Search results for: shoot regeneration

Authors: Fang Bin Guo, Emma Roberts, Haibin Du, Yonggang Wang, Yu Chen, Xiuli Ge

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This paper explores the findings so far from a major externally-funded project which operates internationally in China, Germany and the UK. The research team is working in the context of the redevelopment of post-industrial sites in China and how these might be platforms for creative enterprises and thereby, the economy and welfare to flourish. Results from the project are anticipated to inform urban design policies in China and possibly farther afield. The research has utilised ethnographic studies and participatory design methods to investigate alternative strategies for sustainable urban renewal of China’s post-industrial areas. Additionally, it has undertaken comparative studies of successful examples of European and Chinese urban regeneration cases. The international cross-disciplinary team has been seeking different opportunities for developing relevant creative industries whilst retaining cultural and industrial heritage. This paper will explore the research conducted so far by the team and offer initial findings. Findings point out the development challenges of cities respecting the protection of local culture/heritages, history of the industries and transformation of the local economies. The preliminary results and pilot analysis of the current research have demonstrated that local government policyholders, business investors/developers and creative industry practitioners are the three major stakeholders that will impact city revitalisations. These groups are expected to work together with asynchronous vision in order for redevelopments to be successful. Meanwhile, local geography, history, culture, politics, economy and ethnography have been identified as important factors that impact on project design and development during urban transformations. Data is being processed from the team’s research conducted across the focal Western and Chinese cities. This has provided theoretical guidance and practical support to the development of significant experimental projects. Many were re-examined with a more international perspective, and adjustments have been based on the conclusions of the research. The observations and research are already generating design solutions in terms of ascertaining essential site components, layouts, visual design and practical facilities for regenerated sites. Two significant projects undertaken by this project team have been nominated by the central Chinese government as the most successful exemplars. They have been listed as outstanding national industry heritage projects; in particular, one of them was nominated by ArchDaily as Building of the Year 2019, and so this project outcome has made a substantial contribution to research and innovation. In summary, this paper will outline the funded project, discuss the work conducted so far, and pinpoint the initial discoveries. It will detail the future steps and indicate how these will impact on national and local governments in China, designers, local citizens and building users.

Keywords: cultural & industrial heritages, ethnographic research, participatory design, regeneration of post-industrial sites, sustainable

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Authors: N. Kotabin, Y. Tahara, K. Issakul, O. Chunhachart

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Cadmium (II) (Cd) is one of the major toxic elemental pollutants which is hazardous for humans, animals and plants. γ-Polyglutamic acid (γ-PGA) is an extracellular biopolymer produced by several species of Bacillus which has been reported to be an effective biosorbent for metal ions. The effect of γ-PGA on growth of rice grown under laboratory conditions was investigated. Rice seeds were germinated and then grown at 30±1°C on filter paper soaked with Cd solution and γ-PGA for 7 days. The result showed that Cd significantly inhibited the growth of roots and shoots by reducing root and shoot lengths. Fresh and dry weights also decreased compared with control; however, the addition of 500 mg•L-1 γ-PGA alleviated rice seedlings from the adverse effects of Cd. The analysis of physiological traits revealed that Cd caused a decrease in the total chlorophyll and soluble protein contents and amylase activities in all treatments. The Cd content in seedling tissues increased for the Cd 250 μM treatment (P < 0.05) but the addition of 500 mg•L-1 γ-PGA resulted in a noticeable decrease in Cd (P < 0.05).

Keywords: polyglutamic acid, cadmium, rice, bacillus subtilis

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Authors: H. Kishor, G. Prabhuling, D. S. Ambika, D. P. Prakash

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The chemical mutagens have become important tool to enhance agronomic traits of banana crop. It is being used to develop fusarium resistance lines in various susceptible banana cultivars. There are several mutagens like EMS and NaN3 available for banana crop improvement and each mutagen has its own important role as positive or negative effects on growth and development of banana plants. Explants from shoot tip culture were treated with various EMS (0.30, 0.60, 0.90 and 0.12%) and NaN3 (0.01, 0.02 and 0.03%) concentrations. The putative mutants obtained after in vitro rooting were subjected for artificial inoculation of Fusarium oxysporum f.sp. cubense. Screening putative mutants resistance to Panama disease was carried out by using syringe method of inoculation. It was observed that, EMS treated mutants were more susceptible compared to NaN3 treatment. Among the NaN3 doses 0.01% found to produce 3 resistant lines during preliminary screening under greenhouse conditions.

Keywords: Nanjangud rasabale, EMS, NaN3, putative mutants

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Authors: Saeid H. Mobini, Monika Lulsdorf, Thomas D. Warkentin

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The length of the breeding cycle from seed to seed is a limiting factor in the development of improved homozygous lines for breeding or recombinant inbred lines (RILs) for genetic analysis. The objective of this research was to accelerate the production of field pea RILs through application of rapid generation technology (RGT). RGT is based on the principle of growing miniature plants in an artificial medium under controlled conditions, and allowing them to produce a few flowers which develop seeds that are harvested prior to normal seed maturity. We aimed to maintain population size and genetic diversity in regeneration cycles. The effects of flurprimidol (a gibberellin synthesis inhibitor), plant density, hydroponic system, scheduled fertilizer applications, artificial light spectrum, photoperiod, and light/dark temperature were evaluated in the development of RILs from a cross between cultivars CDC Dakota and CDC Amarillo. The main goal was to accelerate flowering while reducing maintenance and space costs. In addition, embryo rescue of immature seeds was tested for shortening the seed fill period. Data collected over seven generations included plant height, the percentage of plant survival, flowering rate, seed setting rate, the number of seeds per plant, and time from seed to seed. Applying 0.6 µM flurprimidol reduced the internode length. Plant height was decreased to approximately 32 cm allowing for higher plant density without a delay in flowering and seed setting rate. The three light systems (T5 fluorescent bulbs, LEDs, and High Pressure Sodium +Metal-halide lamp) evaluated did not differ significantly in terms of flowering time in field pea. Collectively, the combination of 0.6 µM flurprimidol, 217 plant. m-2, 20 h photoperiod, 21/16 oC light/dark temperature in a hydroponic system with vermiculite substrate, applying scheduled fertilizer application based on growth stage, and 500 µmole.m-2.s-1 light intensity using T5 bulbs resulted in 100% of plants flowering within 34 ± 3 days and 96.5% of plants completed seed setting in 68.2 ± 3.6 days, i.e., 30-45 days/generation faster than conventional single seed descent (SSD) methods. These regeneration cycles were reproducible consistently. Hence, RGT could double (5.3) generations per year, using 3% occupying space, compared to SSD (2-3 generation/year). Embryo rescue of immature seeds at 7-8 mm stage, using commercial fertilizer solutions (Holland’s Secret™) showed seed setting rate of 95%, while younger embryos had lower germination rate. Mature embryos had a seed setting rate of 96.5% without either hormones or sugar added. So, considering the higher cost of embryo rescue using a procedure which requires skill, additional materials, and expenses, it could be removed from RGT with a further cost saving, and the process could be stopped between generations if required.

Keywords: field pea, flowering, rapid regeneration, recombinant inbred lines, single seed descent

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Authors: Dong-An Wang

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

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

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Authors: H. Jung, N. Kim, B. Kang, J. Choe

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History matching is a crucial procedure for predicting reservoir performances and making future decisions. However, it is difficult due to uncertainties of initial reservoir models. Therefore, it is important to have reliable initial models for successful history matching of highly heterogeneous reservoirs such as channel reservoirs. In this paper, we proposed a novel scheme for regenerating geological models using support vector machine (SVM) and principal component analysis (PCA). First, we perform PCA for figuring out main geological characteristics of models. Through the procedure, permeability values of each model are transformed to new parameters by principal components, which have eigenvalues of large magnitude. Secondly, the parameters are projected into two-dimensional plane by multi-dimensional scaling (MDS) based on Euclidean distances. Finally, we train an SVM classifier using 20% models which show the most similar or dissimilar well oil production rates (WOPR) with the true values (10% for each). Then, the other 80% models are classified by trained SVM. We select models on side of low WOPR errors. One hundred channel reservoir models are initially generated by single normal equation simulation. By repeating the classification process, we can select models which have similar geological trend with the true reservoir model. The average field of the selected models is utilized as a probability map for regeneration. Newly generated models can preserve correct channel features and exclude wrong geological properties maintaining suitable uncertainty ranges. History matching with the initial models cannot provide trustworthy results. It fails to find out correct geological features of the true model. However, history matching with the regenerated ensemble offers reliable characterization results by figuring out proper channel trend. Furthermore, it gives dependable prediction of future performances with reduced uncertainties. We propose a novel classification scheme which integrates PCA, MDS, and SVM for regenerating reservoir models. The scheme can easily sort out reliable models which have similar channel trend with the reference in lowered dimension space.

Keywords: history matching, principal component analysis, reservoir modelling, support vector machine

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Authors: Kishor Prasad Bhatta, Suman Ghimire, Durga Prasad Joshi

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Invasive Alien Species (IAS) are considered waste forest resources in Nepal. The rapid expansion of IAS is one of the nine main drivers of forest degradation, though the extent and distribution of this species are not well known. Further, the knowledge of the impact of IAS removal on forest plant diversity is hardly known, and the possibilities of income generation from them at the grass-root communities are rarely documented. Systematic sampling of 1% with nested circular plots of 500 square meters was performed in IAS removed and non-removed area, each of 30 hectares in Udayapur Community Forest User Group (CFUG), Chitwan, central Nepal to observe whether the removal of IAS contributed to an increase in plant diversity. In addition, ten entrepreneurs of Udaypur CFUG, involved in the charcoal production, briquette making and marketing were interviewed and interacted as well as their record keeping booklets were reviewed to understand if the charcoal production contributed to their income and employment. The average annual precipitation and temperature of the study area is 2100 mm and 34 degree Celsius respectively with Shorea robusta as main tree species and Eupatorium odoratum as dominant IAS. All the interviewed households were from the ̔below-poverty-line’ category as per Community Forestry Guidelines. A higher Shannon-Weiner plant diversity index at regeneration level was observed in IAS removed areas (2.43) than in control site (1.95). Furthermore, the number of tree seedlings and saplings in the IAS harvested blocks were significantly higher (p < 0.005) compared to the unharvested one. The sale of charcoal produced through the pyrolysis of IAS in ̔ Bio-energy kilns’ contributed for an average increased income of 30.95 % (Nepalese rupees 31,000) of the involved households. Despite above factors, some operational policy hurdles related to charcoal transport and taxation existed at field level. This study suggests that plant diversity could be increased through the removal of IAS, and considerable economic benefits could be achieved if charcoal is substantially produced and utilized.

Keywords: briquette, economic benefits, pyrolysis, regeneration

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Authors: Mozhgan Farzami Sepehr, Farhad Nourozi

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In a greenhouse study, decontamination capacity of the species Polypogon monspoliensis, for detoxification of petroleum-polluted soils caused by sewage and waste materials of Tehran Petroleum Refinery. For this purpose, the amount of total oil and grease before and 45 days after transplanting one-month-old seedlings in the soils of five different treatments in which pollution-free agricultural soil and contaminated soil were mixed together with the weight ratio of respectively 1 to 9 (% 10), 2 to 8 (%20), 3 to 7 (%30) , 4 to 6 (%40), and 5 to 5 (%50) were evaluated and compared with the amounts obtained from control treatment without vegetation, but with the same concentration of pollution. Findings demonstrated that the maximum reduction in the petroleum rate ,as much as 84.85 percent, is related to the treatment 10% containing the plant. Increasing the shoot height in treatments 10% and 20% as well as the root dry and fresh weight in treatments 10% , 20% , and 30% shows that probably activity of more rhizosphere microorganisms of the plant in these treatments has led to the improvement in growth of plant organs comparing to the treatments without pollution.

Keywords: phytoremediation, total oil and grease, rhizosphere, microorganisms, petroleum-contaminated soil

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Authors: Mubashar Iqbal, Iqra Anjum, Muhammad Dildar Gogi, Muhammad Jalal Arif

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The present study was carried to screen out the effective entomopathogenic fungi (EPF) inoculation method in maize and to evaluate pathogenicity and oviposition-choice in C. partellus. Three entomopathogenic fungi (EPF) formulations Pacer® (Metarhizium anisopliae), Racer® (Beauveria bassiana) and Meailkil® (Verticillium lecanii) were evaluated at three concentrations (5000, 10000 and 20000 ppm) for their endophytism in maize and pathogenicity in C. partellus. The stock solution of the highest concentration (20,000 ppm) was prepared and next lower from stock solution. In the first experiment, three EPF was inoculated in maize plant by four methods, i.e., leaf-inoculation (LI), whorl-inoculation (WI), shoot-inoculation (SI) and root-inoculation (RI). Leaf-discs and stem-cutting were sampled in all four inoculation methods and placed on fungus growth media in Petri dishes. In the second experiment, pathogenicity, pupal formation, adult emergence, sex ratio, oviposition-choice, and growth index of C. partellus were calculated. The leaves and stem of the inoculated plants were given to the counted number of larvae of C. Partellus. The mortality of larvae was recorded on daily basis till the pupation. The result shows that maximum percent mortality (86.67%) was recorded at high concentration (20000ppm) of Beauveria bassiana by leaf inoculation method. For oviposition choice bioassay, the newly emerged adults were fed on diet (water, honey and yeast in 9:1:1) for 48 hours. One pair of C. Partellus were aspirated from the rearing cages and were detained in large test tube plugged with diet soaked cotton. A set of four plants for each treatment were prepared and randomized inside the large oviposition chamber. The test tubes were opened and fitted in the hole made in the wall of oviposition chamber in front of each treatment. The oviposition chamber was placed in a completely dark laboratory to eliminate the effect of light on moth’s behavior. The plants were removed from the oviposition chamber after the death of adults. The number of eggs deposited on the plant was counted. The results of 2nd experiment revealed that in all EPF and inoculation methods, the fecundity, egg fertility and growth index of C. partellus decreased with the increase in concentration being significantly higher at low concentration (5000ppm) and lower at higher concentration (20000ppm). Application of B. bassiana demonstrated that minimum fecundity (126.83), egg fertility (119.52) and growth index (15%) in C. partellus followed by M. anisopliae with fecundity (135.93), egg fertility (132.29) and growth index (17.50%) while V. lecanii show higher values of fecundity (137.37), egg fertility (1135.42) and growth index (20%). Overall leaf inoculation method showed least fecundity (123.89) with egg fertility (115.36) and growth index (14%) followed by whorl, shoot inoculation method and root inoculation method show higher values of fecundity, egg fertility and growth index.

Keywords: Beauveria bassiana, Chilo partellus, entomopathoganic, Metarhizium anisopliae, Verticillium lecanii

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Authors: Alena O. Stepanova, Vera S. Chernonosova, Tatyana S. Godovikova, Konstantin A. Bulatov, Andrey Y. Patrushev, Pavel P. Laktionov

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Trauma and arthrosis, not to mention cartilage destruction in overweight and elders put hyaline cartilage lesion among the most frequent diseases of locomotor system. These problems combined with low regeneration potential of the cartilage make regeneration of articular cartilage a high-priority task of tissue engineering. Many types of matrices, the procedures of their installation and autologous chondrocyte implantation protocols were offered, but certain aspects including adhesion of the implant with surrounding cartilage/bone, prevention of the ossification and fibrosis were not resolved. Simplification and acceleration of the procedures resulting in restoration of normal cartilage are also required. We have demonstrated that human chondroblasts can be successfully cultivated at the surface of electrospun scaffolds and produce extracellular matrix components in contrast to chondroblasts grown in homogeneous hydrogels. To restore cartilage we offer to use stacks of electrospun scaffolds fixed with photopolymerized solution of prepared from gelatin and chondroitin-4-sulfate both modified by glycidyl methacrylate and non-toxic photoinitator Darocur 2959. Scaffolds were prepared from nylon 6, polylactide-co-glicolide and their mixtures with modified gelatin. Illumination of chondroblasts in photopolymerized solution using 365 nm LED light had no effect on cell viability at compressive strength of the gel less than0,12 MPa. Stacks of electrospun scaffolds provide good compressive strength and have the potential for substitution with cartilage when biodegradable scaffolds are used. Vascularization can be prevented by introduction of biostable scaffolds in the layers contacting the subchondral bone. Studies of two-component materials (2-3 sheets of electrospun scaffold) implanted in the knee-joints of rabbits and fixed by photopolymerization demonstrated good crush resistance, biocompatibility and good adhesion of the implant with surrounding cartilage. Histological examination of the implants 3 month after implantation demonstrates absence of any inflammation and signs of replacement of the biodegradable scaffolds with normal cartilage. The possibility of intraoperative population of the implants with autologous cells is being investigated.

Keywords: chondroblasts, electrospun scaffolds, hyaline cartilage, photopolymerized gel

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Authors: Babak Moradi, Hassan Zare-Maivan

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A greenhouse investigation has been conducted to study the effect of crude oil on morphology and protein content of Avicennia marina plant. Avicennia marina seeds were sown in different concentrations of the crude oil mixed soil (i.e., 2.5, 5, 7.5, and 10 w/w). Controls and replicates were also set up. Morphological traits were recorded 4 months after plantation. Avicennia marina seedlings could tolerate up to 10% (w/w). Results demonstrated that there was a reduction in plant shoot and root biomass with the increase of crude oil concentration. Plant height, total leaf number and length reduced significantly with increase of crude oil contamination. Investigation revealed that there is a great impact of crude oil contamination on protein content of the roots of the experimental plant. Protein content of roots grown in different concentrations of crude oil were more than those of the control plant. Further, results also showed that protein content was increased with increased concentration of crude oil.

Keywords: Avicennia marina, morphology, oil contamination, protein content

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Authors: C. Chaiyaraksa, T. Khamko

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This study investigated the influence of limestone and sepiolite on heavy metals accumulation in the soil and soybean. The soil was synthesized to contaminate with zinc 150 mg/kg, copper 100 mg/kg, and cadmium 1 mg/kg. The contaminated soil was mixed with limestone and sepiolite at the ratio of 1:0, 0:1, 1:1, and 2:1. The amount of soil modifier added to soil was 0.2%, 0.4%, and 0.8%. The metals determination was performed on soil both before and after soybean planting and in the root, shoot, and seed of soybean after harvesting. The study was also on metal translocate from root to seed and on bioaccumulation factor. Using of limestone and sepiolite resulted in a reduction of metals accumulated in soybean. For soil containing a high concentration of copper, cadmium, and zinc, a mixture of limestone and sepiolite (1:1) was recommended to mix with soil with the amount of 0.2%. Zinc could translocate from root to seed more than copper, and cadmium. From studying the movement of metals from soil to accumulate in soybean, the result was that soybean could absorb the highest amount of cadmium, followed by zinc, and copper, respectively.

Keywords: heavy metals, limestone, sepiolite, soil, soybean

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Authors: Shanna Swart, Caryn Fenner, Athanasios Kotsiopoulos, Susan Harrison

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Linear alkanes are produced as by-products from the increasing use of gas-to-liquid fuel technologies for synthetic fuel production and offer great potential for value addition. Their current use as low-value fuels and solvents do not maximize this potential. Therefore, attention has been drawn towards direct activation of these aliphatic alkanes to more useful products such as alcohols, aldehydes, carboxylic acids and derivatives. Cytochrome P450 monooxygenases (P450s) can be used for activation of these aliphatic alkanes using whole-cells or cell-free systems. Some limitations of whole-cell systems include reduced mass transfer, stability and possible side reactions. Since the P450 systems are little studied as cell-free systems, they form the focus of this study. Challenges of a cell-free system include co-factor regeneration, substrate availability and enzyme stability. Enzyme immobilization offers a positive outlook on this dilemma, as it may enhance stability of the enzyme. In the present study, 2 different P450s (CYP153A6 and CYP102A1) as well as the relevant accessory enzymes required for electron transfer (ferredoxin and ferredoxin reductase) and co-factor regeneration (glucose dehydrogenase) have been expressed in E. coli and purified by metal affinity chromatography. Glucose dehydrogenase (GDH), was used as a model enzyme to assess the potential of various enzyme immobilization strategies including; surface attachment on MagReSyn® microspheres with various functionalities and on electrospun nanofibers, using self-assembly based methods forming Cross Linked Enzymes (CLE), Cross Linked Enzyme Aggregates (CLEAs) and spherezymes as well as in a sol gel. The nanofibers were synthesized by electrospinning, which required the building of an electrospinning machine. The nanofiber morphology has been analyzed by SEM and binding will be further verified by FT-IR. Covalent attachment based methods showed limitations where only ferredoxin reductase and GDH retained activity after immobilization which were largely attributed to insufficient electron transfer and inactivation caused by the crosslinkers (60% and 90% relative activity loss for the free enzyme when using 0.5% glutaraldehyde and glutaraldehyde/ethylenediamine (1:1 v/v), respectively). So far, initial experiments with GDH have shown the most potential when immobilized via their His-tag onto the surface of MagReSyn® microspheres functionalized with Ni-NTA. It was found that Crude GDH could be simultaneously purified and immobilized with sufficient activity retention. Immobilized pure and crude GDH could be recycled 9 and 10 times, respectively, with approximately 10% activity remaining. The immobilized GDH was also more stable than the free enzyme after storage for 14 days at 4˚C. This immobilization strategy will also be applied to the P450s and optimized with regards to enzyme loading and immobilization time, as well as characterized and compared with the free enzymes. It is anticipated that the proposed immobilization set-up will offer enhanced enzyme stability (as well as reusability and easy recovery), minimal mass transfer limitation, with continuous co-factor regeneration and minimal enzyme leaching. All of which provide a positive outlook on this robust multi-enzyme system for efficient activation of linear alkanes as well as the potential for immobilization of various multiple enzymes, including multimeric enzymes for different bio-catalytic applications beyond alkane activation.

Keywords: alkane activation, cytochrome P450 monooxygenase, enzyme catalysis, enzyme immobilization

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Authors: Zanele D. Ngwenya, Mustapha Mohammed, Felix D. Dakora

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Legumes are a major source of biological nitrogen, and therefore play a crucial role in maintaining soil productivity in smallholder agriculture in southern Africa. Through their ability to fix atmospheric nitrogen in root nodules, legumes are a better option for sustainable nitrogen supply in cropping systems than chemical fertilisers. For decades, farmers have been highly receptive to the use of rhizobial inoculants as a source of nitrogen due mainly to the availability of elite rhizobial strains at a much lower compared to chemical fertilisers. To improve the efficiency of the legume-rhizobia symbiosis in African soils would require the use of highly effective rhizobia capable of nodulating a wide range of host plants. This study assessed the morphogenetic diversity, photosynthetic functioning and relative symbiotic effectiveness (RSE) of groundnut, jack bean and soybean microsymbionts in Eswatini soils as a first step to identifying superior isolates for inoculant production. According to the manufacturer's instructions, rhizobial isolates were cultured in yeast-mannitol (YM) broth until the late log phase and the bacterial genomic DNA was extracted using GenElute bacterial genomic DNA kit. The extracted DNA was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and a dendrogram constructed from the band patterns to assess rhizobial diversity. To assess the N2-fixing efficiency of the authenticated rhizobia, photosynthetic rates (A), stomatal conductance (gs), and transpiration rates (E) were measured at flowering for plants inoculated with the test isolates. The plants were then harvested for nodulation assessment and measurement of plant growth as shoot biomass. The results of ERIC-PCR fingerprinting revealed the presence of high genetic diversity among the microsymbionts nodulating each of the three test legumes, with many of them showing less than 70% ERIC-PCR relatedness. The dendrogram generated from ERIC-PCR profiles grouped the groundnut isolates into 5 major clusters, while the jack bean and soybean isolates were grouped into 6 and 7 major clusters, respectively. Furthermore, the isolates also elicited variable nodule number per plant, nodule dry matter, shoot biomass and photosynthetic rates in their respective host plants under glasshouse conditions. Of the groundnut isolates tested, 38% recorded high relative symbiotic effectiveness (RSE >80), while 55% of the jack bean isolates and 93% of the soybean isolates recorded high RSE (>80) compared to the commercial Bradyrhizobium strains. About 13%, 27% and 83% of the top N₂-fixing groundnut, jack bean and soybean isolates, respectively, elicited much higher relative symbiotic efficiency (RSE) than the commercial strain, suggesting their potential for use in inoculant production after field testing. There was a tendency for both low and high N₂-fixing isolates to group together in the dendrogram from ERIC-PCR profiles, which suggests that RSE can differ significantly among closely related microsymbionts.

Keywords: genetic diversity, relative symbiotic effectiveness, inoculant, N₂-fixing

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Authors: Wasif Mughees, Malik Al-Ahmad, Muhammad Naeem

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This research involves the design and analysis of pinch-based water/wastewater networks to minimize water utility in the petrochemical and petroleum industries. A study has been done on Tehran Oil Refinery to analyze feasibilities of regeneration, reuse and recycling of water network. COD is considered as a single key contaminant. Amount of freshwater was reduced about 149m3/h (43.8%) regarding COD. Re-design (or retrofitting) of water allocation in the networks was undertaken. The results were analyzed through graphical method and mathematical programming technique which clearly demonstrated that amount of required water would be determined by mass transfer of COD.

Keywords: minimization, water pinch, water management, pollution prevention

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Authors: Madhavi T., Sireesha Sakhamuri, Hema Sri A., Harika K.

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Today as technological advancements are taking place, these advancements are being used by the armed forces to reduce the risk of their losses and to defeat their enemies. The development of sophisticated technology relies mostly on the use of high- tech weapons or machinery. Robotics is one of the hot spheres of the modern age in which nations concentrate on the state of war and peace for military purposes. They have been in use for demining and rescue operations for some time now but are being propelled by using them for combat and spy missions. This project focuses on creating a LoRa-based spying robot with a wireless IP camera attached to it that can rising the human target. This robot transmits the signal via an IP camera to the base station. One of this project’s major applications can be analyzed using a PC that can be used to control the robot’s movement. The robot sends the signal through the LoRa transceiver at the base station to the LoRa transceiver mounted on the robot. With this function, the, robot can relay videos in real- time along with anti-collision capabilities and the enemies in the war zone cannot recognize them. More importantly, this project focuses on increasing communication using LoRa.

Keywords: lora, IP cam, metal detector, laser shoot

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Authors: Pham-Thi Huong, Byeong-Kyu Lee, Chi-Hyeon Lee, JiTae Kim

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This study analyzed the removal of p-nitrophenol from wastewater using Fe-nano zeolite synthesized. The basic physical-chemical properties of Fe-nano zeolite was determined by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy. We focus on finding out the optimum conditions in adsorption and desorption processes for removal of p-nitrophenol by using Fe-nano zeolite in wastewater. The optimum pH for p-nitrophenol removal in wastewater was 5.0. Adsorption isotherms were better fitted with the Langmuir isotherm than with the Freundlich with 165.58 mg/g adsorption capacity of p-nitrophenol. These findings support potential of Fe-nano zeolite as an effective adsorbent for p-nitrophenol removal from wastewater.

Keywords: Fe-nano zeolite, adsorption, wastewater, regeneration

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Authors: Ramesh Joshi, Nisha Khatik

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Murraya koenigii (L.) Spreng locally known as “Curry patta” or “Meetha neem” belonging to the family Rutaceae that grows wildly in Southern Asia. Its aromatic leaves are commonly used as the raw material for traditional medicinal formulations in India. The leaves contain essential oil and also used as a condiment. Several monomeric and binary carbazol alkaloids present in the various plant parts. These alkaloids have been reported to possess anti-microbial, mosquitocidal, topo-isomerase inhibition and antioxidant properties. Some of the alkaloids reported in this plant have showed anti carcinogenic and anti-diabetic properties. The conventional method of propagation of this tree is limited to seeds only, which retain their viability for only a short period. Hence, a biotechnological approach might have an advantage edging over traditional breeding as well as the genetic improvement of M. koenigii within a short period. The development of a reproducible regeneration protocol is the prerequisite for ex situ conservation and micropropagation. An efficient protocol for high frequency regeneration of in vitro plants of Murraya koenigii via different explants such as- nodal segments, intermodal segments, leaf, root segments, hypocotyle, cotyledons and cotyledonary node explants is described. In the present investigation, assessment of clonal fidelity in the micropropagated plantlets of Murraya koenigii was attempted using RAPD and ISSR markers at different pathways of plant tissue culture technique. About 20 ISSR and 40 RAPD primers were used for all the samples. Genomic DNA was extracted by CTAB method. ISSR primer were found to be more suitable as compared to RAPD for the analysis of clonal fidelity of M. koenigii. The amplifications however, were finally performed using RAPD, ISSR markers owing to their better performance in terms of generation of amplification products. In RAPD primer maximum 75% polymorphism was recorded in OPU-2 series which exhibited out of 04 scorable bands, three bands were polymorphic with a band range of size 600-1500 bp. In ISSR primers the UBC 857 showed 50% polymorphism with 02 band were polymorphic of band range size between 400-1000 bp.

Keywords: genetic fidelity, Murraya koenigii, aromatic plants, ISSR primers

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Authors: May Newisar, Richard Kingston, Philip Black

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Heritage-led regeneration developments have been considered as one of the cornerstones of the economic and social revival of historic towns and cities in the UK. However, this approach has proved its deficiency within the development of Liverpool World Heritage site. This is due to the conflict between sustaining the tangible and intangible values as well as achieving the aimed economic developments. Accordingly, the development of such areas is influenced by a top-down approach which considers heritage as consumable experience and urban regeneration as the economic development for it. This neglects the heritage sites characteristics and values as well as the design criteria for public open spaces that overlap with the heritage sites. Currently, knowledge regarding the relationship between the physical built environment of the heritage sites including the intangible values and the design development of the public open spaces is limited. Public open spaces have been studied from different perspectives such as increasing walkability, a source of social cohesion, provide a good quality of life as well as understanding users’ perception. While heritage sites have been discussed heavily on how to maintain the physical environment, understanding the courses of threats and how to be protected. In addition to users’ experiences and motivations of visiting such areas. Furthermore, new approaches tried to overcome the gap such as the historic urban landscape approach. This approach is focusing on the entire human environment with all its tangible and intangible qualities. However, this research aims to understand the relationship between the heritage sites and public open spaces and how the overlap of the design and development of both could be used as a quality to enhance the heritage sites and improve users’ experience. A meaning-making approach will be used in order to understand and articulate how the development of Liverpool World Heritage site and its value could influence and shape the design of public open space Pier Head in order to attract a different level of tourists to be used as a tool for economic development. Consequently, this will help in bridging the gap between the planning and conservation areas’ policies through an understanding of how flexible is the system in order to adopt alternative approaches for the design and development strategies for those areas.

Keywords: historic urban landscape, environmental psychology, urban governance, identity

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Authors: Manal M. Kame, Zeinab A. Demerdash, Hanan G. El-Baz, Salwa M. Hassan, Faten M. Salah, Wafaa Mansour, Olfat Hammam

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Cord blood (CB) derived Unrestricted Somatic Stem Cells (USSCs) with their multipotentiality hold great promise in liver regeneration. This work aims at evaluation of the therapeutic potentiality of USSCs in two experimental models of chronic liver injury induced either by S. mansoni infection in balb/c mice or CCL4 injection in hamsters. Isolation, propagation, and characterization of USSCs from CB samples were performed. USSCs were induced to differentiate into osteoblasts, adipocytes and hepatocyte-like cells. Cells of the third passage were transplanted in two models of liver fibrosis: (1) Twenty hamsters were induced to liver fibrosis by repeated i. p. injection of 100 μl CCl4 /hamster for 8 weeks. This model was designed as; 10 hamsters with liver fibrosis and treated with i.h. injection of 3x106 USSCs (USSCs transplanted group), 10 hamsters with liver fibrosis (pathological control group), and 10 hamsters with healthy livers (normal control group). (2) Murine chronics S.mansoni model: twenty mice were induced to liver fibrosis with S. mansoni ceracariae (60 cercariae/ mouse) using the tail immersion method and left for 12 weeks. This model was designed as; 10 mice with liver fibrosis were transplanted with i. v. injection of 1×106 USCCs (USSCs transplanted group). Other 2 groups were designed as in hamsters model. Animals were sacrificed 12 weeks after USSCs transplantation, and their liver sections were examined for detection of human hepatocyte-like cells by immunohistochemistry staining. Moreover, liver sections were examined for fibrosis level, and fibrotic indices were calculated. Sera of sacrificed animals were tested for liver functions. CB USSCs, with fibroblast-like morphology, expressed high levels of CD44, CD90, CD73 and CD105 and were negative for CD34, CD45, and HLA-DR. USSCs showed high expression of transcripts for Oct4 and Sox2 and were in vitro differentiated into osteoblasts, adipocytes. In both animal models, in vitro induced hepatocyte-like cells were confirmed by cytoplasmic expression of glycogen, alpha-fetoprotein, and cytokeratin18. Livers of USSCs transplanted group showed engraftment with human hepatocyte-like cells as proved by cytoplasmic expression of human alpha-fetoprotein, cytokeratin18, and OV6. In addition, livers of this group showed less fibrosis than the pathological control group. Liver functions in the form of serum AST & ALT level and serum total bilirubin level were significantly lowered in USSCs transplanted group than pathological control group (p < 0.001). Moreover, the fibrotic index was significantly lower (p< 0.001) in USSCs transplanted group than pathological control group. In addition liver sections, of i. v. injection of 1×106 USCCs of mice, stained with either H&E or sirius red showed diminished granuloma size and a relative decrease in hepatic fibrosis. Our experimental liver fibrosis models transplanted with CB-USSCs showed liver engraftment with human hepatocyte-like cells as well as signs of liver regeneration in the form of improvement in liver function assays and fibrosis level. These data provide hope that human CB- derived USSCs are introduced as multipotent stem cells with great potentiality in regenerative medicine & strengthens the concept of cellular therapy for the treatment of liver fibrosis.

Keywords: cord blood, liver fibrosis, stem cells, transplantation

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Authors: Kinnsley Travis, Camerron M. Crowder

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Mapk8ip3 (Mitogen-Activated Protein Kinase 8 Interacting Protein 3) is a gene that codes for the JIP3 protein, which is a part of the JIP scaffolding protein family. This protein is involved in axonal vesicle transport, elongation and regeneration. Variants in the Mapk8ip3 gene are associated with a rare-genetic condition that results in a neurodevelopmental disorder that can cause a range of phenotypes including global developmental delay and intellectual disability. Currently, there are 18 known individuals diagnosed to have sequenced confirmed Mapk8ip3 genetic disorders. This project focuses on examining the impact of a subset of missense patient variants on the Jip3 protein function by overexpressing the mRNA of these variants in a zebrafish knockout model for Jip3. Plasmids containing cDNA with individual missense variants were reverse transcribed, purified, and injected into single-cell zebrafish embryos (Wild Type, Jip3 -/+, and Jip3 -/-). At 6-days post mRNA microinjection, morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Morphologically, we compared the size and shape of the zebrafish during their development over a 5-day period. Total locomotive activity was assessed using the Microtracker assay and patterns of movement over time were examined using the DanioVision assay. Lastly, we used confocal microscopy to examine sensory axons for swelling and shortened length, which are phenotypes observed in the loss-of-function knockout Jip3 zebrafish model. Using these assays during embryonic development, we determined the impact of various missense variants on Jip3 protein function, compared to knockout and wild-type zebrafish embryo models. Variants in the gene Mapk8ip3 cause rare-neurodevelopmental disorders due to an essential role in axonal vesicle transport, elongation and regeneration. A subset of missense variants was examined by overexpressing the mRNA of these variants in a Jip3 knock-out zebrafish. Morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Using these assays, the spectrum of disorders can be phenotypically determined and the impact of variant location can be compared to knockout and wild-type zebrafish embryo models.

Keywords: rare disease, neurodevelopmental disorders, mrna overexpression, zebrafish research

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Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

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Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

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Authors: Heide Imai

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The paper will introduce and discuss specific examples of urban practices which take place within the dynamic urban landscape of contemporary Tokyo. The rising interest and importance of derelict places as resilient and creative clusters will be analysed, before relating this to the rediscovery of small urban niches and the emergence of different forms of social entrepreneurs. Secondly, two different case study areas will be introduced before discussing different forms of hybrid lifestyles, social micro scale enterprises and social innovations, understanding the concept of ‘small places of resilience’ as zones of human interaction, desire and care in which spontaneous practices take place.

Keywords: entrepreneurship, social innovation, Tokyo, urban regeneration

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Authors: Snehal G. Teli, R. J. Shelke

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CNN and MultiUNet models are the framework for the proposed method for enhancing and reconstructing underwater images. Multiscale merging of features and regeneration are both performed by the MultiUNet. CNN collects relevant features. Extensive tests on benchmark datasets show that the proposed strategy performs better than the latest methods. As a result of this work, underwater images can be represented and interpreted in a number of underwater applications with greater clarity. This strategy will advance underwater exploration and marine research by enhancing real-time underwater image processing systems, underwater robotic vision, and underwater surveillance.

Keywords: convolutional neural network, image enhancement, machine learning, multiunet, underwater images

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Authors: Pawan Kumar, Pitamber Singh Negi

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Many of the regeneration failures in conifers are often being attributed to heavy insect attack and pathogens during the period of seed formation and under storage conditions. Conifer berries and seed insects occur throughout the known range of the hosts and also limit the production of seed for nursery stock. On occasion, even entire seed crops are lost due to insect attacks. The berry and seeds of both the species have been found to be infected with insects. Recently, heavy damage to the berry and seeds of Juniper and Chilgoza Pine was observed in the field as well as in stored conditions, leading to reduction in the viability of seeds to germinate. Both the species are under great threat and regeneration of the species is very low. Due to lack of adequate literature, the study on the damage potential of seed insects was urgently required to know the exact status of the insect-pests attacking seeds/berries of both the pine species so as to develop pest management practices against the insect pests attack. As both the species are also under threat and are fighting for survival, so the study is important to develop management practices for the insect-pests of seeds/berries of Juniper and Chilgoza pine so as to evaluate in the nursery, as these species form major vegetation of their distribution zones. A six-year study on the management of insect pests of seeds of Chilgoza revealed that seeds of this species are prone to insect pests mainly borers. During present investigations, it was recorded that cones of are heavily attacked only by Dioryctria abietella (Lepidoptera: Pyralidae) in natural conditions, but seeds which are economically important are heavily infected, (sometimes up to 100% damage was also recorded) by insect borer, Plodia interpunctella (Lepidoptera: Pyralidae) and is recorded for the first time ‘to author’s best knowledge’ infesting the stored Chilgoza seeds. Similarly, Juniper berries and seeds were heavily attacked only by a single borer, Homaloxestis cholopis (Lepidoptera: Lecithoceridae) recorded as a new report in natural habitat as well as in stored conditions. During the present investigation details of insect pest attack on Juniper and Chilgoza pine seeds and berries was observed and suitable management practices were also developed to contain the insect-pests attack.

Keywords: borer, chilgozapine, cones, conifer, Lepidoptera, juniper, management, seed

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Authors: Kashif Jaeel, Bingru Huang

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Stress-induced accumulation of ethylene exacerbates drought damages in plants, and suppressing stress induction of ethylene may promote plant tolerance to heat stress. The objective of this study was to investigate the effects of endophytic bacteria (Paraburkholderia aspalathi) with 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzymes in suppressing ethylene production on plant tolerance to heat stress and underlying physiological mechanisms of P. aspalathi-regulation in creeping bentgrass (Agrostis stolonifera). A novel strain of P. aspalathi, ‘WSF23’, with ACC deaminase activity was used to inoculate the roots of plants (cv. ‘Penncross’) subjected to heat stress in controlled-environment chambers. Inoculation with WSF23 bacteria resulted in improved shoot and root growth during heat stress. The differential changes in metabolite regulation due to the bacterial inoculation could contribute to ACC deamination bacteria-improved heat tolerance in cool-season grass species.

Keywords: rhizobacteria, grass, heat, plant metabolism, soil bacteria

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Authors: Djamel Boussaa

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Cultural heritage and tourism have become during the last two decades dynamic areas of development in the world. The idea of heritage is crucial to the critical decision-making process as to how irreplaceable resources are to be utilized by people of the present or conserved for future generations in a fast changing world. In view of the importance of ‘heritage’ to the development of a tourist destination the emphasis on developing appropriate adaptive reuse strategies cannot be overemphasized. In October 1999, the 12th general assembly of the ICOMOS in Mexico stated, that in the context of sustainable development, two interrelated issues need urgent attention, cultural tourism and historic towns and cities. These two issues underscore the fact that historic resources are non-renewable, belonging to all of humanity. Without adequate adaptive reuse actions to ensure a sustainable future for these historic resources, may lead to their complete vanishing. The growth of tourism and its role in dispersing cultural heritage to everyone is developing rapidly. According to the World Tourism Organization, natural and cultural heritage resources are and will remain motivating factors for travel in the foreseeable future. According to the experts, people choose travel destinations where they can learn about traditional and distinct cultures in their historic context. The Qatar rich urban heritage is now being recognized as a valuable resource for future development. This paper discusses the role of cultural heritage and tourism in regenerating Souk Waqif, and consequently the city of Doha. Therefore, in order to use cultural heritage wisely, it will be necessary to position heritage as an essential element of sustainable development, giving particular attention to cultural heritage and tourism. The research methodology is based on an empirical survey of the situation, based on several visits, meetings and interviews with the local heritage players. The rehabilitation project initiated since 2004 will be examined and assessed. Therefore, there is potential to assess the situation and propose directions for a sustainable future to this historic landmark. Conservation for the sake of conservation appears to be an outdated concept. Many irreplaceable natural and cultural sites are being compromised because local authorities are not giving economic consideration to the value of rehabilitating such sites. The question to be raised here is 'How can cultural heritage be used wisely for tourism without compromising its social sustainability within the emerging global world?'

Keywords: cultural heritage, tourism, regeneration, economy, social sustainability

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Authors: O. P. Tripathi, P. L. Verma

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Coronal Mass Ejections (CMEs) are mostly reached on Earth from 1 to 5 days from the Sun. As a consequence, slow CMEs are accelerated toward the speed of solar wind and fast CMEs are decelerated toward the speed of the solar wind. Coronal mass ejections (CMEs) are bursts of solar material i.e. clouds of plasma and magnetic fields that shoot off the sun’s surface. Other solar events include solar wind streams that come from the coronal holes on the Sun and solar energetic particles that are primarily released by CMEs. We have studied geomagnetic storms (DST ≤ - 80nT) during 1997-2012 with halo and partial halo coronal mass ejections and found that 73.28% CMEs (halo and partial halo coronal mass ejections) are associated with geomagnetic storms. The association rate of halo and partial halo coronal mass ejections are found 67.06% and 32.94% with geomagnetic storms respectively. We have also determined positive co-relation between magnitude of geomagnetic storms and speed of coronal mass ejection with correlation co-efficient 0.23.

Keywords: geomagnetic storms, coronal mass ejections (CMEs), disturbance storm time (Dst), interplanetary magnetic field (IMF)

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Authors: Payman A. A. Zibari, Mosleh M. S. Duhoky

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These experiments were conducted at Tissue Culture Laboratory/ Faculty of Agriculture / University of Duhok during the period from January 2011 to May 2013. The objectives of this study were to study the effects of interaction between colchcine concentrations and treatment time duration of Creps capilaris (with and without 2B chromosome) on chromosome polyploidy during fifteen passages until regeneration of plants from the callus. Data showed that high percentage of chromosome polyploidy approximately can be obtained from high concentration of colchicin and long time of duration.

Keywords: polyploidy, Crepis capilaris, colchicine, B chromosome

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Authors: Azieva A. M., Yastremsky E. V., Kirillova D. A., Patsaev T. D., Sharikov R. V., Kamyshinsky R. A., Lukanina K. I., Sharikova N. A., Grigoriev T. E., Vasiliev A. L.

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Adhesive properties of scaffolds, which predominantly depend on the chemical and structural features of their surface, play the most important role in tissue engineering. The basic requirements for such scaffolds are biocompatibility, biodegradation, high cell adhesion, which promotes cell proliferation and differentiation. In many cases, synthetic polymers scaffolds have proven advantageous because they are easy to shape, they are tough, and they have high tensile properties. The regeneration of nerve tissue still remains a big challenge for medicine, and neural stem cells provide promising therapeutic potential for cell replacement therapy. However, experiments with stem cells have their limitations, such as low level of cell viability and poor control of cell differentiation. Whereas the study of already differentiated neuronal cell culture obtained from newborn mouse brain is limited only to cell adhesion. The growth and implantation of neuronal culture requires proper scaffolds. Moreover, the polymer scaffolds implants with neuronal cells could demand specific morphology. To date, it has been proposed to use numerous synthetic polymers for these purposes, including polystyrene, polylactic acid (PLA), polyglycolic acid, and polylactide-glycolic acid. Tissue regeneration experiments demonstrated good biocompatibility of PLA scaffolds, despite the hydrophobic nature of the compound. Problem with poor wettability of the PLA scaffold surface could be overcome in several ways: the surface can be pre-treated by poly-D-lysine or polyethyleneimine peptides; roughness and hydrophilicity of PLA surface could be increased by plasma treatment, or PLA could be combined with natural fibers, such as collagen or chitosan. This work presents a study of adhesion of both induced pluripotent stem cells (iPSCs) and mouse primary neuronal cell culture on the polylactide scaffolds of various types: oriented and non-oriented fibrous nonwoven materials and sponges – with and without the effect of plasma treatment and composites with collagen and chitosan. To evaluate the effect of different types of PLA scaffolds on the neuronal differentiation of iPSCs, we assess the expression of NeuN in differentiated cells through immunostaining. iPSCs more effectively differentiate into neurons on PLA scaffolds with high adhesive properties for primary neuronal cells.

Keywords: PLA scaffold, neurons, neuronal differentiation, stem cells, polylactid

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