Search results for: alternative media

Authors: Miyoko Taniguchi

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Conflict and violence have prevailed over the last four decades in conflict-affected areas in Muslim Mindanao, despite the signing of several peace agreements between the Philippine government and Islamic separatist insurgents (the Moro National Liberation Front (MNLF) and the Moro Islamic Liberation Front (MILF)), and peacebuilding activities on the ground. In the meantime, the peace talks had been facilitated and mediated by international actors such as the Organization of Islamic Cooperation (OIC) and its member countries such as Indonesia, and Malaysia, and Japan. In 2014, both the Government of the Philippines (GPH) and the MILF finally reached a Comprehensive Peace Agreement (CAB) in 2014 under the Aquino III administration, though a Bangsamoro Basic Law (BBL) based on the CAB was not enacted at the Catholic-majority of the Philippine Congress. After a long process of deliberations at the Congress, Republic Act 11054, known as the Bangsamoro Organic Law (BOL), was enacted in 2018 under the Duterate administration. In the beginning, President Duterte adopted an 'inclusive approach' that involves the MILF, all factions of the MNLF, non-Islamized indigenous peoples, and other influential clan leaders to align all peace processes under a single Bangsamoro peace process. A notable difference from past administrations, there is an explicit recognition of all agreements and legislations based on the rights of each stakeholder. This created a new identity as 'Bangsamoro', the residents of Muslim Mindanao, enhancing political legitimacy. Besides, it should be noted an important role of 'insider mediators' -a platform for the Bangsamoro from diverse sectors attempting to work within their respective organizations in Moro society. Give the above background, this paper aims at probing the effectiveness of insider mediation as one of the alternative approaches for mediation in the peace process. For the objectives, this research uses qualitative methods such as process-tracing and semi-structured interviews from diverse groups of stakeholders at from the state to the regional level, including the government officials involved in peace process under the Presidential Office, rebels (MILF and MNLF), civil society organizations involved in lobbying and facilitating peace process, especially in the legislative process. The key outcomes and findings are that the Insider Mediators Group, formed in 2016, had taken on a significant role in facilitating the achievement of a wider consensus among stakeholders on major Moro issues such as BBL’s passing during the last administration to call for unity among the Bangsamoro. Most of its members are well-educated professionals affiliated with the MILF, the MNLF, and influential clans. One of the group’s biggest achievements has been the lobbying and provision of legal advice to legislators who were not necessarily knowledgeable about the peace process during the deliberation of the bicameral conference of the BBL, which eventually led to its passage. It can be concluded that in the long run, strengthening vertical and horizontal relations between the Moro society and the State and among the Moro peoples that can be viewed as a means to sustainable peace.

Keywords: insider mediation, Mindanao, peace process, Moro Islamic liberation front

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Authors: Asegid Belay Kebede, Getachew Biru Worku

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The Earth and its inhabitants have faced an existential threat as a result of severe manmade actions. Global warming and climate change have been the most apparent manifestations of this threat throughout the world, with increasingly intense heat waves, temperature rises, flooding, sea-level rise, ice sheet melting, and so on. One of the major contributors to this disaster is the ever-increasing production and consumption of energy, which is still primarily fossil-based and emits billions of tons of hazardous GHG. The transportation industry is recognized as the biggest actor in terms of emissions, accounting for 24% of direct CO2 emissions and being one of the few worldwide sectors where CO2 emissions are still growing. Rail transportation, which includes all from light rail transit to high-speed rail services, is regarded as one of the most efficient modes of transportation, accounting for 9% of total passenger travel and 7% of total freight transit. Nonetheless, there is still room for improvement in the transportation sector, which might be done by incorporating alternative and/or renewable energy sources. As a result of these rapidly changing global energy situations and rapidly dwindling fossil fuel supplies, we were driven to analyze the possibility of renewable energy sources for traction applications. Even a small achievement in energy conservation or harnessing might significantly influence the total railway system and have the potential to transform the railway sector like never before. As a result, the paper begins by assessing the potential for photovoltaic (PV) power generation on train rooftops and existing infrastructure such as railway depots, passenger stations, traction substation rooftops, and accessible land along rail lines. As a result, a method based on a Google Earth system (using Helioscopes software) is developed to assess the PV potential along rail lines and on train station roofs. As an example, the Addis Ababa light rail transit system (AA-LRTS) is utilized. The case study examines the electricity-generating potential and economic performance of photovoltaics installed on AALRTS. As a consequence, the overall capacity of solar systems on all stations, including train rooftops, reaches 72.6 MWh per day, with an annual power output of 10.6 GWh. Throughout a 25-year lifespan, the overall CO2 emission reduction and total profit from PV-AA-LRTS can reach 180,000 tons and 892 million Ethiopian birrs, respectively. The PV-AA-LRTS has a 200% return on investment. All PV stations have a payback time of less than 13 years, and the price of solar-generated power is less than $0.08/kWh, which can compete with the benchmark price of coal-fired electricity. Our findings indicate that PV-AA-LRTS has tremendous potential, with both energy and economic advantages.

Keywords: sustainable development, global warming, energy crisis, photovoltaic energy conversion, techno-economic analysis, transportation system, light rail transit

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Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

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Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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Authors: Eman AbdelSabour

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Sustainability is increasingly being considered as a critical aspect in shaping the urban environment. It works as an invention development basis for global urban growth. Currently, different models and structures impact the means of interpreting the criteria that would be included in defining a sustainable city. There is a collective need to improve the growth path to an extremely durable path by presenting different suggestions regarding multi-scale initiatives. The global rise in urbanization has led to increased demand and pressure for better urban planning choice and scenarios for a better sustainable urban alternative. The need for an assessment tool at the urban scale was prompted due to the trend of developing increasingly sustainable urban development (SUD). The neighborhood scale is being managed by a growing research committee since it seems to be a pertinent scale through which economic, environmental, and social impacts could be addressed. Although neighborhood design is a comparatively old practice, it is in the initial years of the 21st century when environmentalists and planners started developing sustainable assessment at the neighborhood level. Through this, urban reality can be considered at a larger scale whereby themes which are beyond the size of a single building can be addressed, while it still stays small enough that concrete measures could be analyzed. The neighborhood assessment tool has a crucial role in helping neighborhood sustainability to perform approach and fulfill objectives through a set of themes and criteria. These devices are also known as neighborhood assessment tool, district assessment tool, and sustainable community rating tool. The primary focus of research has been on sustainability from the economic and environmental aspect, whereas the social, cultural issue is rarely focused. Therefore, this research is based on Doha, Qatar, the current urban conditions of the neighborhoods is discussed in this study. The research problem focuses on the spatial features in relation to the socio-cultural aspects. This study is outlined in three parts; the first section comprises of review of the latest use of wellbeing assessment methods to enhance decision process of retrofitting physical features of the neighborhood. The second section discusses the urban settlement development, regulations and the process of decision-making rule. An analysis of urban development policy with reference to neighborhood development is also discussed in this section. Moreover, it includes a historical review of the urban growth of the neighborhoods as an atom of the city system present in Doha. Last part involves developing quantified indicators regarding subjective well-being through a participatory approach. Additionally, applying GIS will be utilized as a visualizing tool for the apparent Quality of Life (QoL) that need to develop in the neighborhood area as an assessment approach. Envisaging the present QoL situation in Doha neighborhoods is a process to improve current condition neighborhood function involves many days to day activities of the residents, due to which areas are considered dynamic.

Keywords: neighborhood, subjective wellbeing, decision support tools, Doha, retrofiring

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Authors: Sara Parker, Kay Standing

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This paper will share insights into how menstruators bodies in Nepal are viewed and controlled in Nepal due to the deeply held stigmas and taboos that exist that frame menstrual blood as impure and polluting. It draws on a British Academy Global Challenges Research (BA/GCRF) funded project, ‘Dignity Without Danger,’ that ran from December 2019 to 2022. In Nepal, beliefs and myths around menstrual related practices prevail and vary in accordance to time, generation, caste and class. Physical seclusion and/or restrictions include the consumption of certain foods, the ability to touch certain people and objects, and restricted access to water sources. These restrictions not only put women at risk of poor health outcomes, but they also promote discrimination and challenge fundamental human rights. Despite the pandemic, a wealth of field research and creative outputs have been generated to help break the silence that surrounds menstruation and also highlights the complexity of addressing the harms associated with the exclusion from sacred and profane spaces that menstruators face. Working with locally recruited female research assistants, NGOS and brining together academics from the UK and Nepal, we explore the intersecting factors that impact on menstrual experiences and how they vary throughout Nepal. WE concur with Tamang that there is no such thing as a ‘Nepali Woman’, and there is no one narrative that captures the experiences of menstruators in Nepal. These deeply held beliefs and practices mean that menstruators are denied their right to a dignified menstruation. By being excluded from public and private spaces, such as temples and religious sites, as well as from kitchens and your own bedroom in your own home, these beliefs impact on individuals in complex and interesting ways. Existing research in Nepal by academics and activists demonstrates current programmes and initiatives do not fully address the misconceptions that underpin the exclusionary practices impacting on sexual and reproductive health, a sense of well being and highlight more work is needed in this area. Research has been conducted in all 7 provinces and through exploring and connecting disparate stories, artefacts and narratives, we will deepen understanding of the complexity of menstrual practices enabling local stakeholders to challenge exclusionary practices. By using creative methods to engage with stakeholders and share our research findings as well as highlighting the wealth of activism in Nepal. We highlight the importance of working with local communities, leaders and cutting across disciplines and agencies to promote menstrual justice and dignity. Our research findings and creative outputs that we share on social media channels such as Dignity Without Danger Facebook, Instagram and you tube stress the value of employing a collaborative action research approach to generate material which helps local people take control of their own narrative and change social relations that lead to harmful practices.

Keywords: menstruation, Nepal, stigma, social norms

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Authors: Alastair Hales, Xi Jiang, Siming Zhang

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Numerical methods are used to generate vortices in a domain. Through considered design, two counter-rotating vortices may interact and effectively drive one another downstream. This phenomenon is comparable to the vortex interaction that occurs in a region immediately downstream from two counter-oscillating piezoelectric (PE) fan blades. PE fans are small blades clamped at one end and driven to oscillate at their first natural frequency by an extremely low powered actuator. In operation, the high oscillation amplitude and frequency generate sufficient blade tip speed through the surrounding air to create downstream air flow. PE fans are considered an ideal solution for low power hot spot cooling in a range of small electronic devices, but a single blade does not typically induce enough air flow to be considered a direct alternative to conventional air movers, such as axial fans. The development of face-to-face PE fan arrays containing multiple blades oscillating in counter-phase to one another is essential for expanding the range of potential PE fan applications regarding the cooling of power electronics. Even in an unoptimised state, these arrays are capable of moving air volumes comparable to axial fans with less than 50% of the power demand. Replicating the airflow generated by face-to-face PE fan arrays without including the actual blades in the model reduces the process’s computational demands and enhances the rate of innovation and development in the field. Vortices are generated at a defined inlet using a time-dependent velocity profile function, which pulsates the inlet air velocity magnitude. This induces vortex generation in the considered domain, and these vortices are shown to separate and propagate downstream in a regular manner. The generation and propagation of a single vortex are compared to an equivalent vortex generated from a PE fan blade in a previous experimental investigation. Vortex separation is found to be accurately replicated in the present numerical model. Additionally, the downstream trajectory of the vortices’ centres vary by just 10.5%, and size and strength of the vortices differ by a maximum of 10.6%. Through non-dimensionalisation, the numerical method is shown to be valid for PE fan blades with differing parameters to the specific case investigated. The thorough validation methods presented verify that the numerical model may be used to replicate vortex formation from an oscillating PE fans blade. An investigation is carried out to evaluate the effects of varying the distance between two PE fan blade, pitch. At small pitch, the vorticity in the domain is maximised, along with turbulence in the near vicinity of the inlet zones. It is proposed that face-to-face PE fan arrays, oscillating in counter-phase, should have a minimal pitch to optimally cool nearby heat sources. On the other hand, downstream airflow is maximised at a larger pitch, where the vortices can fully form and effectively drive one another downstream. As such, this should be implemented when bulk airflow generation is the desired result.

Keywords: piezoelectric fans, low energy cooling, vortex formation, computational fluid dynamics

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Authors: Angelica Alvarez-Lee, Sergio Martinez-Diaz, Jose Luis Garcia-Corona, Humberto Lanz-Mendoza

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World aquaculture is always looking for improvements to achieve productions with high yields avoiding the infection by pathogenic agents. The best way to achieve this is to know the biological model to create alternative treatments that could be applied in the hatcheries, which results in greater economic gains and improvements in human public health. In the last decade, immunomodulation in shrimp culture with probiotics, organic acids and different carbon sources has gained great interest, mainly in larval and juvenile stages. Immune priming is associated with a strong protective effect against a later pathogen challenge. This work provides another perspective about immunostimulation from spawning until hatching. The stimulation happens during development embryos and generates resistance to infection by pathogenic bacteria. Massive spawnings of white shrimp L. vannamei were obtained and placed in experimental units with 700 mL of sterile seawater at 30 °C, salinity of 28 ppm and continuous aeration at a density of 8 embryos.mL⁻¹. The immunostimulating effect of three death strains of non-pathogenic bacterial (Escherichia coli, Staphylococcus aureus and Bacillus subtilis) and a pathogenic strain for white shrimp (Vibrio parahaemolyticus) was evaluated. The strains killed by heat were adjusted to O.D. 0.5, at A 600 nm, and directly added to the seawater of each unit at a ratio of 1/100 (v/v). A control group of embryos without inoculum of dead bacteria was kept under the same physicochemical conditions as the rest of the treatments throughout the experiment and used as reference. The duration of the stimulus was 12 hours, then, the larvae that hatched were collected, counted and transferred to a new experimental unit (same physicochemical conditions but at a salinity of 28 ppm) to carry out a challenge of infection against the pathogen V. parahaemolyticus, adding directly to seawater an amount 1/100 (v/v) of the live strain adjusted to an OD 0.5; at A 600 nm. Subsequently, 24 hrs after infection, nauplii survival was evaluated. The results of this work shows that, after 24 hrs, the hatching rates of immunostimulated shrimp embryos with the dead strains of B. subtillis and V. parahaemolyticus are significantly higher compared to the rest of the treatments and the control. Furthermore, survival of L. vanammei after a challenge of infection of 24 hrs against the live strain of V. parahaemolyticus is greater (P < 0.05) in the larvae immunostimulated during the embryonic development with the dead strains B. subtillis and V. parahaemolyticus, followed by those that were treated with E. coli. In summary superficial antigens can stimulate the development cells to promote hatching and can have normal development in agreeing with the optical observations, plus exist a differential response effect between each treatment post-infection. This research provides evidence of the immunostimulant effect of death pathogenic and non-pathogenic bacterial strains in the rate of hatching and oversight of shrimp L. vannamei during embryonic and larval development. This research continues evaluating the effect of these death strains on the expression of genes related to the defense priming in larvae of L. vannamei that come from massive spawning in hatcheries before and after the infection challenge against V. parahaemolyticus.

Keywords: immunostimulation, L. vannamei, hatching, survival

Procedia PDF Downloads 117

Authors: Nadeen Felemban, Hamsa Banjer, Rabaah Jaafari

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One of the most common hazards in the pharmaceutical industry is the chemical hazard, which can cause harm or develop occupational health diseases/illnesses due to chronic exposures to hazardous substances. Therefore, a chemical agent management system is required, including hazard identification, risk assessment, controls for specific hazards and inspections, to keep your workplace healthy and safe. However, routine management monitoring is also required to verify the effectiveness of the control measures. Moreover, Betamethasone Valerate and Clobetasol Propionate are some of the APIs (Active Pharmaceutical Ingredients) with highly hazardous classification-Occupational Hazard Category (OHC 4), which requires a full containment (ECA-D) during handling to avoid chemical exposure. According to Safety Data Sheet, those chemicals are reproductive toxicants (reprotoxicant H360D), which may affect female workers’ health and cause fatal damage to an unborn child, or impair fertility. In this study, qualitative (chemical Risk assessment-qCRA) was conducted to assess the chemical exposure during handling of Betamethasone Valerate and Clobetasol Propionate in pharmaceutical laboratories. The outcomes of qCRA identified that there is a risk of potential chemical exposure (risk rating 8 Amber risk). Therefore, immediate actions were taken to ensure interim controls (according to the Hierarchy of controls) are in place and in use to minimize the risk of chemical exposure. No open handlings should be done out of the Steroid Glove Box Isolator (SGB) with the required Personal Protective Equipment (PPEs). The PPEs include coverall, nitrile hand gloves, safety shoes and powered air-purifying respirators (PAPR). Furthermore, a quantitative assessment (personal air sampling) was conducted to verify the effectiveness of the engineering controls (SGB Isolator) and to confirm if there is chemical exposure, as indicated earlier by qCRA. Three personal air samples were collected using an air sampling pump and filter (IOM2 filters, 25mm glass fiber media). The collected samples were analyzed by HPLC in the BV lab, and the measured concentrations were reported in (ug/m3) with reference to Occupation Exposure Limits, 8hr OELs (8hr TWA) for each analytic. The analytical results are needed in 8hr TWA (8hr Time-weighted Average) to be analyzed using Bayesian statistics (IHDataAnalyst). The results of the Bayesian Likelihood Graph indicate (category 0), which means Exposures are de "minimus," trivial, or non-existent Employees have little to no exposure. Also, these results indicate that the 3 samplings are representative samplings with very low variations (SD=0.0014). In conclusion, the engineering controls were effective in protecting the operators from such exposure. However, routine chemical monitoring is required every 3 years unless there is a change in the processor type of chemicals. Also, frequent management monitoring (daily, weekly, and monthly) is required to ensure the control measures are in place and in use. Furthermore, a Similar Exposure Group (SEG) was identified in this activity and included in the annual health surveillance for health monitoring.

Keywords: occupational health and safety, risk assessment, chemical exposure, hierarchy of control, reproductive

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Authors: Domenico Carmelo Mongelli

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The context of this research is that of the Italian reality, which, in order to adapt to the EU Directives that prohibit the production of internal combustion engines in favor of electric mobility from 2035, is extremely concerned about the significant loss of jobs resulting from the difficulty of the automotive industry in converting in such a short time and due to the reticence of potential buyers in the face of such an epochal change. The aim of the research is to evaluate for Italy the potential of the most valid alternative to this transition to electric: leaving the current production of diesel engines unchanged, no longer powered by gasoil, imported and responsible for greenhouse gas emissions, but powered entirely by a nationally produced and eco-sustainable fuel such as biodiesel. Today in Italy, the percentage of biodiesel mixed with gasoil for diesel engines is too low (around 10%); for this reason, this research aims to evaluate the functioning of current diesel engines powered 100% by biodiesel and the ability of the Italian production system to cope to this hypothesis. The research geographically identifies those abandoned lands in Italy, now out of the food market, which is best suited to an energy crop for the final production of biodiesel. The cultivation of oilseeds is identified, which for the Italian agro-industrial reality allows maximizing the agricultural and industrial yields of the transformation of the agricultural product into a final energy product and minimizing the production costs of the entire agro-industrial chain. To achieve this objective, specific databases are used, and energy and economic balances are prepared for the different agricultural product alternatives. Solutions are proposed and tested that allow the optimization of all production phases in both the agronomic and industrial phases. The biodiesel obtained from the most feasible of the alternatives examined is analyzed, and its compatibility with current diesel engines is identified, and from the evaluation of its thermo-fluid-dynamic properties, the engineering measures that allow the perfect functioning of current internal combustion engines are examined. The results deriving from experimental tests on the engine bench are evaluated to evaluate the performance of different engines fueled with biodiesel alone in terms of power, torque, specific consumption and useful thermal efficiency and compared with the performance of engines fueled with the current mixture of fuel on the market. The results deriving from experimental tests on the engine bench are evaluated to evaluate the polluting emissions of engines powered only by biodiesel and compared with current emissions. At this point, we proceed with the simulation of the total replacement of gasoil with biodiesel as a fuel for the current fleet of diesel vehicles in Italy, drawing the necessary conclusions in technological, energy, economic, and environmental terms and in terms of social and employment implications. The results allow us to evaluate the potential advantage of a total replacement of diesel fuel with biodiesel for powering road vehicles with diesel cycle internal combustion engines without significant changes to the current vehicle fleet and without requiring future changes to the automotive industry.

Keywords: biodiesel, economy, engines, environment

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Authors: Sumeet Rai, Anuj Tyagi, B. T. Naveen Kumar, Shubhkaramjeet Kaur, Niraj K. Singh

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Since their discovery, indiscriminate use of antibiotics in human, veterinary and aquaculture systems has resulted in global emergence/spread of multidrug-resistant bacterial pathogens. Thus, the need for alternative approaches to control bacterial infections has become utmost important. High selectivity/specificity of bacteriophages (phages) permits the targeting of specific bacteria without affecting the desirable flora. In this study, a lytic phage (Ahp1) specific to Aeromonas hydrophila subsp. hydrophila was isolated from finfish aquaculture pond. The host range of Ahp1 range was tested against 10 isolates of A. hydrophila, 7 isolates of A. veronii, 25 Vibrio cholerae isolates, 4 V. parahaemolyticus isolates and one isolate each of V. harveyi and Salmonella enterica collected previously. Except the host A. hydrophila subsp. hydrophila strain, no lytic activity against any other bacterial was detected. During the adsorption rate and one-step growth curve analysis, 69.7% of phage particles were able to get adsorbed on host cell followed by the release of 93 ± 6 phage progenies per host cell after a latent period of ~30 min. Phage nucleic acid was extracted by column purification methods. After determining the nature of phage nucleic acid as dsDNA, phage genome was subjected to next-generation sequencing by generating paired-end (PE, 2 x 300bp) reads on Illumina MiSeq system. De novo assembly of sequencing reads generated circular phage genome of 42,439 bp with G+C content of 58.95%. During open read frame (ORF) prediction and annotation, 22 ORFs (out of 49 total predicted ORFs) were functionally annotated and rest encoded for hypothetical proteins. Proteins involved in major functions such as phage structure formation and packaging, DNA replication and repair, DNA transcription and host cell lysis were encoded by the phage genome. The complete genome sequence of Ahp1 along with gene annotation was submitted to NCBI GenBank (accession number MF683623). Stability of Ahp1 preparations at storage temperatures of 4 °C, 30 °C, and 40 °C was studied over a period of 9 months. At 40 °C storage, phage counts declined by 4 log units within one month; with a total loss of viability after 2 months. At 30 °C temperature, phage preparation was stable for < 5 months. On the other hand, phage counts decreased by only 2 log units over a period of 9 during storage at 4 °C. As some of the phages have also been reported as glycerol sensitive, the stability of Ahp1 preparations in (0%, 15%, 30% and 45%) glycerol stocks were also studied during storage at -80 °C over a period of 9 months. The phage counts decreased only by 2 log units during storage, and no significant difference in phage counts was observed at different concentrations of glycerol. The Ahp1 phage discovered in our study had a very narrow host range and it may be useful for phage typing applications. Moreover, the endolysin and holin genes in Ahp1 genome could be ideal candidates for recombinant cloning and expression of antimicrobial proteins.

Keywords: Aeromonas hydrophila, endolysin, phage, narrow host range

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Authors: Shubhangi Roy

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Relational Contracts (informal understandings not enforceable by law) are a common feature of most economies. However, their dominance is higher in developing countries. Such informality of economic sectors is often co-related to lower economic growth. The aim of this paper is to investigate whether informal arrangements i.e. relational contracts are a cause or symptom of lower levels of economic and/or institutional development. The methodology followed involves an initial survey of 150 test subjects in Northern India. The subjects are all members of occupations where they frequently transact ensuring uniformity in transaction volume. However, the subjects are from varied socio-economic backgrounds to ensure sufficient variance in transaction values allowing us to understand the relationship between the amount of money involved to the method of transaction used, if any. Questions asked are quantitative and qualitative with an aim to observe both the behavior and motivation behind such behavior. An overarching similarity observed during the survey across all subjects’ responses is that in an economy like India with pervasive corruption and delayed litigation, economy participants have created alternative social sanctions to deal with non-performers. In a society that functions predominantly on caste, class and gender classifications, these sanctions could, in fact, be more cumbersome for a potential rule-breaker than the legal ramifications. It, therefore, is a symptom of weak formal regulatory enforcement and dispute settlement mechanism. Additionally, the study bifurcates such informal arrangements into two separate systems - a) when it exists in addition to and augments a legal framework creating an efficient socio-legal equilibrium or; b) in conflict with the legal system in place. This categorization is an important step in regulating informal arrangements. Instead of considering the entire gamut of such arrangements as counter-development, it helps decision-makers understand when to dismantle (latter) and when to pivot around existing informal systems (former). The paper hypothesizes that those social arrangements that support the formal legal frameworks allow for cheaper enforcement of regulations with lower enforcement costs burden on the state mechanism. On the other hand, norms which contradict legal rules will undermine the formal framework. Law infringement, in presence of these norms, will have no impact on the reputation of the business or individual outside of the punishment imposed under the law. It is especially exacerbated in the Indian legal system where enforcement of penalties for non-performance of contracts is low. In such a situation, the social norm will be adhered to more strictly by the individuals rather than the legal norms. This greatly undermines the role of regulations. The paper concludes with recommendations that allow policy-makers and legal systems to encourage the former category of informal arrangements while discouraging norms that undermine legitimate policy objectives. Through this investigation, we will be able to expand our understanding of tools of market development beyond regulations. This will allow academics and policymakers to harness social norms for less disruptive and more lasting growth.

Keywords: distribution of income, emerging economies, relational contracts, sample survey, social norms

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Authors: Maximilian Engelhardt, Stephan Seeck

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The COVID-19 pandemic is accelerating the already growing field of e-commerce. The resulting urban freight transport volume leads to traffic and negative environmental impact. Furthermore, the service level of parcel logistics service provider is lacking far behind the expectations of consumer. These challenges can be solved by radically reorganize the urban last mile distribution structure: parcels could be consolidated in a micro hub within the inner city and delivered within time windows by cargo bike. This approach leads to a significant improvement of consumer satisfaction with their overall delivery experience. However, this approach also leads to significantly increased costs per parcel. While there is a relevant share of online shoppers that are willing to pay for such a delivery service there are no deeper insights about this target group available in the literature. Being aware of the importance of knowing target groups for businesses, the aim of this paper is to elaborate the most important factors that determine the willingness to pay for sustainable and service-oriented parcel delivery (factor analysis) and to derive customer segments (cluster analysis). In order to answer those questions, a data set is analyzed using quantitative methods of multivariate statistics. The data set was generated via an online survey in September and October 2020 within the five largest cities in Germany (n = 1.071). The data set contains socio-demographic, living-related and value-related variables, e.g. age, income, city, living situation and willingness to pay. In a prior work of the author, the data was analyzed applying descriptive and inference statistical methods that only provided limited insights regarding the above-mentioned research questions. The analysis in an exploratory way using factor and cluster analysis promise deeper insights of relevant influencing factors and segments for user behavior of the mentioned parcel delivery concept. The analysis model is built and implemented with help of the statistical software language R. The data analysis is currently performed and will be completed in December 2021. It is expected that the results will show the most relevant factors that are determining user behavior of sustainable and service-oriented parcel deliveries (e.g. age, current service experience, willingness to pay) and give deeper insights in characteristics that describe the segments that are more or less willing to pay for a better parcel delivery service. Based on the expected results, relevant implications and conclusions can be derived for startups that are about to change the way parcels are delivered: more customer-orientated by time window-delivery and parcel consolidation, more environmental-friendly by cargo bike. The results will give detailed insights regarding their target groups of parcel recipients. Further research can be conducted by exploring alternative revenue models (beyond the parcel recipient) that could compensate the additional costs, e.g. online-shops that increase their service-level or municipalities that reduce traffic on their streets.

Keywords: customer segmentation, e-commerce, last mile delivery, parcel service, urban logistics, willingness-to-pay

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Authors: B. Shaw, K. Doherty

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The Cultural Communication and Computing Research Institute (C3RI) is a diverse multidisciplinary research institute including art, design, media production, communication studies, computing and engineering. Across these disciplines it can seem like there are enormous differences of research practice and convention, including differing positions on objectivity and subjectivity, certainty and evidence, and different political and ethical parameters. These differences sit within, often unacknowledged, histories, codes, and communication styles of specific disciplines, and it is all these aspects that can make understanding of research practice across disciplines difficult. To explore this, a one day event was orchestrated, testing how a PhD community might communicate and share research in progress in a multi-disciplinary context. Instead of presenting results at a conference, research students were tasked to articulate their method of inquiry. A working party of students from across disciplines had to design a conference call, visual identity and an event framework that would work for students across all disciplines. The process of establishing the shape and identity of the conference was revealing. Even finding a linguistic frame that would meet the expectations of different disciplines for the conference call was challenging. The first abstracts submitted either resorted to reporting findings, or only described method briefly. It took several weeks of supported intervention for research students to get ‘inside’ their method and to understand their research practice as a process rich with philosophical and practical decisions and implications. In response to the abstracts the conference committee generated key methodological categories for conference sessions, including sampling, capturing ‘experience’, ‘making models’, researcher identities, and ‘constructing data’. Each session involved presentations by visual artists, communications students and computing researchers with inter-disciplinary dialogue, facilitated by alumni Chairs. The apparently simple focus on method illuminated research process as a site of creativity, innovation and discovery, and also built epistemological awareness, drawing attention to what is being researched and how it can be known. It was surprisingly difficult to limit students to discussing method, and it was apparent that the vocabulary available for method is sometimes limited. However, by focusing on method rather than results, the genuine process of research, rather than one constructed for approval, could be captured. In unlocking the twists and turns of planning and implementing research, and the impact of circumstance and contingency, students had to reflect frankly on successes and failures. This level of self – and public- critique emphasised the degree of critical thinking and rigour required in executing research and demonstrated that honest reportage of research, faults and all, is good valid research. The process also revealed the degree that disciplines can learn from each other- the computing students gained insights from the sensitive social contextualizing generated by communications and art and design students, and art and design students gained understanding from the greater ‘distance’ and emphasis on application that computing students applied to their subjects. Finding the means to develop dialogue across disciplines makes researchers better equipped to devise and tackle research problems across disciplines, potentially laying the ground for more effective collaboration.

Keywords: interdisciplinary, method, research student, training

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Authors: Sandeep Kumar Agrawal, Aditi Bhattacharya, Janvie Manhas, Krushna Bhatt, Yatin Kholakiya, Nupur Khera, Ajoy Roychoudhury, Sudip Sen

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Autologous cultured explants of human oral mucosal epithelial cells (OMEC) are a potential therapeutic modality for limbal stem cell deficiency (LSCD). Injury or inflammation of the ocular surface in the form of burns, chemicals, Stevens Johnson syndrome, ocular cicatricial pemphigoid etc. can lead to destruction and deficiency of limbal stem cells. LSCD manifests in the form of severe ocular surface diseases (OSD) characterized by persistent and recurrent epithelial defects, conjuntivalisation and neovascularisation of the corneal surface, scarring and ultimately opacity and blindness. Most of the cases of OSD are associated with severe dry eye pertaining to diminished mucin and aqueous secretion. Mycophenolate mofetil (MMF) has been shown to upregulate the mucin expression in conjunctival goblet cells in vitro. The aim of this study was to evaluate the effects of MMF on mucin expression in primary cultures of oral mucosal epithelial cells. With institutional ethics committee approval and written informed consent, thirty oral mucosal epithelial tissue samples were obtained from patients undergoing oral surgery for non-malignant conditions. OMEC were grown on human amniotic membrane (HAM, obtained from expecting mothers undergoing elective caesarean section) scaffold for 2 weeks in growth media containing DMEM & Ham’s F12 (1:1) with 10% FBS and growth factors. In vitro dosage of MMF was standardised by MTT assay. Analysis of stem cell markers was done using RT-PCR while mucin mRNA expression was quantified using RT-PCR and q-PCR before and after treating cultured OMEC with graded concentrations of MMF for 24 hours. Protein expression was validated using immunocytochemistry. Morphological studies revealed a confluent sheet of proliferating, stratified oral mucosal epithelial cells growing over the surface of HAM scaffold. The presence of progenitor stem cell markers (p63, p75, β1-Integrin and ABCG2) and cell surface associated mucins (MUC1, MUC15 and MUC16) were elucidated by RT-PCR. The mucin mRNA expression was found to be upregulated in MMF treated primary cultures of OMEC, compared to untreated controls as quantified by q-PCR with β-actin as internal reference gene. Increased MUC1 protein expression was validated by immunocytochemistry on representative samples. Our findings conclude that OMEC have the ability to form a multi-layered confluent sheet on the surface of HAM similar to a cornea, which is important for the reconstruction of the damaged ocular surface. Cultured OMEC has stem cell properties as demonstrated by stem cell markers. MMF can be a novel enhancer of mucin production in OMEC. It has the potential to improve dry eye in patients undergoing OMEC transplantation for bilateral OSD. Further clinical trials are required to establish the role of MMF in patients undergoing OMEC transplantation.

Keywords: limbal stem cell deficiency, mycophenolate mofetil, mucin, ocular surface disease

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Authors: Çağrı Bekir Baysal, Göktuğ Burak Çalık

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Armored vehicles may have different traction layouts as a result of terrain capabilities and mobility needs. Two main categories of layouts can be separated as wheeled and tracked. Tracked vehicles have superior off-road capabilities but what they gain on terrain performance they lose on mobility front. Wheeled vehicles on the other hand do not have as good terrain capabilities as tracked vehicles but they have superior mobility capabilities such as top speed, range and agility with respect to tracked vehicles. Conventional armored vehicles employ a diesel ICE as main power source. In these vehicles ICE is mechanically connected to the powertrain. This determines the ICE rpm as a result of speed and torque requested by the driver. ICE efficiency changes drastically with torque and speed required and conventional vehicles suffer in terms of fuel consumption because of this. Hybrid electric vehicles employ at least one electric motor in order to improve fuel efficiency. There are different types of hybrid vehicles but main types are Series Hybrid, Parallel Hybrid and Series-Parallel Hybrid. These vehicles introduce an electric motor for traction and also can have a generator electric motor for range extending purposes. Having an electric motor as the traction power source brings the flexibility of either using the ICE as an alternative traction source while it is in efficient range or completely separating the ICE from traction and using it solely considering efficiency. Hybrid configurations have additional advantages for armored vehicles in addition to fuel efficiency. Heat signature, silent operation and prolonged stationary missions can be possible with the help of the high-power battery pack that will be present in the vehicle for hybrid drivetrain. Because of the reasons explained, hybrid armored vehicles are becoming a target area for military and also for vehicle suppliers. In order to have a better idea and starting point when starting a hybrid armored vehicle design, hybrid drivetrain configuration has to be selected after performing a trade-off study. This study has to include vehicle mobility simulations, integration level, vehicle level and performance level criteria. In this study different hybrid traction configurations possible for an 8x8 vehicle is compared using above mentioned criteria set. In order to compare hybrid traction configurations ease of application, cost, weight advantage, reliability, maintainability, redundancy and performance criteria have been used. Performance criteria points have been defined with the help of vehicle simulations and tests. Results of these simulations and tests also help determining required tractive power for an armored vehicle including conditions like trench and obstacle crossing, gradient climb. With the method explained in this study, each configuration is assigned a point for each criterion. This way, correct configuration can be selected objectively for every application. Also, key aspects of armored vehicles, mine protection and ballistic protection will be considered for hybrid configurations. Results are expected to vary for different types of vehicles but it is observed that having longitudinal differential locking capability improves mobility and having high motor count increases complexity in general.

Keywords: armored vehicles, electric drivetrain, electric mobility, hybrid vehicles

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Authors: Ashima Sharma

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Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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Authors: Santos Maza, Enzo Aldoradin, Carlos Pariona, Eliud Arpi, Maria Rosales

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The objective of this study was to investigate the effect of flaying then bleeding anchovy (Engraulis ringens) immersed within a sodium citrate solution. Anchovy is a pelagic fish that readily deteriorates due to its high content of polyunsaturated fatty acids. As such, within the Peruvian food industry, the shelf life of frozen anchovy is explicitly 6 months, this short duration imparts a barrier to use for direct consumption human. Thus, almost all capture of anchovy by the fishing industry is eventually used in the production of fishmeal. We offer this an alternative to its typical production process in order to increase shelf life. In the present study, 100 kg of anchovies were captured and immediately mixed with ice on ship, maintaining a high quality sensory metric (e.g., with color blue in back) while still arriving for processing less than 2 h after capture. Anchovies with fat content of 3% were immediately flayed (i.e., reducing subcutaneous fat), beheaded, gutted and bled (i.e., removing hemoglobin) by immersion in water (Control) or in a solution of 2.5% sodium citrate (treatment), then subsequently frozen at -30 °C for 8 h in 2 kg batches. Subsequent glazing and storage at -25 °C for 14 months completed the experiments parameters. The peroxide value (PV), acidity (A), fatty acid profile (FAP), thiobarbituric acid reactive substances (TBARS), heme iron (HI), pH and sensory attributes of the samples were evaluated monthly. The results of the PV, TBARS, A, pH and sensory analyses displayed significant differences (p<0.05) between treatment and control sample; where the sodium citrate treated samples showed increased preservation features. Specifically, at the beginning of the study, flayed, beheaded, gutted and bled anchovies displayed low content of fat (1.5%) with moderate amount of PV, A and TBARS, and were not rejected by sensory analysis. HI values and FAP displayed varying behavior, however, results of HI did not reveal a decreasing trend. This result is indicative of the fact that levels of iron were maintained as HI and did not convert into no heme iron, which is known to be the primary catalyst of lipid oxidation in fish. According to the FAP results, the major quantity of fatty acid was of polyunsaturated fatty acid (PFA) followed by saturated fatty acid (SFA) and then monounsaturated fatty acid (MFA). According to sensory analysis, the shelf life of flayed, beheaded and gutted anchovy (control and treatment) was 14 months. This shelf life was reached at laboratory level because high quality anchovies were used and immediately flayed, beheaded, gutted, bled and frozen. Therefore, it is possible to maintain the shelf life of anchovies for a long time. Overall, this method displayed a large increase in shelf life relative to that commonly seen for anchovies in this industry. However, these results should be extrapolated at industrial scales to propose better processing conditions and improve the quality of anchovy for direct human consumption.

Keywords: citrate sodium solution, heme iron, polyunsaturated fatty acids, shelf life of frozen anchovy

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Authors: Laura Alvarez-Alonso, Francisco Garcia-Carro, Jorge Loredo

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Population growth and industrial development imply an increase in the energy demands and the problems caused by emissions of greenhouse effect gases, which has inspired the search for clean sources of energy. Hydrogen (H₂) is expected to play a key role in the world’s energy future by replacing fossil fuels. The properties of H₂ make it a green fuel that does not generate pollutants and supplies sufficient energy for power generation, transportation, and other applications. Supercritical Water Gasification (SCWG) represents an attractive alternative for the recovery of energy from wastes. SCWG allows conversion of a wide range of raw materials into a fuel gas with a high content of hydrogen and light hydrocarbons through their treatment at conditions higher than those that define the critical point of water (temperature of 374°C and pressure of 221 bar). Methane used as a transport fuel is another important gasification product. The number of different uses of gas and energy forms that can be produced depending on the kind of material gasified and type of technology used to process it, shows the flexibility of SCWG. This feature allows it to be integrated with several industrial processes, as well as power generation systems or waste-to-energy production systems. The final aim of this work is to study which conditions and equipment are the most efficient and advantageous to explore the possibilities to obtain streams rich in H₂ from oily wastes, which represent a major problem both for the environment and human health throughout the world. In this paper, the relative complexity of technology needed for feasible gasification process cycles is discussed with particular reference to the different feedstocks that can be used as raw material, different reactors, and energy recovery systems. For this purpose, a review of the current status of SCWG technologies has been carried out, by means of different classifications based on key features as the feed treated or the type of reactor and other apparatus. This analysis allows to improve the technology efficiency through the study of model calculations and its comparison with experimental data, the establishment of kinetics for chemical reactions, the analysis of how the main reaction parameters affect the yield and composition of products, or the determination of the most common problems and risks that can occur. The results of this work show that SCWG is a promising method for the production of both hydrogen and methane. The most significant choices of design are the reactor type and process cycle, which can be conveniently adopted according to waste characteristics. Regarding the future of the technology, the design of SCWG plants is still to be optimized to include energy recovery systems in order to reduce costs of equipment and operation derived from the high temperature and pressure conditions that are necessary to convert water to the SC state, as well as to find solutions to remove corrosion and clogging of components of the reactor.

Keywords: hydrogen production, organic wastes, supercritical water gasification, system integration, waste-to-energy

Procedia PDF Downloads 121

Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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Authors: G. M. Vlasceanu, H. Iovu, E. Vasile, M. Ionita

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Herein, the synthesis and characterization of chitosan-gelatin highly porous scaffold reinforced with graphene oxide, and hydroxyapatite (HAp), crosslinked with genipin was targeted. In tissue engineering, chitosan and gelatin are two of the most robust biopolymers with wide applicability due to intrinsic biocompatibility, biodegradability, low antigenicity properties, affordability, and ease of processing. HAp, per its exceptional activity in tuning cell-matrix interactions, is acknowledged for its capability of sustaining cellular proliferation by promoting bone-like native micro-media for cell adjustment. Genipin is regarded as a top class cross-linker, while graphene oxide (GO) is viewed as one of the most performant and versatile fillers. The composites with natural bone HAp/biopolymer ratio were obtained by cascading sonochemical treatments, followed by uncomplicated casting methods and by freeze-drying. Their structure was characterized by Fourier Transform Infrared Spectroscopy and X-ray Diffraction, while overall morphology was investigated by Scanning Electron Microscopy (SEM) and micro-Computer Tomography (µ-CT). Ensuing that, in vitro enzyme degradation was performed to detect the most promising compositions for the development of in vivo assays. Suitable GO dispersion was ascertained within the biopolymer mix as nanolayers specific signals lack in both FTIR and XRD spectra, and the specific spectral features of the polymers persisted with GO load enhancement. Overall, correlations between the GO induced material structuration, crystallinity variations, and chemical interaction of the compounds can be correlated with the physical features and bioactivity of each composite formulation. Moreover, the HAp distribution within follows an auspicious density gradient tuned for hybrid osseous/cartilage matter architectures, which were mirrored in the mice model tests. Hence, the synthesis route of a natural polymer blend/hydroxyapatite-graphene oxide composite material is anticipated to emerge as influential formulation in bone tissue engineering. Acknowledgement: This work was supported by the project 'Work-based learning systems using entrepreneurship grants for doctoral and post-doctoral students' (Sisteme de invatare bazate pe munca prin burse antreprenor pentru doctoranzi si postdoctoranzi) - SIMBA, SMIS code 124705 and by a grant of the National Authority for Scientific Research and Innovation, Operational Program Competitiveness Axis 1 - Section E, Program co-financed from European Regional Development Fund 'Investments for your future' under the project number 154/25.11.2016, P_37_221/2015. The nano-CT experiments were possible due to European Regional Development Fund through Competitiveness Operational Program 2014-2020, Priority axis 1, ID P_36_611, MySMIS code 107066, INOVABIOMED.

Keywords: biopolymer blend, ectopic osteoinduction, graphene oxide composite, hydroxyapatite

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Authors: A. Knezović

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21st century has transformed the labor market landscape in a way of posing new and different demands on university graduates as well as university lecturers, which means that the knowledge and academic skills students acquire in the course of their studies should be applicable and transferable from the higher education context to their future professional careers. Given the context of the Languages for Specific Purposes (LSP) classroom, the teachers’ objective is not only to teach the language itself, but also to prepare students to use that language as a medium to develop generic skills and competences. These include media and information literacy, critical and creative thinking, problem-solving and analytical skills, effective written and oral communication, as well as collaborative work and social skills, all of which are necessary to make university graduates more competitive in everyday professional environments. On the other hand, due to limitations of time and large numbers of students in classes, the frequently topic-centered syllabus of LSP courses places considerable focus on acquiring the subject matter and specialist vocabulary instead of sufficient development of skills and competences required by students’ prospective employers. This paper intends to explore some of those issues as viewed both by LSP lecturers and by business professionals in their respective surveys. The surveys were conducted among more than 50 LSP lecturers at higher education institutions in Croatia, more than 40 HR professionals and more than 60 university graduates with degrees in economics and/or business working in management positions in mainly large and medium-sized companies in Croatia. Various elements of LSP course content have been taken into consideration in this research, including reading and listening comprehension of specialist texts, acquisition of specialist vocabulary and grammatical structures, as well as presentation and negotiation skills. The ability to hold meetings, conduct business correspondence, write reports, academic texts, case studies and take part in debates were also taken into consideration, as well as informal business communication, business etiquette and core courses delivered in a foreign language. The results of the surveys conducted among LSP lecturers will be analyzed with reference to what extent those elements are included in their courses and how consistently and thoroughly they are evaluated according to their course requirements. Their opinions will be compared to the results of the surveys conducted among professionals from a range of industries in Croatia so as to examine how useful and important they perceive the same elements of the LSP course content in their working environments. Such comparative analysis will thus show to what extent the syllabi of LSP courses meet the demands of the employment market when it comes to the students’ language skills and competences, as well as transferable skills. Finally, the findings will also be compared to the observations based on practical teaching experience and the relevant sources that have been used in this research. In conclusion, the ideas and observations in this paper are merely open-ended questions that do not have conclusive answers, but might prompt LSP lecturers to re-evaluate the content and objectives of their course syllabi.

Keywords: languages for specific purposes (LSP), language skills, topic-centred syllabus, transferable skills

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Authors: Nitza Davidovitch, Yael Yossel-Eisenbach

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The purpose of the study was to inquire whether there was a change in the shaping of undergraduate students’ digitally-oriented study pattern in the pre-Covid (2016-2017) versus post-Covid period (2022-2023), as affected by three factors: social background characteristics, high school, and academic background characteristics. These two-time points were cauterized by dramatic changes in teaching and learning at institutions of higher education. The data were collected via cross-sectional surveys at two-time points, in the 2016-2017 academic school year (N=443) and in the 2022-2023 school year (N=326). The questionnaire was distributed on social media and it includes questions on demographic background characteristics, previous studies in high school and present academic studies, and questions on learning and reading habits. Method of analysis: A. Statistical descriptive analysis, B. Mean comparison tests were conducted to analyze the variations in the mean score for the digitally-oriented learning pattern variable at two-time points (pre- and post-Covid) in relation to each of the independent variables. C. Analysis of variance was performed to test the main effects and the interactions. D. Applying linear regression, the research aimed to examine the combined effect of the independent variables on shaping students' digitally-oriented learning habits. The analysis includes four models. In all four models, the dependent variable is students’ perception of digitally oriented learning. The first model included social background variables; the second model included scholastic background as well. In the third model, the academic background variables were added, and the fourth model includes all the independent variables together with the variable of period (pre- and post-COVID). E. Factor analysis confirms using the principal component method with varimax rotation; the variables were constructed by a weighted mean of all the relevant statements merged to form a single variable denoting a shared content world. The research findings indicate a significant rise in students’ perceptions of digitally-oriented learning in the post-COVID period. From a gender perspective, the impact of COVID on shaping a digital learning pattern was much more significant for female students. The socioeconomic status perspective is eliminated when controlling for the period, and the student’s job is affected - more than all other variables. It may be assumed that the student’s work pattern mediates effects related to the convenience offered by digital learning regarding distance and time. The significant effect of scholastic background on shaping students’ digital learning patterns remained stable, even when controlling for all explanatory variables. The advantage that universities had over colleges in shaping a digital learning pattern in the pre-COVID period dissipated. Therefore, it can be said that after COVID, there was a change in how colleges shape students’ digital learning patterns in such a way that no institutional differences are evident with regard to shaping the digital learning pattern. The study shows that period has a significant independent effect on shaping students’ digital learning patterns when controlling for the explanatory variables.

Keywords: learning pattern, COVID, socioeconomic status, digital learning

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Authors: Iman Farasat, Howard M. Salis

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Engineered genetic circuits reprogram cellular behavior to act as living computers with applications in detecting cancer, creating self-controlling artificial tissues, and dynamically regulating metabolic pathways. Phenemenological models are often used to simulate and design genetic circuit behavior towards a desired behavior. While such models assume that each circuit component’s function is modular and independent, even small changes in a circuit (e.g. a new promoter, a change in transcription factor expression level, or even a new media) can have significant effects on the circuit’s function. Here, we use statistical thermodynamics to account for the several factors that control transcriptional regulation in bacteria, and experimentally demonstrate the model’s accuracy across 825 measurements in several genetic contexts and hosts. We then employ our first principles model to design, experimentally construct, and characterize a family of signal amplifying genetic circuits (genetic OpAmps) that expand the dynamic range of cell sensors. To develop these models, we needed a new approach to measuring the in vivo binding free energies of transcription factors (TFs), a key ingredient of statistical thermodynamic models of gene regulation. We developed a new high-throughput assay to measure RNA polymerase and TF binding free energies, requiring the construction and characterization of only a few constructs and data analysis (Figure 1A). We experimentally verified the assay on 6 TetR-homolog repressors and a CRISPR/dCas9 guide RNA. We found that our binding free energy measurements quantitatively explains why changing TF expression levels alters circuit function. Altogether, by combining these measurements with our biophysical model of translation (the RBS Calculator) as well as other measurements (Figure 1B), our model can account for changes in TF binding sites, TF expression levels, circuit copy number, host genome size, and host growth rate (Figure 1C). Model predictions correctly accounted for how these 8 factors control a promoter’s transcription rate (Figure 1D). Using the model, we developed a design framework for engineering multi-promoter genetic circuits that greatly reduces the number of degrees of freedom (8 factors per promoter) to a single dimensionless unit. We propose the Ptashne (Pt) number to encapsulate the 8 co-dependent factors that control transcriptional regulation into a single number. Therefore, a single number controls a promoter’s output rather than these 8 co-dependent factors, and designing a genetic circuit with N promoters requires specification of only N Pt numbers. We demonstrate how to design genetic circuits in Pt number space by constructing and characterizing 15 2-repressor OpAmp circuits that act as signal amplifiers when within an optimal Pt region. We experimentally show that OpAmp circuits using different TFs and TF expression levels will only amplify the dynamic range of input signals when their corresponding Pt numbers are within the optimal region. Thus, the use of the Pt number greatly simplifies the genetic circuit design, particularly important as circuits employ more TFs to perform increasingly complex functions.

Keywords: transcription factor, synthetic biology, genetic circuit, biophysical model, binding energy measurement

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Authors: Tesfaye Dagne, Dagmawit Solomon, Firmaye Bogale, Yosef Gebreyohannes, Samson Mideksa, Mamuye Hadis, Desalegn Ararso, Ermias Woldie, Tsegaye Getachew, Sabit Ababor, Zelalem Kebede

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Globally, road traffic accident (RTA) is causing millions of deaths and injuries every year. It is one of the leading causes of death among people of all age groups and the problem is worse among young reproductive age group. Moreover the problem is increasing with an increasing number of vehicles. The majority of the problem happen in low and middle income countries (LMIC), even if the number of vehicles in these countries is low compared to their population. So, the objective of this paper is to summarize the best available evidence on interventions that can reduce road traffic accidents in low and middle income countries (LMIC). Method: A rapid evidence synthesis approach adapted from the SURE Rapid Response Service was applied to search, appraise and summarize the best available evidence on effective intervention in reducing road traffic injury. To answer the question under review, we searched for relevant studies from databases including PubMed, the Cochrane Library, TRANSPORT, Health system evidence, Epistemonikos, and SUPPORT summary. The following key terms were used for searching: Road traffic accident, RTA, Injury, Reduc*, Prevent*, Minimiz*, “Low and middle-income country”, LMIC. We found 18 articles through a search of different databases mentioned above. After screening for the titles and abstracts of the articles, four of them which satisfy the inclusion criteria were included in the final review. Then we appraised and graded the methodological quality of systematic reviews that are deemed to be highly relevant using AMSTAR. Finding: The identified interventions to reduce road traffic accidents were legislation and enforcement, public awareness/education, speed control/ rumble strips, road improvement, mandatory motorcycle helmet, graduated driver license, street lighting. Legislation and Enforcement: Legislation focusing on mandatory motorcycle helmet usage, banning cellular phone usage when driving, seat belt laws, decreasing the legal blood alcohol content (BAC) level from 0.06 g/L to 0.02 g/L bring the best result where enforcement is there. Public Awareness/Education: focusing on seat belt use, child restraint use, educational training in health centers and schools/universities, and public awareness with media through the distribution of videos, posters/souvenirs, and pamphlets are effective in the short run. Speed Control: through traffic calming bumps, or speed bumps, rumbled strips are effective in reducing accidents and fatality. Mandatory Motorcycle Helmet: is associated with reduction in mortality. Graduated driver’s license (GDL): reduce road traffic injury by 19%. Street lighting: is a low-cost intervention which may reduce road traffic accidents.

Keywords: evidence synthesis, injury, rapid review, reducing, road traffic accident

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Authors: Chee Wai Ku, Pui See Chin

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As an emerging alternative to hysterectomy, uterine artery embolization (UAE) has been widely used in the management of fibroids and in controlling postpartum hemorrhage (PPH) unresponsive to other therapies. Research has shown UAE to be a safe, minimally invasive procedure with few complications and minimal effects on future fertility. We present two cases highlighting the use of UAE in preventing PPH in a patient with a large fibroid at the time of cesarean section and in the treatment of secondary PPH refractory to other therapies in another patient. We present a 36-year primiparous woman who booked at 18+6 weeks gestation with a 13.7 cm subserosal fibroid at the lower anterior wall of the uterus near the cervix and a 10.8 cm subserosal fibroid in the left wall. Prophylactic internal iliac artery occlusion balloons were placed prior to the planned classical midline cesarean section. The balloons were inflated once the baby was delivered. Bilateral uterine arteries were embolized subsequently. The estimated blood loss (EBL) was 400 mls and hemoglobin (Hb) remained stable at 10 g/DL. Ultrasound scan 2 years postnatally showed stable uterine fibroids 10.4 and 7.1 cm, which was significantly smaller than before. We present the second case of a 40-year-old G2P1 with a previous cesarean section for failure to progress. There were no antenatal problems, and the placenta was not previa. She presented with term labour and underwent an emergency cesarean section for failed vaginal birth after cesarean. Intraoperatively extensive adhesions were noted with bladder drawn high, and EBL was 300 mls. Postpartum recovery was uneventful. She presented with secondary PPH 3 weeks later complicated by hypovolemic shock. She underwent an emergency examination under anesthesia and evacuation of the uterus, with EBL 2500mls. Histology showed decidua with chronic inflammation. She was discharged well with no further PPH. She subsequently returned one week later for secondary PPH. Bedside ultrasound showed that the endometrium was thin with no evidence of retained products of conception. Uterotonics were administered, and examination under anesthesia was performed, with uterine Bakri balloon and vaginal pack insertion after. EBL was 1000 mls. There was no definite cause of PPH with no uterine atony or products of conception. To evaluate a potential cause, pelvic angiogram and super selective left uterine arteriogram was performed which showed profuse contrast extravasation and acute bleeding from the left uterine artery. Superselective embolization of the left uterine artery was performed. No gross contrast extravasation from the right uterine artery was seen. These two cases demonstrated the superior efficacy of UAE. Firstly, the prophylactic use of intra-arterial balloon catheters in pregnant patients with large fibroids, and secondly, in the diagnosis and management of secondary PPH refractory to uterotonics and uterine tamponade. In both cases, the need for laparotomy hysterectomy was avoided, resulting in the preservation of future fertility. UAE should be a consideration for hemodynamically stable patients in centres with access to interventional radiology.

Keywords: fertility preservation, secondary postpartum hemorrhage, uterine embolization, uterine fibroids

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Authors: Sonia P. Marrero Duran, Lilian Noya Dominguez, Lisandra Quintana Alvarez, Evert Martinez Perez, Ana Julia Acevedo Urquiaga

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Cuban business and economic policy are in the constant update as well as facing a client ever more knowledgeable and demanding. For that reason become fundamental for companies competitiveness through the optimization of its processes and services. One of the Cuban’s pillars, which has been sustained since the triumph of the Cuban Revolution back in 1959, is the free health service to all those who need it. This service is offered without any charge under the concept of preserving human life, but it implied costly management processes and logistics services to be able to supply the necessary medicines to all the units who provide health services. One of the key actors on the medicine supply chain is the Havana Distribution Center (HDC), which is responsible for the delivery of medicines in the province; as well as the acquisition of medicines from national and international producers and its subsequent transport to health care units and pharmacies in time, and with the required quality. This HDC also carries for all distribution centers in the country. Given the eminent need to create an actor in the supply chain that specializes in the medicines supply, the possibility of centralizing this operation in a logistics service provider is analyzed. Based on this decision, pharmacies operate as clients of the logistic service center whose main function is to centralize all logistics operations associated with the medicine supply chain. The HDC is precisely the logistic service provider in Havana and it is the center of this research. In 2017 the pharmacies had affectations in the availability of medicine due to deficiencies in the distribution routes. This is caused by the fact that they are not based on routing studies, besides the long distribution cycle. The distribution routs are fixed, attend only one type of customer and there respond to a territorial location by the municipality. Taking into consideration the above-mentioned problem, the objective of this research is to optimize the routes system in the Havana Distribution Center. To accomplish this objective, the techniques applied were document analysis, random sampling, statistical inference and tools such as Ishikawa diagram and the computerized software’s: ArcGis, Osmand y MapIfnfo. As a result, were analyzed four distribution alternatives; the actual rout, by customer type, by the municipality and the combination of the two last. It was demonstrated that the territorial location alternative does not take full advantage of the transportation capacities or the distance of the trips, which leads to elevated costs breaking whit the current ways of distribution and the currents characteristics of the clients. The principal finding of the investigation was the optimum option distribution rout is the 4th one that is formed by hospitals and the join of pharmacies, stomatology clinics, polyclinics and maternal and elderly homes. This solution breaks the territorial location by the municipality and permits different distribution cycles in dependence of medicine consumption and transport availability.

Keywords: computerized geographic software, distribution, distribution routs, vehicle problem routing (VPR)

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Authors: Jo Anne Rey

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Caring for Country (Ngurrain Dharug language) is core to Aboriginal identity, Law/Lore, practice, and resilience within the continent called ‘Australia’. It is the basis of thousands of years of sustainability. However, when Ngurra is a city known as Sydney, due to 235 years of colonial impact, caring for the Country is limited, being controlled by the State and private ownership of the land title. Recent research indicates that localised Indigenous activism is most successful when community members are geographically proximate to the presences and places of connection, caring, and belonging. This article frames these findings through the cadence that proximity provides. This presentation is centred on the proximate agency that is being exercised by Dharug community through three significant sites within the Sydney basin. Those sites include, firstly, Shaw’s Creek Aboriginal Place, at the foot of the Blue Mountains in far western Sydney. Second inclusion is the site of Blacktown Native Institution, that was the part of the authoritarian colonial governance of British Governor Lachlan Macquarie (after who Macquarie University is named), which saw the beginnings of the removal of children from their families and culture to ‘civilize’ them. The third site is that of the so-called Brown’s Waterhole in the State government administered Lane Cove National Park. Each of these sites is being activated through Dharug and, more broadly, Aboriginalways of knowing, doing, and being. These ways involvethe land, water, wind, and star-based ecologies interwoven with traditional transgenerational storying of the presences (Ancestral and spiritual) creating them. Activations include, but are not limited to, the return of cultural fire for reviving plants, soils, animals, and birds. These fire practices have traditionally been at the basis of sustainable, regenerative biodiversity. These practices involve the literacy of reading Ngurra and the seasonal interactions across the ecologies. Together, they both care for the Country and support humanity, and have done so across thousands of years. However, when the cost of real-estate and rental accommodation prevents community members from being able to live on Dharug Ngurra when bureaucratic governance restricts and/or excludes traditional custodial relationships, and when private treaty land title destroys the presences and places while disconnecting people from their Ancestral practices, it becomes clear that caring for Country is only possible when the community can afford to live nearby. Recognising the cadence of proximityas the agency that underpinscaring for Country-in-the-city, sustainable change opportunities don’t have to only focus on regional and remote areas. Urban-based Aboriginal relationality offers an alternative to the unsustainable practices that underpin human-centric disconnection. Weaving Indigenous cadence offers opportunities for sustainable futures even when facing the extremes of climate changing catastrophes.

Keywords: australian aboriginal, biocultural knowledges, climate change, dharug ngurra, sustainability, resilience

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Authors: Same Noel Ngando, Yakub Abdulfatai Olatunji

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Renewable energy micro-grids, such as those powered by solar or wind energy, are often intermittent in nature. This means that the amount of energy generated by these systems can vary depending on weather conditions or other factors, which can make it difficult to ensure a steady supply of power. To address this issue, energy storage systems have been developed to increase the reliability of renewable energy micro-grids. Battery systems have been the dominant energy storage technology for renewable energy micro-grids. Batteries can store large amounts of energy in a relatively small and compact package, making them easy to install and maintain in a micro-grid setting. Additionally, batteries can be quickly charged and discharged, allowing them to respond quickly to changes in energy demand. However, the process involved in recycling batteries is quite costly and difficult. An alternative energy storage system that is gaining popularity is hydrogen storage. Hydrogen is a versatile energy carrier that can be produced from renewable energy sources such as solar or wind. It can be stored in large quantities at low cost, making it suitable for long-distance mass storage. Unlike batteries, hydrogen does not degrade over time, so it can be stored for extended periods without the need for frequent maintenance or replacement, allowing it to be used as a backup power source when the micro-grid is not generating enough energy to meet demand. When hydrogen is needed, it can be converted back into electricity through a fuel cell. Energy consumption data is got from a particular residential area in Daegu, South Korea, and the data is processed and analyzed. From the analysis, the total energy demand is calculated, and different hybrid energy system configurations are designed using HOMER Pro (Hybrid Optimization for Multiple Energy Resources) and MATLAB software. A techno-economic and environmental comparison and life cycle assessment (LCA) of the different configurations using battery and hydrogen as storage systems are carried out. The various scenarios included PV-hydrogen-grid system, PV-hydrogen-grid-wind, PV-hydrogen-grid-biomass, PV-hydrogen-wind, PV-hydrogen-biomass, biomass-hydrogen, wind-hydrogen, PV-battery-grid-wind, PV- battery -grid-biomass, PV- battery -wind, PV- battery -biomass, and biomass- battery. From the analysis, the least cost system for the location was the PV-hydrogen-grid system, with a net present cost of about USD 9,529,161. Even though all scenarios were environmentally friendly, taking into account the recycling cost and pollution involved in battery systems, all systems with hydrogen as a storage system produced better results. In conclusion, hydrogen is becoming a very prominent energy storage solution for renewable energy micro-grids. It is easier to store compared with electric power, so it is suitable for long-distance mass storage. Hydrogen storage systems have several advantages over battery systems, including flexibility, long-term stability, and low environmental impact. The cost of hydrogen storage is still relatively high, but it is expected to decrease as more hydrogen production, and storage infrastructure is built. With the growing focus on renewable energy and the need to reduce greenhouse gas emissions, hydrogen is expected to play an increasingly important role in the energy storage landscape.

Keywords: renewable energy systems, microgrid, hydrogen production, energy storage systems

Procedia PDF Downloads 65

Authors: Tamar Trop

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Bike sharing systems can be important elements of smart cities as they have the potential for impact on multiple levels. These systems can add a significant alternative to other modes of mass transit in cities that are continuously looking for measures to become more livable and maintain their attractiveness for citizens, businesses and tourism. Bike-sharing began in Europe in 1965, and a viable format emerged in the mid-2000s thanks to the introduction of information technology. The rate of growth in bike-sharing schemes and fleets has been very rapid since 2008 and has probably outstripped growth in every other form of urban transport. Today, public bike-sharing systems are available on five continents, including over 700 cities, operating more than 800,000 bicycles at approximately 40,000 docking stations. Since modern bike sharing systems have become prevalent only in the last decade, the existing literature analyzing these systems and their sustainability is relatively new. The purpose of the presented study is to assess the sustainability of these newly emerging transportation systems, by using the Envision™ rating system as a methodological framework and the Israeli 'Tel -O-Fun' – bike sharing project as a case study. The assessment was conducted by project team members. Envision™ is a new guidance and rating system used to assess and improve the sustainability of all types and sizes of infrastructure projects. This tool provides a holistic framework for evaluating and rating the community, environmental, and economic benefits of infrastructure projects over the course of their life cycle. This evaluation method has 60 sustainability criteria divided into five categories: Quality of life, leadership, resource allocation, natural world, and climate and risk. 'Tel -O-Fun' project was launched in Tel Aviv-Yafo on 2011 and today provides about 1,800 bikes for rent, at 180 rental stations across the city. The system is based on a complex computer terminal that is located in the docking stations. The highest-rated sustainable features that the project scored include: (a) Improving quality of life by: offering a low cost and efficient form of public transit, improving community mobility and access, enabling the flexibility of travel within a multimodal transportation system, saving commuters time and money, enhancing public health and reducing air and noise pollution; (b) improving resource allocation by: offering inexpensive and flexible last-mile connectivity, reducing space, materials and energy consumption, reducing wear and tear on public roads, and maximizing the utility of existing infrastructure, and (c) reducing of greenhouse gas emissions from transportation. Overall, 'Tel -O-Fun' project was highly scored as an environmentally sustainable and socially equitable infrastructure. The use of this practical framework for evaluation also yielded various interesting insights on the shortcoming of the system and the characteristics of good solutions. This can contribute to the improvement of the project and may assist planners and operators of bike sharing systems to develop a sustainable, efficient and reliable transportation infrastructure within smart cities.

Keywords: bike sharing, Envision™, sustainability rating system, sustainable infrastructure

Procedia PDF Downloads 317

Authors: Isaac Martin, Abigail Lark, Sandra Morales, Eric W. Alton, Jane C. Davies

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Objectives: The cystic fibrosis (CF) lung is a unique microbiological niche, wherein harmful bacteria persist for many years despite antibiotic therapy. Pseudomonas aeruginosa (Pa), the major culprit leading to lung decline and increased mortality, thrives in the lungs of patients with CF due to several factors that have been linked with poor antibiotic performance. Our group is investigating alternative therapies including bacteriophage cocktails with which we have previously demonstrated efficacy against planktonic organisms. In this study, we explored the effects of a 4-phage cocktail on Pa grown in two different conditions, intended to mirror the CF lung: a) alongside standard antibiotic treatment in pre-formed biofilms (structures formed by Pa-secreted exopolysaccharides which provide both physical and cell division barriers to antimicrobials and host defenses and b) in an acidic environment postulated to be present in the CF airway due both to the primary defect in bicarbonate secretion and secondary effects of inflammation. Methods: 16 Pa strains from CF patients at the Royal Brompton Hospital were selected based on sensitivity to a) ceftazidime/ tobramycin and b) the phage cocktail in a conventional plaque assay. To assess efficacy of phage in biofilms, 96 well plates with Pa (5x10⁷ CFU/ ml) were incubated in static conditions, allowing adherent bacterial colonies to form for 24 hr. Ceftazidime and tobramycin (both at 2 × MIC) were added, +/- bacteriophage (4x10⁸ PFU/mL) for a further 24 hr. Cell viability and biomass were estimated using fluorescent resazurin and crystal violet assays, respectively. To evaluate the effect of pH, strains were grown planktonically in shaking 96 well plates at pH 6.0, 6.6, 7.0 and 7.5 with tobramycin or phage, at varying concentrations. Cell viability was quantified by fluorescent resazurin assay. Results: For the biofilm assay, treatment groups were compared with untreated controls and expressed as percent reduction in cell viability and biomass. Addition of the 4-phage cocktail resulted in a 1.3-fold reduction in cell viability and 1.7-fold reduction in biomass (p < 0.001) when compared to standard antibiotic treatment alone. Notably, there was a 50 ± 15% reduction in cell viability and 60 ± 12% reduction in biomass (95% CI) for the 4 biofilms demonstrating the most resistance to antibiotic treatment. 83% of strains tested (n=6) showed decreased bacterial killing by tobramycin at acidic pHs (p < 0.01). However, 25% of strains (n=12) showed improved phage killing at acidic pHs (p < 0.05), with none showing the pattern of reduced efficacy at acidic pH demonstrated by tobramycin. Conclusion: The 4-phage anti-Pa cocktail tested against Pa performs well in pre-formed biofilms and in acidic environments; two conditions intended to mimic the CF lung. To our knowledge, these are the first data looking at the effects of subtle pH changes on phage-mediated bacterial killing in the context of Pa infection. These findings contribute to a growing body of evidence supporting the use of nebulised lytic bacteriophage as a treatment in the context of lung infection.

Keywords: biofilm, cystic fibrosis, pH, Pseudomonas aeruginosa, lytic bacteriophage

Procedia PDF Downloads 154