Search results for: Total Productive Maintenance (TPM)
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 10462

Search results for: Total Productive Maintenance (TPM)

6592 Use of Geoinformatics and Mathematical Equations to Assess Erosion and Soil Fertility in Cassava Growing Areas in Maha Sarakham Province, Thailand

Authors: Sasirin Srisomkiew, Sireewan Ratsadornasai, Tanomkwan Tipvong, Isariya Meesing

Abstract:

Cassava is an important food source in the tropics and has recently gained attention as a potential source of biofuel that can replace limited fossil fuel sources. As a result, the demand for cassava production to support industries both within the country and abroad has increased. In Thailand, most farmers prefer to grow cassava in sandy and sandy loam areas where the soil has low natural fertility. Cassava is a tuber plant that has large roots to store food, resulting in the absorption of large amounts of nutrients from the soil, such as nitrogen, phosphorus, and potassium. Therefore, planting cassava in the same area for a long period causes soil erosion and decreases soil fertility. The loss of soil fertility affects the economy, society, and food and energy security of the country. Therefore, it is necessary to know the level of soil fertility and the amount of nutrients in the soil. To address this problem, this study applies geo-informatics technology and mathematical equations to assess erosion and soil fertility and to analyze factors affecting the amount of cassava production in Maha Sarakham Province. The results show that the area for cassava cultivation has increased in every district of Maha Sarakham Province between 2015-2022, with the total area increasing to 180,922 rai or 5.47% of the province’s total area during this period. Furthermore, it was found that it is possible to assess areas with soil erosion problems that had a moderate level of erosion in areas with high erosion rates ranging from 5-15 T/rai/year. Soil fertility assessment and information obtained from the soil nutrient map for 2015–2023 reveal that farmers in the area have improved the soil by adding chemical fertilizers along with organic fertilizers, such as manure and green manure, to increase the amount of nutrients in the soil. This is because the soil resources of Maha Sarakham Province mostly have relatively low agricultural potential due to the soil texture being sand and sandy loam. In this scenario, the ability to absorb nutrients is low, and the soil holds little water, so it is naturally low in fertility. Moreover, agricultural soil problems were found, including the presence of saline soil, sandy soil, and acidic soil, which is a serious restriction on land use because it affects the release of nutrients into the soil. The results of this study may be used as a guideline for managing soil resources and improving soil quality to prevent soil degradation problems that may occur in the future.

Keywords: Cassava, geoinformatics, soil erosion, soil fertility, land use change

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6591 Protective Effects of Ethanolic Purslane Extracts on Doxorubicin-Induced Hepatotoxicity in Albino Rats

Authors: Osama M. Ahmed, Walaa G. Hozayen, Haidy Tamer Abo Sree

Abstract:

The effect of doxorubicin (4 mg/kg b.w.week) without or with oral administration of ethanolic purslane (Portulaca oleracea) shoot (leaves and stems) extract (50 mg/kg b.w.day) or ethanolic purslane seeds extract (50 mg/kg b.w.day) co-treatments for 6 weeks was evaluated in adult male rats. There was an increase in serum levels of ALT, AST, ALP, GGT and total bilirubin. In addition, hepatic glutathine, glutathione transferase, peroxidase, SOD, CAT activities were decreased while lipid peroxidation in the liver was increased. Co-administration of ethanolic purslane and seed extracts successfully improved the adverse changes in the liver functions with an increase in antioxidants activities and reduction of lipid peroxidation.

Keywords: antioxidants, doxorubicin, hepatotoxicity, purslane

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6590 Developmental Delays among Children with Neonatal Hyperbilirubinemia

Authors: Simplejit Kaur Dhanoa, Manmohan Singh

Abstract:

This study was done with the primary objective to evaluate the motor and mental developmental delays among children having neonatal Jaundice. A total sample of 300 neonates were collected; out of them, 150 were preterm neonates, and 150 were full term neonates from the hospital setting and follow up study was done with the help of the Developmental Assessment scale of the Indian Infant. The registered samples were assessed up to 2.6 years with a gap of 6 months. The outcomes of this study reveal that developmental delays were present among children who had preterm neonatal jaundice as compare to full term normal babies. Further, It was reported that both motor and mental development is affected due to neonatal hyperbilirubinemia in addition to preterm birth.

Keywords: hyperbinirubinemia, preterm neonates, developmental delays, preterm

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6589 Survival of Micro-Encapsulated Probiotic Lactic Acid Bacteria in Mutton Nuggets and Their Assessments in Simulated Gastro-Intestinal Conditions

Authors: Rehana Akhter, Sajad A. Rather, F. A. Masoodi, Adil Gani, S. M. Wani

Abstract:

During recent years probiotic food products receive market interest as health-promoting, functional foods, which are believed to contribute health benefits. In order to deliver the health benefits by probiotic bacteria, it has been recommended that they must be present at a minimum level of 106 CFU/g to 107 CFU/g at point of delivery or be eaten in sufficient amounts to yield a daily intake of 108 CFU. However a major challenge in relation to the application of probiotic cultures in food matrix is the maintenance of viability during processing which might lead to important losses in viability as probiotic cultures are very often thermally labile and sensitive to acidity, oxygen or other food constituents for example, salts. In this study Lactobacillus plantarum and Lactobacillus casei were encapsulated in calcium alginate beads with the objective of enhancing their survivability and preventing exposure to the adverse conditions of the gastrointestinal tract and where then inoculated in mutton nuggets. Micro encapsulated Lactobacillus plantarum and Lactobacillus casei were resistant to simulated gastric conditions (pH 2, 2h) and bile solution (3%, 2 h) resulting in significantly (p ≤ 0.05) improved survivability when compared with free cell counterparts. A high encapsulation yield was found due to the encapsulation procedure. After incubation at low pH-values, micro encapsulation yielded higher survival rates compared to non-encapsulated probiotic cells. The viable cell numbers of encapsulated Lactobacillus plantarum and Lactobacillus casei were 107-108 CFU/g higher compared to free cells after 90 min incubation at pH 2.5. The viable encapsulated cells were inoculated into mutton nuggets at the rate of 108 to 1010 CFU/g. The micro encapsulated Lactobacillus plantarum and Lactobacillus casei achieved higher survival counts (105-107 CFU/g) than the free cell counterparts (102-104 CFU/g). Thus micro encapsulation offers an effective means of delivery of viable probiotic bacterial cells to the colon and maintaining their survival during simulated gastric, intestinal juice and processing conditions during nugget preparation.

Keywords: survival, Lactobacillus plantarum, Lactobacillus casei, micro-encapsulation, nugget

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6588 Determination of the Phytochemicals Composition and Pharmacokinetics of whole Coffee Fruit Caffeine Extract by Liquid Chromatography-Tandem Mass Spectrometry

Authors: Boris Nemzer, Nebiyu Abshiru, Z. B. Pietrzkowski

Abstract:

Coffee cherry is one of the most ubiquitous agricultural commodities which possess nutritional and human health beneficial properties. Between the two most widely used coffee cherries Coffea arabica (Arabica) and Coffea canephora (Robusta), Coffea arabica remains superior due to its sensory properties and, therefore, remains in great demand in the global coffee market. In this study, the phytochemical contents and pharmacokinetics of Coffeeberry® Energy (CBE), a commercially available Arabica whole coffee fruit caffeine extract, are investigated. For phytochemical screening, 20 mg of CBE was dissolved in an aqueous methanol solution for analysis by mass spectrometry (MS). Quantification of caffeine and chlorogenic acids (CGAs) contents of CBE was performed using HPLC. For the bioavailability study, serum samples were collected from human subjects before and after 1, 2 and 3 h post-ingestion of 150mg CBE extract. Protein precipitation and extraction were carried out using methanol. Identification of compounds was performed using an untargeted metabolomic approach on Q-Exactive Orbitrap MS coupled to reversed-phase chromatography. Data processing was performed using Thermo Scientific Compound Discover 3.3 software. Phytochemical screening identified a total of 170 compounds, including organic acids, phenolic acids, CGAs, diterpenoids and hydroxytryptamine. Caffeine & CGAs make up more than, respectively, 70% & 9% of the total CBE composition. For serum samples, a total of 82 metabolites representing 32 caffeine- and 50 phenolic-derived metabolites were identified. Volcano plot analysis revealed 32 differential metabolites (24 caffeine- and 8 phenolic-derived) that showed an increase in serum level post-CBE dosing. Caffeine, uric acid, and trimethyluric acid isomers exhibited 4- to 10-fold increase in serum abundance post-dosing. 7-Methyluric acid, 1,7-dimethyluric acid, paraxanthine and theophylline exhibited a minimum of 1.5-fold increase in serum level. Among the phenolic-derived metabolites, iso-feruloyl quinic acid isomers (3-, 4- and 5-iFQA) showed the highest increase in serum level. These compounds were essentially absent in serum collected before dosage. More interestingly, the iFQA isomers were not originally present in the CBE extract, as our phytochemical screen did not identify these compounds. This suggests the potential formation of the isomers during the digestion and absorption processes. Pharmacokinetics parameters (Cmax, Tmax and AUC0-3h) of caffeine- and phenolic-derived metabolites were also investigated. Caffeine was rapidly absorbed, reaching a maximum concentration (Cmax) of 10.95 µg/ml in just 1 hour. Thereafter, caffeine level steadily dropped from the peak level, although it did not return to baseline within the 3-hour dosing period. The disappearance of caffeine from circulation was mirrored by the rise in the concentration of its methylxanthine metabolites. Similarly, serum concentration of iFQA isomers steadily increased, reaching maximum (Cmax: 3-iFQA, 1.54 ng/ml; 4-iFQA, 2.47 ng/ml; 5-iFQA, 2.91 ng/ml) at tmax of 1.5 hours. The isomers remained well above the baseline during the 3-hour dosing period, allowing them to remain in circulation long enough for absorption into the body. Overall, the current study provides evidence of the potential health benefits of a uniquely formulated whole coffee fruit product. Consumption of this product resulted in a distinct serum profile of bioactive compounds, as demonstrated by the more than 32 metabolites that exhibited a significant change in systemic exposure.

Keywords: phytochemicals, mass spectrometry, pharmacokinetics, differential metabolites, chlorogenic acids

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6587 Activation-TV® to Reduce Elderly Loneliness and Insecurity

Authors: Hannele Laaksonen, Seija Nyqvist, Kari Nurmes

Abstract:

Objectives: In the year 2011 the City of Vaasa started to develop know-how in the technology and the introduction of services for aging people in cooperation with the Polytechnic Novia University of Applied Sciences and VAMK, University of Applied Sciences. The project´s targets included: to help elderly people to maintain their ability to function, to provide them social and physical activities, to prevent their social exclusion, to decrease their feelings of loneliness and insecurity and to develop their technical know-how. Methods: The project was built based on open source code, tailor-made service system and user interface for the elderly living at home and their families, based on the users´ expectations and experiences of services. Activation-TV®-project vas carried out 1.4.2011-31.3.2014. A pilot group of eight elderly persons, who were living at home, were selected to the project. All necessary technical means as well as guidance and teaching equipment were provided to the pilot group. The students of University of Applied Sciences (VAMK, Novia) and employees of Center of Ageing were made all programs to the Activation-TV®. The project group were interviewed after and before intervention. The data were evaluated both qualitatively and quantitatively. Results: The built service includes a video library, a group room for interactive programs and a personal room for bilateral meetings and direct shipment. The program is bilingual and produced in both national languages. The Activation TV® reduced elderly peoples´ (n=8) feelings of emptiness, added mental well-being and quality of life with social contacts. Relatives felt, that they were able to get in to older peoples´ everyday life with Activation TV®. Discussion: The built application was tailored to the model that has not been developed elsewhere in Finland. This model can be copied from one server to another and thus transferred to other municipalities but the program requires its own personnel system management and maintenance as well as program production cooperation between the different actors. This service can be used for the elderly who are living at home without dementia.

Keywords: mental well-being, quality of life, elderly people, Finland

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6586 Application of Geotube® Method for Sludge Handling in Adaro Coal Mine

Authors: Ezman Fitriansyah, Lestari Diah Restu, Wawan

Abstract:

Adaro coal mine in South Kalimantan-Indonesia maintains catchment area of approximately 15,000 Ha for its mine operation. As an open pit surface coal mine with high erosion rate, the mine water in Adaro coal mine contains high TSS that needs to be treated before being released to rivers. For the treatment process, Adaro operates 21 Settling Ponds equipped with combination of physical and chemical system to separate solids and water to ensure the discharged water complied with regional environmental quality standards. However, the sludge created from the sedimentation process reduces the settling ponds capacity gradually. Therefore regular maintenance activities are required to recover and maintain the ponds' capacity. Trucking system and direct dredging had been the most common method to handle sludge in Adaro. But the main problem in applying these two methods is excessive area required for drying pond construction. To solve this problem, Adaro implements an alternative method called Geotube®. The principle of Geotube® method is the sludge contained in the Settling Ponds is pumped into Geotube® containers which have been designed to release water and retain mud flocks. During the pumping process, an amount of flocculants chemicals are injected into the sludge to form bigger mud flocks. Due to the difference in particle size, the mud flocks are settled in the container whilst the water continues to flow out through the container’s pores. Compared to the trucking system and direct dredging method, this method provides three advantages: space required to operate, increasing of overburden waste dump volume, and increasing of water treatment process speed and quality. Based on the evaluation result, Geotube® method only needs 1:8 of space required by the other methods. From the geotechnical assessment result conducted by Adaro, the potential loss of waste dump volume capacity prior to implementation of the Geotube® method was 26.7%. The water treatment process of TSS in well maintained ponds is 16% more optimum.

Keywords: geotube, mine water, settling pond, sludge handling, wastewater treatment

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6585 Establishment of Nursing School in the Backward Region of Nepal

Authors: Shyam lamsal

Abstract:

Introduction: Karnali Academy of Health Sciences (KAHS) has been established in 2011, by an Act of parliament of Nepal, in Jumla, to provide health services in easy way in backward areas, to produce skilled health professionals & conduct research. The backward areas mentioned in act of KAHS are Humla, Jumla, Kalikot, Dolpa, Mugu districts of Karnali zone, Jajarkot district of Bheri zone & Bajura, Baghang & Achham districts of Seti zone in Nepal occupying around 25 % of the total national geography. Backward area of Nepal is specific to having worst health indicators with life expectancy (47 years), HDI (0.35), Literacy rate (58%), global acute malnutrition (13%), crude birth rate (33.6), crude death rate (9.6), Total fertility rate (4.2), infant mortality rate (61.5 per 1000 live births), under five mortality rate (59 per 1000 live births) and maternal mortality ratio (400 per 1000 live births). History of health facilities in backward region: All the nine districts of this region have a district hospital with very few grass root level health manpower. Government of Nepal regularly deploys one or two medical officers to each district who generally are not regular to their care. Jumla district itself was having one medical officer before the establishment of KAHS. Development activities: Establishment of 100 bedded specialty teaching hospital with 10 medical officers and five specialists, accredited its own nursing school for running diploma nursing programme, started “Karnali health survey” which covers 55 thousand households of backward region, started community care and school health camps, planning phase completed for 300 bedded teaching hospital construction. Future Plan: Expansion of the teaching hospital to 300 beds within 3 years, start health assistant and bachelor midwifery course in 2015 AD, start bachelor in laboratory and bachelor in public health course in 2016 AD and start MBBS course in 2018 AD. Deploy the medical officers and family physicians to all the district hospitals within 3 years. KAHS provides reservation up to 45% students from backward region with the commitment to stay for at least five years of their service period. Conclusion: This institution may be the example for the rest of the world in providing nursing care, education in remote areas as well as the best model for nursing manpower retention in remote areas of developing countries.

Keywords: backward area, nursing school

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6584 Forecasting Market Share of Electric Vehicles in Taiwan Using Conjoint Models and Monte Carlo Simulation

Authors: Li-hsing Shih, Wei-Jen Hsu

Abstract:

Recently, the sale of electrical vehicles (EVs) has increased dramatically due to maturing technology development and decreasing cost. Governments of many countries have made regulations and policies in favor of EVs due to their long-term commitment to net zero carbon emissions. However, due to uncertain factors such as the future price of EVs, forecasting the future market share of EVs is a challenging subject for both the auto industry and local government. This study tries to forecast the market share of EVs using conjoint models and Monte Carlo simulation. The research is conducted in three phases. (1) A conjoint model is established to represent the customer preference structure on purchasing vehicles while five product attributes of both EV and internal combustion engine vehicles (ICEV) are selected. A questionnaire survey is conducted to collect responses from Taiwanese consumers and estimate the part-worth utility functions of all respondents. The resulting part-worth utility functions can be used to estimate the market share, assuming each respondent will purchase the product with the highest total utility. For example, attribute values of an ICEV and a competing EV are given respectively, two total utilities of the two vehicles of a respondent are calculated and then knowing his/her choice. Once the choices of all respondents are known, an estimate of market share can be obtained. (2) Among the attributes, future price is the key attribute that dominates consumers’ choice. This study adopts the assumption of a learning curve to predict the future price of EVs. Based on the learning curve method and past price data of EVs, a regression model is established and the probability distribution function of the price of EVs in 2030 is obtained. (3) Since the future price is a random variable from the results of phase 2, a Monte Carlo simulation is then conducted to simulate the choices of all respondents by using their part-worth utility functions. For instance, using one thousand generated future prices of an EV together with other forecasted attribute values of the EV and an ICEV, one thousand market shares can be obtained with a Monte Carlo simulation. The resulting probability distribution of the market share of EVs provides more information than a fixed number forecast, reflecting the uncertain nature of the future development of EVs. The research results can help the auto industry and local government make more appropriate decisions and future action plans.

Keywords: conjoint model, electrical vehicle, learning curve, Monte Carlo simulation

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6583 Development of Microsatellite Markers for Dalmatian Pyrethrum Using Next-Generation Sequencing

Authors: Ante Turudic, Filip Varga, Zlatko Liber, Jernej Jakse, Zlatko Satovic, Ivan Radosavljevic, Martina Grdisa

Abstract:

Microsatellites (SSRs) are highly informative repetitive sequences of 2-6 base pairs, which are the most used molecular markers in assessing the genetic diversity of plant species. Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip) is an outcrossing diploid (2n = 18) endemic to the eastern Adriatic coast and source of the natural insecticide pyrethrin. Due to the high repetitiveness and large size of the genome (haploid genome size of 9,58 pg), previous attempts to develop microsatellite markers using the standard methods were unsuccessful. A next-generation sequencing (NGS) approach was applied on genomic DNA extracted from fresh leaves of Dalmatian pyrethrum. The sequencing was conducted using NovaSeq6000 Illumina sequencer, after which almost 400 million high-quality paired-end reads were obtained, with a read length of 150 base pairs. Short reads were assembled by combining two approaches; (1) de-novo assembly and (2) joining of overlapped pair-end reads. In total, 6.909.675 contigs were obtained, with the contig average length of 249 base pairs. Of the resulting contigs, 31.380 contained one or multiple microsatellite sequences, in total 35.556 microsatellite loci were identified. Out of detected microsatellites, dinucleotide repeats were the most frequent, accounting for more than half of all microsatellites identifies (21,212; 59.7%), followed by trinucleotide repeats (9,204; 25.9%). Tetra-, penta- and hexanucleotides had similar frequency of 1,822 (5.1%), 1,472 (4.1%), and 1,846 (5.2%), respectively. Contigs containing microsatellites were further filtered by SSR pattern type, transposon occurrences, assembly characteristics, GC content, and the number of occurrences against the draft genome of T. cinerariifolium published previously. After the selection process, 50 microsatellite loci were used for primer design. Designed primers were tested on samples from five distinct populations, and 25 of them showed a high degree of polymorphism. The selected loci were then genotyped on 20 samples belonging to one population resulting in 17 microsatellite markers. Availability of codominant SSR markers will significantly improve the knowledge on population genetic diversity and structure as well as complex genetics and biochemistry of this species. Acknowledgment: This work has been fully supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir/ Sch. Bip.) insecticidal potential’ - (PyrDiv) (IP-06-2016-9034).

Keywords: genome assembly, NGS, SSR, Tanacetum cinerariifolium

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6582 Emergency Surgery in the Elderly, What Particularities

Authors: Mekroud Amel

Abstract:

Introduction The rate of use by the elderly of emergency departments, operating rooms and intensive care units has increased worldwide. Emergency surgery is a context where evaluation is often insufficient, with incomplete information gathering. The aim of this work is to shed light on the frequent use of emergency surgeries by the elderly and their characteristics, as well as on the lack of geriatric assessment scores in the emergency room. Material : Prospective, observational and descriptive, monocentric study. Patients aged 65 and over, admitted for emergency surgery in the operating room, were counted. Emergency operating room including visceral surgery, urology, traumatology and neurosurgery. Parameters studied: Patient characteristics, degree of autonomy, type of surgical pathology, operative management times, preoperative evaluation, postoperative outcome Results : 192 patients were identified over 12 months, from 09.01.2017 to 08.31.2018 Age from 65 to 101 years, 79.81 years +/- 8.38. With predominance of the age group between [65-75 years] 41.1% Female predominance, Sexratio = 0.81 Elderly subjects with total motor autonomy are in the majority at 57.8% Subjects without pathological ATCD represent 12.5% of cases Those who are on only one type of medication or without any treatment are at 36.9% Discussion : The emergency operative care of the elderly patient for a surgical or traumatological pathology is characterized by many specificities linked first to the emergency context, where the evaluation is often insufficient, besides the fact that the elderly patient has particularities requiring reception in centers with experience in the care of this category of patient, or, failing that, a center which uses the minimum of geriatric evaluation scores which are simplified for the emergency departments. In our hospital, we have not yet made this evaluation routine in the emergency room and this delay in the introduction of these scores can be directly attributed to the covid 19 pandemic. Besides the standard preoperative assessment, only 43.2% of patients were assessed in the preoperative period by an anesthesiologist. Traumatological emergencies come first 68.2% followed by visceral emergencies 19.2% (including proctological, urological emergencies), neurosurgical emergencies 7.8% and finally peripheral emergency surgery all acts combined 4.7%. Hospital stay at 9.6 +/- 16.8 days, average operability time of 4.5 +/- 3 days. Death rate at 7.29% Conclusion This work has demonstrated the major impact of emergency surgery, which remains curable for the most part, on the elderly patient despite total motor and cognitive autonomy preoperatively. The improvement of the preoperative evaluation, the reduction of the operating time and enhanced recovery after surgery, with personalized protocols, are the only guarantee for the resumption of preoperative autonomy in these patients.

Keywords: emergency surgery, elderly patients, preoperative geriatric scores, curable emergency surgical pathologies

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6581 Creating Renewable Energy Investment Portfolio in Turkey between 2018-2023: An Approach on Multi-Objective Linear Programming Method

Authors: Berker Bayazit, Gulgun Kayakutlu

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The World Energy Outlook shows that energy markets will substantially change within a few forthcoming decades. First, determined action plans according to COP21 and aim of CO₂ emission reduction have already impact on policies of countries. Secondly, swiftly changed technological developments in the field of renewable energy will be influential upon medium and long-term energy generation and consumption behaviors of countries. Furthermore, share of electricity on global energy consumption is to be expected as high as 40 percent in 2040. Electrical vehicles, heat pumps, new electronical devices and digital improvements will be outstanding technologies and innovations will be the testimony of the market modifications. In order to meet highly increasing electricity demand caused by technologies, countries have to make new investments in the field of electricity production, transmission and distribution. Specifically, electricity generation mix becomes vital for both prevention of CO₂ emission and reduction of power prices. Majority of the research and development investments are made in the field of electricity generation. Hence, the prime source diversity and source planning of electricity generation are crucial for improving the wealth of citizen life. Approaches considering the CO₂ emission and total cost of generation, are necessary but not sufficient to evaluate and construct the product mix. On the other hand, employment and positive contribution to macroeconomic values are important factors that have to be taken into consideration. This study aims to constitute new investments in renewable energies (solar, wind, geothermal, biogas and hydropower) between 2018-2023 under 4 different goals. Therefore, a multi-objective programming model is proposed to optimize the goals of minimizing the CO₂ emission, investment amount and electricity sales price while maximizing the total employment and positive contribution to current deficit. In order to avoid the user preference among the goals, Dinkelbach’s algorithm and Guzel’s approach have been combined. The achievements are discussed with comparison to the current policies. Our study shows that new policies like huge capacity allotment might be discussible although obligation for local production is positive. The improvements in grid infrastructure and re-design support for the biogas and geothermal can be recommended.

Keywords: energy generation policies, multi-objective linear programming, portfolio planning, renewable energy

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6580 Transformation of ectA Gene From Halomonas elongata in Tomato Plant

Authors: Narayan Moger, Divya B., Preethi Jambagi, Krishnaveni C. K., Apsana M. R., B. R. Patil, Basvaraj Bagewadi

Abstract:

Salinity is one of the major threats to world food security. Considering the requirement for salt tolerant crop plants in the present study was undertaken to clone and transferred the salt tolerant ectA gene from marine ecosystem into agriculture crop system to impart salinity tolerance. Ectoine is the compatible solute which accumulates in the cell membrane, is known to be involved in salt tolerance activity in most of the Halophiles. The present situation is insisting to development of salt tolerant transgenic lines to combat abiotic stress. In this background, the investigation was conducted to develop transgenic tomato lines by cloning and transferring of ectA gene is an ectoine derivative capable of enzymatic action for the production of acetyl-diaminobutyric acid. The gene ectA is involved in maintaining the osmotic balance of plants. The PCR amplified ectA gene (579bp) was cloned into T/A cloning vector (pTZ57R/T). The construct pDBJ26 containing ectA gene was sequenced by using gene specific forward and reverse primers. Sequence was analyzed using BLAST algorithm to check similarity of ectA gene with other isolates. Highest homology of 99.66 per cent was found with ectA gene sequences of isolates Halomonas elongata with the available sequence information in NCBI database. The ectA gene was further sub cloned into pRI101-AN plant expression vector and transferred into E. coli DH5α for its maintenance. Further pDNM27 was mobilized into A. tumefaciens LBA4404 through tri-parental mating system. The recombinant Agrobacterium containing pDNM27 was transferred into tomato plants through In planta plant transformation method. Out of 300 seedlings, co-cultivated only twenty-seven plants were able to well establish under the greenhouse condition. Among twenty-seven transformants only twelve plants showed amplification with gene specific primers. Further work must be extended to evaluate the transformants at T1 and T2 generations for ectoine accumulation, salinity tolerance, plant growth and development and yield.

Keywords: salinity, computable solutes, ectA, transgenic, in planta transformation

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6579 Circular Tool and Dynamic Approach to Grow the Entrepreneurship of Macroeconomic Metabolism

Authors: Maria Areias, Diogo Simões, Ana Figueiredo, Anishur Rahman, Filipa Figueiredo, João Nunes

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It is expected that close to 7 billion people will live in urban areas by 2050. In order to improve the sustainability of the territories and its transition towards circular economy, it’s necessary to understand its metabolism and promote and guide the entrepreneurship answer. The study of a macroeconomic metabolism involves the quantification of the inputs, outputs and storage of energy, water, materials and wastes for an urban region. This quantification and analysis representing one opportunity for the promotion of green entrepreneurship. There are several methods to assess the environmental impacts of an urban territory, such as human and environmental risk assessment (HERA), life cycle assessment (LCA), ecological footprint assessment (EF), material flow analysis (MFA), physical input-output table (PIOT), ecological network analysis (ENA), multicriteria decision analysis (MCDA) among others. However, no consensus exists about which of those assessment methods are best to analyze the sustainability of these complex systems. Taking into account the weaknesses and needs identified, the CiiM - Circular Innovation Inter-Municipality project aims to define an uniform and globally accepted methodology through the integration of various methodologies and dynamic approaches to increase the efficiency of macroeconomic metabolisms and promoting entrepreneurship in a circular economy. The pilot territory considered in CiiM project has a total area of 969,428 ha, comprising a total of 897,256 inhabitants (about 41% of the population of the Center Region). The main economic activities in the pilot territory, which contribute to a gross domestic product of 14.4 billion euros, are: social support activities for the elderly; construction of buildings; road transport of goods, retailing in supermarkets and hypermarkets; mass production of other garments; inpatient health facilities; and the manufacture of other components and accessories for motor vehicles. The region's business network is mostly constituted of micro and small companies (similar to the Central Region of Portugal), with a total of 53,708 companies identified in the CIM Region of Coimbra (39 large companies), 28,146 in the CIM Viseu Dão Lafões (22 large companies) and 24,953 in CIM Beiras and Serra da Estrela (13 large companies). For the construction of the database was taking into account data available at the National Institute of Statistics (INE), General Directorate of Energy and Geology (DGEG), Eurostat, Pordata, Strategy and Planning Office (GEP), Portuguese Environment Agency (APA), Commission for Coordination and Regional Development (CCDR) and Inter-municipal Community (CIM), as well as dedicated databases. In addition to the collection of statistical data, it was necessary to identify and characterize the different stakeholder groups in the pilot territory that are relevant to the different metabolism components under analysis. The CIIM project also adds the potential of a Geographic Information System (GIS) so that it is be possible to obtain geospatial results of the territorial metabolisms (rural and urban) of the pilot region. This platform will be a powerful visualization tool of flows of products/services that occur within the region and will support the stakeholders, improving their circular performance and identifying new business ideas and symbiotic partnerships.

Keywords: circular economy tools, life cycle assessment macroeconomic metabolism, multicriteria decision analysis, decision support tools, circular entrepreneurship, industrial and regional symbiosis

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6578 ‘Made of Glass’: Women Seafarers and the International Cargo Shipping Industry

Authors: Iris Lavalle Acejo, Helen Sampson

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Drawing from the perspectives of female seafarers, this article offers an analysis of an empirical investigation of the mechanisms by which women are excluded from working on board cargo ships. Female seafarers currently make up about 1% of seafarers in the global cargo shipping industry. As female presence onboard continues to constitute a small proportion of the total crew, we consider their professional advancement in comparison with their male counterparts and the ways in which they are discouraged from entering, or remining within, the industry. With interactions at work severely constrained by the traditionally masculine working culture of the ship, the study hopes to provide insights on the mechanisms in which they face and navigate barriers as they both accommodate and resist gendered differences onboard.

Keywords: female seafarers, glasswalls, genderbarriers, genderedworkplaces

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6577 Distributed Framework for Pothole Detection and Monitoring Using Federated Learning

Authors: Ezil Sam Leni, Shalen S.

Abstract:

Transport service monitoring and upkeep are essential components of smart city initiatives. The main risks to the relevant departments and authorities are the ever-increasing vehicular traffic and the conditions of the roads. In India, the economy is greatly impacted by the road transport sector. In 2021, the Ministry of Road Transport and Highways Transport, Government of India, produced a report with statistical data on traffic accidents. The data included the number of fatalities, injuries, and other pertinent criteria. This study proposes a distributed infrastructure for the monitoring, detection, and reporting of potholes to the appropriate authorities. In a distributed environment, the nodes are the edge devices, and local edge servers, and global servers. The edge devices receive the initial model to be employed from the global server. The YOLOv8 model for pothole detection is used in the edge devices. The edge devices run the pothole detection model, gather the pothole images on their path, and send the updates to the nearby edge server. The local edge server selects the clients for its aggregation process, aggregates the model updates and sends the updates to the global server. The global server collects the updates from the local edge servers, performs aggregation and derives the updated model. The updated model has the information about the potholes received from the local edge servers and notifies the updates to the local edge servers and concerned authorities for monitoring and maintenance of road conditions. The entire process is implemented in FedCV distributed environment with the implementation using the client-server model and aggregation entities. After choosing the clients for its aggregation process, the local edge server gathers the model updates and transmits them to the global server. After gathering the updates from the regional edge servers, the global server aggregates them and creates the updated model. Performance indicators and the experimentation environment are assessed, discussed, and presented. Accelerometer data may be taken into consideration for improved performance in the future development of this study, in addition to the images captured from the transportation routes.

Keywords: federated Learning, pothole detection, distributed framework, federated averaging

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6576 Gender Supportive Systems-Key to Good Governance in Agriculture: Challenges and Strategies

Authors: Padmaja Kaja, Kiran Kumar Gellaboina

Abstract:

A lion’s share of agricultural work is contributed by women in India as it is the case in many developing countries, yet women are not securing the pride as a farmer. Many policies are supporting women empowerment in India, especially in agriculture sector considering the importance of sustainable food security. However these policies many times failed to achieve the targeted results of mainstreaming gender. Implementing the principles of governance would lead to gender equality in agriculture. This paper deals with the social norms and obligations prevailed with reference to Indian context which abstain women from having resources. This paper is formulated by using primary research done in eight districts of Telangana and Andhra Pradesh states of India supported by secondary research. Making amendments to Hindu Succession Act in united Andhra Pradesh much prior to the positioning of the amended act in the whole country lead to a better land holding a share of women in Andhra Pradesh. The policies like registering government distributed lands in the name of women in the state also have an added value. However, the women participation in decision-making process in agriculture is limited in elite families when compared to socially under privileged families, further too it was higher in drought affected districts like Mahbubnagar in Telangana when compared to resource-rich East Godavari district in Andhra Pradesh. Though National Gender Resource Centre for Agriculture (NGRCA) at centre and Gender Cells in the states were established a decade ago, extension reach to the women farmers is still lagging behind. Capturing the strength of women self groups in India especially in Andhra Pradesh to link up with agriculture extension might improve the extension reach of women farmers. Maintenance of micro level women data sets, creating women farmers networks with government departments like agriculture, irrigation, revenue and formal credit institutes would result in good governance to mainstream gender in agriculture. Further to add that continuous monitoring and impact assessments of the programmes and projects for gender inclusiveness would reiterate the government efforts.

Keywords: food security, gender, governance, mainstreaming

Procedia PDF Downloads 236
6575 Study of Eatable Aquatic Invertebrates in the River Dhansiri, Dimapur, Nagaland, India

Authors: Dilip Nath

Abstract:

A study has been conducted on the available aquatic invertebrates in the river Dhansiri at Dimapur site. The study confirmed that the river body composed of aquatic macroinvertebrate community under two phyla viz., Arthropods and Molluscs. Total 10 species have been identified from there as the source of alternative protein food for the common people. Not only the protein source, they are also the component of aquatic food chain and indicators of aquatic ecosystem. Proper management and strategies to promote the edible invertebrates can be considered as the alternative protein and alternative income source for the common people for sustainable livelihood improvement.

Keywords: Dhansiri, Dimapur, invertebrates, livelihood improvement, protein

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6574 Association Between Renewable Energy and Community Forest User Group of Siranchowk Rural Municipality, Nepal

Authors: Prem Bahadur Giri, Mathinee Yucharoen

Abstract:

Community Forest User Groups (CFUGs) have been the core stone of forest management efforts in Nepal. Due to the lack of a smooth transition into the local governance structure in 2017, policy instruments have not been effectively cascaded to the local level, creating ambiguity and inconsistency in forest governance. Descriptive mixed-method research was performed with community users and stakeholders of Tarpakha community forest, Siranchowk Rural Municipality, to understand the role of the political economy in CFUG management. A household survey was conducted among 100 households (who also are existing members of the Tarpakha CFUG) to understand and document their energy consumption preferences and practices. Likewise, ten key informant interviews and five focus group discussions with the municipality and forest management officials were also conducted to have a wider overview of the factors and political, socio-economic, and religious contexts behind the utilization of renewable energy for sustainable development. Findings from our study suggest only 3% of households use biogas as the main source of energy. The rest of the households mention liquid petroleum gas (LPG), electricity and firewood as major sources of energy for domestic purposes. Community members highlighted the difficulty in accessing firewood due to strict regulations from the CFUG, lack of cattle and manpower to rear cattle to produce cow dung (for biogas), lack of technical expertise at the community level for the operation and maintenance of solar energy, among others as challenges of resource. Likewise, key informants have mentioned policy loopholes at both the federal and local levels, especially with regard to the promotion of alternative or renewable energy, as there are no clear mandates and provisions to regulate the renewable energy industry. The study recommends doing an in-depth study on the feasibility of renewable energy sources, especially in the context of CFUGs, where biodiversity conservation aspects need to be equally taken into consideration while thinking of the promotion and expansion of renewable energy sources.

Keywords: community forest, renewable energy, sustainable development, Nepal

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6573 Modeling SET Effect on Charge Pump Phase Locked Loop

Authors: Varsha Prasad, S. Sandya

Abstract:

Cosmic Ray effects in microelectronics such as single event effect (SET) and total dose ionization (TID) have been of major concern in space electronics since 1970. Advanced CMOS technologies have demonstrated reduced sensitivity to TID effect. However, charge pump Phase Locked Loop is very much vulnerable to single event transient effect. This paper presents an SET analysis model, where the SET is modeled as a double exponential pulse. The time domain analysis reveals that the settling time of the voltage controlled oscillator (VCO) depends on the SET pulse strength, setting the time constant and the damping factor. The analysis of the proposed SET analysis model is confirmed by the simulation results.

Keywords: charge pump, phase locked loop, SET, VCO

Procedia PDF Downloads 422
6572 Microbiological Quality and Safety of Meatball Sold in Payakumbuh City, West Sumatra, Indonesia

Authors: Ferawati, H. Purwanto, Y. F. Kurnia, E. Purwati

Abstract:

The aim of this study was to evaluate the microbiological quality and safety of meatball obtained from five different manufacturers around Payakumbuh City, West Sumatra, Indonesia. Microbiological analysis of meatball sample resulted in aerobic plate count range from 7 log CFU/gr to 8.623 log CFU/gr, respectively. Total coliform ranges from 1.041 log Most Probable Number (MPN)/gr to 3.380 log MPN/gr, respectively. Chemical analysis of meatball sample consisted of borax and formalin content. The result of qualitative detection of borax and formalin content on all meatball samples were not detected. Thus, it remains essential to include the significance of effective hygiene practices as an important safety measure in consumer education programmes.

Keywords: borax, formalin, meatball, microbiological quality

Procedia PDF Downloads 277
6571 Isothermal Vapour-Liquid Equilibria of Binary Mixtures of 1, 2-Dichloroethane with Some Cyclic Ethers: Experimental Results and Modelling

Authors: Fouzia Amireche-Ziar, Ilham Mokbel, Jacques Jose

Abstract:

The vapour pressures of the three binary mixtures: 1, 2- dichloroethane + 1,3-dioxolane, + 1,4-dioxane or + tetrahydropyrane, are carried out at ten temperatures ranging from 273 to 353.15 K. An accurate static device was employed for these measurements. The VLE data were reduced using the Redlich-Kister equation by taking into consideration the vapour pressure non-ideality in terms of the second molar virial coefficient. The experimental data were compared to the results predicted with the DISQUAC and Dortmund UNIFAC group contribution models for the total pressures P and the excess molar Gibbs energies GE.

Keywords: disquac model, dortmund UNIFAC model, excess molar Gibbs energies GE, VLE

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6570 Effect of Waste Dumping on Groundwater Quality at Guntun Layi Funtua, Katsina State

Authors: Isiya Aminu Dabai, Adebola Kayode, Adeosun Kayode Daniel

Abstract:

Rural water supply relies mainly on groundwater exploitation, because it is more accessible, reliable, cheaper to develop and maintain, also with good quality compared to the surface water. Despite these advantages, groundwater has come under pollution threats like waste dumps, mineral exploitation, industrialization etc. This study investigates the effects of an open dumping to the surrounding groundwater. Ten hand dug well water samples were collected from the surroundings and tested. The average result shows that temperature, colour and turbidity to be 8.50 c, 6.1 TCU and 3.1 NTU respectively and pH, conductivity, total dissolved solids, chloride content and hardness to be 7.2, 4.78, 1.8, 5.7, and 3.4 respectively while in the bacteriological test well no. 1, 2, 3, and 5 shows the presence of coliforms and E. Coli bacteria.

Keywords: groundwater, pollution, waste, dump site, unsafe, quality

Procedia PDF Downloads 666
6569 Brazilian Constitution and the Fundamental Right to Sanitation

Authors: Michely Vargas Delpupo, José Geraldo Romanello Bueno

Abstract:

The right to basic sanitation, was elevated to the category of fundamental right by the Brazilian Constitution of 1988 to protect the ecologically balanced environment, ensuring social rights to health and adequate housing warranting dignity of the human person as a principle of the Brazilian Democratic State. Because of their essentiality to the Brazilian population, this article seeks to understand why universal access to basic sanitation is a goal so difficult to achieve in Brazil. Therefore, this research uses the deductive and analytical method. Given the nature of the research literature, research techniques were centered in specialized books on the subject, journals, theses and dissertations, laws, relevant law case and raising social indicators relating to the theme. The relevance of the topic stems, among other things, the fact that sanitation services are essential for a dignified life, i.e. everyone is entitled to the maintenance of the necessary existence conditions are satisfied. However, the effectiveness of this right is undermined in society, since Brazil has huge deficit in sanitation services, denying thus a worthy life to most of the population. Thus, it can be seen that the provision of water and sewage services in Brazil is still characterized by a large imbalance, since the municipalities with lower population index have greater disability in the sanitation service. The truth is that the precariousness of water and sewage services in Brazil is still very concentrated in the North and Northeast regions, limiting the effective implementation of the Law 11.445/2007 in the country. Therefore, there is urgent need for a positive service by the State in the provision of sanitation services in order to prevent and control disease, improve quality of life and productivity of individuals, besides preventing contamination of water resources. More than just social and economic necessity, there is even a an obligation of the government to implement such services. In this sense, given the current scenario, to achieve universal access to basic sanitation imposes many hurdles. These are mainly in the field of properly formulated and implemented public policies, i.e. it requires an excellent institutional organization, management services, strategic planning, social control, in order to provide answers to complex challenges.

Keywords: fundamental rights, health, sanitation, universal access

Procedia PDF Downloads 395
6568 Investigation of the Effects of Sampling Frequency on the THD of 3-Phase Inverters Using Space Vector Modulation

Authors: Khattab Al Qaisi, Nicholas Bowring

Abstract:

This paper presents the simulation results of the effects of sampling frequency on the total harmonic distortion (THD) of three-phase inverters using the space vector pulse width modulation (SVPWM) and space vector control (SVC) algorithms. The relationship between the variables was studied using curve fitting techniques, and it has been shown that, for 50 Hz inverters, there is an exponential relation between the sampling frequency and THD up to around 8500 Hz, beyond which the performance of the model becomes irregular, and there is an negative exponential relation between the sampling frequency and the marginal improvement to the THD. It has also been found that the performance of SVPWM is better than that of SVC with the same sampling frequency in most frequency range, including the range where the performance of the former is irregular.

Keywords: DSI, SVPWM, THD, DC-AC converter, sampling frequency, performance

Procedia PDF Downloads 468
6567 Implementation of Total Quality Management in Public Sector: Case of Tunisia

Authors: Rafla Hchaichi

Abstract:

The public administration is currently experiencing in the field of quality unprecedented effervescence. However, in a globalized world more and more competitive, public services are confronted with the need to improve their performances which push public companies to implement quality approaches. Quality approaches have taken diverse forms such as service commitment, labels, certifications and the Common Assessment Framework. This paper provides an overview on the strategy for administrative development in Tunisia since the Carthaginian civilization until today. It outlines the evolution of quality management in the Tunisian public context while focusing on the National Referential of Quality of Administrative Services.

Keywords: quality approach, the common assessment framework, service commitment, label, certification, quality of public service, performance of public service, Tunisian Public Service

Procedia PDF Downloads 529
6566 Assessing the Contribution of Informal Buildings to Energy Inefficiency in Kenya: A Case of Mukuru Slums

Authors: Bessy Thuranira

Abstract:

Buildings, as they are designed and used, may contribute to serious environmental problems because of excessive consumption of energy and other natural resources. Buildings in the informal settlements particularly, due to their unplanned physical structure and design, have significantly contributed the global energy problematic scenario typified by high-level inefficiencies. Energy used in buildings in Africa is estimated to be the highest of the total national electricity consumption. Over the last decade, assessments of energy consumption and efficiency/inefficiency has focused on formal and modern buildings. This study seeks to go off the beaten path, by focusing on energy use in informal settlements. Operationally, it sought to establish the contribution of informal buildings in the overall energy consumption in the city and the country at large. This study was carried out in Mukuru kwa Reuben informal settlement where there is distinct manifestation of different settlement morphologies within a small locality. The research narrowed down to three villages (Mombasa, Kosovo and Railway villages) within the settlement, that were representative of the different slum housing typologies. Due to the unpredictability nature and informality in slums, this study takes a multi-methodology approach. Detailed energy audits and measurements are carried out to predict total building consumption, and document building design and envelope, typology, materials and occupancy levels. Moreover, the study uses semi-structured interviews and to access energy supply, cost, access and consumption patterns. Observations and photographs are also used to shed more light on these parameters. The study reveals the high energy inefficiencies in slum buildings mainly related to sub-standard equipment and appliances, building design and settlement layout, poor access and utilization/consumption patterns of energy. The impacts of this inefficiency are high economic burden to the poor, high levels of pollution, lack of thermal comfort and emissions to the environment. The study highlights a set of urban planning and building design principles that can be used to retrofit slums into more energy efficient settlements. The study explores principles of responsive settlement layouts/plans and appropriate building designs that use the beneficial elements of nature to achieve natural lighting, natural ventilation, and solar control to create thermally comfortable, energy efficient, and environmentally responsive buildings/settlements. As energy efficiency in informal settlements is a relatively less explored area of efficiency, it requires further research and policy recommendations, for which this paper will set a background.

Keywords: energy efficiency, informal settlements, renewable energy, settlement layout

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6565 Congenital Diaphragmatic Hernia Outcomes in a Low-Volume Center

Authors: Michael Vieth, Aric Schadler, Hubert Ballard, J. A. Bauer, Pratibha Thakkar

Abstract:

Introduction: Congenital diaphragmatic hernia (CDH) is a condition characterized by the herniation of abdominal contents into the thoracic cavity requiring postnatal surgical repair. Previous literature suggests improved CDH outcomes at high-volume regional referral centers compared to low-volume centers. The purpose of this study was to examine CDH outcomes at Kentucky Children’s Hospital (KCH), a low-volume center, compared to the Congenital Diaphragmatic Hernia Study Group (CDHSG). Methods: A retrospective chart review was performed at KCH from 2007-2019 for neonates with CDH, and then subdivided into two cohorts: those requiring ECMO therapy and those not requiring ECMO therapy. Basic demographic data and measures of mortality and morbidity including ventilator days and length of stay were compared to the CDHSG. Measures of morbidity for the ECMO cohort including duration of ECMO, clinical bleeding, intracranial hemorrhage, sepsis, need for continuous renal replacement therapy (CRRT), need for sildenafil at discharge, timing of surgical repair, and total ventilator days were collected. Statistical analysis was performed using IBM SPSS Statistics version 28. One-sample t-tests and one-sample Wilcoxon Signed Rank test were utilized as appropriate.Results: There were a total of 27 neonatal patients with CDH at KCH from 2007-2019; 9 of the 27 required ECMO therapy. The birth weight and gestational age were similar between KCH and the CDHSG (2.99 kg vs 2.92 kg, p =0.655; 37.0 weeks vs 37.4 weeks, p =0.51). About half of the patients were inborn in both cohorts (52% vs 56%, p =0.676). KCH cohort had significantly more Caucasian patients (96% vs 55%, p=<0.001). Unadjusted mortality was similar in both groups (KCH 70% vs CDHSG 72%, p =0.857). Using ECMO utilization (KCH 78% vs CDHSG 52%, p =0.118) and need for surgical repair (KCH 95% vs CDHSG 85%, p =0.060) as proxy for severity, both groups’ mortality were comparable. No significant difference was noted for pulmonary outcomes such as average ventilator days (KCH 43.2 vs. CDHSG 17.3, p =0.078) and home oxygen dependency (KCH 44% vs. CDHSG 24%, p =0.108). Average length of hospital stay for patients treated at KCH was similar to CDHSG (64.4 vs 49.2, p=1.000). Conclusion: Our study demonstrates that outcome in CDH patients is independent of center’s case volume status. Management of CDH with a standardized approach in a low-volume center can yield similar outcomes. This data supports the treatment of patients with CDH at low-volume centers as opposed to transferring to higher-volume centers.

Keywords: ECMO, case volume, congenital diaphragmatic hernia, congenital diaphragmatic hernia study group, neonate

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6564 Fuzzy Vehicle Routing Problem for Extreme Environment

Authors: G. Sirbiladze, B. Ghvaberidze, B. Matsaberidze

Abstract:

A fuzzy vehicle routing problem is considered in the possibilistic environment. A new criterion, maximization of expectation of reliability for movement on closed routes is constructed. The objective of the research is to implement a two-stage scheme for solution of this problem. Based on the algorithm of preferences on the first stage, the sample of so-called “promising” routes will be selected. On the second stage, for the selected promising routes new bi-criteria problem will be solved - minimization of total traveled distance and maximization of reliability of routes. The problem will be stated as a fuzzy-partitioning problem. Two possible solutions of this scheme are considered.

Keywords: vehicle routing problem, fuzzy partitioning problem, multiple-criteria optimization, possibility theory

Procedia PDF Downloads 533
6563 Real-Time Quantitative Polymerase Chain Reaction Assay for the Detection of microRNAs Using Bi-Directional Extension Sequences

Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

Abstract:

MicroRNAs (miRNA) are a class of endogenous, single-stranded, small, and non-protein coding RNA molecules typically 20-25 nucleotides long. They are thought to regulate the expression of other genes in a broad range by binding to 3’- untranslated regions (3’-UTRs) of specific mRNAs. The detection of miRNAs is very important for understanding of the function of these molecules and in the diagnosis of variety of human diseases. However, detection of miRNAs is very challenging because of their short length and high sequence similarities within miRNA families. So, a simple-to-use, low-cost, and highly sensitive method for the detection of miRNAs is desirable. In this study, we demonstrate a novel bi-directional extension (BDE) assay. In the first step, a specific linear RT primer is hybridized to 6-10 base pairs from the 3’-end of a target miRNA molecule and then reverse transcribed to generate a cDNA strand. After reverse transcription, the cDNA was hybridized to the 3’-end which is BDE sequence; it played role as the PCR template. The PCR template was amplified in an SYBR green-based quantitative real-time PCR. To prove the concept, we used human brain total RNA. It could be detected quantitatively in the range of seven orders of magnitude with excellent linearity and reproducibility. To evaluate the performance of BDE assay, we contrasted sensitivity and specificity of the BDE assay against a commercially available poly (A) tailing method using miRNAs for let-7e extracted from A549 human epithelial lung cancer cells. The BDE assay displayed good performance compared with a poly (A) tailing method in terms of specificity and sensitivity; the CT values differed by 2.5 and the melting curve showed a sharper than poly (A) tailing methods. We have demonstrated an innovative, cost-effective BDE assay that allows improved sensitivity and specificity in detection of miRNAs. Dynamic range of the SYBR green-based RT-qPCR for miR-145 could be represented quantitatively over a range of 7 orders of magnitude from 0.1 pg to 1.0 μg of human brain total RNA. Finally, the BDE assay for detection of miRNA species such as let-7e shows good performance compared with a poly (A) tailing method in terms of specificity and sensitivity. Thus BDE proves a simple, low cost, and highly sensitive assay for various miRNAs and should provide significant contributions in research on miRNA biology and application of disease diagnostics with miRNAs as targets.

Keywords: bi-directional extension (BDE), microRNA (miRNA), poly (A) tailing assay, reverse transcription, RT-qPCR

Procedia PDF Downloads 150