenced by Next Generation Sequencing service using an Illumina HiSeq2500 platform with 50 million reads per sample. Unigenes were assembled using Trinity v2.4.0 package and transcript abundance was obtained using edgeR. Genes were identified using BLASTN and BLASTX locally installed in a Unix system and based on our own Tortricidae database. Those Unigenes related to ORs were characterized using ORFfinder and protein Blastp server. Finally, a phylogenetic analysis was performed with the candidate amino acid sequences for LbotORs including amino acid sequences of other moths ORs, such as Bombyx mori, Cydia pomonella, among others. Our findings suggest 61 genes encoding ORs and one gene encoding an ORCO in both sexes, where the greatest difference was found in the OR6 because of the transcript abundance according to the value of FPKM in females and males was 1.48 versus 324.00. In addition, according to phylogenetic analysis OR6 is closely related to OR1 in Cydia pomonella and OR6, OR7 in Epiphyas postvittana, which have been described as pheromonal receptors (PRs). These results represent the first evidence of ORs present in the antennae of L. botrana and a suitable starting point for further functional studies with selected ORs, such as OR6, which is potentially related to pheromonal recognition.

Keywords: antennal transcriptome, lobesia botrana, odorant receptors (ORs), phylogenetic analysis

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3240 Fractal: Formative Reflective Assessment and Critical Thinking in Learning

Authors: Yannis Stavrakakis, Damian Gordon

Abstract:

Critical Thinking and Reflective Practice are two vital skills that students undertaking postgraduate studies should ideally possess. To help students develop and enhance these skills, this research developed several authentic activities to be undertaken as part of a module that is delivered early in a taught MSc to enhance these skills. One of the challenges of these topics is that they are somewhat ill-defined in terms of precisely what they mean, and also, there is no clear route to operationalizing the teaching of these skills. This research focuses on identifying suitable models of these skills and delivering them in a manner that is both clear and highly motivating. To achieve this, a class of 22 Master's students was divided into two groups, one was provided with a presentation and checklist about critical thinking skills, and the other group was given the same materials on the reflective practice process. The groups were given two scenarios each to analyze using their respective checklists and were asked to present their outcomes to each other and give peer review. The results were coded and compared, and key differences were noted, including the fact that the Critical Thinking outcomes were more future-focused, and the Reflective Practice outcomes were more past-focused and present-focused, as well as the fact that the Reflective Practice process generated a significantly wider range of perspectives on the scenarios.

Keywords: critical thinking, ethical scenarios, formative assessment, reflective practice

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3239 The Mediation Impact of Demographic and Clinical Characteristics on the Relationship between Trunk Control and Quality of Life among the Sub-Acute Stroke Population: A Cross-Sectional Study

Authors: Kumar Gular, Viswanathan S., Mastour Saeed Alshahrani, Ravi Shankar Reddy, Jaya Shanker Tedla, Snehil Dixit, Ajay Prasad Gautam, Venkata Nagaraj Kakaraparthi, Devika Rani Sangadala

Abstract:

Background: Despite trunk control’s significant contribution to improving various functional activity components, the independent effect of trunk performance on quality of life is yet to be estimated in stroke survivors. Ascertaining the correlation between trunk control and self-reported quality of life while evaluating the effect of demographic and clinical characteristics on their relationship will guide concerned healthcare professionals in designing ideal rehabilitation protocols during the late sub-acute stroke stage of recovery. The aims of the present research were to (1) investigate the associations of trunk performance with self-rated quality of life and (2) evaluate if age, body mass index (BMI), and clinical characteristics mediate the relationship between trunk motor performance and perceived quality of life in the sub-acute stroke population. Methods: Trunk motor functions and quality of life among the late sub-acute stroke population aged 57.53 ± 6.42 years were evaluated through the trunk Impairment Scale (TIS) and Stroke specific quality of life (SSQOL) questionnaire, respectively. Pearson correlation coefficients and mediation analysis were performed to elucidate the relationship of trunk motor function with quality of life and determine the mediation impact of demographic and clinical characteristics on their association, respectively. Results: The current study observed significant correlations between trunk motor functions (TIS) and quality of life (SSQOL) with r=0.68 (p<0.001). Age, BMI, and type of stroke were detected as potential mediating factors in the association between trunk performance and quality of life. Conclusion: Validated associations between trunk motor functions and perceived quality of life among the late sub-acute stroke population emphasize the importance of comprehensive evaluation of trunk control. Rehabilitation specialists should focus on appropriate strategies to enhance trunk performance anticipating the potential effects of age, BMI, and type of stroke to improve health-related quality of life in stroke survivors.

Keywords: sub-acute stroke, quality of life, functional independence, trunk control

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3238 Peach as a Potential Functional Food: Biological Activity and Important Phenolic Compound Source

Authors: Luís R. Silva, Catarina Bento, Ana C. Gonçalves, Fábio Jesus, Branca M. Silva

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Nowadays, the general population is more and more concerned about nutrition and the health implications of an unbalanced diet. Current knowledge regarding the health benefits and antioxidant properties of certain foods such as fruits and vegetables has gained the interest of both the general public and scientific community. Peach (Prunus persica (L.) Batsch) is one of the most consumed fruits worldwide, with low sugar contents and a broad range of nutrients essential to the normal functioning of the body. Six different peach cultivars from the Fundão region in Portugal were evaluated regarding their phenolic composition by LC-DAD and biological activity. The prepared extracts’ capacity to scavenge free-radicals was tested through the stable free radical DPPH• and nitric oxide (•NO). Additionally, antidiabetic potential and protective effects against peroxyl radical (ROO•) induced damage to erythrocytes were also tested. LC-DAD analysis allowed the identification of 17 phenolic compounds, among which 5-O-caffeoylquinic acids and 3-O-caffeoylquinic acids are pointed out as the most abundant. Regarding the antioxidant activity, all cultivars displayed concentration-dependent free-radical scavenging activity against both nitrogen species and DPPH•. In respect to α-glucosidase inhibitory activity, Royal Magister and Royal Glory presented the highest inhibitory activity (IC50 = 11.7 ± 1.4 and 17.1 ± 1.7 μg/mL, respectively), nevertheless all six cultivars presented higher activity than the control acarbose. As for the protective effect of Royal Lu extract on the oxidative damage induced in erythrocytes by ROO•, the results were quite promising showing inhibition IC50 values of 110.0 ± 4.5 μg/mL and 83.8 ± 6.5 μg/mL for hemolysis and hemoglobin oxidation, respectively. The demonstrated activity is of course associated to the peaches’ phenolic profile, rich in phenolic acids and flavonoids with high hydrogen donating capacity. These compounds have great industrial interest for the manufacturing of natural products. The following step would naturally be the extraction and isolation from the plant tissues and large-scale production through biotechnology techniques.

Keywords: antioxidants, functional food, phenolic compounds, peach

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3237 Traditional Terms, Spaces, Forms and Artifacts in Cultural Semiotics of Southwest Nigeria

Authors: Ajibade Adeyemo

Abstract:

The paper examined local terms used for spaces, forms and building practices in southwest Nigeria as cultural semiotics. Housing has more cultural meaning than mere shelter as shown in building terms such as ‘roof over my head’. The study is significant in the study area because its people were traditionally orally centered until ‘culture contact’ led to graphical presentation and appreciation in the form of drawings which is a modern language of architecture. This semiotic study will facilitate the understanding of the wholesomeness of traditional building practices and thoughts. This is in the culture of the traditional multi-sensory appreciation of architecture, urban design and the arts. It will analyze traditional aphoristic words and terms which are like proverbs which are significant in language because of their metaphorical essence. Many of such terms in the dominant Yoruba language of the study area are oftentimes phenomenal reducing universal terms like the earth and heaven to the simple module of housing. These words could be worth investigating because they are symbolic serve as codes which are cultural tool of regional ethnic significance. Sassure’s and Pierce’s concepts of Semiotics in line with Eco’s concept of semiotics of metaphor shall be deployed.

Keywords: traditional terms, spaces, forms, artifacts, cultural semiotics, southwest

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3236 In vitro Effects of Porcine Follicular Fluid Proteins on Cell Culture Growth in Luteal Phase Porcine Oviductal Epithelial Cells

Authors: Mayuva Youngsabanant, Chanikarn Srinark, Supanyika Sengsai, Soratorn Kerdkriangkrai, Nongnuch Gumlungpat, Mayuree Pumipaiboon

Abstract:

The follicular fluid proteins of healthy medium size follicles (4-6 mm in diameters) and large size follicles (7-8 mm in diameter) of large white pig ovaries were collected by using sterile technique. They were used for testing the effect on primary in vitro cell culture growth of porcine oviductal epithelial cells (pOEC). Porcine oviductal epithelial cells of luteal phase was culture in M199 and added with 10% fetal calf serum 2.2 mg/mL, NaHCO₃, 0.25 mM pyruvate, 15 µg/mL and 50 µg/mL, gentamycin sulfate at high humidified atmosphere with 5% CO₂ in 95% air atmosphere at 37°C for 96 h before testing. The optimized concentration of pFF of two follicle sizes (at concentration of 2, 4, 20, 40, 200, 400, 500, and 600 µg proteins) in culture medium was observed for 24 h using MTT assay. Results were analyzed with a one-way ANOVA in SPSS statistic. Moreover, pOEC was also studied in morphological characteristic on long-term culture. The results of long-term study revealed that pOEC showed 70-80 percentage of healthy morphology on epithelial-like character and contained 30 percentage of an elongated shape (fibroblast-like morphology) at 4 weeks of culture time. MTT assay reviewed an increase in the percentage of viability of pOEC in 2 treated of follicular fluid groups. Two treatment concentration groups were higher than control group (p < 0.05) but not in positive control group. Interestingly, at 200 µg protein of 2 treated follicular fluid groups were reached the highest cell viability which is higher than a positive control and it is significantly different form control group (P < 0.05). These cells are developed and had fibroblast elongate shape which is longer than the cells in control group and positive control group. This report implies that pFF of medium follicle size at 200 µg proteins and large follicle size at 200 and 500 µg proteins could be optimized concentration for using as a supplement in culture medium to promote cell growth and development instead of growth hormone from fetal calf serum. It could be applied in cell biotechnology researches. Acknowledgements: The project was funded by a grant from Silpakorn University Research and Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.

Keywords: in vitro, porcine follicular fluid protein (pFF), porcine oviductal epithelial cells (pOEC), MTT

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3235 Control of Lymphatic Remodelling by miR-132

Authors: Valeria Arcucci, Musarat Ishaq, Steven A. Stacker, Greg J. Goodall, Marc G. Achen

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Metastasis is the lethal aspect of cancer for most patients. Remodelling of lymphatic vessels associated with a tumour is a key initial step in metastasis because it facilitates the entry of cancer cells into the lymphatic vasculature and their spread to lymph nodes and distant organs. Although it is clear that vascular endothelial growth factors (VEGFs), such as VEGF-C and VEGF-D, are key drivers of lymphatic remodelling, the means by which many signaling pathways in endothelial cells are coordinately regulated to drive growth and remodelling of lymphatics in cancer is not understood. We seek to understand the broader molecular mechanisms that control cancer metastasis, and are focusing on microRNAs, which coordinately regulate signaling pathways involved in complex biological responses in health and disease. Here, using small RNA sequencing, we found that a specific microRNA, miR-132, is upregulated in expression in lymphatic endothelial cells (LECs) in response to the lymphangiogenic growth factors. Interestingly, ectopic expression of miR-132 in LECs in vitro stimulated proliferation and tube formation of these cells. Moreover, miR-132 is expressed in lymphatic vessels of a subset of human breast tumours which were previously found to express high levels of VEGF-D by immunohistochemical analysis on tumour tissue microarrays. In order to dissect the complexity of regulation by miR-132 in lymphatic biology, we performed Argonaute HITS-CLIP, which led us to identify the miR-132-mRNA interactome in LECs. We found that this microRNA in LECs is involved in the control of many different pathways mainly involved in cell proliferation and regulation of the extracellular matrix and cell-cell junctions. We are now exploring the functional significance of miR-132 targets in the biology of LECs using biochemical techniques, functional in vitro cell assays and in vivo lymphangiogenesis assays. This project will ultimately define the molecular regulation of lymphatic remodelling by miR-132, and thereby identify potential therapeutic targets for drugs designed to restrict the growth and remodelling of tumour lymphatics resulting in metastatic spread.

Keywords: argonaute HITS-CLIP, cancer, lymphatic remodelling, miR-132, VEGF

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