Search results for: broth micro dilution assay

Authors: Adil Mustafa, Ahmet Erten, Alper Kiraz, Melikhan Tanyeri

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Work presented here is based on a novel experimental technique used to hydrodynamically trap oil microdroplets inside a microfluidic chip at the junction of microchannels known as stagnation point. Hydrodynamic trapping has been recently used to trap and manipulate a number of particles starting from microbeads to DNA and single cells. Benzyl Benzoate (BB) is used as droplet material. The microdroplets are trapped individually at stagnation point and their dissolution was observed. Experiments are performed for two concentrations (10mM or 10µM) of AOT surfactant (Docusate Sodium Salt) and two flow rates for each case. Moreover, experimental data is compared with Zhang-Yang-Mao (ZYM) model which studies dissolution of liquid microdroplets in the presence of a host fluid experiencing extensional creeping flow. Industrial processes like polymer blending systems in which heat or mass transport occurs experience extensional flow and an insight into these phenomena is of significant importance to many industrial processes. The experimental technique exploited here gives an insight into the dissolution of liquid microdroplets under extensional flow regime. The comparison of our experimental results with ZYM model reveals that dissolution of microdroplets at lower surfactant concentration (10µM) fits the ZYM model at saturation concentration (Cs) value reported in literature (Cs = 15×10⁻³Kg\m³) while for higher surfactant concentration (10mM) which is also above the critical micelle concentration (CMC) of surfactant (5mM) the data fits ZYM model at (Cs = 45×10⁻³Kg\m³) which is 3X times the value reported in literature. The difference in Cs value from the literature shows enhancement in dissolution rate of sparingly soluble BB microdroplets at surfactant concentrations higher than CMC. Enhancement in the dissolution of sparingly soluble materials is of great importance in pharmaceutical industry. Enhancement in the dissolution of sparingly soluble drugs is a key research area for drug design industry. The experimental method is also advantageous because it is robust and has no mechanical contact with droplets under study are freely suspended in the fluid as compared existing methods used for testing dissolution of drugs. The experiments also give an insight into CMC measurement for surfactants.

Keywords: extensional flow, hydrodynamic trapping, Zhang-Yang-Mao, CMC

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Authors: Roberto Rosenhaim, Marcos Antonio Crus Moreira, Robson da Cunha, Gerson Gomes Cunha

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This work provides details on the wind speed behavior within cities of Arraial do Cabo and São Pedro da Aldeia located in the Lakes Region of the State of Rio de Janeiro, Brazil. This region has one of the best potentials for wind power generation. In interurban layer, wind conditions are very complex and depend on physical geography, size and orientation of buildings and constructions around, population density, and land use. In the same context, the fundamental surface parameter that governs the production of flow turbulence in urban canyons is the surface roughness. Such factors can influence the potential for power generation from the wind within the cities. Moreover, the use of wind on a small scale is not fully utilized due to complexity of wind flow measurement inside the cities. It is difficult to accurately predict this type of resource. This study demonstrates how fuzzy logic can facilitate the assessment of the complexity of the wind potential inside the cities. It presents a decision support tool and its ability to deal with inaccurate information using linguistic variables created by the heuristic method. It relies on the already published studies about the variables that influence the wind speed in the urban environment. These variables were turned into the verbal expressions that are used in computer system, which facilitated the establishment of rules for fuzzy inference and integration with an application for smartphones used in the research. In the first part of the study, challenges of the sustainable development which are described are followed by incentive policies to the use of renewable energy in Brazil. The next chapter follows the study area characteristics and the concepts of fuzzy logic. Data were collected in field experiment by using qualitative and quantitative methods for assessment. As a result, a map of the various points is presented within the cities studied with its wind viability evaluated by a system of decision support using the method multivariate classification based on fuzzy logic.

Keywords: behavior of winds, wind power, fuzzy logic, sustainable development

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Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra

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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.

Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA

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Authors: E. Obreque-Slier, P. Osorio-Umaña, G. Vidal-Acevedo, A. Peña-Neira, M. Medel-Marabolí

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Sensory and nutraceutical characteristics of a wine are determined by different chemical compounds, such as organic acids, sugars, alcohols, polysaccharides, aromas, and polyphenols. The polyphenols correspond to secondary metabolites that are associated with the prevention of several pathologies, and those are responsible for color, aroma, bitterness, and astringency in wines. These compounds come from grapes and wood during aging in barrels, which correspond to the format of wood most widely used in wine production. However, the barrels is a high-cost input with a limited useful life (3-4 years). For this reason, some oenological products have been developed in order to renew the barrels and increase their useful life in some years. These formats are being used slowly because limited information exists about the effect on the wine chemical characteristics. The objective of the study was to evaluate the effect of different laubarrel renewal systems (staves and zigzag) on the polyphenolic characteristics of a Carménère wine (Vitis vinifera), an emblematic cultivar of Chile. For this, a completely randomized experimental design with 5 treatments and three replicates per treatment was used. The treatments were: new barrels (T0), used barrels during 4 years (T1), scraped used barrels (T2), used barrels with staves (T3) and used barrels with zigzag (T4). The study was performed for 12 months, and different spectrophotometric parameters (phenols, anthocyanins, and total tannins) and HPLC-DAD (low molecular weight phenols) were evaluated. The wood inputs were donated by Toneleria Nacional and corresponded to products from the same production batch. The total phenols content increased significantly after 40 days, while the total tannin concentration decreased gradually during the study. The anthocyanin concentration increased after 120 days of the assay in all treatments. Comparatively, it was observed that the wine of T2 presented the lowest values of these polyphenols, while the T0 and T4 presented the highest total phenol contents. Also, T1 presented the highest values of total tannins in relation to the rest of the treatments in some samples. The low molecular weight phenolic compounds identified by HPLC-DAD were 7 flavonoids (epigallocatechin, catechin, procyanidin gallate, epicatechin, quercetin, rutin and myricetin) and 14 non-flavonoids (gallic, protocatechuic, hydroxybenzoic, trans-cutaric, vanillinic, caffeic, syringic, p-coumaric and ellagic acids; tyrosol, vanillin, syringaldehyde, trans-resveratrol and cis-resveratrol). Tyrosol was the most abundant compound, whereas ellagic acid was the lowest in the samples. Comparatively, it was observed that the wines of T2 showed the lowest concentrations of flavonoid and non-flavonoid phenols during the study. In contrast, wines of T1, T3, and T4 presented the highest contents of non-flavonoid polyphenols. In summary, the use of barrel renovators (zig zag and staves) is an interesting alternative which would emulate the contribution of polyphenols from the barrels to the wine.

Keywords: barrels, oak wood aging, polyphenols, red wine

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Authors: Bao Chen, Frederic Kuznik, Matthieu Horgnies, Kevyn Johannes, Vincent Morin, Edouard Gengembre

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The contradiction between energy consumption, environment protection, and social development is increasingly intensified during recent decade years. At the same time, as avoiding fossil-fuels-thirsty, people turn their view on the renewable green energy, such as solar energy, wind power, hydropower, etc. However, due to the unavoidable mismatch on geography and time for production and consumption, energy storage seems to be one of the most reasonable solutions to enlarge the use of renewable energies. Thermal energy storage (TES), a branch of energy storage solution, mainly concerns the capture, storage and consumption of thermal energy for later use in different scales (individual house, apartment, district, and city). In TES research field, sensible heat and latent heat storage have been widely studied and presented at an advanced stage of development. Compared with them, thermochemical energy storage is still at initial phase but provides a relatively higher theoretical energy density and a long shelf life without heat dissipation during storage. Among thermochemical energy storage materials, inorganic pure or composite compounds like micro-porous silica gel, SrBr₂ hydrate and MgSO₄-Zeolithe have been reported as promising to be integrated into thermal energy storage systems. However, the cost of these materials, one of main obstacles, may hinder the wide use of energy storage systems in real application scales (individual house, apartment, district and even city). New studies on ettringite show promising application for thermal energy storage since its high energy density and large resource from cementitious materials. Ettringite, or calcium trisulfoaluminate hydrate, of which chemical formula is 3CaO∙Al₂O₃∙3CaSO₄∙32H₂O, or C₆AS̅₃H₃₂ as known in cement chemistry notation, is one of the most important members of AFt group. As a common compound in hydrated cements, ettringite has been widely studied for its performances in construction but barely known as a thermochemical material. For this study, we summarize available data about the structure and properties of ettringite and its metastable phase (meta-ettringite), including the processes of hydration, thermal conversion and carbonation durability for thermal energy storage.

Keywords: building materials, ettringite, meta-ettringite, thermal energy storage

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Authors: Ugur Fidan, Naim Karasekreter

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Healthy and clean water should not contain disease-causing micro-organisms and toxic chemicals and must contain the necessary minerals in a balanced manner. Today, water resources have a limited and strategic importance, necessitating the management of water reserves. Water tanks meet the water needs of people and should be regularly chlorinated to prevent waterborne diseases. For this purpose, automatic chlorination systems placed in water tanks for killing bacteria. However, the regular operation of automatic chlorination systems depends on refilling the chlorine tank when it is empty. For this reason, there is a need for a stock control system, in which chlorine levels are regularly monitored and supplied. It has become imperative to take urgent measures against epidemics caused by the fact that most of our country is not aware of the end of chlorine. The aim of this work is to rehabilitate existing water tanks and to provide a method for a modern water storage system in which chlorination is digitally monitored by turning the newly established water tanks into a closed system. A sensor network structure using GSM/GPRS communication infrastructure has been developed in the study. The system consists of two basic units: hardware and software. The hardware includes a chlorine level sensor, an RFID interlock system for authorized personnel entry into water tank, a motion sensor for animals and other elements, and a camera system to ensure process safety. It transmits the data from the hardware sensors to the host server software via the TCP/IP protocol. The main server software processes the incoming data through the security algorithm and informs the relevant unit responsible (Security forces, Chlorine supply unit, Public health, Local Administrator) by e-mail and SMS. Since the software is developed base on the web, authorized personnel are also able to monitor drinking water tank and report data on the internet. When the findings and user feedback obtained as a result of the study are evaluated, it is shown that closed drinking water tanks are built with GRP type material, and continuous monitoring in digital environment is vital for sustainable health water supply for people.

Keywords: wireless sensor networks (WSN), monitoring, chlorine, water tank, security

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Authors: Noriyuki Suyama

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This paper looked into consumer behavioral changes by analyzing the data collected by ASMARKS Co., one of a research companies in Japan. The purpose of the paper is to understand the two differences of before vs. after COVID-19 pandemic and Japanese living in Japan. Subsequently, examining the analysis results helped obtain useful insights into new business models for business parties in Japan as a microlevel perspective. The paper also tried to explore future conditions of globalization by taking into consideration nation’s political and economic changes as a macro-level perspective. The COVID-19 has been continuing its spread across the world with more than 60 million confirmed cases in 190 countries. This pandemic with restricted scopes of behavior mandates have disrupted the consumer habits of their lifestyles. Consumers have tendency to learn new ways when they have trouble in taking routine action. For example, the government forces people to refrain from going out, they try to telecommute at home. If the situation come back to normal, people still change their lifestyles to fit in the best. Some of data show typical effects of COVID-19; forceful exposure to digitalized work-life styles; more flexible time at home; importance of trustful and useful information gathering between what's good and bad;etc. in comparison with before vs. after COVID-19 pandemic. In addition, Japanese have less changed their lifestyles than Expatriates living in Japan. For example, while 94% of the expatriates have decreased their outgo because of self-quarantine, only 55% of the Japanese have done. There are more differences in both comparisons in the analysis results. The economic downtrend resulting from COVID-19 is supposed to be at least as devastating if not more so than that of the financial crisis. With unemployment levels in the US taking two weeks to reach what took 6 months in the 2008 crisis, there is no doubt of a global recession some predict could reach 10% or above of GDP. As a result, globalization in the global supply chain of goods and services will end up with negative impact. A lot of governmental financial and economic policies are supposed to focus on their own profits and interests, exclusing other countries interests as is the case with the Recovery Act just after the global financial crisis from 2007 to 2008. Both micro- and macro-levels analysis successfully reveal important connotations and managerial implications of business in Japan for Japanese consumers as well as after COVID-19 global business.

Keywords: COVID-19, lifestyle in Japan, expatriates, consumer behavior

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Authors: Shwu-Ling Peng, Chiung-Man Tsai, Chia-Jui Weng

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Chronic micro-inflammation is a hallmark of many aging-related neurodegenerative and metabolic syndrome-driven diseases. In high glucose (HG) environment, reactive oxygen species (ROS) is generated and the ROS induced inflammation, cytokines secretion, DNA damage, and cell cycle arrest to lead to cellular senescence. Water chestnut shell (WCS) is a plant hull which containing polyphenolic compounds and showed antioxidant and anticancer activities. Orchid, which containing a natural polysaccharide compound, possesses many physiological activities including anti-inflammatory and neuroprotective effects. These agricultural plants might be able to reduce oxidative stress and inflammation. This study was used HG-induced human normal dermal fibroblasts (HG-HNDFs) as an in vitro model to disclose the effects of water extract of Phalaenopsis orchid flower (WEPF) and ethanol extract of water chestnut shell (EEWCS) on the anti-aging and their underlying molecular mechanisms. The toxicity of extracts on human normal dermal fibroblasts (HNDFs) was determined by MTT method. The senescence of cells was assayed by β-galactosidase (SA-β-gal) kit. ROS and nitrate production was analyzed by Intracellular ROS contents and ELISA, respectively. Western blotting was used to detect the proteins in cells. The results showed that the exposure of HNDFs to HG (30 mM) for 72 h were caused cellular senescence and arrested cells at G0/G1 phase. Indeed, the treatment of HG-HNDFs with WEPF (200 μg/ml) and EEWCS (10 μg/ml) significantly released cell cycle arrest and promoted cell proliferation. The G1/S phase transition regulatory proteins such as protein retinoblastoma (pRb), p53, and p16ᴵᴺᴷ⁴ᵃ depressed by WEPF and EEWCS were also observed. Additionally, the treatment of WEPF and EEWCS increased the activity of HO-1 through upregulating Nrf2 as well as decreased the ROS and NO of HG-HNDFs. Therefore, the senescence marker protein-30 (SMP30) in cells was diminished. In conclusion, the WEPF and EEWCS might inhibit HG-induced aging of HNDFs by reducing oxidative stress and free radicals.

Keywords: agricultural plant extract, anti-aging, high glucose, Phalaenopsis orchid flower, water chestnut shell

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Authors: Elisabetta Ceretti, Silvia Bonizzoni, Alberto Bonetti, Milena Villarini, Marco Verani, Maria Antonella De Donno, Sara Bonetta, Umberto Gelatti

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Introduction: Air pollution is a global problem. In 2013, the International Agency for Research on Cancer (IARC) classified air pollution and particulate matter as carcinogenic to human. The study of the health effects of air pollution in children is very important because they are a high-risk group in terms of the health effects of air pollution and early exposure during childhood can increase the risk of developing chronic diseases in adulthood. The MAPEC_LIFE (Monitoring Air Pollution Effects on Children for supporting public health policy) is a project founded by EU Life+ Programme which intends to evaluate the associations between air pollution and early biological effects in children and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children. Methods: The study was carried out on 6-8-year-old children living in five Italian towns in two different seasons. Two biomarkers of early biological effects, primary DNA damage detected with the comet assay and frequency of micronuclei, were investigated in buccal cells of children. Details of children diseases, socio-economic status, exposures to other pollutants and life-style were collected using a questionnaire administered to children’s parents. Child exposure to urban air pollution was assessed by analysing PM0.5 samples collected in the school areas for PAHs and nitro-PAHs concentration, lung toxicity and in vitro genotoxicity on bacterial and human cells. Data on the chemical features of the urban air during the study period were obtained from the Regional Agency for Environmental Protection. The project created also the opportunity to approach the issue of air pollution with the children, trying to raise their awareness on air quality, its health effects and some healthy behaviors by means of an educational intervention in the schools. Results: 1315 children were recruited for the study and participate in the first sampling campaign in the five towns. The second campaign, on the same children, is still ongoing. The preliminary results of the tests on buccal mucosa cells of children will be presented during the conference as well as the preliminary data about the chemical composition and the toxicity and genotoxicity features of PM0.5 samples. The educational package was tested on 250 children of the primary school and showed to be very useful, improving children knowledge about air pollution and its effects and stimulating their interest. Conclusions: The associations between levels of air pollutants, air mutagenicity and biomarkers of early effects will be investigated. A tentative model to calculate the global absolute risk of having early biological effects for air pollution and other variables together will be proposed and may be useful to support policy-making and community interventions to protect children from possible health effects of air pollutants.

Keywords: air pollution exposure, biomarkers of early effects, children, public health policy

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Authors: Pradip Choudhury, Tapobrata Sanyal

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Effectiveness of Jute Geotextiles (JGT) in hill slope management now stands substantiated. The reasons of its efficacy are attributed to its bio-degradability, hygroscopic property and its thickness. Usually open weave JGT is used for slope management. Thickness of JGT helps in reducing the velocity of surface run-off, thus curbing the extent of migration of soil particles detached as a result of kinetic energy of rain-drops and also of wind effects. Initially JGT acts as cover of the surface of slope thus protect movement of loose soil particles. Hygroscopic property of jute effects overland storage of the flow. JGT acts as mulch and creates a congenial micro-climate that fosters quick growth of vegetation on bio-degradation. In fact JGT plays an important role in bio-remediation of slope-erosion problems. Considering the environmental aftermath, JGT is the preferred option in developed countries for surface soil conservation against erosion. In India JGT has not been tried in low temperature zones at high altitudes where temperature goes below the freezing point (even below - 25° Celsius). The behavior of JGT in such low-temperature zones is not precisely known. The 16th BRTF of Project Himank of Border Roads Organization (BRO) has recently taken the initiative to try two varieties of JGT , ie, 292 gsm and 500 gsm at two different places for hill slope management in Leh, a high altitude place of about 2,660 mtrs and 4900 mtrs above MSL respectively in Jammu & Kashmir where erosion is caused more as a result of rapid movement of sand particles due to high wind (wind erosion. Soil particles of the region formed naturally by weathering of fragile rocks are usually loosely bonded (non-cohesive), undergo dissociation with the rise in wind force and kinetic energy of rain drops and are blown away by wind. Open weave JGT interestingly was observed to contain the dissociated soil particles within its pores and lend stability the affected soil mass to a great extent thus preventing its movement by extraneous agents such as wind. The paper delineates about climatic factors, type of JGT used and the prevailing site conditions with an attempt to analyze the mechanism of functioning of JGT in low temperature zones.

Keywords: climate, erosion, jutegeotextile, stabilize

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Authors: Jan Masak, Eva Kvasnickova, Vladimir Scholtz, Olga Matatkova, Marketa Valkova, Alena Cejkova

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Microbial colonization of medical instruments, catheters, implants, etc. is a serious problem in the spread of nosocomial infections. Biofilms exhibit enormous resistance to environment. The resistance of biofilm populations to antibiotic or biocides often increases by two to three orders of magnitude in comparison with suspension populations. Subjects of interests are substances or physical processes that primarily cause the destruction of biofilm, while the released cells can be killed by existing antibiotics. In addition, agents that do not have a strong lethal effect do not cause such a significant selection pressure to further enhance resistance. Non-thermal plasma (NTP) is defined as neutral, ionized gas composed of particles (photons, electrons, positive and negative ions, free radicals and excited or non-excited molecules) which are in permanent interaction. In this work, the effect of NTP generated by the cometary corona with a metallic grid on the formation and stability of biofilm and metabolic activity of cells in biofilm was studied. NTP was applied on biofilm populations of Staphylococcus epidermidis DBM 3179, Pseudomonas aeruginosa DBM 3081, DBM 3777, ATCC 15442 and ATCC 10145, Escherichia coli DBM 3125 and Candida albicans DBM 2164 grown on solid media on Petri dishes and on the titanium alloy (Ti6Al4V) surface used for the production joint replacements. Erythromycin (for S. epidermidis), polymyxin B (for E. coli and P. aeruginosa), amphotericin B (for C. albicans) and ceftazidime (for P. aeruginosa) were used to study the combined effect of NTP and antibiotics. Biofilms were quantified by crystal violet assay. Metabolic activity of the cells in biofilm was measured using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) colorimetric test based on the reduction of MTT into formazan by the dehydrogenase system of living cells. Fluorescence microscopy was applied to visualize the biofilm on the surface of the titanium alloy; SYTO 13 was used as a fluorescence probe to stain cells in the biofilm. It has been shown that biofilm populations of all studied microorganisms are very sensitive to the type of used NTP. The inhibition zone of biofilm recorded after 60 minutes exposure to NTP exceeded 20 cm², except P. aeruginosa DBM 3777 and ATCC 10145, where it was about 9 cm². Also metabolic activity of cells in biofilm differed for individual microbial strains. High sensitivity to NTP was observed in S. epidermidis, in which the metabolic activity of biofilm decreased after 30 minutes of NTP exposure to 15% and after 60 minutes to 1%. Conversely, the metabolic activity of cells of C. albicans decreased to 53% after 30 minutes of NTP exposure. Nevertheless, this result can be considered very good. Suitable combinations of exposure time of NTP and the concentration of antibiotic achieved in most cases a remarkable synergic effect on the reduction of the metabolic activity of the cells of the biofilm. For example, in the case of P. aeruginosa DBM 3777, a combination of 30 minutes of NTP with 1 mg/l of ceftazidime resulted in a decrease metabolic activity below 4%.

Keywords: anti-biofilm activity, antibiotic, non-thermal plasma, opportunistic pathogens

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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: Luvindran Sugumaran, Mohd Nashrul Mohd Zubir, Kazi Md Salim Newaz, Tuan Zaharinie Tuan Zahari, Suazlan Mt Aznam, Aiman Mohd Halil

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With the fast growth of integrated circuits and the trend towards making electronic devices smaller, the heat dissipation load of electronic devices has continued to go over the limit. The high heat flux element would not only harm the operation and lifetime of the equipment but would also impede the performance upgrade brought about by the iteration of technological updates, which would have a direct negative impact on the economic and production cost benefits of rising industries. Hence, in high-tech industries like radar, information and communication, electromagnetic power, and aerospace, the development and implementation of effective heat dissipation technologies were urgently required. Pool boiling is favored over other cooling methods because of its capacity to dissipate a high heat flux at a low wall superheat without the usage of mechanical components. Enhancing the pool boiling performance by increasing the heat transfer coefficient via surface modification techniques has received a lot of attention. There are several surface modification methods feasible today, but the stability and durability of surface modification are the greatest priority. Thus, laser machining is an interesting choice for surface modification due to its low production cost, high scalability, and repeatability. In this study, different patterns of laser-processed copper surfaces are fabricated to investigate the nucleate pool boiling heat transfer performance of distilled water. The investigation showed that there is a significant enhancement in the pool boiling heat transfer performance of the laser-processed surface compared to the reference surface due to the notable increase in nucleation frequency and nucleation site density. It was discovered that the heat transfer coefficients increased when both the surface area ratio and the ratio of peak-to-valley height of the microstructure were raised. It is believed that the development of microstructures on the surface as a result of laser processing is the primary factor in the enhancement of heat transfer performance.

Keywords: heat transfer coefficient, laser processing, micro structured surface, pool boiling

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Authors: Ashraf Nawaz Khan, R. Alagirusamy, Apurba Das, Puneet Mahajan

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The thermoplastic-based fibre composites are acquiring a market sector of conventional as well as thermoset composites. However, replacing the thermoset with a thermoplastic composite has never been an easy task. The inherent high viscosity of thermoplastic resin reveals poor interface properties. In this work, a homo-polymer towpreg is produced through an electrostatic powder spray coating methodology. The produced flexible towpreg offers a low melt-flow distance during the consolidation of the laminate. The reduced melt-flow distance demonstrates a homogeneous fibre/matrix distribution (and low void content) on consolidation. The composite laminate has been fabricated with two manufacturing techniques such as conventional film stack (FS) and powder-coated (PC) technique. This helps in understanding the distinct response of produced laminates on applying load since the laminates produced through the two techniques are comprised of the same constituent fibre and matrix (constant fibre volume fraction). The changed behaviour is observed mainly due to the different fibre/matrix configurations within the laminate. The interface adhesion influences the load transfer between the fibre and matrix. Therefore, it influences the elastic, plastic, and failure patterns of the laminates. Moreover, the effect of preform geometries (plain weave and satin weave structure) are also studied for corresponding composite laminates in terms of various mechanical properties. The fracture analysis is carried out to study the effect of resin at the interlacement points through micro-CT analysis. The PC laminate reveals a considerably small matrix-rich and deficient zone in comparison to the FS laminate. The different load tensile, shear, fracture toughness, and drop weight impact test) is applied to the laminates, and corresponding damage behaviour is analysed in the successive stage of failure. The PC composite has shown superior mechanical properties in comparison to the FS composite. The damage that occurs in the laminate is captured through the SEM analysis to identify the prominent mode of failure, such as matrix cracking, fibre breakage, delamination, debonding, and other phenomena.

Keywords: composite, damage, fibre, manufacturing

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Authors: Anne Öman

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Today in Sweden and in other countries, a variety of digital artefacts, such as laptops, tablets, interactive whiteboards, are being used at all school levels. From an educational perspective, digital artefacts challenge traditional teaching because they provide a range of modes for expression and communication and are not limited to the traditional medium of paper. Digital technologies offer new opportunities for representations and physical interactions with objects, which put forward the role of the body in interaction and learning. From a multimodal perspective the emphasis is on the use of multiple semiotic resources for meaning- making and the study presented here has examined the differential use of semiotic resources by pupils interacting in a digitally designed task in a primary school context. The instances analyzed in this paper come from a case study where the learning task was to create an advertising film in a film-software. The study in focus involves the analysis of a single case with the emphasis on the examination of the classroom setting. The research design used in this paper was based on a micro ethnographic perspective and the empirical material was collected through video recordings of small-group work in order to explore pupils’ communication within the group activity. The designed task described here allowed students to build, share, collaborate upon and publish the redesigned products. The analysis illustrates the variety of communicative modes such as body position, gestures, visualizations, speech and the interaction between these modes and the representations made by the pupils. The findings pointed out the importance of embodied communication during the small- group processes from a learning perspective as well as a pedagogical understanding of pupils’ representations, which were similar from a cultural literacy perspective. These findings open up for discussions with further implications for the school practice concerning the small- group processes as well as the redesigned products. Wider, the findings could point out how multimodal interactions shape the learning experience in the meaning-making processes taking into account that language in a globalized society is more than reading and writing skills.

Keywords: communicative learning, interactive learning environments, pedagogical issues, primary school education

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Authors: Siyang Wang, Yujin Hu, Xuelin Wang, Wenqian Zhang

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Austenitic stainless steels are widely used in nuclear industry to manufacture critical components owing to their excellent corrosion resistance at high temperatures. Almost all the components used in nuclear power plants are produced by surface finishing (surface cold work) such as milling, grinding and so on. The change of surface states induced by machining has great influence on the corrosion behavior. In the present study, long time oxidation behavior of machined 316 austenitic stainless steel exposed to simulated pressure water reactor environment was investigated considering different surface states. Four surface finishes were produced by electro-polishing (P), grinding (G), and two milling (M and M1) processes respectively. Before oxidation, the surface Vickers micro-hardness, surface roughness of each type of sample was measured. Corrosion behavior of four types of sample was studied by using oxidation weight gain method for six oxidation periods. The oxidation time of each period was 120h, 216h, 336h, 504h, 672h and 1344h, respectively. SEM was used to observe the surface morphology of oxide film in several period. The results showed that oxide film on austenitic stainless steel has a duplex-layer structure. The inner oxide film is continuous and compact, while the outer layer is composed of oxide particles. The oxide particle consisted of large particles (nearly micron size) and small particles (dozens of nanometers to a few hundred nanometers). The formation of oxide particle could be significantly affected by the machined surface states. The large particle on cold worked samples (grinding and milling) appeared earlier than electro-polished one, and the milled sample has the largest particle size followed by ground one and electro-polished one. For machined samples, the large particles were almost distributed along the direction of machining marks. Severe exfoliation was observed on one milled surface (M) which had the most heavily cold worked layer, while rare local exfoliation occurred on the ground sample (G) and the other milled sample (M1). The electro-polished sample (P) entirely did not exfoliate.

Keywords: austenitic stainless steel, oxidation, machining, SEM

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Authors: Nourhan Hussein Fanaki, Hoda Mohamed Gamal El-Din Omar, Nihal Kadry Moussa, Eva Adel Edward Farid

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The resistance of staphylococci to various antibiotics has become a major concern for health care professionals. The efficacy of the combinations of selected glycopeptides (vancomycin and teicoplanin) with gentamicin or rifampicin, as well as that of gentamicin/rifampicin combination, was studied against selected pathogenic staphylococcus isolated from Egypt. The molecular distribution of genes conferring resistance to these four antibiotics was detected among tested clinical isolates. Antibiotic combinations were studied using the checkerboard technique and the time-kill assay (in both the stationary and log phases). Induction of resistance to glycopeptides in staphylococci was tried in the absence and presence of diclofenac sodium as inducer. Transmission electron microscopy was used to study the effect of glycopeptides on the ultrastructure of the cell wall of staphylococci. Attempts were made to cure gentamicin resistance plasmids and to study the transfer of these plasmids by conjugation. Trials for the transformation of the successfully isolated gentamicin resistance plasmid to competent cells were carried out. The detection of genes conferring resistance to the tested antibiotics was performed using the polymerase chain reaction. The studied antibiotic combinations proved their efficacy, especially when tested during the log phase. Induction of resistance to glycopeptides in staphylococci was more promising in presence of diclofenac sodium, compared to its absence. Transmission electron microscopy revealed the thickening of bacterial cell wall in staphylococcus clinical isolates due to the presence of tested glycopeptides. Curing of gentamicin resistance plasmids was only successful in 2 out of 9 tested isolates, with a curing rate of 1 percent for each. Both isolates, when used as donors in conjugation experiments, yielded promising conjugation frequencies ranging between 5.4 X 10-2 and 7.48 X 10-2 colony forming unit/donor cells. Plasmid isolation was only successful in one out of the two tested isolates. However, low transformation efficiency (59.7 transformants/microgram plasmid DNA) of such plasmids was obtained. Negative regulators of autolysis, such as arlR, lytR and lrgB, as well as cell-wall associated genes, such as pbp4 and/or pbp2, were detected in staphylococcus isolates with reduced susceptibility to the tested glycopeptides. Concerning rifampicin resistance genes, rpoBstaph was detected in 75 percent of the tested staphylococcus isolates. It could be concluded that in vitro studies emphasized the usefulness of the combination of vancomycin or teicoplanin with gentamicin or rifampicin, as well as that of gentamicin with rifampicin, against staphylococci showing varying resistance patterns. However, further in vivo studies are required to ensure the safety and efficacy of such combinations. Diclofenac sodium can act as an inducer of resistance to glycopeptides in staphylococci. Cell-wall thickness is a major contributor to such resistance among them. Gentamicin resistance in these strains could be chromosomally or plasmid mediated. Multiple mutations in the rpoB gene could mediate staphylococcus resistance to rifampicin.

Keywords: glycopeptides, combinations, induction, diclofenac, transmission electron microscopy, polymerase chain reaction

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Authors: Garima Sharma, R. N. Sharma

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Integrated watershed management programme based on 'Four Water Concept' was implemented in Sali village, in Jaipur District, Rajasthan State of India . The latitude 26.7234486 North and longitude 75.023876 East are the geocoordinate of the Sali. 'Four Waters Concept' is evolved by integrating the 'Four Waters', viz. rain water, soil moisture, ground water and surface water This methodology involves various water harvesting techniques to prevent the runoff of water by treatment of catchment, proper utilization of available water harvesting structures, renovation of the non-functional water harvesting structures and creation of new water harvesting structures. The case study included questionnaire survey from farmers and continuous study of village for two years. The total project area is 6153 Hac, and the project cost is Rs. 92.25 million. The sanctioned area of Sali Micro watershed is 2228 Hac with an outlay of Rs. 10.52 million. Watershed treatment activities such as water absorption trench, continuous contour trench, field bunding, check dams, were undertaken on agricultural lands for soil and water conservation. These measures have contributed in preventing runoff and increased the perennial availability of water in wells. According to the survey, water level in open wells in the area has risen by approximately 5 metres after the introduction of water harvesting structures. The continuous availability of water in wells has increased the area under irrigation and helped in crop diversification. Watershed management activities have brought the changes in cropping patterns and crop productivity. It helped in transforming 567 Hac culturable waste land into culturable arable land in the village. The farmers of village have created an additional income from the increased crop production. The programme also assured the availability of water during peak summers for the day to day activities of villagers. The outcomes indicate that there is positive impact of watershed management practices on the water resource potential as well the crop production of the area. This suggests that persistent efforts in this direction may lead to sustainability of the watershed.

Keywords: four water concept, groundwater potential, irrigation potential, watershed management

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Authors: Obiajulu Anthony Ugochukwu Nnedum, Bernard Chukwukelue Chine, Jerome Ogochukwu Ezisi

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A notable challenge of organizational citizenship behavior in Nigerian organizations is the prevalence of individualistic work cultures among employees, as this mindset can result in employees being less willing to go beyond their formal job requirements to contribute to the organization overall success. However, the dearth and scarce research on the antecedents of organizational citizenship behavior, such as paternalistic leadership and employee loyalty to supervisors in sub-Saharan African cultures such as Nigeria, motivated the current study to take a deep investigation into the moderating role of employee loyalty to supervisor on the relationship between paternalistic leadership and organizational citizenship behavior. The relevance of the current study ensures that when employees are loyal to their paternalistic leaders who show care and support, they are more likely to exhibit organizational citizenship behavior. The current study employed a sample size of four hundred and twenty participants (one hundred and five managers and three hundred and five subordinates) from eleven large organizations randomly selected through lucky dip from twenty-two large organizations from the directory of the Chamber of Commerce and Industry in Anambra state, south-eastern Nigeria. Also, a twelve-item organizational citizenship behavior scale, a thirty-nine-item paternalistic leadership scale, and a six-item loyalty to supervisor scale were employed for the collection of data for the current study. Adopting a one manager/Leader by triad subordinates cross-sectional survey design, Hayes process micro model and statistical package for social sciences (SPSS) version twenty-five, the findings from the result of the analysis of the hypotheses demonstrated that loyalty to supervisor moderated the relationship between paternalistic leadership and organizational citizenship behavior-conscientiousness. Also, the findings from the result revealed that loyalty to the supervisor moderated the relationship between authoritative leadership and organizational citizenship behavior identification. Furthermore, the findings from the result showed that loyalty to the supervisor moderated the relationship between moral leadership and organizational citizenship behavior. Accordingly, the result from the analysis implies that when employees are loyal to their supervisors, they are more likely to exhibit organizational citizenship behavior by going above and beyond their formal job requirements, as this loyalty can be fostered through a paternalistic leadership style that emphasizes a supportive and caring relationship between supervisors and subordinates.

Keywords: authoritative leadership, moral leadership, loyalty to supervisor, organizational citizenship behavior

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Authors: Hameed Ullah, Shujaul Mulk Khan, Zahid Ullah, Zeeshan Ahmad Sadia Jahangir, Abdullah, Amin Ur Rahman, Muhammad Suliman, Dost Muhammad

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The interrelationship between vegetation and abiotic variables inside an ecosystem is one of the main jobs of plant scientists. This study was designed to investigate the vegetation structure and species diversity along with the environmental variables in the Yakhtangay hill district Shangla of the Himalayan Mountain series Pakistan by using multivariate statistical analysis. Quadrat’s method was used and a total of 171 Quadrats were laid down 57 for Tree, Shrubs and Herbs, respectively, to analyze the phytosociological attributes of the vegetation. The vegetation of the selected area was classified into different Life and leaf-forms according to Raunkiaer classification, while PCORD software version 5 was used to classify the vegetation into different plants communities by Two-way indicator species Analysis (TWINSPAN). The CANOCCO version 4.5 was used for DCA and CCA analysis to find out variation directories of vegetation with different environmental variables. A total of 114 plants species belonging to 45 different families was investigated inside the area. The Rosaceae (12 species) was the dominant family followed by Poaceae (10 species) and then Asteraceae (7 species). Monocots were more dominant than Dicots and Angiosperms were more dominant than Gymnosperms. Among the life forms the Hemicryptophytes and Nanophanerophytes were dominant, followed by Therophytes, while among the leaf forms Microphylls were dominant, followed by Leptophylls. It is concluded that among the edaphic factors such as soil pH, the concentration of soil organic matter, Calcium Carbonates concentration in soil, soil EC, soil TDS, and physiographic factors such as Altitude and slope are affecting the structure of vegetation, species composition and species diversity at the significant level with p-value ≤0.05. The Vegetation of the selected area was classified into four major plants communities and the indicator species for each community was recorded. Classification of plants into 4 different communities based upon edaphic gradients favors the individualistic hypothesis. Indicator Species Analysis (ISA) shows the indicators of the study area are mostly indicators to the Himalayan or moist temperate ecosystem, furthermore, these indicators could be considered for micro-habitat conservation and respective ecosystem management plans.

Keywords: species richness, edaphic gradients, canonical correspondence analysis (CCA), TWCA

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Authors: N. G. Klivleyeva, T. I. Glebova, G. V. Lukmanova, S. B. Bayseit, S. Z. Taubaeva, M. K. Kalkozhaeva

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The role of viral diseases in the general infectious disease incidence increases every year and requires special attention to the problem of interpreting the etiology of infectious agents. Influenza and acute respiratory viral infections are one of the most pressing public health issues. In the period 2012-2014, collection of 419 nasal swabs and 150 blood sera has been carried out in the patient care institutions of the various Kazakhstan regions from patients with symptoms of ARVI and pneumonia. Primary identification of biosamples for the presence of influenza viral antigens in enzyme immunoassay on nitrocellulose membrane gave positive results in 125 swabs (29.8%). Biosample screening in immunofluorescence test revealed the presence of influenza viral antigens against A/H1 in 63 samples (15.0%), A/H3 – in 70 samples (16.7%) and type B – in 9 samples (2.1%). As a result of primary infection, and successive passages in chick embryos and MDCK cell cultures, 38 HAAg were isolated from 419 samples with a clear cytopathic effect and hemagglutination titre in MDCK cell culture within 1:2-1:4, in CE - 1:8-1:256. The infectivity of isolates in chicken embryos were 3.5-6.5 lg EID50/0.2, in MDCK cell culture – 2.5-6.5 lg PFU/ml. Identification of 28 isolates was carried out in inhibition reactions of hemagglutinating activity and neuraminidase activity, showed their belonging to the influenza virus: 26 strains to A/H1N1, one - to A/H3N2, and one - to type B. Serological examination of blood sera for the presence of specific antibodies being an indirect evidence of the performed isolation and contributing to the timely interpretation of the disease etiology in the epidemics takes an important place in the comprehensive study of influenza viruses circulating among people. Serological analyzes were carried out in HAI assay using a kit consisting of 12 reference strains obtained from the WHO centre for reference and research on Influenza (CDC, Atlanta, USA) and three Kazakhstan (A/Almaty/347/09 (H1N1v), A/Almaty/462/11 (H3N2) and B/Almaty/414/10) human influenza viruses that are stored in the laboratory collection. The results of serological analysis of 150 blood sera showed that antihaemagglutinins against the A/H3N2 virus serosubtype were found in 46 samples (49.4%) out of 93 sera collected in 2012-2013. The antibody titres were within 1:160-1:320. 19 sera (20.4%) were seropositive against influenza A/H1N1 virus, the antibodies were observed in titres of 1:20-1:40. Six sera (6.4%) were positive against the influenza A/H1N1+A/H3N2 virus (mixed infection); the antibodies were recorded in titres of 1:20-1:40. Antihaemagglutinins against influenza type B virus were detected only in five sera (5.4%). The results of analysis of 57 sera collected in 2014 showed that antihaemagglutinins against A/H3N2 virus subtype were detected in 32 blood sera (56.1%) in titres of 1:160-1:640. Ten sera (17.5%) were seropositive against A/H1N1 virus; antihaemagglutinins against influenza type B virus were not detected. Therefore, virological and serological studies have shown that in Kazakhstan, as well as in the world, the influenza viruses A/H1N1, A/H3N2 and influenza B viruses were actively circulating during the epidemic seasons in 2012-2014.

Keywords: influenza, MDCK cell, serological analysis, virus

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Authors: C. Zimmermann, E. Lackner, M. Ebner

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Massive Open Online Courses (MOOCs) are increasingly popular learning hubs that are boasting considerable participant numbers, innovative technical features, and a multitude of instructional resources. Still, there is a high level of evidence showing that almost all MOOCs suffer from a declining frequency of participant activity and fairly low completion rates. In this paper, we would like to share the lessons learned in implementing several design patterns that have been suggested in order to foster participant activity. Our conclusions are based on experiences with the ‘Dr. Internet’ MOOC, which was created as an xMOOC to raise awareness for a more critical approach to online health information: participants had to diagnose medical case studies. There is a growing body of recommendations (based on Learning Analytics results from earlier xMOOCs) as to how the decline in participant activity can be alleviated. One promising focus in this regard is instructional design patterns, since they have a tremendous influence on the learner’s motivation, which in turn is a crucial trigger of learning processes. Since Medieval Age storytelling, micro-learning units and specific comprehensible, narrative structures were chosen to animate the audience to follow narration. Hence, MOOC participants are not likely to abandon a course or information channel when their curiosity is kept at a continuously high level. Critical aspects that warrant consideration in this regard include shorter course duration, a narrative structure with suspense peaks (according to the ‘storytelling’ approach), and a course schedule that is diversified and stimulating, yet easy to follow. All of these criteria have been observed within the design of the Dr. Internet MOOC: 1) the standard eight week course duration was shortened down to six weeks, 2) all six case studies had a special quiz format and a corresponding resolution video which was made available in the subsequent week, 3) two out of six case studies were split up in serial video sequences to be presented over the span of two weeks, and 4) the videos were generally scheduled in a less predictable sequence. However, the statistical results from the first run of the MOOC do not indicate any strong influences on the retention rate, so we conclude with some suggestions as to why this might be and what aspects need further consideration.

Keywords: case study, Dr. internet, experience, MOOCs, design patterns

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Authors: Hanjun Hu, Jinxiang Zhang

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With the decline of "growthism", China's urbanization process has shifted from the stage of spatial expansion to the stage of optimization of built-up spaces, and urban renewal has gradually become a new wave of China's urban movement in recent years. The ongoing urban renewal movement in China not only needs to generate new motivation for urban development but also solve the backlog of social problems caused by rapid urbanization, which provides an opportunity for the transformation of China's urban governance model. Unlike previous approaches that focused on physical space and functional renewal, such as urban reconstruction, redevelopment, and reuse, the key challenge of urban renewal in the post-growth era lies in coordinating the complex interest relationships between multiple stakeholders. The traditional theoretical frameworks that focus on the structural relations between social groups are insufficient to explain the behavior logic and mutual cooperation mechanism of various groups and individuals in the current urban renewal practices. Therefore, based on the long-term tracking of the urban renewal practices in the Old City of Nanjing (OCN), this paper introduces the "collective action" theory to deeply analyze changes in the urban renewal governance model in OCN and tries to summarize the governance strategies that promote the formation of collective action within recent practices from a micro-scale. The study found that the practice in OCN experienced three different stages "government-led", "growth coalition" and "asymmetric game". With the transformation of government governance concepts, the rise of residents' consciousness of rights, and the wider participation of social organizations in recent years, the urban renewal in OCN is entering a new stage of "collective renewal action". Through the establishment of the renewal organization model, incentive policies, and dynamic negotiation mechanism, urban renewal in OCN not only achieves a relative balance between individual interests and collective interests but also makes the willingness of residents the dominant factor in formulating urban renewal policies. However, the presentation of "collective renewal action" in OCN is still mainly based on typical cases. Although the government is no longer the dominant role, a large number of resident-led collective actions have not yet emerged, which puts forward new research needs for a sustainable governance policy innovation in this action.

Keywords: urban renewal, collective action theory, governance, cooperation mechanism, China

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Authors: Zane Brough, Claudio Paoloni

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Modern low earth orbit (LEO) satellites that require multi-mission flexibility are highly likely to be repositioned between different operational orbits. While executing this process the satellite may experience high levels of vibration and environmental hazards, exposing the deployed solar panel to dangerous stress levels, fatigue and space debris, hence it is desirable to retract the solar array before satellite repositioning to avoid damage or failure. Furthermore, to accommodate for today's technological world, the power demand of a modern LEO satellite is rapidly increasing, which consequently provides pressure upon the design of the satellites solar array system to conform to the strict volume and mass limitations. A novel concept of deployable/retractable hybrid solar array system, aimed to provide a greater power to volume ratio while dramatically reducing the disadvantages of system mass and cost is proposed. Taking advantage of the new lightweight technology in solar panels, a mechanical system composed of both rigid and flexible solar panels arranged within a petal formation is proposed to yield a stowed to deployment area ratio up to at least 1:7, which improves the power density dramatically. The system consists of five subsystems, the outer ones based on a novel eight-petal configuration that provides a large surface and supports the flexible solar panels. A single cable and spool based hinge mechanism were designed to synchronously deploy/retract the panels in a safe, simple and efficient manner while the mass compared to the previous systems is considerably reduced. The relevant challenge to assure a smooth movement is resolved by a proper minimization of the gearing system and the use of a micro-controller system. A prototype was designed by 3D simulators and successfully constructed and tested. Further design works are in progress to implement an epicyclical gear hinge mechanism, which will further reduce the volume, mass and complexity of the system significantly. The proposed system due to an effective and reliable mechanism provides a large active surface, whilst being very compact. It could be extremely advantageous for use as ground portable solar panel system.

Keywords: mechatronic engineering, satellite, solar panel, deployable/retractable mechanism

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Authors: O. Gsib, U. Peirera, C. Egles, S. A. Bencherif

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In skin regenerative medicine, one of the critical issues is to produce a three-dimensional scaffold with optimized porosity for dermal fibroblast infiltration and neovascularization, which exhibits high mechanical properties and displays sufficient wound healing characteristics. In this study, we report on the synthesis and characterization of macroporous sequential interpenetrating polymer networks (IPNs) combining skin wound healing properties of fibrin with the excellent physical properties of polyethylene glycol (PEG). Fibrin fibers serve as a provisional biologically active network to promote cell adhesion and proliferation while PEG provides the mechanical stability to maintain the entire 3D construct. After having modified both PEG and Serum Albumin (used for promoting enzymatic degradability) by adding methacrylate residues (PEGDM and SAM, respectively), Fibrin/PEGDM-SAM sequential IPNs were synthesized as follows: Macroporous sponges were first produced from PEGDM-SAM hydrogels by a freeze-drying technique and then rehydrated by adding the fibrin precursors. Environmental Scanning Electron Microscopy (ESEM) and Confocal Laser Scanning Microscopy (CLSM) were used to characterize their microstructure. Human dermal fibroblasts were cultivated during one week in the constructs and different cell culture parameters (viability, morphology, proliferation) were evaluated. Subcutaneous implantations of the scaffolds were conducted on five-week old male nude mice to investigate their biocompatibility in vivo. We successfully synthesized interconnected and macroporous Fibrin/PEGDM-SAM sequential IPNs. The viability of primary dermal fibroblasts was well maintained (above 90%) after 2 days of culture. Cells were able to adhere, spread and proliferate in the scaffolds suggesting the suitable porosity and intrinsic biologic properties of the constructs. The fibrin network adopted a spider web shape that covered partially the pores allowing easier cell infiltration into the macroporous structure. To further characterize the in vitro cell behavior, cell proliferation (EdU incorporation, MTS assay) is being studied. Preliminary histological analysis of animal studies indicated the persistence of hydrogels even after one-month post implantation and confirmed the absence of inflammation response, good biocompatibility and biointegration of our scaffolds within the surrounding tissues. These results suggest that our Fibrin/PEGDM-SAM IPNs could be considered as potential candidates for dermis regenerative medicine. Histological analysis will be completed to further assess scaffold remodeling including de novo extracellular matrix protein synthesis and early stage angiogenesis analysis. Compression measurements will be conducted to investigate the mechanical properties.

Keywords: fibrin, hydrogels for dermal reconstruction, polyethylene glycol, semi-interpenetrating polymer network

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Authors: Suehyun Cho, Darrell Harrington, Florent Cros, Olin Palmer, John Caputo, Michael Kardosh, Eran Oren, William Loudon, Alex Kiselyov, Michael Shpigelmacher

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The Dandy-Walker malformation (DWM) represents a clinical syndrome manifesting as a combination of posterior fossa cyst, hypoplasia of the cerebellar vermis, and obstructive hydrocephalus. Anatomic hallmarks include hypoplasia of the cerebellar vermis, enlargement of the posterior fossa, and cystic dilatation of the fourth ventricle. Current treatments of DWM, including shunting of the cerebral spinal fluid ventricular system and endoscopic third ventriculostomy (ETV), are frequently clinically insufficient, require additional surgical interventions, and carry risks of infections and neurological deficits. Bionaut Labs develops an alternative way to treat Dandy-Walker Malformation (DWM) associated with non-communicating hydrocephalus. We utilize our discreet microsurgical Bionaut™ particles that are controlled externally and remotely to perform safe, accurate, effective fenestration of the Dandy-Walker cyst, specifically in the posterior fossa of the brain, to directly normalize intracranial pressure. Bionaut™ allows for complex non-linear trajectories not feasible by any conventional surgical techniques. The microsurgical particle safely reaches targets in the lower occipital section of the brain. Bionaut™ offers a minimally invasive surgical alternative to highly involved posterior craniotomy or shunts via direct fenestration of the fourth ventricular cyst at the locus defined by the individual anatomy. Our approach offers significant advantages over the current standards of care in patients exhibiting anatomical challenge(s) as a manifestation of DWM, and therefore, is intended to replace conventional therapeutic strategies. Current progress, including platform optimization, Bionaut™ control, and real-time imaging and in vivo safety studies of the Bionauts™ in large animals, specifically the spine and the brain of ovine models, will be discussed.

Keywords: Bionaut™, cerebral spinal fluid, CSF, cyst, Dandy-Walker, fenestration, hydrocephalus, micro-robot

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Authors: Yuvraj Singh Dangi, Brajesh Kumar Tiwari, Ashok Kumar Jain, Kamta Prasad Namdeo

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The potential use of liposomes as drug carriers by i.v. injection is limited by their low stability in blood stream. Firstly, phospholipid exchange and transfer to lipoproteins, mainly HDL destabilizes and disintegrates liposomes with subsequent loss of content. To avoid the pain associated with injection and to obtain better patient compliance studies concerning various dosage forms, have been developed. Conventional liposomes (unilamellar and multilamellar) have certain drawbacks like low entrapment efficiency, stability and release of drug after single breach in external membrane, have led to the new type of liposomal systems. The challenge has been successfully met in the form of Double Liposomes (DL). DL is a recently developed type of liposome, consisting of smaller liposomes enveloped in lipid bilayers. The outer lipid layer of DL can protect inner liposomes against various enzymes, therefore DL was thought to be more effective than ordinary liposomes. This concept was also supported by in vitro release characteristics i.e. DL formation inhibited the release of drugs encapsulated in inner liposomes. DL consists of several small liposomes encapsulated in large liposomes, i.e., multivesicular vesicles (MVV), therefore, DL should be discriminated from ordinary classification of multilamellar vesicles (MLV), large unilamellar vesicles (LUV), small unilamellar vesicles (SUV). However, for these liposomes, the volume of inner phase is small and loading volume of water-soluble drugs is low. In the present study, the potential of phosphatidylethanolamine (PE) lipid anchored double liposomes (DL) to incorporate two drugs in a single system is exploited as a tool to augment the H. pylori eradication rate. Preparation of DL involves two steps, first formation of primary (inner) liposomes by thin film hydration method containing one drug, then addition of suspension of inner liposomes on thin film of lipid containing the other drug. The success of formation of DL was characterized by optical and transmission electron microscopy. Quantitation of DL-bacterial interaction was evaluated in terms of percent growth inhibition (%GI) on reference strain of H. pylori ATCC 26695. To confirm specific binding efficacy of DL to H. pylori PE surface receptor we performed an agglutination assay. Agglutination in DL treated H. pylori suspension suggested selectivity of DL towards the PE surface receptor of H. pylori. Monotherapy is generally not recommended for treatment of a H. pylori infection due to the danger of development of resistance and unacceptably low eradication rates. Therefore, combination therapy with amoxicillin trihydrate (AMOX) as anti-H. pylori agent and ranitidine bismuth citrate (RBC) as antisecretory agent were selected for the study with an expectation that this dual-drug delivery approach will exert acceptable anti-H. pylori activity.

Keywords: Helicobacter pylorI, amoxicillin trihydrate, Ranitidine Bismuth citrate, phosphatidylethanolamine, multi vesicular systems

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Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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

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

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

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Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

Abstract:

Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

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