Search results for: regeneration
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 501

Search results for: regeneration

81 Properties of Adipose Tissue Derived Mesenchymal Stem Cells with Long-Term Cryopreservation

Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

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Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: mesenchymal stem cells, cryopreservation, stemness, senescence

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80 Adsorptive Media Selection for Bilirubin Removal: An Adsorption Equilibrium Study

Authors: Vincenzo Piemonte

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The liver is a complex, large-scale biochemical reactor which plays a unique role in the human physiology. When liver ceases to perform its physiological activity, a functional replacement is required. Actually, liver transplantation is the only clinically effective method of treating severe liver disease. Anyway, the aforementioned therapeutic approach is hampered by the disparity between organ availability and the number of patients on the waiting list. In order to overcome this critical issue, research activities focused on liver support device systems (LSDs) designed to bridging patients to transplantation or to keep them alive until the recovery of native liver function. In recirculating albumin dialysis devices, such as MARS (Molecular Adsorbed Recirculating System), adsorption is one of the fundamental steps in albumin-dialysate regeneration. Among the albumin-bound toxins that must be removed from blood during liver-failure therapy, bilirubin and tryptophan can be considered as representative of two different toxin classes. The first one, not water soluble at physiological blood pH and strongly bounded to albumin, the second one, loosely albumin bound and partially water soluble at pH 7.4. Fixed bed units are normally used for this task, and the design of such units requires information both on toxin adsorption equilibrium and kinetics. The most common adsorptive media used in LSDs are activated carbon, non-ionic polymeric resins and anionic resins. In this paper, bilirubin adsorption isotherms on different adsorptive media, such as polymeric resin, albumin-coated resin, anionic resin, activated carbon and alginate beads with entrapped albumin are presented. By comparing all the results, it can be stated that the adsorption capacity for bilirubin of the five different media increases in the following order: Alginate beads < Polymeric resin < Albumin-coated resin < Activated carbon < Anionic resin. The main focus of this paper is to provide useful guidelines for the optimization of liver support devices which implement adsorption columns to remove albumin-bound toxins from albumin dialysate solutions.

Keywords: adsorptive media, adsorption equilibrium, artificial liver devices, bilirubin, mathematical modelling

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79 Regulation of the Regeneration of Epidermal Langerhans Cells by Stress Hormone

Authors: Junichi Hosoi

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Epidermal Langerhans cells reside in upper layer of epidermis and play a role in immune surveillance. The finding of the close association of nerve endings to Langerhans cells triggered the research on systemic regulation of Langerhans cells. They disappear from epidermis after exposure to environmental and internal stimuli and reappear about a week later. Myeloid progenitor cells are assumed to be one of the sources of Langerhans cells. We examined the effects of cortisol on the reappearance of Langerhans cells in vitro. Cord-blood derived CD34-positive cells were cultured in the medium supplemented with stem cell factor/Flt3 ligand/granulocyte macrophage-colony stimulating factor/tumor necrosis factor alpha/bone morphologic protein 7/transforming growth factor beta in the presence or absence of cortisol. Cells were analyzed by flow cytometry for CD1a (cluster differentiation 1a), a marker of Langerhans cells and dermal dendritic cells, and CD39 (cluster differentiation factor 39), extracellular adenosine triphosphatase. Both CD1a-positive cells and CD39-positive cells were decreased by treatment with cortisol (suppression by 35% and 22% compared to no stress hormone, respectively). Differentiated Langerhans cells are attracted to epidermis by chemokines that are secreted from keratinocytes. Epidermal keratinocytes were cultured in the presence or absence of cortisol and analyzed for the expression of CCL2 (C-C motif chemokine ligand 2) and CCL20 (C-C motif chemokine ligand 20), which are typical attractants of Langerhans cells, by quantitative reverse transcriptase polymerase chain reaction. The expression of both chemokines, CCL2 and CCL20, were suppressed by treatment with cortisol (suppression by 38% and 48% compared to no stress hormone, respectively). We examined the possible regulation of the suppression by cortisol with plant extracts. The extracts of Ganoderma lucidum and Iris protected the suppression of the differentiation to CD39-positive cells and also the suppression of the gene expression of LC-chemoattractants. These results suggest that cortisol, which is either systemic or locally produced, blocks the supply of epidermal Langerhans cells at 2 steps, differentiation from the precursor and attraction to epidermis. The suppression is possibly blocked by some plant extracts.

Keywords: Langerhans cell, stress, CD39, chemokine

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78 Electrospun Fibre Networks Loaded with Hydroxyapatite and Barium Titanate as Smart Scaffolds for Tissue Regeneration

Authors: C. Busuioc, I. Stancu, A. Nicoara, A. Zamfirescu, A. Evanghelidis

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The field of tissue engineering has expanded its potential due to the use of composite biomaterials belonging to increasingly complex systems, leading to bone substitutes with properties that are continuously improving to meet the patient's specific needs. Furthermore, the development of biomaterials based on ceramic and polymeric phases is an unlimited resource for future scientific research, with the final aim of restoring the original tissue functionality. Thus, in the first stage, composite scaffolds based on polycaprolactone (PCL) or polylactic acid (PLA) and inorganic powders were prepared by employing the electrospinning technique. The targeted powders were: commercial and laboratory synthesized hydroxyapatite (HAp), as well as barium titanate (BT). By controlling the concentration of the powder within the precursor solution, together with the processing parameters, different types of three-dimensional architectures were achieved. In the second stage, both the mineral powders and hybrid composites were investigated in terms of composition, crystalline structure, and microstructure so that to demonstrate their suitability for tissue engineering applications. Regarding the scaffolds, these were proven to be homogeneous on large areas and loaded with mineral particles in different proportions. The biological assays demonstrated that the addition of inorganic powders leads to modified responses in the presence of simulated body fluid (SBF) or cell cultures. Through SBF immersion, the biodegradability coupled with bioactivity were highlighted, with fiber fragmentation and surface degradation, as well as apatite layer formation within the testing period. Moreover, the final composites represent supports accepted by the cells, favoring implant integration. Concluding, the purposed fibrous materials based on bioresorbable polymers and mineral powders, produced by the electrospinning technique, represent candidates with considerable potential in the field of tissue engineering. Future improvements can be attained by optimizing the synthesis process or by simultaneous incorporation of multiple inorganic phases with well-defined biological action in order to fabricate multifunctional composites.

Keywords: barium titanate, electrospinning, fibre networks, hydroxyapatite, smart scaffolds

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77 Understanding the Processwise Entropy Framework in a Heat-powered Cooling Cycle

Authors: P. R. Chauhan, S. K. Tyagi

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Adsorption refrigeration technology offers a sustainable and energy-efficient cooling alternative over traditional refrigeration technologies for meeting the fast-growing cooling demands. With its ability to utilize natural refrigerants, low-grade heat sources, and modular configurations, it has the potential to revolutionize the cooling industry. Despite these benefits, the commercial viability of this technology is hampered by several fundamental limiting constraints, including its large size, low uptake capacity, and poor performance as a result of deficient heat and mass transfer characteristics. The primary cause of adequate heat and mass transfer characteristics and magnitude of exergy loss in various real processes of adsorption cooling system can be assessed by the entropy generation rate analysis, i. e. Second law of Thermodynamics. Therefore, this article presents the second law of thermodynamic-based investigation in terms of entropy generation rate (EGR) to identify the energy losses in various processes of the HPCC-based adsorption system using MATLAB R2021b software. The adsorption technology-based cooling system consists of two beds made up of silica gel and arranged in a single stage, while the water is employed as a refrigerant, coolant, and hot fluid. The variation in process-wise EGR is examined corresponding to cycle time, and a comparative analysis is also presented. Moreover, the EGR is also evaluated in the external units, such as the heat source and heat sink unit used for regeneration and heat dump, respectively. The research findings revealed that the combination of adsorber and desorber, which operates across heat reservoirs with a higher temperature gradient, shares more than half of the total amount of EGR. Moreover, the EGR caused by the heat transfer process is determined to be the highest, followed by a heat sink, heat source, and mass transfer, respectively. in case of heat transfer process, the operation of the valve is determined to be responsible for more than half (54.9%) of the overall EGR during the heat transfer. However, the combined contribution of the external units, such as the source (18.03%) and sink (21.55%), to the total EGR, is 35.59%. The analysis and findings of the present research are expected to pinpoint the source of the energy waste in HPCC based adsorption cooling systems.

Keywords: adsorption cooling cycle, heat transfer, mass transfer, entropy generation, silica gel-water

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76 Public Art as Social Critique to Shape Urban-Scape

Authors: Po-Ching Wang

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Public art may be regarded as a social agenda. It is assumed that public art acts as an intermediate form that contributes significantly to community resurgence. That is, public art may be regarded as a verb/process or social intervention. It functions as a vanguard form, attacking boundaries and providing a sensibility for social strategy. Public art in tradition is generally expected to bring aesthetic pleasure to public. Contemporary public art, however, not only focuses on art installation, but it also often offers a process that aims to comment on, question, and challenge the socio-cultural status quo. During the last few decades, accelerated changes in the values and expectations brought to bear on varied urban issues, together with the destruction of the hegemony of traditional art and of museum authorities, has begun to contribute to freer and more democratic representations of public art. It is said that part of a public artwork’s role is to ruffle sacred feathers. In many cases, public art is created to address the dynamic social contradictions and mutability of public life; and artists and community participants approach public art from a variety of social critical perspectives and methodologies. Urban issues, such as social and environmental justice, health problems, violence, and political statements, provide plentiful source materials that fuel the performance of public art in many different settings. Further, public artworks have been extensively adopted to express social identity, make political statements, and/or to remedy social and environmental crises. Many murals on urban walls, for instance, reflect social conflicts and address civic rights, and these projects are usually the work of artists who though denied access to traditional gallery and museum channels are supported by community engagement and involvement. Public art as a social practice challenges the traditional western view of artistic practice. Art in the public realm creates a new media that provides a platform for a dialogical exchange between diverse social groups. It seems that public art has evolved as an arena for activism that addresses wide-ranging and highly controversial social issues and civilian concerns. The findings of this study indicate that public artworks are capable of playing a role of activist in facilitating community evolution via social progress.

Keywords: aesthetics, community regeneration, city development, publicness, public participation, social progress

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75 Influence of Genotype, Explant, and Hormone Treatment on Agrobacterium-Transformation Success in Salix Callus Culture

Authors: Lukas J. Evans, Danilo D. Fernando

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Shrub willows (Salix spp.) have many characteristics which make them suitable for a variety of applications such as riparian zone buffers, environmental contaminant sequestration, living snow fences, and biofuel production. In some cases, these functions are limited due to physical or financial obstacles associated with the number of individuals needed to reasonably satisfy that purpose. One way to increase the efficiency of willows is to bioengineer them with the genetic improvements suitable for the desired use. To accomplish this goal, an optimized in vitro transformation protocol via Agrobacterium tumefaciens is necessary to reliably express genes of interest. Therefore, the aim of this study is to observe the influence of tissue culture with different willow cultivars, hormones, and explants on the percentage of calli expressing reporter gene green florescent protein (GFP) to find ideal transformation conditions. Each callus was produced from 1 month old open-pollinated seedlings of three Salix miyabeana cultivars (‘SX61’, ‘WT1’, and ‘WT2’) from three different explants (lamina, petiole, and internodes). Explants were cultured for 1 month on an MS media with different concentrations of 6-Benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA) (No hormones, 1 mg⁻¹L BAP only, 3 mg⁻¹L NAA only, 1 mg⁻¹L BAP and 3 mg⁻¹L NAA, and 3 mg⁻¹L BAP and 1 mg⁻¹L NAA) to produce a callus. Samples were then treated with Agrobacterium tumefaciens at an OD600 of 0.6-0.8 to insert the transgene GFP for 30 minutes, co-cultivated for 72 hours, and selected on the same media type they were cultured on with added 7.5 mg⁻¹L of Hygromycin for 1 week before GFP visualization under a UV dissecting scope. Percentage of GFP expressing calli as well as the average number of fluorescing GFP units per callus were recorded and results were evaluated through an ANOVA test (α = 0.05). The WT1 internode-derived calli on media with 3 mg-1L NAA+1 mg⁻¹L BAP and mg⁻¹L BAP alone produced a significantly higher percentage of GFP expressing calli than each other group (19.1% and 19.4%, respectively). Additionally, The WT1 internode group cultured with 3 mg⁻¹L NAA+1 mg⁻¹L BAP produced an average of 2.89 GFP units per callus while the group cultivated with 1 mg⁻¹L BAP produced an average of 0.84 GFP units per callus. In conclusion, genotype, explant choice, and hormones all play a significant role in increasing successful transformation in willows. Future studies to produce whole callus GFP expression and subsequent plantlet regeneration are necessary for a complete willow transformation protocol.

Keywords: agrobacterium, callus, Salix, tissue culture

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74 Filling the Policy Gap for Coastal Resources Management: Case of Evidence-Based Mangrove Institutional Strengthening in Cameroon

Authors: Julius Niba Fon, Jean Hude E. Moudingo

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Mangrove ecosystems in Cameroon are valuable both in services and functions as they play host to carbon sinks, fishery breeding grounds and natural coastal barriers against storms. In addition to the globally important biodiversity that they contain, they also contribute to local livelihoods. Despite these appraisals, a reduction of about 30 % over a 25 years period due to anthropogenic and natural actions has been recorded. The key drivers influencing mangrove change include population growth, climate change, economic and political trends and upstream habitat use. Reversing the trend of mangrove loss and growing vulnerability of coastal peoples requires a real commitment by the government to develop and implement robust level policies. It has been observed in Cameroon that special ecosystems like mangroves are insufficiently addressed by forestry and/or environment programs. Given these facts, the Food Agriculture Organization (FAO) in partnership with the Government of Cameroon and other development actors have put in place the project for sustainable community-based management and conservation of mangrove ecosystems in Cameroon. The aim is to address two issues notably the present weak institutional and legal framework for mangrove management, and the unrestricted and unsustainable harvesting of mangrove resources. Civil society organizations like the Cameroon Wildlife Conservation Society, Cameroon Ecology and Organization for the Environment and Development have been working to reduce the deforestation and degradation trend of Cameroon mangroves and also bringing the mangrove agenda to the fore in national and international arenas. Following a desktop approach, we found out that in situ and ex situ initiatives on mangrove management and conservation exist on propagation of improved fish smoke ovens to reduce fuel wood consumption, mangrove forest regeneration, shrimps farming and mangrove protected areas management. The evidence generated from the field experiences are inputs for processes of improving the legal and institutional framework for mangrove management in Cameroon, such as the elaboration of norms for mangroves management engaged by the government.

Keywords: mangrove ecosystem, legal and institutional framework, climate change, civil society organizations

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73 Finding a Redefinition of the Relationship between Rural and Urban Knowledge

Authors: Bianca Maria Rulli, Lenny Valentino Schiaretti

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The considerable recent urbanization has increasingly sharpened environmental and social problems all over the world. During the recent years, many answers to the alarming attitudes in modern cities have emerged: a drastic reduction in the rate of growth is becoming essential for future generations and small scale economies are considered more adaptive and sustainable. According to the concept of degrowth, cities should consider surpassing the centralization of urban living by redefining the relationship between rural and urban knowledge; growing food in cities fundamentally contributes to the increase of social and ecological resilience. Through an innovative approach, this research combines the benefits of urban agriculture (increase of biological diversity, shorter and thus more efficient supply chains, food security) and temporary land use. They stimulate collaborative practices to satisfy the changing needs of communities and stakeholders. The concept proposes a coherent strategy to create a sustainable development of urban spaces, introducing a productive green-network to link specific areas in the city. By shifting the current relationship between architecture and landscape, the former process of ground consumption is deeply revised. Temporary modules can be used as concrete tools to create temporal areas of innovation, transforming vacant or marginal spaces into potential laboratories for the development of the city. The only permanent ground traces, such as foundations, are minimized in order to allow future land re-use. The aim is to describe a new mindset regarding the quality of space in the metropolis which allows, in a completely flexible way, to bring back the green and the urban farming into the cities. The wide possibilities of the research are analyzed in two different case-studies. The first is a regeneration/connection project designated for social housing, the second concerns the use of temporary modules to answer to the potential needs of social structures. The intention of the productive green-network is to link the different vacant spaces to each other as well as to the entire urban fabric. This also generates a potential improvement of the current situation of underprivileged and disadvantaged persons.

Keywords: degrowth, green network, land use, temporary building, urban farming

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72 Batch and Dynamic Investigations on Magnesium Separation by Ion Exchange Adsorption: Performance and Cost Evaluation

Authors: Mohamed H. Sorour, Hayam F. Shaalan, Heba A. Hani, Eman S. Sayed

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Ion exchange adsorption has a long standing history of success for seawater softening and selective ion removal from saline sources. Strong, weak and mixed types ion exchange systems could be designed and optimized for target separation. In this paper, different types of adsorbents comprising zeolite 13X and kaolin, in addition to, poly acrylate/zeolite (AZ), poly acrylate/kaolin (AK) and stand-alone poly acrylate (A) hydrogel types were prepared via microwave (M) and ultrasonic (U) irradiation techniques. They were characterized using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The developed adsorbents were evaluated on bench scale level and based on assessment results, a composite bed has been formulated for performance evaluation in pilot scale column investigations. Owing to the hydrogel nature of the partially crosslinked poly acrylate, the developed adsorbents manifested a swelling capacity of about 50 g/g. The pilot trials have been carried out using magnesium enriched Red Seawater to simulate Red Seawater desalination brine. Batch studies indicated varying uptake efficiencies, where Mg adsorption decreases according to the following prepared hydrogel types AU>AM>AKM>AKU>AZM>AZU, being 108, 107, 78, 69, 66 and 63 mg/g, respectively. Composite bed adsorbent tested in the up-flow mode column studies indicated good performance for Mg uptake. For an operating cycle of 12 h, the maximum uptake during the loading cycle approached 92.5-100 mg/g, which is comparable to the performance of some commercial resins. Different regenerants have been explored to maximize regeneration and minimize the quantity of regenerants including 15% NaCl, 0.1 M HCl and sodium carbonate. Best results were obtained by acidified sodium chloride solution. In conclusion, developed cation exchange adsorbents comprising clay or zeolite support indicated adequate performance for Mg recovery under saline environment. Column design operated at the up-flow mode (approaching expanded bed) is appropriate for such type of separation. Preliminary cost indicators for Mg recovery via ion exchange have been developed and analyzed.

Keywords: batch and dynamic magnesium separation, seawater, polyacrylate hydrogel, cost evaluation

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71 Thermal Analysis of Adsorption Refrigeration System Using Silicagel–Methanol Pair

Authors: Palash Soni, Vivek Kumar Gaba, Shubhankar Bhowmick, Bidyut Mazumdar

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Refrigeration technology is a fast developing field at the present era since it has very wide application in both domestic and industrial areas. It started from the usage of simple ice coolers to store food stuffs to the present sophisticated cold storages along with other air conditioning system. A variety of techniques are used to bring down the temperature below the ambient. Adsorption refrigeration technology is a novel, advanced and promising technique developed in the past few decades. It gained attention due to its attractive property of exploiting unlimited natural sources like solar energy, geothermal energy or even waste heat recovery from plants or from the exhaust of locomotives to fulfill its energy need. This will reduce the exploitation of non-renewable resources and hence reduce pollution too. This work is aimed to develop a model for a solar adsorption refrigeration system and to simulate the same for different operating conditions. In this system, the mechanical compressor is replaced by a thermal compressor. The thermal compressor uses renewable energy such as solar energy and geothermal energy which makes it useful for those areas where electricity is not available. Refrigerants normally in use like chlorofluorocarbon/perfluorocarbon have harmful effects like ozone depletion and greenhouse warming. It is another advantage of adsorption systems that it can replace these refrigerants with less harmful natural refrigerants like water, methanol, ammonia, etc. Thus the double benefit of reduction in energy consumption and pollution can be achieved. A thermodynamic model was developed for the proposed adsorber, and a universal MATLAB code was used to simulate the model. Simulations were carried out for a different operating condition for the silicagel-methanol working pair. Various graphs are plotted between regeneration temperature, adsorption capacities, the coefficient of performance, desorption rate, specific cooling power, adsorption/desorption times and mass. The results proved that adsorption system could be installed successfully for refrigeration purpose as it has saving in terms of power and reduction in carbon emission even though the efficiency is comparatively less as compared to conventional systems. The model was tested for its compliance in a cold storage refrigeration with a cooling load of 12 TR.

Keywords: adsorption, refrigeration, renewable energy, silicagel-methanol

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70 Kinetic Energy Recovery System Using Spring

Authors: Mayuresh Thombre, Prajyot Borkar, Mangirish Bhobe

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New advancement of technology and never satisfying demands of the civilization are putting huge pressure on the natural fuel resources and these resources are at a constant threat to its sustainability. To get the best out of the automobile, the optimum balance between performance and fuel economy is important. In the present state of art, either of the above two aspects are taken into mind while designing and development process which puts the other in the loss as increase in fuel economy leads to decrement in performance and vice-versa. In-depth observation of the vehicle dynamics apparently shows that large amount of energy is lost during braking and likewise large amount of fuel is consumed to reclaim the initial state, this leads to lower fuel efficiency to gain the same performance. Current use of Kinetic Energy Recovery System is only limited to sports vehicles only because of the higher cost of this system. They are also temporary in nature as power can be squeezed only during a small time duration and use of superior parts leads to high cost, which results on concentration on performance only and neglecting the fuel economy. In this paper Kinetic Energy Recovery System for storing the power and then using the same while accelerating has been discussed. The major storing element in this system is a Flat Spiral Spring that will store energy by compression and torsion. The use of spring ensure the permanent storage of energy until used by the driver unlike present mechanical regeneration system in which the energy stored decreases with time and is eventually lost. A combination of internal gears and spur gears will be used in order to make the energy release uniform which will lead to safe usage. The system can be used to improve the fuel efficiency by assisting in overcoming the vehicle’s inertia after braking or to provide instant acceleration whenever required by the driver. The performance characteristics of the system including response time, mechanical efficiency and overall increase in efficiency are demonstrated. This technology makes the KERS (Kinetic Energy Recovery System) more flexible and economical allowing specific application while at the same time increasing the time frame and ease of usage.

Keywords: electric control unit, energy, mechanical KERS, planetary gear system, power, smart braking, spiral spring

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69 Comparative Analysis of Chemical Composition and Biological Activities of Ajuga genevensis L. in in vitro Culture and Intact Plants

Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

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68 Identification of Cocoa-Based Agroforestry Systems in Northern Madagascar: Pillar of Sustainable Management

Authors: Marizia Roberta Rasoanandrasana, Hery Lisy Tiana. Ranarijaona, Herintsitohaina Razakamanarivo, Eric Delaitre, Nandrianina Ramifehiarivo

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Madagascar is one of the producer’s countries of world's fine cocoa. Cocoa-based agroforestry systems (CBAS) plays a very important economic role for over 75% of the population in the north of Madagascar, the island's main cocoa-producing area. It is also viewed as a key factor in the deforestation of local protected areas. It is therefore urgent to establish a compromise between cocoa production and forest conservation in this region which is difficult due to a lack of accurate cocoa agro-systems data. In order to fill these gaps and to response to these socio-economic and environmental concerns, this study aims to describe CBAS by providing precise data on their characteristics and to establish a typology. To achieve this, 150 farms were surveyed and observed to characterize CBAS based on 11 agronomic and 6 socio-economic data. Also, 30 representative plots of CBAS among the 150 farms were inventoried for providing accurate ecological data (6 variables) as an additional data for the typology determination. The results showed that Madagascar’s CBAS systems are generally extensive and practiced by smallholders. Four types of cocoa-based agroforestry system were identified, with significant differences between the following variables: yield, planting age, cocoa density, density of associated trees, preceding crop, associated crops, Shannon-Wiener indices and species richness in the upper stratum. Type 1 is characterized by old systems (>45 years) with low crop density (425 cocoa trees/ha), installed after conversion of crops other than coffee (> 50%) and giving low yields (427 kg/ha/year). Type 2 consists of simple agroforestry systems (no associated crop 0%), fairly young (20 years) with low density of associated trees (77 trees/ha) and low species diversity (H'=1.17). Type 3 is characterized by high crop density (778 trees/ha and 175 trees/ha for cocoa and associated trees respectively) and a medium level of species diversity (H'=1.74, 8 species). Type 4 is particularly characterized by orchard regeneration method involving replanting and tree lopping (100%). Analysis of the potential of these four types has identified Type 4 as a promising practice for sustainable agriculture.

Keywords: conservation, practices, productivity, protect areas, smallholder, trade-off, typology

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67 Socio-Spatial Transformations in Obsolete Port Regions: A Case for Istanbul-Karaköy District

Authors: Umut Tuğlu Karslı

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While, port function had a major role during the antiquity and medieval times, it has started to lose its significance in 19th century. In many port cities, while heavy industrial functions and ports have been moved out of the former port districts, the resulting derelict spaces have been transformed to new waterfront quarters to accommodate commercial, tourism, cultural, residential and public uses. Primary aim of these operations is to revitalize abandoned spaces of historical potential and re-establish a relationship between the city and the coast. Karakoy Port, field of this study, located in the Bosphorus, was surrounded by the city centre in time due to the transformation of urban functions, and as a result it has lost its former significance. While Karakoy has 24 hours lively residential and commercial uses in old times; in early 1980s, became a district of mechanical, plumbing and electronic parts suppliers during the day and a place for homeless at night. Today, activities for revitalization of this region continue in two forms and scales. First of these activities is the "planned transformation projects," which also includes the most important one “Galataport project”, and the second one is "spontaneous transformation," which consists of individual interventions. Galataport project that based on the idea of arranging the area specifically for tourists was prepared in 2005 and became a topic of tremendous public debate. On the other hand, the "spontaneous transformation" that is observed in the Karakoy District starts in 2004 with the foundation of “Istanbul Modern Museum”. Istanbul Modern, the first contemporary arts museum of the city, allowed the cultural integration of old naval warehouses of the port to the daily life. Following this adaptive reuse intervention, the district started to accommodate numerous art galleries, studios, café-workshops and design stores. In this context, this paper briefly examines revitalization studies in obsolete port regions, analyzes the planned and ongoing socio-spatial transformations in the specific case of Karakoy under the subjects of "planned transformation projects" and "spontaneous transformation", and realizes a critical review of the sustainability of the proposals on how to reinstate the district in the active life of Istanbul.

Keywords: port cities, socio-spatial transformation, urban regeneration, urban revitalization

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66 725 Arcadia Street in Pretoria: A Pretoria Case Study Focusing on Urban Acupuncture

Authors: Konrad Steyn, Jacques Laubscher

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South African urban design solutions are mostly aligned with European and North American models that are often not appropriate in addressing some of this country’s challenges such as multiculturalism and decaying urban areas. Sustainable urban redevelopment in South Africa should be comprehensive in nature, sensitive in its manifestation, and should be robust and inclusive in order to achieve social relevance. This paper argues that the success of an urban design intervention is largely dependent on the public’s perceptions and expectations, and the way people participate in shaping their environments. The concept of sustainable urbanism is thus more comprehensive than – yet should undoubtedly include – methods of construction, material usage and climate control principles. The case study is a central element of this research paper. 725 Arcadia Street in Pretoria, was originally commissioned as a food market structure. A starkly contrasting existing modernist adjacent building forms the morphological background. Built in 1969, it is a valuable part of Pretoria’s modernist fabric. It was realised early on that the project should not be a mere localised architectural intervention, but rather an occasion to revitalise the neighbourhood through urban regeneration. Because of the complex and comprehensive nature of the site and rich cultural diversity of the area, a multi-faceted approach seemed the most appropriate response. The methodology for collating data consisted of a combination of literature reviews (regarding the historic original fauna and flora and current plants, observation (frequent site visits) and physical surveying on the neighbourhood level (physical location, connectivity to surrounding landmarks as well as movement systems and pedestrian flows). This was followed by an exploratory design phase, culminating in the present redevelopment proposal. Since built environment interventions are increasingly based on generalised normative guidelines, an approach focusing of urban acupuncture could serve as an alternative. Celebrating the specific urban condition, urban acupuncture offers an opportunity to influence the surrounding urban fabric and achieve urban renewal through physical, social and cultural mediation.

Keywords: neighbourhood, urban renewal, South African urban design solutions, sustainable urban redevelopment

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65 Conflict around the Brownfield Reconversion of the Canadian Forces Base Rockcliffe in Ottawa: A Clash of Ambitions and Visions in Canadian Urban Sustainability

Authors: Kenza Benali

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Over the past decade, a number of remarkable projects in urban brownfield reconversion emerged across Canada, including the reconversion of former military bases owned by the Canada Lands Company (CLC) into sustainable communities. However, unlike other developments, the regeneration project of the former Canadian Forces Base Rockcliffe in Ottawa – which was announced as one of the most ambitious Smart growth projects in Canada – faced serious obstacles in terms of social acceptance by the local community, particularly urban minorities composed of Francophones, Indigenous and vulnerable groups who live near or on the Base. This turn of events led to the project being postponed and even reconsidered. Through an analysis of its press coverage, this research aims to understand the causes of this urban conflict which lasted for nearly ten years. The findings reveal that the conflict is not limited to the “standard” issues common to most conflicts related to urban mega-projects in the world – e.g., proximity issues (threads to the quality of the surrounding neighbourhoods; noise, traffic, pollution, New-build gentrification) often associated with NIMBY phenomena. In this case, the local actors questioned the purpose of the project (for whom and for what types of uses is it conceived?), its local implementation (to what extent are the local history and existing environment taken into account?), and the degree of implication of the local population in the decision-making process (with whom is the project built?). Moreover, the interests of the local actors have “jumped scales” and transcend the micro-territorial level of their daily life to take on a national and even international dimension. They defined an alternative view of how this project, considered strategic by his location in the nation’s capital, should be a reference as well as an international showcase of Canadian ambition and achievement in terms of urban sustainability. This vision promoted, actually, a territorial and national identity approach - in which some cultural values are highly significant (respect of social justice, inclusivity, ethnical diversity, cultural heritage, etc.)- as a counterweight to planners’ vision which is criticized as a normative/ universalist logic that ignore the territorial peculiarities.

Keywords: smart growth, brownfield reconversion, sustainable neighborhoods, Canada Lands Company, Canadian Forces Base Rockcliffe, urban conflicts

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64 Prospects of Regenerative Medicine with Human Allogeneic Adipose Tissue-Derived Mesenchymal Stem Cell Sheets: Achievements and Future Outlook in Clinical Trials for Myopic Chorioretinal Atrophy

Authors: Norimichi Nagano, Yoshio Hirano, Tsutomu Yasukawa

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Mesenchymal stem cells are thought to confer neuroprotection, facilitate tissue regeneration and exert their effects on retinal degenerative diseases, however, adverse events such as proliferative vitreoretinopathy and preretinal membrane disease associated with cell suspension transplantation have also been reported. We have recently developed human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) sheets through our proprietary sheet transformation technique, which could potentially mitigate these adverse events. To clarify the properties of our adMSC sheets named PAL-222, we performed in vitro studies such as viability testing, cytokine secretions by ELISA, immunohistochemical study, and migration assay. The viability of the cells exceeded 70%. Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium-Derived Factor (PEDF), which are quite important cytokines for the retinal area, were observed. PAL-222 expressed type I collagen, a strength marker, type IV collagen, a marker of the basement membrane, and elastin, an elasticity marker. Finally, the migration assay was performed and showed negative, which means that PAL-222 is stably kept in the topical area and does not come to pieces. Next, to evaluate the efficacy in vivo, we transplanted PAL-222 into the subretinal space of the eye of Royal College of Surgeons rats with congenital retinal degeneration and assessed it for three weeks after transplantation. We confirmed that PAL-222 suppressed the decrease in the thickness of the outer nuclear layer, which means that the photoreceptor protective effect treated with PAL-222 was significantly higher than that in the sham group. (p < 0.01). This finding demonstrates that PAL-222 showed their retinoprotective effect in a model of congenital retinal degeneration. As the study suggested the efficacy of PAL-222 in both in vitro and in vivo studies, we are presently engaged in clinical trials of PAL-222 for myopic chorioretinal atrophy, which is one of the retinal degenerative diseases, for the purpose of regenerative medicine.

Keywords: cell sheet, clinical trial, mesenchymal stem cell, myopic chorioretinal atrophy

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63 New Recombinant Netrin-a Protein of Lucilia Sericata Larvae by Bac to Bac Expression Vector System in Sf9 Insect Cell

Authors: Hamzeh Alipour, Masoumeh Bagheri, Abbasali Raz, Javad Dadgar Pakdel, Kourosh Azizi, Aboozar Soltani, Mohammad Djaefar Moemenbellah-Fard

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Background: Maggot debridement therapy is an appropriate, effective, and controlled method using sterilized larvae of Luciliasericata (L.sericata) to treat wounds. Netrin-A is an enzyme in the Laminins family which secreted from salivary gland of L.sericata with a central role in neural regeneration and angiogenesis. This study aimed to production of new recombinant Netrin-A protein of Luciliasericata larvae by baculovirus expression vector system (BEVS) in SF9. Material and methods: In the first step, gene structure was subjected to the in silico studies, which were include determination of Antibacterial activity, Prion formation risk, homology modeling, Molecular docking analysis, and Optimization of recombinant protein. In the second step, the Netrin-A gene was cloned and amplified in pTG19 vector. After digestion with BamH1 and EcoR1 restriction enzymes, it was cloned in pFastBac HTA vector. It was then transformed into DH10Bac competent cells, and the recombinant Bacmid was subsequently transfected into insect Sf9 cells. The expressed recombinant Netrin-A was thus purified in the Ni-NTA agarose. This protein evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay method. Results: The Bacmid vector structure with Netrin-A was successfully constructed and then expressed as Netrin-A protein in the Sf9 cell lane. The molecular weight of this protein was 52 kDa with 404 amino acids. In the in silico studies, fortunately, we predicted that recombinant LSNetrin-A have Antibacterial activity and without any prion formation risk.This molecule hasa high binding affinity to the Neogenin and a lower affinity to the DCC-specific receptors. Signal peptide located between amino acids 24 and 25. The concentration of Netrin-A recombinant protein was calculated to be 48.8 μg/ml. it was confirmed that the characterized gene in our previous study codes L. sericata Netrin-A enzyme. Conclusions: Successful generation of the recombinant Netrin-A, a secreted protein in L.sericata salivary glands, and because Luciliasericata larvae are used in larval therapy. Therefore, the findings of the present study could be useful to researchers in future studies on wound healing.

Keywords: blowfly, BEVS, gene, immature insect, recombinant protein, Sf9

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62 Alveolar Ridge Preservation in Post-extraction Sockets Using Concentrated Growth Factors: A Split-Mouth, Randomized, Controlled Clinical Trial

Authors: Sadam Elayah

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Background: One of the most critical competencies in advanced dentistry is alveolar ridge preservation after exodontia. The aim of this clinical trial was to assess the impact of autologous concentrated growth factor (CGF) as a socket-filling material and its ridge preservation properties following the lower third molar extraction. Materials and Methods: A total of 60 sides of 30 participants who had completely symmetrical bilateral impacted lower third molars were enrolled. The short-term outcome variables were wound healing, swelling and pain, clinically assessed at different time intervals (1st, 3rd & 7th days). While the long-term outcome variables were bone height & width, bone density and socket surface area in the coronal section. Cone beam computed tomography images were obtained immediately after surgery and three months after surgery as a temporal measure. Randomization was achieved by opaque, sealed envelopes. Follow-up data were compared to baseline using Paired & Unpaired t-tests. Results: The wound healing index was significantly better in the test sides (P =0.001). Regarding the facial swelling, the test sides had significantly fewer values than the control sides, particularly on the 1st (1.01±.57 vs 1.55 ±.56) and 3rd days (1.42±0.8 vs 2.63±1.2) postoperatively. Nonetheless, the swelling disappeared within the 7th day on both sides. The pain scores of the visual analog scale were not a statistically significant difference between both sides on the 1st day; meanwhile, the pain scores were significantly lower on the test sides compared with the control sides, especially on the 3rd (P=0.001) and 7th days (P˂0.001) postoperatively. Regarding long-term outcomes, CGF sites had higher values in height and width when compared to Control sites (Buccal wall 32.9±3.5 vs 29.4±4.3 mm, Lingual wall 25.4±3.5 vs 23.1±4 mm, and Alveolar bone width 21.07±1.55vs19.53±1.90 mm) respectively. Bone density showed significantly higher values in CGF sites than in control sites (Coronal half 200±127.3 vs -84.1±121.3, Apical half 406.5±103 vs 64.2±158.6) respectively. There was a significant difference between both sites in reducing periodontal pockets. Conclusion: CGF application following surgical extraction provides an easy, low-cost, and efficient option for alveolar ridge preservation. Thus, dentists may encourage using CGF during dental extractions, particularly when alveolar ridge preservation is required.

Keywords: platelet, extraction, impacted teeth, alveolar ridge, regeneration, CGF

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61 Urban Rehabilitation Assessment: Buildings' Integrity and Embodied Energy

Authors: Joana Mourão

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Transition to a low carbon economy requires changes in consumption and production patterns, including the improvement of existing buildings’ environmental performance. Urban rehabilitation is a top policy priority in Europe, creating an opportunity to increase this performance. However, urban rehabilitation comprises different typologies of interventions with distinct levels of consideration for cultural urban heritage values and for environmental values, thus with different impacts. Cities rely on both material and non-material forms of heritage that are deep-rooted and resilient. One of the most relevant parts of that urban heritage is the historical pre-industrial housing stock, with an extensive presence in many European cities, as Lisbon. This stock is rehabilitated and transformed at the framework of urban management and local governance traditions, as well as the framework of the global economy, and in that context, faces opportunities and threats that need evaluation and control. The scope of this article is to define methodological bases and research lines for the assessment of impacts that urban rehabilitation initiatives set on the vulnerable and historical pre-industrial urban housing stock, considering it as an environmental and cultural unreplaceable material value and resource. As a framework, this article reviews the concepts of urban regeneration, urban renewal, current buildings conservation and refurbishment, and energy refurbishment of buildings, seeking to define key typologies of urban rehabilitation that represent different approaches to the urban fabric, in terms of scope, actors, and priorities. Moreover, main types of interventions - basing on a case-study in a XVIII century neighborhood in Lisbon - are defined and analyzed in terms of the elements lost in each type of intervention, and relating those to urbanistic, architectonic and constructive values of urban heritage, as well as to environmental and energy efficiency. Further, the article overviews environmental cultural heritage assessment and life-cycle assessment tools, selecting relevant and feasible impact assessment criteria for urban buildings rehabilitation regulation, focusing on multi-level urban heritage integrity. Urbanistic, architectonic, constructive and energetic integrity are studied as criteria for impact assessment and specific indicators are proposed. The role of these criteria in sustainable urban management is discussed. Throughout this article, the key challenges for urban rehabilitation planning and management, concerning urban built heritage as a resource for sustainability, are discussed and clarified.

Keywords: urban rehabilitation, impact assessment criteria, buildings integrity, embodied energy

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60 Resourcing Remote Rural Social Enterprises to Foster Resilience and Regional Development

Authors: Heather Fulford, Melanie Liddell

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The recruitment and retention of high quality employees can prove to be challenging for social enterprises, particularly in some of the core business support functions such as marketing, communications, IT and finance. This holds true for social enterprises in urban contexts, where roles with more attractive remuneration in these business functions can often be found quite readily in the private sector. For social enterprises situated in rural locations, the challenges of staff recruitment and retention are even more acute. Such challenges can lead to a skills deficit in rural social enterprises, which can, at best, hinder their growth potential, and worse, jeopardise their chances of survival. This in turn, can have a negative impact on the sustainability and resilience of the surrounding rural community in which the social enterprise is located. The purpose of this paper is to report on aspects of a collaborative initiative established to stimulate innovation and business growth in remote rural businesses in Scotland. Launched in 2010, this initiative was designed to attract young students and graduates from the region to stay in the region upon completion of their studies, and to attract others from outside the region to re-locate there post-university. To facilitate this, SMEs in the region were offered wage subsidies to encourage them to recruit a student or graduate on a work placement for up to one year to participate in an innovation or business growth-oriented project. A number of the employers offering work placements were social enterprises. Through analysis of the placement project and role specifications devised by the participating social enterprises, an overview is provided of their business development needs and the skills they require to stimulate innovation and growth. Scrutiny of the reflective accounts compiled by the students and graduates at the close of their work placements highlights the benefits they derived from being able to put their academic knowledge and skills into action within a social enterprise. Examination of interviews conducted with a sample of placement employers reveals the contribution the students and graduates made during the business development projects with the social enterprises. The challenges of hosting such placements are also discussed. The paper concludes with indications of the lessons learned and an outline of the wider implications for other remote rural locations in which social enterprises play an important role in the local economy and life of the community.

Keywords: resilience, rural development, regeneration, regional development, recruitment, resource management, retention, remuneration

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59 Understanding Neuronal and Glial Cell Behaviour in Multi-Layer Nanofibre Systems to Support the Development of an in vitro Model of Spinal Cord Injury and Personalised Prostheses for Repair

Authors: H. Pegram, R. Stevens, L. De Girolamo

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Aligned electrospun nanofibres act as effective neuronal and glial cell scaffolds that can be layered to contain multiple sheets harboring different cell populations. This allows personalised biofunctional prostheses to be manufactured with both acellular and cellularised layers for the treatment of spinal cord injury. Additionally, the manufacturing route may be configured to produce in-vitro 3D cell based model of spinal cord injury to aid drug development and enhance prosthesis performance. The goal of this investigation was to optimise the multi-layer scaffold design parameters for prosthesis manufacture, to enable the development of multi-layer patient specific implant therapies. The work has also focused on the fabricating aligned nanofibre scaffolds that promote in-vitro neuronal and glial cell population growth, cell-to-cell interaction and long-term survival following trauma to mimic an in-vivo spinal cord lesion. The approach has established reproducible lesions and has identified markers of trauma and regeneration marked by effective neuronal migration across the lesion with glial support. The investigation has advanced the development of an in-vitro model of traumatic spinal cord injury and has identified a route to manufacture prostheses which target the repair spinal cord injury. Evidence collated to investigate the multi-layer concept suggests that physical cues provided by nanofibres provide both a natural extra-cellular matrix (ECM) like environment and controls cell proliferation and migration. Specifically, aligned nanofibre layers act as a guidance system for migrating and elongating neurons. On a larger scale, material type in multi-layer systems also has an influence in inter-layer migration as cell types favour different material types. Results have shown that layering nanofibre membranes create a multi-level scaffold system which can enhance or prohibit cell migration between layers. It is hypothesised that modifying nanofibre layer material permits control over neuronal/glial cell migration. Using this concept, layering of neuronal and glial cells has become possible, in the context of tissue engineering and also modelling in-vitro induced lesions.

Keywords: electrospinning, layering, lesion, modeling, nanofibre

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58 A Minimally Invasive Approach Using Bio-Miniatures Implant System for Full Arch Rehabilitation

Authors: Omid Allan

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The advent of ultra-narrow diameter implants initially offered an alternative to wider conventional implants. However, their design limitations have restricted their applicability primarily to overdentures and cement-retained fixed prostheses, often with unpredictable long-term outcomes. The introduction of the new Miniature Implants has revolutionized the field of implant dentistry, leading to a more streamlined approach. The utilization of Miniature Implants has emerged as a promising alternative to the traditional approach that entails the traumatic sequential bone drilling procedures and the use of conventional implants for full and partial arch restorations. The innovative "BioMiniatures Implant System serves as a groundbreaking bridge connecting mini implants with standard implant systems. This system allows practitioners to harness the advantages of ultra-small implants, enabling minimally invasive insertion and facilitating the application of fixed screw-retained prostheses, which were only available to conventional wider implant systems. This approach streamlines full and partial arch rehabilitation with minimal or even no bone drilling, significantly reducing surgical risks and complications for clinicians while minimizing patient morbidity. The ultra-narrow diameter and self-advancing features of these implants eliminate the need for invasive and technically complex procedures such as bone augmentation and guided bone regeneration (GBR), particularly in cases involving thin alveolar ridges. Furthermore, the absence of a microcap between the implant and abutment eliminates the potential for micro-leakage and micro-pumping effects, effectively mitigating the risk of marginal bone loss and future peri-implantitis. The cumulative experience of restoring over 50 full and partial arch edentulous cases with this system has yielded an outstanding success rate exceeding 97%. The long-term success with a stable marginal bone level in the study firmly establishes these implants as a dependable alternative to conventional implants, especially for full arch rehabilitation cases. Full arch rehabilitation with these implants holds the promise of providing a simplified solution for edentulous patients who typically present with atrophic narrow alveolar ridges, eliminating the need for extensive GBR and bone augmentation to restore their dentition with fixed prostheses.

Keywords: mini-implant, biominiatures, miniature implants, minimally invasive dentistry, full arch rehabilitation

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57 Cord Blood Hematopoietic Stem Cell Expansion Ability of Mesenchymal Stem Cells Isolated From Different Sources

Authors: Ana M. Lara, Manuela Llano, Felipe Gaitán, Rosa H. Bustos, Ana Maria Perdomo-Arciniegas, Ximena Bonilla

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Umbilical cord blood is used as a source of progenitor and stem cells for the regeneration of the hematopoietic and immune system to treat patients with different hematological or non-hematological diseases. This stem cell source represents an advantage over the use of bone marrow or mobilized peripheral blood because it has a lower incidence rate of graft-versus-host disease, probably due to fewer immunological compatibility restrictions. However, its low cellular dose limits its use in pediatric patients. This work proposes the standardization of a cell expansion technique to compensate for the dose of infused cells through the ex-vivo manipulation of hematopoietic progenitor cells from umbilical cord blood before transplantation. The expansion model is carried out through co-cultures with mesenchymal stem cells (MSC) from bone marrow (BM) and less explored fetal tissues such as Wharton's jelly (WJ) and umbilical cord blood (UCB). Initially, a master cell bank of primary mesenchymal stem cells isolated from different sources was established and characterized following International Society of Cell Therapies (ISCT) indications. Additionally, we assessed the effect of a short 25 Gy cycle of gamma irradiation on cell cycle arrest of mesenchymal cells over the support capacity for the expansion of hematopoietic stem cells from umbilical cord blood was evaluated. The results show that co-cultures with MSC from WJ and UCB allow the cellular dose of HSPC to be maximized between 5 and 16 times having a similar support capacity as BM. In addition, was evaluated the hematopoietic stem progenitor cell's HSPC functionality through the evaluation of migration capacity, their differentiation capacity during culture time by flow cytometry to evaluate the expression of membrane markers associated with lineage-committed progenitors, their clonogenic potential, and the evaluation of secretome profile in the expansion process was evaluated. So far, the treatment with gamma irradiation maintains the hematopoietic support capacity of mesenchymal stem cells from the three sources studied compared to treatments without irradiation, favoring the use of fetal tissues that are generally waste to obtain mesenchymal cell lines for ex-vivo expansion systems. With the results obtained, a standardized protocol that will contribute to the development of ex-vivo expansion with MSC on a larger scale will be achieved, enabling its clinical use and expanding its application in adults.

Keywords: ex-vivo expansion, hematopoietic stem cells, hematopoietic stem cell transplantation, mesenchymal stem cells, umbilical cord blood

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56 Precursor Muscle Cell’s Phenotype under Compression in a Biomimetic Mechanical Niche

Authors: Fatemeh Abbasi, Arne Hofemeier, Timo Betz

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Muscle growth and regeneration critically depend on satellite cells (SCs) which are muscle stem cells located between the basal lamina and myofibres. Upon damage, SCs become activated, enter the cell cycle, and give rise to myoblasts that form new myofibres, while a sub-population self-renew and re-populate the muscle stem cell niche. In aged muscle as well as in certain muscle diseases such as muscular dystrophy, some of the SCs lose their regenerative ability. Although it is demonstrated that the chemical composition of SCs quiescent niche is different from the activated niche, the mechanism initially activated in the SCs remains unknown. While extensive research efforts focused on potential chemical activation, no such factor has been identified to the author’s best knowledge. However, it is substantiated that niche mechanics affects SCs behaviors, such as stemness and engraftment. We hypothesize that mechanical stress in the healthy niche (homeostasis) is different from the regenerative niche and that this difference could serve as an early signal activating SCs upon fiber damage. To investigate this hypothesis, we develop a biomimetic system to reconstitute both, the mechanical and the chemical environment of the SC niche. Cells will be confined between two elastic polyacrylamide (PAA) hydrogels with controlled elastic moduli and functionalized surface chemistry. By controlling the distance between the PAA hydrogel surfaces, we vary the compression forces exerted by the substrates on the cells, while the lateral displacement of the upper hydrogel will create controlled shear forces. To establish such a system, a simplified system is presented. We engineered a sandwich-like configuration of two elastic PAA layer with stiffnesses between 1 and 10 kPa and confined a precursor myoblast cell line (C2C12) in between these layers. Our initial observations in this sandwich model indicate that C2C12 cells show different behaviors under mechanical compression if compared to a control one-layer gel without compression. Interestingly, this behavior is stiffness-dependent. While the shape of C2C12 cells in the sandwich consisting of two stiff (10 kPa) layers was much more elongated, showing almost a neuronal phenotype, the cell shape in a sandwich situation consisting of one stiff and one soft (1 kPa) layer was more spherical. Surprisingly, even in proliferation medium and at very low cell density, the sandwich situation stimulated cell differentiation with increased striation and myofibre formation. Such behavior is commonly found for confluent cells in differentiation medium. These results suggest that mechanical changes in stiffness and applied pressure might be a relevant stimulation for changes in muscle cell behavior.

Keywords: C2C12 cells, compression, force, satellite cells, skeletal muscle

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

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

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

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

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54 The Comparison of Bird’s Population between Naturally Regenerated Acacia Forest with Adjacent Secondary Indigenous Forest in Universiti Malaysia Sabah

Authors: Jephte Sompud, Emily A. Gilbert, Andy Russel Mojiol, Cynthia B. Sompud, Alim Biun

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Naturally regenerated acacia forest and secondary indigenous forest forms some of the urban forests in Sabah. Naturally regenerated acacia trees are usually seen along the road that exists as forest islands. Acacia tree is not an indigenous tree species in Sabah that was introduced in the 1960’s as fire breakers that eventually became one of the preferred trees for forest plantation for paper and pulp production. Due to its adaptability to survive even in impoverished soils and poor-irrigated land, this species has rapidly spread throughout Sabah through natural regeneration. Currently, there is a lack of study to investigate the bird population in the naturally regenerated acacia forest. This study is important because it shed some light on the role of naturally regenerated acacia forest on bird’s population, as bird is known to be a good bioindicator forest health. The aim of this study was to document the bird’s population in naturally regenerated acacia forest with that adjacent secondary indigenous forest. The study site for this study was at Universiti Malaysia Sabah (UMS) Campus. Two forest types in the campus were chosen as a study site, of which were naturally regenerated Acacia Forest and adjacent secondary indigenous forest, located at the UMS Hill. A total of 21 sampling days were conducted in each of the forest types. The method used during this study was solely mist nets with three pockets. Whenever a bird is caught, it is extracted from the net to be identified and measurements were recorded in a standard data sheet. Mist netting was conducted from 6 morning until 5 evening. This study was conducted between February to August 2014. Birds that were caught were ring banded to initiate a long-term study on the understory bird’s population in the Campus The data was analyzed using descriptive analysis, diversity indices, and t-test. The bird population diversity at naturally regenerated Acacia forest with those at the secondary indigenous forest was calculated using two common indices, of which were Shannon-Wiener and Simpson diversity index. There were 18 families with 33 species that were recorded from both sites. The number of species recorded at the naturally regenerated acacia forest was 26 species while at the secondary indigenous forest were 19 species. The Shannon diversity index for Naturally Regenerated Acacia Forest and secondary indigenous forests were 2.87 and 2.46. The results show that there was very significantly higher species diversity at the Naturally Regenerated Acacia Forest as opposed to the secondary indigenous forest (p<0.001). This suggests that Naturally Regenerated Acacia forest plays an important role in urban bird conservation. It is recommended that Naturally Regenerated Acacia Forests should be considered as an established urban forest conservation area as they do play a role in biodiversity conservation. More future studies in Naturally Regenerated Acacia Forest should be encouraged to determine the status and value of biodiversity conservation of this ecosystem.

Keywords: naturally regenerated acacia forest, bird population diversity, Universiti Malaysia Sabah, biodiversity conservation

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53 The Church of San Paolo in Ferrara, Restoration and Accessibility

Authors: Benedetta Caglioti

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The ecclesiastical complex of San Paolo in Ferrara represents a monument of great historical, religious and architectural importance. Its long and articulated story, over time, is already manifested by the mere reading of its planimetric and altimetric configuration, apparently unitary but, in reality, marked by modifications and repeated additions, even of high quality. It follows, in terms of protection, restoration and enhancement, a commitment of due respect for how the ancient building was built and enriched over its centuries of life. Hence a rigorous methodological approach, while being aware of the fact that every monument, in order to live and make use of the indispensable maintenance, must always be enjoyed and visited, therefore it must enjoy, in the right measure and compatibly with its nature, the possibility of improvements and functional, distributive, technological adjustments and related to the safety of people and things. The methodological approach substantiates the different elements of the project (such as distribution functionality, safety, structural solidity, environmental comfort, the character of the site, building and urban planning regulations, financial resources and materials, the same organization methods of the construction site) through the guiding principles of restoration, defined for a long time: the 'minimum intervention,' the 'recognisability' or 'distinguishability' of old and new, the Physico-chemical and figurative 'compatibility,' the 'durability' and the, at least potential, 'reversibility' of what is done, leading to the definition of appropriate "critical choices." The project tackles, together with the strictly functional ones, also the directly conservative and restoration issues, of a static, structural and material technology nature, with special attention to precious architectural surfaces, In order to ensure the best architectural quality through conscious enhancement, the project involves a redistribution of the interior and service spaces, an accurate lighting system inside and outside the church and a reorganization of the adjacent urban space. The reorganization of the interior is designed with particular attention to the issue of accessibility for people with disabilities. To accompany the community to regain possession of the use of the church's own space, already in its construction phase, the project proposal has hypothesized a permeability and flexibility in the management of the works such as to allow the perception of the found Monument to gradually become more and more familiar at the citizenship. Once the interventions have been completed, it is expected that the Church of San Paolo, second in importance only to the Cathedral, from which it is a few steps away, will be inserted in an already existing circuit of use of the city which over the years has systematized the different aspects of culture, the environment and tourism for the creation of greater awareness in the perception of what Ferrara can offer in cultural terms.

Keywords: conservation, accessibility, regeneration, urban space

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52 Mapping the Urban Catalytic Trajectory for 'Convention and Exhibition' Projects: A Case of India International Convention and Expo Centre, New Delhi

Authors: Bhavana Gulaty, Arshia Chaudhri

Abstract:

Great civic projects contribute integrally to a city, and every city undergoes a recurring cycle of urban transformations and regeneration by their insertion. The M.I.C.E. (Meetings, Incentives, Convention and Exhibitions) industry is the forbearer of one category of such catalytic civic projects. Through a specific focus on M.I.C.E. destinations, this paper illustrates the multifarious dimensions that urban catalysts impact the city on S.P.U.R. (Seed. Profile. Urbane. Reflections), the theoretical framework of this paper aims to unearth these dimensions in the realm of the COEX (Convention & Exhibition) biosphere. The ‘COEX Biosphere’ is the filter of such catalysts being ecosystems unto themselves. Like a ripple in water, the impact of these strategic interventions focusing on art, culture, trade, and promotion expands right from the trigger; the immediate context to the region and subsequently impacts the global scale. These ripples are known to bring about significant economic, social, and political and network changes. The COEX inventory in the Asian context has one such prominent addition; the proposed India International Convention and Exhibition Centre (IICC) at New Delhi. It is envisioned to be the largest facility in Asia currently and would position India on the global M.I.C.E map. With the first phase of the project scheduled to open for use in the end of 2019, this flagship project of the Government of India is projected to cater to a peak daily footfall of 3,20,000 visitors and estimated to generate 5,00,000 jobs. While the economic benefits are yet to manifest in real time and ‘Good design is good business’ holds true, for the urban transformation to be meaningful, the benefits have to go beyond just a balance sheet for the city’s exchequer. This aspect has been found relatively unexplored in research on these developments. The methodology for investigation will comprise of two steps. The first will be establishing an inventory of the global success stories and associated benefits of COEX projects over the past decade. The rationale for capping the timeframe is the significant paradigm shift that has been observed in their recent conceptualization; for instance ‘Innovation Districts’ conceptualised in the city of Albuquerque that converges into the global economy. The second step would entail a comparative benchmarking of the projected transformations by IICC through a toolkit of parameters. This is posited to yield a matrix that can form the test bed for mapping the catalytic trajectory for projects in the pipeline globally. As a ready reckoner, it purports to be a catalyst to substantiate decision making in the planning stage itself for future projects in similar contexts.

Keywords: catalysts, COEX, M.I.C.E., urban transformations

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