Search results for: STEM and N-STEM undergraduates
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 868

Search results for: STEM and N-STEM undergraduates

628 In vitro Regeneration of Neural Cells Using Human Umbilical Cord Derived Mesenchymal Stem Cells

Authors: Urvi Panwar, Kanchan Mishra, Kanjaksha Ghosh, ShankerLal Kothari

Abstract:

Background: Day-by-day the increasing prevalence of neurodegenerative diseases have become a global issue to manage them by medical sciences. The adult neural stem cells are rare and require an invasive and painful procedure to obtain it from central nervous system. Mesenchymal stem cell (MSCs) therapies have shown remarkable application in treatment of various cell injuries and cell loss. MSCs can be derived from various sources like adult tissues, human bone marrow, umbilical cord blood and cord tissue. MSCs have similar proliferation and differentiation capability, but the human umbilical cord-derived mesenchymal stem cells (hUCMSCs) are proved to be more beneficial with respect to cell procurement, differentiation to other cells, preservation, and transplantation. Material and method: Human umbilical cord is easily obtainable and non-controversial comparative to bone marrow and other adult tissues. The umbilical cord can be collected after delivery of baby, and its tissue can be cultured using explant culture method. Cell culture medium such as DMEMF12+10% FBS and DMEMF12+Neural growth factors (bFGF, human noggin, B27) with antibiotics (Streptomycin/Gentamycin) were used to culture and differentiate mesenchymal stem cells into neural cells, respectively. The characterisations of MSCs were done with Flow Cytometer for surface markers CD90, CD73 and CD105 and colony forming unit assay. The differentiated various neural cells will be characterised by fluorescence markers for neurons, astrocytes, and oligodendrocytes; quantitative PCR for genes Nestin and NeuroD1 and Western blotting technique for gap43 protein. Result and discussion: The high quality and number of MSCs were isolated from human umbilical cord via explant culture method. The obtained MSCs were differentiated into neural cells like neurons, astrocytes and oligodendrocytes. The differentiated neural cells can be used to treat neural injuries and neural cell loss by delivering cells by non-invasive administration via cerebrospinal fluid (CSF) or blood. Moreover, the MSCs can also be directly delivered to different injured sites where they differentiate into neural cells. Therefore, human umbilical cord is demonstrated to be an inexpensive and easily available source for MSCs. Moreover, the hUCMSCs can be a potential source for neural cell therapies and neural cell regeneration for neural cell injuries and neural cell loss. This new way of research will be helpful to treat and manage neural cell damages and neurodegenerative diseases like Alzheimer and Parkinson. Still the study has a long way to go but it is a promising approach for many neural disorders for which at present no satisfactory management is available.

Keywords: bone marrow, cell therapy, explant culture method, flow cytometer, human umbilical cord, mesenchymal stem cells, neurodegenerative diseases, neuroprotective, regeneration

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627 The Impact of Total Parenteral Nutrition on Pediatric Stem Cell Transplantation and Its Complications

Authors: R. Alramyan, S. Alsalamah, R. Alrashed, R. Alakel, F. Altheyeb, M. Alessa

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Background: Nutritional support with total parenteral nutrition (TPN) is usually commenced with hematopoietic stem cell transplantation (HSCT) patients. However, it has its benefits and risks. Complications related to central venous catheter such as infections, and metabolic disturbances, including abnormal liver function, is usually of concern in such patients. Methods: A retrospective charts review of all pediatric patients who underwent HSCT between the period 2015-2018 in a tertiary hospital in Riyadh, Saudi Arabia. Patients' demographics, types of conditioning, type of nutrition, and patients' outcomes were collected. Statistical analysis was conducted using SPSS version 22. Frequencies and percentages were used to describe categorical variables. Mean, and standard deviation were used for continuous variables. A P value of less than 0.05 was considered as statically significant. Results: a total of 162 HSCTs were identified during the period mentioned. Indication of allogenic transplant included hemoglobinopathy in 50 patients (31%), acute lymphoblastic leukemia in 21 patients (13%). TPN was used in 96 patients (59.30%) for a median of 14 days, nasogastric tube feeding (NGT) in 16 (9.90%) patients for a median of 11 days, and 71 of patients (43.80%) were able to tolerate oral feeding. Out of the 96 patients (59.30%) who were dependent on TPN, 64 patients (66.7%) had severe mucositis in comparison to 17 patients (25.8%) who were either on NGT or tolerated oral intake. (P-value= 0.00). Sinusoidal obstruction syndrome (SOS) was seen in 14 patients (14.6%) who were receiving TPN compared to none in non-TPN patients (P=value 0.001). Moreover, majority of patients who had SOS received myeloablative conditioning therapy for non-malignant disease (hemoglobinopathy). However, there were no statistically significant differences in Graft-vs-Host Disease (both acute and chronic), bacteremia, and patient outcome between both groups. Conclusions: Nutritional support using TPN is used in majority of patients, especially post-myeloablative conditioning associated with severe mucositis. TPN was associated with VOD, especially in hemoglobinopathy patients who received myeloablative therapy. This may emphasize on use of preventative measures such as fluid restriction, use of diuretics, or defibrotide in high-risk patients.

Keywords: hematopoeitic stem cell transplant, HSCT, stem cell transplant, sinusoidal obstruction syndrome, total parenteral nutrition

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626 Flexural Properties of Typha Fibers Reinforced Polyester Composite

Authors: Sana Rezig, Yosr Ben Mlik, Mounir Jaouadi, Foued Khoffi, Slah Msahli, Bernard Durand

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Increasing interest in environmental concerns, natural fibers are once again being considered as reinforcements for polymer composites. The main objective of this study is to explore another natural resource, Typha fiber; which is renewable without production cost and available abundantly in nature. The aim of this study was to study the flexural properties of composite resin with and without reinforcing Typha leaf and stem fibers. The specimens were made by the hand-lay-up process using polyester matrix. In our work, we focused on the effect of various treatment conditions (sea water, alkali treatment and a combination of the two treatments), as a surface modifier, on the flexural properties of the Typha fibers reinforced polyester composites. Moreover, weight ratio of Typha leaf or stem fibers was investigated. Besides, both fibers from leaf and stem of Typha plant were used to evaluate the reinforcing effect. Another parameter, which is reinforcement structure, was investigated. In fact, a first composite was made with air-laid nonwoven structure of fibers. A second composite was with a mixture of fibers and resin for each kind of treatment. Results show that alkali treatment and combined process provided better mechanical properties of composites in comparison with fiber treated by sea water. The fiber weight ratio influenced the flexural properties of composites. Indeed, a maximum value of flexural strength of 69.8 and 62,32 MPa with flexural modulus of 6.16 and 6.34 GPawas observed respectively for composite reinforced with leaf and stem fibers for 12.6 % fiber weight ratio. For the different treatments carried out, the treatment using caustic soda, whether alone or after retting seawater, show the best results because it improves adhesion between the polyester matrix and the fibers of reinforcement. SEM photographs were made to ascertain the effects of the surface treatment of the fibers. By varying the structure of the fibers of Typha, the reinforcement used in bulk shows more effective results as that used in the non-woven structure. In addition, flexural strength rises with about (65.32 %) in the case of composite reinforced with a mixture of 12.6% leaf fibers and (27.45 %) in the case of a composite reinforced with a nonwoven structure of 12.6 % of leaf fibers. Thus, to better evaluate the effect of the fiber origin, the reinforcing structure, the processing performed and the reinforcement factor on the performance of composite materials, a statistical study was performed using Minitab. Thus, ANOVA was used, and the patterns of the main effects of these parameters and interaction between them were established. Statistical analysis, the fiber treatment and reinforcement structure seem to be the most significant parameters.

Keywords: flexural properties, fiber treatment, structure and weight ratio, SEM photographs, Typha leaf and stem fibers

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625 Antımıcrobıal Actıvıty of Gırardınıa Heterophılla

Authors: P. S. BEDI* , Neavty Thakur, Balvınder Sıngh

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In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3, NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria eg. Bacillus subtilis, Gram negative bacteria eg. E. coli, K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant posseses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

Keywords: Girardinia heterophylla, leaves and stem extracts, Antibacterial activity, antifungal activity.

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624 Designing Entrepreneurship Education Contents for Entrepreneurial Intention Building among Undergraduates in India

Authors: Sumita Srivastava

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Despite several measures taken by the Government of India, entrepreneurship is still not perceived as a viable career option by the young generation. Although the rate of startups has improved a little after the penetration of e portals as business platforms, still the numbers are not very significant. It is also important to note that entrepreneurial initiatives are mostly taken up by graduates of premier institutions of India like Indian Institute of Technology (IITs) and Indian Institute of Management (IIMs). The scenario is not very satisfactory amongst the masses graduating from mainstream universities of the country. Indian youth at large are not attracted towards entrepreneurship as a career choice. The reason probably lies in the social fabric of the country and inappropriate education system which does not support the entrepreneurship at large amongst youth in the country. Education is critical to the development of an economy from the poverty level to the level of self-sustenance and development. The current curriculum in the majority of business schools in India prepares the average graduate to become employed by the available firms or business owners in society. For graduates in other streams, employment opportunities are very limited. The aim of this study was to identify and design entrepreneurship education contents to encourage undergraduates to pursue entrepreneurship as a career choice. This comprehensive study was conducted in multiple stages. Extensive research was conducted at each stage with an appropriate methodology. These stages of the project study were interconnected with each other, and each preceding stage provided inputs for the following stage of the study. In the first stage of the study, an empirical analysis was conducted to understand the current state of entrepreneurial intentions of undergraduates of Agra city. Various stakeholders were contacted at the stage, including students (n = 500), entrepreneurs (n = 20) and academicians and field experts (n = 10). At the second stage of the project study, a systems science technique, Nominal Group Technique (NGT) was used to identify the critical elements of entrepreneurship education in India based upon the findings of stage 1. The application of the Nominal Group Technique involved a workshop format; 15 domain experts participated in the workshop. Throughout the process, a democratic process was followed to avoid individual dominance and premature focusing on a single idea. The study obtained 63 responses from experts for effective entrepreneurship education in India. The responses were reduced to seven elements after a few thematic iterations. These elements were then segregated into content (knowledge, skills and attitude) and learning interaction on the basis of experts’ responses. After identifying critical elements of entrepreneurship education in the previous stage, the course was designed and validated at stage 3 of the project. Scientific methods were used at this stage to validate the curriculum contents and training interventions experimentally. The educational and training interventions designed through this study would not only help in developing entrepreneurial intentions but also creating skills relevant to the local entrepreneurial opportunities in the vicinity.

Keywords: curriculum design, entrepreneurial intention, entrepreneuship education, nominal group technique

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623 Extraction and Characterization of Ethiopian Hibiscus macranthus Bast Fiber

Authors: Solomon Tilahun Desisa, Muktar Seid Hussen

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Hibiscus macranthus is one of family Malvaceae and genus Hibiscus plant which grows mainly in western part of Ethiopia. Hibiscus macranthus is the most adaptable and abundant plant in the nation, which are used as an ornamental plant often a hedge or fence plant, and used as a firewood after harvesting the stem together with the bark, and used also as a fiber for trying different kinds of things by forming the rope. However, Hibiscus macranthus plant fibre has not been commercially exploited and extracted properly. This study of work describes the possibility of mechanical and retting methods of Hibiscus macranthus fibre extraction and characterization. Hibiscus macranthus fibre is a bast fibre which obtained naturally from the stem or stalks of the dicotyledonous plant since it is a natural cellulose plant fiber. And the fibre characterized by studying its physical and chemical properties. The physical characteristics were investigated as follows, including the length of 100-190mm, fineness of 1.0-1.2Tex, diameter under X100 microscopic view 16-21 microns, the moisture content of 12.46% and dry tenacity of 48-57cN/Tex along with breaking extension of 0.9-1.6%. Hibiscus macranthus fiber productivity was observed that 12-18% of the stem out of which more than 65% is primary long fibers. The fiber separation methods prove to decrease of non-cellulose ingredients in the order of mechanical, water and chemical methods. The color measurement also shows the raw Hibiscus macranthus fiber has a natural golden color according to YID1925 and paler look under both retting methods than mechanical separation. Finally, it is suggested that Hibiscus macranthus fibre can be used for manufacturing of natural and organic crop and coffee packages as well as super absorbent, fine and high tenacity textile products.

Keywords: Hibiscus macranthus, bast fiber, extraction, characterization

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622 Growth and Differentiation of Mesenchymal Stem Cells on Titanium Alloy Ti6Al4V and Novel Beta Titanium Alloy Ti36Nb6Ta

Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

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Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

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621 ReactorDesign App: An Interactive Software for Self-Directed Explorative Learning

Authors: Chia Wei Lim, Ning Yan

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The subject of reactor design, dealing with the transformation of chemical feedstocks into more valuable products, constitutes the central idea of chemical engineering. Despite its importance, the way it is taught to chemical engineering undergraduates has stayed virtually the same over the past several decades, even as the chemical industry increasingly leans towards the use of software for the design and daily monitoring of chemical plants. As such, there has been a widening learning gap as chemical engineering graduates transition from university to the industry since they are not exposed to effective platforms that relate the fundamental concepts taught during lectures to industrial applications. While the success of technology enhanced learning (TEL) has been demonstrated in various chemical engineering subjects, TELs in the teaching of reactor design appears to focus on the simulation of reactor processes, as opposed to arguably more important ideas such as the selection and optimization of reactor configuration for different types of reactions. This presents an opportunity for us to utilize the readily available easy-to-use MATLAB App platform to create an educational tool to aid the learning of fundamental concepts of reactor design and to link these concepts to the industrial context. Here, interactive software for the learning of reactor design has been developed to narrow the learning gap experienced by chemical engineering undergraduates. Dubbed the ReactorDesign App, it enables students to design reactors involving complex design equations for industrial applications without being overly focused on the tedious mathematical steps. With the aid of extensive visualization features, the concepts covered during lectures are explicitly utilized, allowing students to understand how these fundamental concepts are applied in the industrial context and equipping them for their careers. In addition, the software leverages the easily accessible MATLAB App platform to encourage self-directed learning. It is useful for reinforcing concepts taught, complementing homework assignments, and aiding exam revision. Accordingly, students are able to identify any lapses in understanding and clarify them accordingly. In terms of the topics covered, the app incorporates the design of different types of isothermal and non-isothermal reactors, in line with the lecture content and industrial relevance. The main features include the design of single reactors, such as batch reactors (BR), continuously stirred tank reactors (CSTR), plug flow reactors (PFR), and recycle reactors (RR), as well as multiple reactors consisting of any combination of ideal reactors. A version of the app, together with some guiding questions to aid explorative learning, was released to the undergraduates taking the reactor design module. A survey was conducted to assess its effectiveness, and an overwhelmingly positive response was received, with 89% of the respondents agreeing or strongly agreeing that the app has “helped [them] with understanding the unit” and 87% of the respondents agreeing or strongly agreeing that the app “offers learning flexibility”, compared to the conventional lecture-tutorial learning framework. In conclusion, the interactive ReactorDesign App has been developed to encourage self-directed explorative learning of the subject and demonstrate the industrial applications of the taught design concepts.

Keywords: explorative learning, reactor design, self-directed learning, technology enhanced learning

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620 Hypoglycaemic and Hypolipidemic Activity of Cassia occidentalis Linn. Stem Bark Extract in Streptozotocin Induced Diabetes

Authors: Manjusha Choudhary

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Objective: Cassia occidentalis Linn. belongs to Family Caesalpiniaceae is a common weed scattered from the foothills of Himalayas to West Bengal, South India, Burma, and Sri Lanka. It is used widely in folklore medicine in India as laxative, expectorant, analgesic, anti-malarial, hepatoprotective, relaxant, anti-inflammatory and antidiabetic. The present study was carried out to investigate the hypoglycaemic and hypolipidemic activities of ethanolic extract of Cassia occidentalis stem bark. Methods: Stem bark extract of Cassia occidentalis (SBCO) was administered orally at 250 and 500 mg/kg doses to normal and streptozotocin (STZ) induced type-2 diabetic mice. Various parameters like fasting blood glucose (FBG) level, serum cholesterol, high density lipoprotein (HDL) cholesterol, triglycerides (TG), total protein, urea, creatinine, serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) levels and physical parameters like change in body weight, food intake, water intake were performed for the evaluation of antidiabetic effects. Results: Both the doses of extract caused a marked decrease in FBG levels in STZ induced type 2 diabetic mice. Administration of SBCO led to the decrease in the blood glucose, food intake, water intake, organ weight, SGOT, SGPT levels with significant value and increased the levels of TG, HDL cholesterol, creatinine, cholesterol, total protein with a significant value (p < 0.05-0.01). The decrease in body weight induced by STZ was restored to normal with a significant value (p < 0.01) at both doses. Conclusion: Present study reveals that SBCO possess potent hypoglycaemic and hypolipidemic activities and supports the folklore use of the stem bark of plant as antidiabetic agent.

Keywords: Cassia occidentalis, diabetes, folklore, herbs, hypoglycemia, streptozotocin

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619 TNF Receptor-Associated Factor 6 (TRAF6) Mediating the Angiotensin-Induced Non-Canonical TGFβ Pathway Activation and Differentiation of c-kit+ Cardiac Stem Cells

Authors: Qing Cao, Fei Wang, Yu-Qiang Wang, Li-Ya Huang, Tian-Tian Sang, Shu-Yan Chen

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Aims: TNF Receptor-Associated Factor 6 (TRAF6) acts as a multifunctional regulator of the Transforming Growth Factor (TGF)-β signaling pathway, and mediates Smad-independent JNK and p38 activation via TGF-β. This study was performed to test the hypothesis that TGF-β/TRAF6 is essential for angiotensin-II (Ang II)-induced differentiation of rat c-kit+ Cardiac Stem Cells (CSCs). Methods and Results: c-kit+ CSCs were isolated from neonatal Sprague Dawley (SD) rats, and their c-kit status was confirmed with immunofluorescence staining. A TGF-β type I receptor inhibitor (SB431542) or the small interfering RNA (siRNA)-mediated knockdown of TRAF6 were used to investigate the role of TRAF6 in TGF-β signaling. Rescue of TRAF6 siRNA transfected cells with a 3'UTR deleted siRNA insensitive construct was conducted to rule out the off target effects of the siRNA. TRAF6 dominant negative (TRAF6DN) vector was constructed and used to infect c-kit+ CSCs, and western blotting was used to assess the expression of TRAF6, JNK, p38, cardiac-specific proteins, and Wnt signaling proteins. Physical interactions between TRAF6 and TGFβ receptors were studied by coimmunoprecipitation. Cardiac differentiation was suppressed in the absence of TRAF6. Forced expression of TRAF6 enhanced the expression of TGF-β-activated kinase1 (TAK1), and inhibited Wnt signaling. Furthermore, TRAF6 increased the expression of cardiac-specific proteins (cTnT and Cx-43) but inhibited the expression of Wnt3a. Conclusions: Our data suggest that TRAF6 plays an important role in Ang II induced differentiation of c-kit+ CSCs via the non-canonical signaling pathway.

Keywords: cardiac stem cells, differentiation, TGF-β, TRAF6, ubiquitination, Wnt

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618 Cellular Technologies in Urology

Authors: R. Zhankina, U. Zhanbyrbekuly, A. Tamadon, M. Askarov, R. Sherkhanov, D. Akhmetov, D. Saipiyeva, N. Keulimzhaev

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Male infertility affects about 15% of couples of reproductive age. Approximately 10–15% have azoospermia who have previously been diagnosed with male infertility. Azoospermia is regarded as the absence of spermatozoa in the ejaculate and is found in 10-15% of infertile men. Non-obstructive azoospermia is considered a cause of male infertility that is not amenable to drug therapy. Patients with non-obstructive azoospermia are unable to have their "own" children and have only options for adoption or use of donor sperm. Advances in assisted reproductive technologies such as intracytoplasmic sperm injection in vitro fertilization have significantly changed the management of patients with non-obstructive azoospermia. Advances in biotechnology have increased the options for treating patients with non-obstructive azoospermia. Mesenchymal stem cell therapy has been recognized as a new option for infertility treatment. Material and methods of the study: After obtaining informed consent, 5 patients diagnosed with non-obstructive azoospermia were included in an open, non-randomized study. The age of the patients ranged from 24 to 35 years. The examination was carried out before the start of treatment, which included biochemical blood tests, hormonal profile levels (luteinizing hormone, follicle-stimulating hormone, testosterone, prolactin, inhibin B); tests for tumor markers; genetic research. All studies were carried out in compliance with the requirements of Protocol No. 8 dated 06/09/20, approved by the Local Ethical Commission of NJSC "Astana Medical University". The control examination of patients was carried out after 6 months, by re-taking the program and hormonal profile (testosterone, luteinizing hormone, follicle-stimulating hormone, prolactin, inhibin B). Before micro-TESE of the testis, all 5 patients underwent myeloexfusion in the operating room. During the micro-TESE, autotransplantation of mesenchymal stem cells into the testicular network, previously cultured in a cell technology laboratory for 2 weeks, was performed. Results of the study: in all patients, the levels of total testosterone increased, the level of follicle-stimulating hormone decreased, the levels of luteinizing hormone returned to normal, the level of inhibin B increased. IVF with a positive result; another patient (20%) had spermatogenesis cells. Non-obstructive azoospermia and mesenchymal stem cells Conclusions: The positive results of this work serve as the basis for the application of a new cellular therapeutic approach for the treatment of non-obstructive azoospermia using mesenchymal stem cells.

Keywords: cell therapy, regenerative medicine, male infertility, mesenchymal stem cells

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617 Effect of Three Sand Types on Potato Vegetative Growth and Yield

Authors: Shatha A. Yousif, Qasim M. Zamil, Hasan Y. Al Muhi, Jamal A. Al Shammari

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Potato (Solanum tuberosum L.) is one of the major vegetable crops that are grown world wide because of its economic importance. This experiment investigated the effect of local sands (River Base, Al-Ekader and Karbala) on number and total weight of mini tubers. Statistical analysis revealed that there were no significant differences among sand cultures in number of stem/plant, chlorophyll index and tubers dry weight. River Base sand had the highest plant height (74.9 cm), leaf number/plant number (39.3), leaf area (84.4 dcm2⁄plant), dry weight/plant (26.31), tubers number/plant (8.5), tubers weight/plant (635.53 gm) and potato tuber yields/trove (28.60 kg), whereas the Karbala sand had lower performance. All the characters had positive and significant correlation with yields except the traits number of stem and tuber dry weight.

Keywords: correlation, potato, sand culture, yield

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616 CCR5 as an Ideal Candidate for Immune Gene Therapy and Modification for the Induced Resistance to HIV-1 Infection

Authors: Alieh Farshbaf, Tayyeb Bahrami

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Introduction: Cc-chemokine receptor-5 (CCR5) is known as a main co-receptor in human immunodeficiency virus type-1 (HIV-1) infection. Many studies showed 32bp deletion (Δ32) in CCR5 gene, provide natural resistance to HIV-1 infection in homozygous individuals. Inducing the resistance mechanism by CCR5 in HIV-1 infected patients eliminated many problems of highly-active-anti retroviral therapy (HAART) drugs like as low safety, side-effects and virus rebounding from latent reservoirs. New treatments solved some restrictions that are based on gene modification and cell therapy. Literature review: The stories of the “Berlin and Boston patients” showed autologous hematopoietic stem cells transplantation (HSCT) could provide effective cure of HIV-1 infected patients. Furthermore, gene modification by zinc finger nuclease (ZFN) demonstrated another successful result again. Despite the other studies for gene therapy by ∆32 genotype, there is another mutation -CCR5 ∆32/m303- that provides HIV-1 resistant. It is a heterozygote genotype for ∆32 and T→A point mutation at nucleotide 303. These results approved the key role of CCR5 gene. Conclusion: Recent studies showed immune gene therapy and cell therapy could provide effective cure for refractory disease like as HIV. Eradication of HIV-1 from immune system was not observed by HAART, because of reloading virus genome from latent reservoirs after stopping them. It is showed that CCR5 could induce natural resistant to HIV-1 infection by the new approaches based on stem cell transplantation and gene modifying.

Keywords: CCR5, HIV-1, stem cell, immune gene therapy, gene modification

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615 The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by miR-375 and Anti-miR-9

Authors: Arefeh Jafarian, Mohammad Taghikani, Saied Abroun, Amir Allahverdi, Masoud Soleimani

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Introduction: The miRNAs have key roles in control of pancreatic islet development and insulin secretion. In this regards, current study investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. Findings: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose as well as extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. In derived IPCs by miR-375 alone are capable to express insulin and other endocrine specific transcription factors, the cells lack the machinery to respond to glucose. The differentiated hMSCs by miR-375 and anti-miR-9 lentiviruses could secrete insulin and c-peptide in a glucose-regulated manner. Conclusion: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.

Keywords: diabetes, differentiation, MSCs, insulin producing cells, miR-375, miR-9

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614 Piled Critical Size Bone-Biomimetic and Biominerizable Nanocomposites: Formation of Bioreactor-Induced Stem Cell Gradients under Perfusion and Compression

Authors: W. Baumgartner, M. Welti, N. Hild, S. C. Hess, W. J. Stark, G. Meier Bürgisser, P. Giovanoli, J. Buschmann

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Perfusion bioreactors are used to solve problems in tissue engineering in terms of sufficient nutrient and oxygen supply. Such problems especially occur in critical size grafts because vascularization is often too slow after implantation ending up in necrotic cores. Biominerizable and biocompatible nanocomposite materials are attractive and suitable scaffold materials for bone tissue engineering because they offer mineral components in organic carriers – mimicking natural bone tissue. In addition, human adipose derived stem cells (ASCs) can potentially be used to increase bone healing as they are capable of differentiating towards osteoblasts or endothelial cells among others. In the present study, electrospun nanocomposite disks of poly-lactic-co-glycolic acid and amorphous calcium phosphate nanoparticles (PLGA/a-CaP) were seeded with human ASCs and eight disks were stacked in a bioreactor running with normal culture medium (no differentiation supplements). Under continuous perfusion and uniaxial cyclic compression, load-displacement curves as a function of time were assessed. Stiffness and energy dissipation were recorded. Moreover, stem cell densities in the layers of the piled scaffold were determined as well as their morphologies and differentiation status (endothelial cell differentiation, chondrogenesis and osteogenesis). While the stiffness of the cell free constructs increased over time caused by the transformation of the a-CaP nanoparticles into flake-like apatite, ASC-seeded constructs showed a constant stiffness. Stem cell density gradients were histologically determined with a linear increase in the flow direction from the bottom to the top of the 3.5 mm high pile (r2 > 0.95). Cell morphology was influenced by the flow rate, with stem cells getting more roundish at higher flow rates. Less than 1 % osteogenesis was found upon osteopontin immunostaining at the end of the experiment (9 days), while no endothelial cell differentiation and no chondrogenesis was triggered under these conditions. All ASCs had mainly remained in their original pluripotent status within this time frame. In summary, we have fabricated a critical size bone graft based on a biominerizable bone-biomimetic nanocomposite with preserved stiffness when seeded with human ASCs. The special feature of this bone graft was that ASC densities inside the piled construct varied with a linear gradient, which is a good starting point for tissue engineering interfaces such as bone-cartilage where the bone tissue is cell rich while the cartilage exhibits low cell densities. As such, this tissue-engineered graft may act as a bone-cartilage interface after the corresponding differentiation of the ASCs.

Keywords: bioreactor, bone, cartilage, nanocomposite, stem cell gradient

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613 Co-Culture of Neonate Mouse Spermatogonial Stem Cells with Sertoli Cells: Inductive Role of Melatonin following Transplantation: Adult Azoospermia Mouse Model

Authors: Mehdi Abbasi, Shadan Navid, Mohammad Pourahmadi, M. Majidi Zolbin

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We have recently reported that melatonin as antioxidant enhances the efficacy of colonization of spermatogonial stem cells (SSCs). Melatonin as an antioxidant plays a vital role in the development of SSCs in vitro. This study aimed to investigate evaluation of sertoli cells and melatonin simultaneously on SSC proliferation following transplantation to testis of adult mouse busulfan-treated azoospermia model. SSCs and sertoli cells were isolated from the testes of three to six-day old male mice.To determine the purity, Flow cytometry technique using PLZF antibody were evaluated. Isolated testicular cells were cultured in αMEM medium in the absence (control group) or presence (experimental group) of sertoli cells and melatonin extract for 2 weeks. We then transplanted SSCs by injection into the azoospermia mice model. Higher viability, proliferation, and Id4, Plzf, expression were observed in the presence of simultaneous sertoli cells and melatonin in vitro. Moreover, immunocytochemistry results showed higher Oct4 expression in this group. Eight weeks after transplantation, injected cells were localized at the base of seminiferous tubules in the recipient testes. The number of spermatogonia and the weight of testis were higher in the experimental group relative to control group. The results of our study suggest that this new protocol can increase the transplantation of these cells can be useful in the treatment of male infertility.

Keywords: colonization, melatonin, spermatogonial stem cell, transplantation

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612 Language Anxiety and Learner Achievement among University Undergraduates in Sri Lanka: A Case Study of University of Sri Jayewardenepura

Authors: Sujeeva Sebastian Pereira

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Language Anxiety (LA) – a distinct psychological construct of self-perceptions and behaviors related to classroom language learning – is perceived as a significant variable highly correlated with Second Language Acquisition (SLA). However, the existing scholarship has inadequately explored the nuances of LA in relation to South Asia, especially in terms of Sri Lankan higher education contexts. Thus, the current study, situated within the broad areas of Psychology of SLA and Applied Linguistics, investigates the impact of competency-based LA and identity-based LA on learner achievement among undergraduates of Sri Lanka. Employing a case study approach to explore the impact of LA, 750 undergraduates of the University of Sri Jayewardenepura, Sri Lanka, thus covering 25% of the student population from all seven faculties of the university, were selected as participants using stratified proportionate sampling in terms of ethnicity, gender, and disciplines. The qualitative and quantitative research inquiry utilized for data collection include a questionnaire consisting a set of structured and unstructured questions, and semi-structured interviews as research instruments. Data analysis includes both descriptive and statistical measures. As per the quantitative measures of data analysis, the study employed Pearson Correlation Coefficient test, Chi-Square test, and Multiple Correspondence Analysis; it used LA as the dependent variable, and two types of independent variables were used: direct and indirect variables. Direct variables encompass the four main language skills- reading, writing, speaking and listening- and test anxiety. These variables were further explored through classroom activities on grammar, vocabulary and individual and group presentations. Indirect variables are identity, gender and cultural stereotypes, discipline, social background, income level, ethnicity, religion and parents’ education level. Learner achievement was measured through final scores the participants have obtained for Compulsory English- a common first-year course unit mandatory for all undergraduates. LA was measured using the FLCAS. In order to increase the validity and reliability of the study, data collected were triangulated through descriptive content analysis. Clearly evident through both the statistical analysis and the qualitative analysis of the results is the significant linear negative correlation between LA and learner achievement, and the significant negative correlation between LA and culturally-operated gender stereotypes which create identity disparities in learners. The study also found that both competency-based LA and identity-based LA are experienced primarily and inescapably due to the apprehensions regarding speaking in English. Most participants who reported high levels of LA were from an urban socio-economic background of lower income families. Findings exemplify the linguistic inequality prevalent in the socio-cultural milieu in Sri Lankan society. This inequality makes learning English a dire need, yet, very much an anxiety provoking process because of many sociolinguistic, cultural and ideological factors related to English as a Second Language (ESL) in Sri Lanka. The findings bring out the intricate interrelatedness of both the dependent variable (LA) and the independent variables stated above, emphasizing that the significant linear negative correlation between LA and learner achievement is connected to the affective, cognitive and sociolinguistic domains of SLA. Thus, the study highlights the promise in linguistic practices such as code-switching, crossing and accommodating hybrid identities as strategies in minimizing LA and maximizing the experience of ESL.

Keywords: language anxiety, identity-based anxiety, competence-based anxiety, TESL, Sri Lanka

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611 Human Dental Pulp Stem Cells Attenuate Streptozotocin-Induced Parotid Gland Injury in Rats

Authors: Gehan ElAkabawy

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Background: Diabetes mellitus causes severe deteriorations of almost all the organs and systems of the body, as well as significant damage to the oral cavity. The oral changes are mainly related to salivary glands dysfunction characterized by hyposalivation and xerostomia, which significantly reduce diabetic patients’ quality of life. Human dental pulp stem cells represent a promising source for cell-based therapies, owing to their easy, minimally invasive surgical access, and high proliferative capacity. It was reported that the trophic support mediated by dental pulp stem cells can rescue the functional and structural alterations of damaged salivary glands. However, potential differentiation and paracrine effects of human dental pulp stem cells in diabetic-induced parotid gland damage have not been previously investigated. Our study aimed to investigate the therapeutic effects of intravenous transplantation of human dental pulp stem cells (hDPSCs) on parotid gland injury in a rat model of streptozotocin (STZ)-induced type 1 diabetes. Methods: Thirty Sprague-Dawley male rats were randomly categorised into three groups: control, diabetic (STZ), and transplanted (STZ+hDPSCs). hDPSCs or vehicle was injected into the tail vein 7 days after STZ injection. The fasting blood glucose levels were monitored weekly. A glucose tolerance test was performed, and the parotid gland weight, salivary flow rate, oxidative stress indices, parotid gland histology, and caspase-3, vascular endothelial growth factor (VEGF), and proliferating cell nuclear antigen (PCNA) expression in parotid tissues were assessed 28 days post-transplantation. Results: Transplantation of hDPSCs downregulated blood glucose, improved the salivary flow rate, and reduced oxidative stress. The cells migrated to, survived, and differentiated into acinar, ductal, and myoepithelial cells in the STZ-injured parotid gland. Moreover, they downregulated the expression of caspase-3 and upregulated the expression of VEGF and PCNA, likely exerting pro-angiogenetic and antiapoptotic effects and promoting endogenous regeneration. In addition, the transplanted cells enhanced the parotid nitric oxide (NO) -tetrahydrobiopterin (BH4) pathway. Conclusions: Our results show that hDPSCs can migrate to and survive within the STZ-injured parotid gland, where they prevent its functional and morphological damage by restoring normal glucose levels, differentiating into parotid cell populations, and stimulating paracrine-mediated regeneration. Thus, hDPSCs may have therapeutic potential in the treatment of diabetes-induced parotid gland injury.

Keywords: dental pulp stem cells, diabetes, streptozotocin, parotid gland

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610 Insulin-Producing Cells from Adult Human Bone Marrow Mesenchymal Stem Cells Control Chemically-Induced Diabetes in Dogs

Authors: Maha Azzam, Mahmoud Gabr, Mahmoud Zakaria, Ayman Refaie, Amani Ismail, Sherry Khater, Sylvia Ashamallah, Mohamed Ghoniem

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Evidence was provided that human bone marrow-derived mesenhymal stem cells (HBM-MSCs) could be differentiated to form insulin-producing cells (IPCs). Transplantation of these cells was able to cure chemically-induced diabetes in nude mice. The efficacy of these cells to control diabetes in large animals was carried out to evaluate the sufficient number of cells needed/Kg body weight and to determine the functional longevity in vivo. Materials/Methods: Ten male mongrel dogs weighing 15-20 Kg were used in this study. Diabetes was chemically-induced in 7 dogs by a mixture of alloxan and streptozotocin. Three non-diabetic served as normal controls. Differentiated HBM-MSCs (5 million/Kg) were encapsulated in theracyte capsules and transplanted beneath the rectus sheath. Each dog received 2 capsules. One dog died 4 days postoperative from inhalation pneumonia. The remaining 6 dogs were followed up for 6-18 months. Results: Four dogs became normoglycemic within 6-8 weeks with normal glucose tolerance curves providing evidence that the transplanted cells were glucose-sensitive and insulin-responsive. In the remaining 2 dogs, fasting blood glucose was reduced but did not reach euglycemic levels. The sera of all transplanted dogs contained human insulin and c-peptide but negligible levels of canine insulin. When the HBM-MSCs loaded capsules were removed, rapid return of diabetic state was noted. The harvested capsules were examined by immunofluorescence. IPCs were seen and co-expression of with c-peptide was confirmed. Furthermore, all the pancreatic endocrine genes were expressed by the transplanted cells. Conclusions: This study provided evidence that theracyte capsules could protect the xenogenic HBM-MSCs from the host immune response. This is an important issue when clinical stem cell therapy is considered for definitive treatment for T1DM.

Keywords: diabetes, mesenchymal stem cells, dogs, Insulin-producing cells

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609 Expansion of Cord Blood Cells Using a Mix of Neurotrophic Factors

Authors: Francisco Dos Santos, Diogo Fonseca-Pereira, Sílvia Arroz-Madeira, Henrique Veiga-Fernandes

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Haematopoiesis is a developmental process that generates all blood cell lineages in health and disease. This relies on quiescent haematopoietic stem cells (HSCs) that are able to differentiate, self renew and expand upon physiological demand. HSCs have great interest in regenerative medicine, including haematological malignancies, immunodeficiencies and metabolic disorders. However, the limited yield from existing HSC sources drives the global need for reliable techniques to expand harvested HSCs at high quality and sufficient quantities. With the extensive use of cord blood progenitors for clinical applications, there is a demand for a safe and efficient expansion protocol that is able to overcome the limitations of the cord blood as a source of HSC. StemCell2MAXTM developed a technology that enhances the survival, proliferation and transplantation efficiency of HSC, leading the way to a more widespread use of HSC for research and clinical purposes. StemCell2MAXTM MIX is a solution that improves HSC expansion up to 20x, while preserving stemness, when compared to state-of-the-art. In a recent study by a leading cord blood bank, StemCell2MAX MIX was shown to support a selective 100-fold expansion of CD34+ Hematopoietic Stem and Progenitor Cells (when compared to a 10-fold expansion of Total Nucleated Cells), while maintaining their multipotent differentiative potential as assessed by CFU assays. The technology developed by StemCell2MAXTM opens new horizons for the usage of expanded hematopoietic progenitors for both research purposes (including quality and functional assays in Cord Blood Banks) and clinical applications.

Keywords: cord blood, expansion, hematopoietic stem cell, transplantation

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608 Cell-Based and Exosome Treatments for Hair Restoration

Authors: Armin Khaghani Boroujeni, Leila Dehghani, Parham Talebi Boroujeni, Sahar Rostamian, Ali Asilian

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Background: Hair loss is a common complaint observed in both genders. Androgenetic alopecia is known pattern for hair loss. To assess new regenerative strategies (PRP, A-SC-BT, conditioned media, exosome-based treatments) compared to conventional therapies for hair loss or hair regeneration, an updated review was undertaken. To address this issue, we carried out this systematic review to comprehensively evaluate the efficacy of cell-based therapies on hair loss. Methods: The available online databases, including ISI Web of Science, Scopus, and PubMed, were searched systematically up to February 2022. The quality assessment of included studies was done using the Cochrane Collaboration's tool. Results: As a result, a total of 90 studies involving 2345 participants were included in the present study. The enrolled studies were conducted between 2010 and 2022. The subjects’ mean age ranged from 19 to 55.11 years old. Approaches using platelet rich plasma (PRP) provide a beneficial impact on hair regrowth. However, other cell-based therapies, including stem cell transplant, stem cell-derived conditioned medium, and stem cell-derived exosomes, revealed conflicting evidence. Conclusion: However, cell-based therapies for hair loss are still in their infancy, and more robust clinical studies are needed to better evaluate their mechanisms of action, efficacy, safety, benefits, and limitations. In this review, we provide the resources to the latest clinical studies and a more detailed description of the latest clinical studies concerning cell-based therapies in hair loss.

Keywords: cell-based therapy, exosome, hair restoration, systematic review

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607 Unleashing Potential in Pedagogical Innovation for STEM Education: Applying Knowledge Transfer Technology to Guide a Co-Creation Learning Mechanism for the Lingering Effects Amid COVID-19

Authors: Lan Cheng, Harry Qin, Yang Wang

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Background: COVID-19 has induced the largest digital learning experiment in history. There is also emerging research evidence that students have paid a high cost of learning loss from virtual learning. University-wide survey results demonstrate that digital learning remains difficult for students who struggle with learning challenges, isolation, or a lack of resources. Large-scale efforts are therefore increasingly utilized for digital education. To better prepare students in higher education for this grand scientific and technological transformation, STEM education has been prioritized and promoted as a strategic imperative in the ongoing curriculum reform essential for unfinished learning needs and whole-person development. Building upon five key elements identified in the STEM education literature: Problem-based Learning, Community and Belonging, Technology Skills, Personalization of Learning, Connection to the External Community, this case study explores the potential of pedagogical innovation that integrates computational and experimental methodologies to support, enrich, and navigate STEM education. Objectives: The goal of this case study is to create a high-fidelity prototype design for STEM education with knowledge transfer technology that contains a Cooperative Multi-Agent System (CMAS), which has the objectives of (1) conduct assessment to reveal a virtual learning mechanism and establish strategies to facilitate scientific learning engagement, accessibility, and connection within and beyond university setting, (2) explore and validate an interactional co-creation approach embedded in project-based learning activities under the STEM learning context, which is being transformed by both digital technology and student behavior change,(3) formulate and implement the STEM-oriented campaign to guide learning network mapping, mitigate the loss of learning, enhance the learning experience, scale-up inclusive participation. Methods: This study applied a case study strategy and a methodology informed by Social Network Analysis Theory within a cross-disciplinary communication paradigm (students, peers, educators). Knowledge transfer technology is introduced to address learning challenges and to increase the efficiency of Reinforcement Learning (RL) algorithms. A co-creation learning framework was identified and investigated in a context-specific way with a learning analytic tool designed in this study. Findings: The result shows that (1) CMAS-empowered learning support reduced students’ confusion, difficulties, and gaps during problem-solving scenarios while increasing learner capacity empowerment, (2) The co-creation learning phenomenon have examined through the lens of the campaign and reveals that an interactive virtual learning environment fosters students to navigate scientific challenge independently and collaboratively, (3) The deliverables brought from the STEM educational campaign provide a methodological framework both within the context of the curriculum design and external community engagement application. Conclusion: This study brings a holistic and coherent pedagogy to cultivates students’ interest in STEM and develop them a knowledge base to integrate and apply knowledge across different STEM disciplines. Through the co-designing and cross-disciplinary educational content and campaign promotion, findings suggest factors to empower evidence-based learning practice while also piloting and tracking the impact of the scholastic value of co-creation under the dynamic learning environment. The data nested under the knowledge transfer technology situates learners’ scientific journey and could pave the way for theoretical advancement and broader scientific enervators within larger datasets, projects, and communities.

Keywords: co-creation, cross-disciplinary, knowledge transfer, STEM education, social network analysis

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606 Antibacterial and Antityrosinase Activity of Isolated Compounds from Stem Bark of Ficus platyphylla Del

Authors: Aminu Muhammad, Mustapha Ya’u, Hasnah Mohd Sirat

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An investigation of the chemical constituents into the stem bark of Ficus platyphylla (Moraceae) has resulted in the isolation of hordenine, epicatechin, lupeol, lupeol acetate and α-amyrin acetate. Their structures were determined using spectroscopic data as well as comparison with literature data. The antibacterial assay has been tested against Gram positive and Gram negative bacteria, while the tyrosinase inhibition assay was examined using L-Dopa as a substrate of mushroom tyrosinase enzyme. hordenine, epicatechin, lupeol, lupeol acetate and α-amyrin acetate showed minimum inhibition concentration (MIC) values in the range of 225-900 µg/mL against the bacterial strains. Lupeol, lupeol acetate and α-amyrin acetate showed significant antityrosinase activity against mushroom tyrosinase enzyme with percent inhibition of 67.7%, 66.2% and 62.2%, respectively.

Keywords: antibacterial, antityrosinase, chemical constituents, Ficus platyphylla

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605 iPSC-derived MSC Mediated Immunosuppression during Mouse Airway Transplantation

Authors: Mohammad Afzal Khan, Fatimah Alanazi, Hala Abdalrahman Ahmed, Talal Shamma, Kilian Kelly, Mohammed A. Hammad, Abdullah O. Alawad, Abdullah Mohammed Assiri, Dieter Clemens Broering

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Lung transplantation is a life-saving surgical replacement of diseased lungs in patients with end-stage respiratory malfunctions. Despite the remarkable short-term recovery, long-term lung survival continues to face several significant challenges, including chronic rejection and severe toxic side-effects due to global immunosuppression. Stem cell-based immunotherapy has been recognized as a crucial immunoregulatory regimen in various preclinical and clinical studies. Despite initial therapeutic outcomes, conventional stem cells face key limitations. The Cymerus™ manufacturing facilitates the production of a virtually limitless supply of consistent human induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells, which could play a key role in selective immunosuppression and graft repair during rejection. Here, we demonstrated the impact of iPSC-derived human MSCs on the development of immune-tolerance and long-term graft survival in mouse orthotopic airway allografts. BALB/c→C57BL/6 allografts were reconstituted with iPSC-derived MSCs (2 million/transplant/ at d0), and allografts were examined for regulatory T cells (Tregs), oxygenation, microvascular blood flow, airway epithelium and collagen deposition during rejection. We demonstrated that iPSC-derived MSC treatment leads to significant increase in tissue expression of hTSG-6 protein, followed by an upregulation of mouse Tregs and IL-5, IL-10, IL-15 cytokines, which augments graft microvascular blood flow and oxygenation, and thereby maintained a healthy airway epithelium and prevented the subepithelial deposition of collagen at d90 post-transplantation. Collectively, these data confirmed that iPSC-derived MSC-mediated immunosuppression has potential to establish immune-tolerance and rescue allograft from sustained hypoxic/ischemic phase and subsequently limits long-term airway epithelial injury and collagen progression, which therapeutically warrant a study of Cymerus iPSC-derived MSCs as a potential management option for immunosuppression in transplant recipients.

Keywords: stem cell therapy, immunotolerance, regulatory T cells, hypoxia and ischemia, microvasculature

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604 Free Radical Scavenging, Antioxidant Activity, Phenolic, Alkaloids Contents and Inhibited Properties against α-Amylase and Invertase Enzymes of Stem Bark Extracts Coula edulis B

Authors: Eric Beyegue, Boris Azantza, Judith Laure Ngondi, Julius E. Oben

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Background: It is clearly that phytochemical constituents of plants in relation exhibit free radical scavenging, antioxidant and glycosylation properties. This study investigated the in vitro antioxidant and free radical scavenging, inhibited activities against α-amylase and invertase enzymes of stem bark extracts C. edulis (Olacaceae). Methods: Four extracts (hexane, dichloromethane, ethanol and aqueous) from the barks of C. edulis were used in this study. Colorimetric in vitro methods were using for evaluate free radical scavenging activity DPPH, ABTS, NO, OH, antioxidant capacity, glycosylation activity, inhibition of α-amylase and invertase activities, phenolic, flavonoid and alkaloid contents. Results: C. edulis extracts (CEE) had a higher scavenging potential on the 2, 2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), nitrite oxide (NO), 2, 2-azinobis (3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radicals and glucose scavenging with the IC50 varied between 41.95 and 36694.43 µg/ml depending on the solvent of extraction. The ethanol extract of C. edulis stem bark (CE EtOH) showed the highest polyphenolic (289.10 + 30.32), flavonoid (1.12 + 0.09) and alkaloids (18.47 + 0.16) content. All the tested extracts demonstrated a relative high inhibition potential against α-amylase and invertase digestive enzymes activities. Conclusion: This study suggests that CEE exhibited higher antioxidant potential and significant inhibition potential against digestive enzymes.

Keywords: Coula edulis, antioxidant, scavenging activity, amylase, invertase

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603 Evaluating the Hepato-Protective Activities of Combination of Aqueous Extract of Roots of Tinospora cordifolia and Rhizomes of Curcuma longa against Paracetamol Induced Hepatic Damage in Rats

Authors: Amberkar Mohanbabu Vittalrao, Avin, Meena Kumari Kamalkishore, Padmanabha Udupa, Vinaykumar Bavimane, Honnegouda

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Objective: To evaluate the hepato-protective activity of Tinospora cordiofolia (Tc) against paracetamol induced hepatic damage in rats. Methods: The plant stem (test drug) was procured locally, shade dried, powdered and extracted with water. Silymarin was used as standard hepatoprotective drugs and 2% gum acacia as a control (vehicle) against paracetamol (PCT) induced hepatotoxicity. Results and Discussion: The hepato-protective activity of aqueous stem extract was assessed by paracetamol induced hepatotoxicity preventive model in rats. Alteration in the levels of biochemical markers of hepatic damage like AST, ALT, ALP and lipid peroxides were tested in both paracetamol treated and untreated groups. Paracetamol (3g/kg) had enhanced the AST, ALT, ALP and the lipid peroxides in the serum. Treatment of silymarin and aqueous stem extract of Tc (200 and 400mg/kg) extract showed significant hepatoprotective activity by altering biochemical marker levels to the near normal. Preliminary phytochemical tests were done. Aqueous Tc extract showed presence of phenolic compound and flavonoids. Our findings suggested that Tc extract possessed hepatoprotective activity in a dose dependent manner. Conclusions: Tc was found to possess significant hepatoprotective property when treated with PCT. This was evident by decreasing the liver enzymes significantly when treated with PCT as compared to PCT only treated group (P < 0.05). Hence Tinospora cardiofolia could be a good, promising, preventive agent against PCT induced hepatotoxicity.

Keywords: Tinospora cardiofolia, hepatoprotection, paracetamol, silymarin

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602 A Mathematical Description of a Growing Cell Colony Based on the Mechanical Bidomain Model

Authors: Debabrata Auddya, Bradley J. Roth

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The mechanical bidomain model is used to describe a colony of cells growing on a substrate. Analytical expressions are derived for the intracellular and extracellular displacements. Mechanotransduction events are driven by the difference between the displacements in the two spaces, corresponding to the force acting on integrins. The equation for the displacement consists of two terms: one proportional to the radius that is the same in the intracellular and extracellular spaces (the monodomain term) and one that is proportional to a modified Bessel function that is responsible for mechanotransduction (the bidomain term). The model predicts that mechanotransduction occurs within a few length constants of the colony’s edge, and an expression for the length constant contains the intracellular and extracellular shear moduli and the spring constant of the integrins coupling the two spaces. The model predictions are qualitatively consistent with experiments on human embryonic stem cell colonies, in which differentiation is localized near the edge.

Keywords: cell colony, integrin, mechanical bidomain model, stem cell, stress-strain, traction force

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601 System for Mechanical Stimulation of the Mesenchymal Stem Cells Supporting Differentiation into Osteogenic Cells

Authors: Jana Stepanovska, Roman Matejka, Jozef Rosina, Marta Vandrovcova, Lucie Bacakova

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The aim of this study was to develop a system for mechanical and also electrical stimulation controlling in vitro osteogenesis under conditions more similar to the in vivo bone microenvironment than traditional static cultivation, which would achieve good adhesion, growth and other specific behaviors of osteogenic cells in cultures. An engineered culture system for mechanical stimulation of the mesenchymal stem cells on the charged surface was designed. The bioreactor allows efficient mechanical loading inducing an electrical response and perfusion of the culture chamber with seeded cells. The mesenchymal stem cells were seeded to specific charged materials, like polarized hydroxyapatite (Hap) or other materials with piezoelectric and ferroelectric features, to create electrical potentials for stimulating of the cells. The material of the matrix was TiNb alloy designed for these purposes, and it was covered by BaTiO3 film, like a kind of piezoelectric material. The process of mechanical stimulation inducing electrical response is controlled by measuring electrical potential in the chamber. It was performed a series of experiments, where the cells were seeded, perfused and stimulated up to 48 hours under different conditions, especially pressure and perfusion. The analysis of the proteins expression was done, which demonstrated the effective mechanical and electrical stimulation. The experiments demonstrated effective stimulation of the cells in comparison with the static culture. This work was supported by the Ministry of Health, grant No. 15-29153A and the Grant Agency of the Czech Republic grant No. GA15-01558S.

Keywords: charged surface, dynamic cultivation, electrical stimulation, ferroelectric layers, mechanical stimulation, piezoelectric layers

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600 In Vitro Propagation in Barleria prionitis L. Via Callus Organogenesis

Authors: Rashmi Ranade, Neelu Joshi

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Barleria prionitis L. is a well explored Indian medicinal plant valued for its stem and leaf which forms an important ingredient of many Ayurvedic formulations. It is used for the treatment of various disorders like toothache, bleeding gums, strengthening gums, whooping cough, inflammation, arthritis, enlargement of scrotum and sciatica etc. The plant is propagated vegetatively through stem cuttings. Frequent harvesting of this plant has led to the shortage of planting material, and it has acquired the status of vulnerable plant species. Plant tissue culture technology offers a very good alternative for propagation and conservation of such plant species. The present investigation was undertaken to develop in vitro regeneration protocol for B. prionitis L. via callus organogenesis pathway. Stem and leaf explants were used for this purpose. Different media and plant growth regulators were optimized to develop the protocol. The problem of phenol secretion and browning and in vitro cultures at the establishment phase was successfully curbed with the usage of antibrowning agents such as ascorbic acid and activated charcoal. Optimum shoot multiplication was achieved by the use of liquid media and incorporation of silver nitrate and TIBA (triiodobenzoic acid) into the media. High percent rooting (76%) was observed on WPM media supplemented with IBA (2.0 mg/l), IAA (0.5 mg/l), GA3(0.5) and activated charcoal(500 mg/l). The rooted plantlets were subjected to in vitro hardening on sterile potting mix (soil:farmyard manure:compost; 1:2:1) and acclimatized under greenhouse conditions. Around 85% survival of plantlets was recorded upon acclimatization. This lab scale protocol would be tested for in vitro scaling up production of B. prionitis L.

Keywords: explant browning, liquid culture, micropropagation, shoot multiplication, phenolic secretion

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599 Determination of Critical Period for Weed Control in the Second Crop Forage Maize (454 Cultivar)

Authors: Farhad Farahvash, Parya Mobaseri

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Weeds control based on their critical period leads to less production costs and risks of wide chemical application of weeds control methods. The present study considered effect of weeds control time (weeds interference after 20, 40 and 60 days, weeds full control, weeds interference and weeds control after 20, 40 and 60 days) on growth and yield of forage maize 454. The experiment based on full-randomized blocks design with three replications was conducted at research farm of Islamic Azad University of Tabriz located at 15th km of East Tabriz in 2013. According to the results, weeds interference after 40 and 60 days as well as weeds control after 20 days prevented from decrease of maize biomass resulted from weeds presence while weeds interference after 20 days, weeds interference and weeds control after 40 and 60 days led respectively to 41.2%, 35%, 25% and 32.5% decrease of forage maize biomass. The weeds-influenced decrease was manifested at different parts of the plant depending on presence period of weeds. Decrease of fresh weight of ear and fresh weight of leaf and stem was observed due to weeds interference after 20 days and weeds interference. If weeds are controlled after 60 days, decrease of ear weight and fresh weight of stem will lead to biomass decrease. Also, if weeds are controlled after 40 days, decrease of fresh weight of maize stems will result in biomass decrease. Ear traits were affected by weeds control treatment. Being affected by treatments of weeds interference after 20 days, weeds non-interference, weeds control after 40 and 60 days, ear length was shortened 29.9 %, 41.4 %, 27.6 % and 37.2 %, respectively. The stem diameter demonstrated a significant decrease although it was only affected by treatments of weeds interference and weeds control after 60 days. Considering results of the present study, generally, it is suggested to control weeds during initial 20-60 days of maize growth in order to prevent undesirable effect of weeds on growth, production and production biomass of maize and decrease of production costs.

Keywords: maize, competition, weed, biomass

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