Search results for: complete genome sequencing

Authors: Rully D. Purwati, Tantri D.A. Anggraeni, Bambang Heliyanto, M. Machfud, Joko Hartono

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The availability of fuel in the world will be reduced in next few years, it is necessary to find alternative energy sources. Jatropha curcas L. is one of oil crops producing non-edible oil which is potential for bio-diesel. Jatropha cultivation and development program in Indonesia is facing several problems especially low seed yield resulting in inefficient crop cultivation cost. To cope with the problem, development of high yielding varieties is necessary. Development of new varieties to improve seed yield was conducted by hybridization and selection and resulted in fourteen potential genotypes. The yield potential of the fourteen genotypes were evaluated and compared with two check varieties. The objective of the evaluation was to find Jatropha hybrids with some characters i.e. their productivity was higher than check varieties, oil content > 40% and harvesting age ≤ 110 days. Hybridization and individual plant selection were carried out from 2010 to 2014. Evaluation of high yield was conducted in Asembagus experimental station, Situbondo, East Java in three years (2015-2017). The experimental designed was Randomized Complete Block Design with three replication, and plot size 10 m x 8 m. The characters observed were number of capsules per plant, dry seed yield (kg/ha) and seed oil content (%). The results of this experiment indicated that all the hybrids evaluated have higher productivity than check variety IP-3A. There were two superior hybrids i.e. HS-49xSP-65/32 and HS-49xSP-19/28 with highest seed yield per hectare and number of capsules per plant for three years.

Keywords: Jatropha, bio energy, hybrid, high seed yield

Procedia PDF Downloads 135

Authors: M. Kazou, A. Gavriil, O. Kalagkatsi, T. Paschos, E. Tsakalidou

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Kopanisti cheese is a Greek soft Protected Designation of Origin (PDO) cheese made of raw cow, sheep or goat milk, or mixtures of them, with similar organoleptic characteristics to that of Roquefort cheese. Traditional manufacturing of Kopanisti cheese is limited in small-scale dairies, without the addition of starter cultures. Instead, an amount of over-mature Kopanisti cheese, called Mana Kopanisti, is used to initiate ripening. Therefore, the selection of proper starter cultures and the understanding of the contribution of various microbial groups to its overall quality is crucial for the production of a high-quality final product with standardized organoleptic and physicochemical characteristics. Taking the above into account, the aim of the present study was the investigation of Kopanisti cheese microbiota and its role in cheese quality. For this purpose, four different types of Kopanisti were produced in triplicates, all with pasteurized cow milk, with the addition of (A) the typical mesophilic species Lactococcus lactis and Lactobacillus paracasei used as starters in the production of soft spread cheeses, (B) strains of Lactobacillus acidipiscis and Lactobacillus rennini previously isolated from Kopanisti and Mana Kopanisti, (C) all the species from (A) and (B) as inoculum, and finally (D) the species from (A) and Mana Kopanisti. Physicochemical and microbiological analysis was performed for milk and cheese samples during ripening. Enumeration was performed for major groups of lactic acid bacteria (LAB), total mesophilic bacteria, yeasts as well as hygiene indicator microorganisms. Bacterial isolates from all the different LAB groups, apart from enterococci, alongside yeasts isolates, were initially grouped using repetitive sequence-based polymerase chain reaction (rep-PCR) and then identified at the species level using 16S rRNA gene and internal transcribed spacer (ITS) DNA region sequencing, respectively. Sensory evaluation was also performed for final cheese samples at the end of the ripening period (35 days). Based on the results of the classical microbiological analysis, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, ranged between 7 and 10 log colony forming unit (CFU) g⁻¹, phychrotrophic bacteria, and yeast extract glucose chloramphenicol (YGC) isolates between 4 and 8 log CFU g⁻¹, while coliforms and enterococci up to 2 log CFU g⁻¹ throughout ripening in cheese samples A, C and D. In contrast, in cheese sample B, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, phychrotrophic bacteria, and YGC isolates ranged between 0 and 10 log CFU g⁻¹ and coliforms and enterococci up to 2 log CFU g⁻¹. Although the microbial counts were not that different among samples, identification of the bacterial and yeasts isolates revealed the complex microbial community structure present in each cheese sample. Differences in the physicochemical characteristics among the cheese samples were also observed, with pH ranging from 4.3 to 5.3 and moisture from 49.6 to 58.0 % in the final cheese products. Interestingly, the sensory evaluation also revealed differences among samples, with cheese sample B ranking first based on the total score. Overall, the combination of these analyses highlighted the impact of different starter cultures on the Kopanisti microbiota as well as on the physicochemical and sensory characteristics of the final product.

Keywords: Kopanisti cheese, microbiota, classical microbiological analysis, physicochemical analysis

Procedia PDF Downloads 133

Authors: Masocorro Gawned, Stephen Myers, Guat Siew Chew

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Nuclear Receptors (NR) are a super family of transcription factors that play a major role in lipid and glucose metabolism in skeletal muscle. Recently, pharmacological evidence supports the view that stimulation of nuclear receptors alleviates Type 2 Diabetes (T2D). The orphan nuclear receptors (ONR) are members of the nuclear receptor (NR) superfamily whose ligands and physiological functions remain unknown. To date, no systematic studies have been carried out to screen for ONRs expressed in insulin resistant (IR) skeletal muscle cells. Therefore, in this study, we have established a model for IR by treating C2C12 skeletal muscle cells with insulin (10nM) for 48 hours. Western Blot analysis of phosphorylated AKT confirmed IR. Real-time quantitative polymerase chain reaction (qPCR) results highlighted key ONRs including NUR77 (NR4A1), NURR1 (NR4A2) and NOR1 (NR4A3) which have been associated with fatty acid oxidation regulation and glucose homeostasis. Increased mRNA expression levels of estrogen-related receptors (ERRs), REV-ERBα, NUR77, NURR1, NOR1, in insulin resistant C2C12 skeletal muscle cells, indicated that these ONRs could potentially play a pivotal regulatory role of insulin secretion in lipid metabolism. Taken together, this study has successfully contributed to the complete analysis of ONR in IR, and has filled in an important void in the study and treatment of T2D.

Keywords: type 2 diabetes, orphan nuclear receptors, transcription receptors, quantitative mRNA expression

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Authors: Priyanka Gupta, Bipin Kumar

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Recent developments in smart materials motivate researchers to create novel textile products for innovative and functional applications, which have several potential uses beyond the conventional. This study investigates the memory behavior of shape memory filaments integrated into a knitted textile structure. The research advances the knowledge of how these intelligent materials respond within textile structures. This integration may also open new avenues for developing smart fabrics with unique sensing and actuation capabilities. A shape memory filament and polyester yarn were knitted to produce a shape memory knitted fabric (SMF). Thermo-mechanical tensile test was carried out to quantify the memory behavior of SMF under different conditions. The experimental findings demonstrate excellent shape recovery (100%) and shape fixity up to 88% at different strains (20% and 60%) and temperatures (30 ℃ and 50 ℃). Experimental results reveal that memory filament behaves differently in a fabric structure than in its pristine condition at various temperatures and strains. The cycle test of SMF under different thermo-mechanical conditions indicated complete shape recovery with an increase in shape fixity. So, the utterly recoverable textile structure was achieved after a few initial cycles. These intelligent textiles are beneficial for the development of novel, innovative, and functional fabrics like elegant curtains, pressure garments, compression stockings, etc. In addition to fashion and medical uses, this unique feature may also be leveraged to build textile-based sensors and actuators.

Keywords: knitting, memory filament, shape memory, smart textiles, thermo-mechanical cycle

Procedia PDF Downloads 85

Authors: Alvarez M., Chapela M. J., Balboa E., Rubianes D., Sinde E., Fernandez de Ana C., Rodríguez-Blanco A.

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The gut microbiota plays an important role as gut inflammation could contribute to colorectal cancer development; however, this role is still not fully understood, and tools able to prevent this progression are yet to be developed. The main objective of this study was to monitor the effects of a mushroom extracts formulation in gut microbial community composition of an Azoxymethane (AOM)/Dextran sodium sulfate (DSS) mice model of inflammation-linked carcinogenesis. For the in vivo study, 41 adult male mice of the C57BL / 6 strain were obtained. 36 of them have been induced in a state of colon carcinogenesis by a single intraperitoneal administration of AOM at a dose of 12.5 mg/kg; the control group animals received instead of the same volume of 0.9% saline. DSS is an extremely toxic polysaccharide sulfate that causes chronic inflammation of the colon mucosa, favoring the appearance of severe colitis and the production of tumors induced by AOM. Induction by AOM/DSS is an interesting platform for chemopreventive intervention studies. This time the model was used to monitor gut microbiota changes as a result of supplementation with a specific mushroom extracts formulation previously shown to have prebiotic activity. The animals have been divided into three groups: (i) Cancer + mushroom extracts formulation experimental group: to which the MicoDigest2.0 mushroom extracts formulation developed by Hifas da Terra S.L has been administered dissolved in drinking water at an estimated concentration of 100 mg / ml. (ii) Control group of animals with Cancer: to which normal water has been administered without any type of treatment. (iii) Control group of healthy animals: these are the animals that have not been induced cancer or have not received any treatment in drinking water. This treatment has been maintained for a period of 3 months, after which the animals were sacrificed to obtain tissues that were subsequently analyzed to verify the effects of the mushroom extract formulation. A microbiological analysis has been carried out to compare the microbial communities present in the intestines of the mice belonging to each of the study groups. For this, the methodology of massive sequencing by molecular analysis of the 16S gene has been used (Ion Torrent technology). Initially, DNA extraction and metagenomics libraries were prepared using the 16S Metagenomics kit, always following the manufacturer's instructions. This kit amplifies 7 of the 9 hypervariable regions of the 16S gene that will then be sequenced. Finally, the data obtained will be compared with a database that makes it possible to determine the degree of similarity of the sequences obtained with a wide range of bacterial genomes. Results obtained showed that, similarly to certain natural compounds preventing colorectal tumorigenesis, a mushroom formulation enriched the Firmicutes and Proteobacteria phyla and depleted Bacteroidetes. Therefore, it was demonstrated that the consumption of the mushroom extracts’ formulation developed could promote the recovery of the microbial balance that is disrupted in the mice model of carcinogenesis. More preclinical and clinical studies are needed to validate this promising approach.

Keywords: carcinogenesis, microbiota, mushroom extracts, inflammation

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Authors: Gyati Shilakari Asthana, Abhay Asthana

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Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1 and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

Procedia PDF Downloads 491

Authors: Chen Chou, Feng-Tyan Lin

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Big Data has been attracted a lot of attentions in many fields for analyzing research issues based on a large number of maternal data. Electronic Toll Collection (ETC) is one of Intelligent Transportation System (ITS) applications in Taiwan, used to record starting point, end point, distance and travel time of vehicle on the national freeway. This study, taking advantage of ETC big data, combined with urban planning theory, attempts to explore various phenomena of inter-city transportation activities. ETC, one of government's open data, is numerous, complete and quick-update. One may recall that living area has been delimited with location, population, area and subjective consciousness. However, these factors cannot appropriately reflect what people’s movement path is in daily life. In this study, the concept of "Living Area" is replaced by "Influence Range" to show dynamic and variation with time and purposes of activities. This study uses data mining with Python and Excel, and visualizes the number of trips with GIS to explore influence range of Tainan city and the purpose of trips, and discuss living area delimited in current. It dialogues between the concepts of "Central Place Theory" and "Living Area", presents the new point of view, integrates the application of big data, urban planning and transportation. The finding will be valuable for resource allocation and land apportionment of spatial planning.

Keywords: Big Data, ITS, influence range, living area, central place theory, visualization

Procedia PDF Downloads 275

Authors: Mahsa Kallhor Mohammadi, Hamideh Reshadatjoo

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This study verified the relationship between in-service training and employees’ feeling of psychological ownership. This research applied a descriptive survey that investigated a correlation between variables. The target population was 140 employees of a Drilling Fluid and Waste Management Service Company, and the sample was 123 employees who were selected randomly and encouraged to complete an electronic questionnaire which was designed based on standard questionnaires for research variables covering 62 questions. The face validity of the questionnaire was supported by an experimental test, and its content validity was approved by the thesis supervisor and consulting advisor. For the descriptive statistics frequency tables and diagrams, measures of central tendency such as mode, median, and mean and measures of variability such as variance, standards deviation, and quartile deviation were used. In the inferential statistics section, the Pearson correlation coefficient was used to verify the relationship between the variables of the research. According to the results, all of the research hypotheses were supported. According to hypothesis 1, there was a positive and significant relationship between training policy-making and employees’ psychological ownership (r=0/408, α=0/05). According to hypothesis 2, there was a positive and significant relationship between training planning and employees’ psychological ownership (r=0/446, α=0/05). According to hypothesis 3, there was a positive and significant relationship between providing the training and employees’ psychological ownership (r=0/512, α=0/05). According to hypothesis 4, there was a positive and significant relationship between training performance management and employees’ psychological ownership (r=0/462, α=0/05). According to hypothesis 5, there was a positive and significant relationship between employees’ motivation and psychological ownership (r=0/694, α=0/05). Therefore, through systematic in-service training, which is in the same line with the strategic goals of an organization and is based on scientific needs analysis, design, implementation, and evaluation, it is possible to improve employees’ sense of psychological ownership toward an organization.

Keywords: in-service training, motivation, organizational behavior, psychological ownership

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Authors: Tamer M. Shehata, Heba S. Elsewedy, Mashel Al Dosary, Alaa Elshehry, Mohamed A. Khedr, Maged E. Mohamed

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Objective: 2,4-Dichlorophenoxy acetic acid (2,4-D) is a well-known herbicide widely used as a weed killer. Recently, 2,4-D was rediscovered as a new anti-inflammatory agent through in silico as well as in-vivo experiments. However, poor solubility of 2,4-D could represent a problems during pharmaceutical development in addition to lower bioavailability. Solid dispersion (SD) refers to a group of solid products consisting of at least two different components, usually a hydrophobic drug and hydrophilic matrix. It is well known technique for enhancing drug solubility. Therefore, selecting SD as a tool for enhancing 2,4-D could be of great interest to the formulator. Method: In our project, several polymers were investigated (such as PEG, HPMC, citric acid and others) in addition to drug polymer ratios and its effect on solubility. Evaluation of drug polymer interaction was investigated through both Fourier Transform Infrared (FTIR) and Differential Scanning Calorimetry (DSC). Finally, in-vivo evaluation was performed for the best selected preparation through inflammatory response of rat induce hind paw. Results: Results indicated that, citric acid 2,4-D and in ratio of 0.75 : 1 showed modified the dissolution profile of the drug. The FTIR resltes indicated no significant chemical interaction, however DSC showed shifting of the drug melting point. Finally, Carragenan induced rat hind paw edema showed significant reduction of the drug solid dispersion in comparison to the pure drug, indicating rapid and complete absorption of the drug in solid dispersion form. Conclusion: Solid dispersion technology can be utilized efficiently to enhance the solubility of 2,4-D.

Keywords: solid dispersion, 2, 4-D solubility, carragenan induced edema

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Authors: Sidika McNeil

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Borderline personality disorder (BPD) is a personality disorder that has been found to have strong origins in childhood trauma. One of the key symptoms of BPD is an association with irregular moods swings, as well as suicidal ideation (SI). Owing to the typically severe trauma patients experience during childhood, it is hard for them to control their emotions and thus makes it hard to emotionally regulate. It is then very common for those suffering from BPD to turn to unhealthy coping mechanisms, such as substance use, unhealthy relationships, and more, often unsuccessfully creating experiences that facilitate safety which leads to further negative experiences. With the high suicide rating among children, adolescents, and teens, and an ever-increasing number of children being diagnosed with BPD, it is very important that more research is done to find further treatments for patients who are currently suffering. Methods: Utilizing data found in prior studies, this paper will analyze the literature to focus on a comprehensive treatment plan for those with DBT. It is currently suggested that with the use of dialectical behavioral therapy (DBT), a therapy that focuses on changing negative thinking patterns and pushes for more positive ones is helpful for treatment for those with BPD. Though this therapy is not a cure to BPD, it does help mitigate the risk; this essay will explore other options that can further the treatment process, such as cognitive analytical therapy (CAT), which focuses on delving into the past to find the root causes of an issue to create coping strategies and harm reduction, a type of therapy used to aid patients in lowering the use of substances without complete cessation. Results: The research provides enough evidence to link between the treatment of BPD with the utilization of CAT.

Keywords: borderline personality disorder, cognitive analytical therapy, dialectical behavioral therapy, harm reduction, suicidal ideation

Procedia PDF Downloads 172

Authors: Nilanchali Singh, Alka Kriplani, Reeta Mahey, Garima Kachhawa

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Objective: Resection of larger uterine septa does improve obstetric performance but whether smaller septa need resection and their impact on obstetric outcome is not clear. We wanted to evaluate the role of septal resection of septa of various sizes in obstetric performance. Methods: This retrospective cohort study comprised of 107 patients with uterine septum. The patients were categorized on the basis of extent of uterine septum into four groups: a) Subsepta (< 1/3rd), b) Septum > 1/3 to ½, c) Septum>1/2 to whole uterine cervix, d) Septum traversing whole of uterine cavity and cervix. Out of these 107 patients, 74 could be contacted telephonically and outcomes recorded. Sensitivity and specificity of investigative modalities were calculated. Results: Infertility was seen in maximum number of cases in complete septa (100%), whereas abortions were seen more commonly, in subsepta (18%). MRI had maximum sensitivity and positive predictive value, followed by hysteron-salpingography. Tubal block, fibroid, endometriosis, pelvic adhesions, ovarian pathologies were seen in some but no definite association of these pathologies was seen with any subgroup of septa. Almost five-year follow-up was recorded in all the subgroups. Significant reduction in infertility was seen in all septal subgroup (p=0.046, 0.032 & 0.05) patients except in subsepta (< 1/3rd uterine cavity) after septum resection. Abortions were significantly reduced (p=0.048) in third subgroup (i.e. septum > ½ to upto internal os) after hysteroscopic septum resection. Take home baby rate was 33% in subsepta and around 50% in the remaining subgroups of septa. Conclusions: Septal resection improves obstetric performance in patients with uterine septa of various sizes. Whether septal resection improves obstetric performance in patients with subsepta or very small septa, is controversial. Larger studies addressing this issue need to be planned.

Keywords: septal resection, obstetric outcome, infertility, septum size

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Authors: Mohammed I.A. Al-Neemi, Mohammed S.B., Al-Hlawee, Ilham N. Ezaddin, Soz A. Faris, Omer E. Fakhry, Heemen S. Mageed

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This objective of this study was to measure the effect of replacing different levels of animal protein concentrate with Red Tiger shrimp meal (RTSM: 60 % crude protein, 2400 M.E kcal/kg and the source of RTSM was imported from china) in the broiler starter diets. A total 300 broiler chicks (Ross-308) were randomly assigned in treatments dietary contained three different levels of RTSM (0.00, 4.16 and 8.32 %) in experimental diet with a completely randomized design (CRD). Each treatment included four replicates (floor pens) and 25 broilers in each replication (Pen). Therefore, floor space for each boilers was 900 cm2. Initially, the broilers where exposed to a continues lighting of 23:30 hours and dark period of 30 minutes in each 24 hours. Feed and water were supplied ad libitum to the broilers throughout the experimental period (1-21 days). The results of this study indicated that body weight (B.W.), body weight gain (B.W.G), conversion ratio of feed, protein and energy (F.CR, P.C.R and E.C.R) were significantly (p ≤ 0.05) decreased by complete substituting (RTSM) for animal protein concentration (third treatment). Mortality percentage significantly (p ≤ 0.05) increased for third dietary treatment. No significant differences were found for feed, protein and energy intake among treatments during the experimental period (three weeks). In conclusion, (RTSM) could be included to 4.16% in the broiler starter diet or substitute the protein Red Tiger shrimp as alternative of protein animal protein concentrate as much as 50%.

Keywords: red tiger shrimp, broiler, starter diet, growth performance, animal protein concentrate

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Authors: Lakmali Anthony, Madeline Gillies

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Purpose: Decreasing mortality has led to increased focus on patient-reported outcomes such as quality of life (QoL) in breast cancer. Breast cancer patients often have decreased QoL even after treatment is complete. This systematic review of the literature aims to identify psychosocial factors associated with decreased QoL in post-treatment breast cancer patients. Methodology: This systematic review was performed in accordance with the 2020 Preferred Reporting Items for Systematic Reviews and Meta-Analyses recommendations. The search was conducted in MEDLINE, EMBASE, and PsychINFO using MeSH headings. The two authors screened studies for relevance and extracted data. Results: Seventeen studies were identified, including 3,150 total participants (mean = 197) with a mean age of 51.9 years. There was substantial heterogeneity in measures of QoL. The most common was the European Organisation for Research and Treatment of Cancer QLQ-C30 (n=7, 41.1%). Most studies (n=12, 70.5%) found that emotional distress correlated with poor QoL, while 3 found no significant association. The most common measure of emotional distress was the Hospital Anxiety and Depression Scale (n=12, 70.5%). Other psychosocial factors associated with QoL were unmet needs, problematic social support, and negative affect. Clinicopathologic determinants included mastectomy without reconstruction, stage IV disease, and adjuvant chemotherapy. Conclusion: This systematic review provides a summary of the psychosocial determinants of poor QoL in post-treatment breast cancer patients, as well as the most commonly reported measures of these. An understanding of these potentially modifiable determinants of poor outcome is pivotal to the provision of quality, patient-centred care in surgical oncology.

Keywords: breast cancer, quality of life, psychosocial determinants, cancer surgery

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Authors: K. H. Oyuntungalag, Scott Kenner, O. Erdenetuya

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The life cycle phases of mining projects are described in this guidance document, and includes initial phases (exploration, feasibility and planning), mine development (construction and operations), closure and reclamation. Initial phases relate to field programs and desktop studies intended to build the data and knowledge base, including the design of water management infrastructure and development during these initial phases. Such a model is essential to demonstrate that the water management plan (WMP) will provide adequate water for the mine operations and sufficient capacity for anticipated flows and volumes, and minimize environmental impacts on the receiving environment. The water and mass balance model must cover the whole mine life cycle, from the start of mine development to a date sufficiently far in the future where the reclaimed landscape is considered self- sustaining following complete closure of the mine (i.e., post- closure). The model simulates the movement of water within the components of the water management infrastructure and project operating areas, and calculates chemical loadings to each mine component. At Erdenet Mining company an initial water balance model reflecting the tailings dam, groundwater seepage and mine process water was developed in collaboration with Dr. Scott Kenner (visiting Fulbright scholar). From this preliminary study the following recommendations were made: 1. Develop a detailed groundwater model to simulate seepage from the tailings dam, 2. Establish an evaporation pan for improving evapotranspiration estimates, and 3. Measure changes in storage of water within the tailings dam and other water storage components within the mine processing.

Keywords: evapotranspiration , monitoring program, Erdenet mining, tailings dam

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Authors: Bhavatharani S., Boopathy V., Kavin S., Naveethkumar R.

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Context: Resistance training with blood flow restriction (BFR) has increased in clinical rehabilitation due to the substantial benefits observed in augmenting muscle mass and strength using low loads. However, there is a great variability of training pressures for clinical populations as well as methods to estimate it. The aim of this study was to estimate the percentage of maximal BFR that could result by applying different methodologies based on arbitrary or individual occlusion levels using a cuff width between 9 and 13 cm. Design: A secondary analysis was performed on the combined databases of 2 previous larger studies using BFR training. Methods: To estimate these percentages, the occlusion values needed to reach complete BFR (100% limb occlusion pressure [LOP]) were estimated by Doppler ultrasound. Seventy-five participants (age 24.32 [4.86] y; weight: 78.51 [14.74] kg; height: 1.77 [0.09] m) were enrolled in the laboratory study for measuring LOP in the thigh, arm, or calf. Results: When arbitrary values of restriction are applied, a supra-occlusive LOP between 120% and 190% LOP may result. Furthermore, the application of 130% resting brachial systolic blood pressure creates a similar occlusive stimulus as 100% LOP. Conclusions: Methods using 100 mm Hg and the resting brachial systolic blood pressure could represent the safest application prescriptions as they resulted in applied pressures between 60% and 80% LOP. One hundred thirty percent of the resting brachial systolic blood pressure could be used to indirectly estimate 100% LOP at cuff widths between 9 and 13 cm. Finally, methodologies that use standard values of 200 and, 300 mm Hg far exceed LOP and may carry additional risk during BFR exercise.

Keywords: lower limb rehabilitation, ESP32, pneumatics for medical, programmed rehabilitation

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Authors: Akaki Qatamidze

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Implementation of the project will be a step forward in terms of reliability in Georgia and the improvement of the construction and the development of construction. Completion of the project is expected to result in a complete knowledge, which is expressed in concrete and stone structures of assessing the technical condition of the processing. This method is based on a detailed examination of the structure, in order to establish the injuries and the elimination of the possibility of changing the structural scheme of the new requirements and architectural preservationists. Reinforced concrete and stone structures research project carried out in a systematic analysis of the important approach is to optimize the process of research and development of new knowledge in the neighboring areas. In addition, the problem of physical and mathematical models of rational consent, the main pillar of the physical (in-situ) data and mathematical calculation models and physical experiments are used only for the calculation model specification and verification. Reinforced concrete and stone construction defects and failures the causes of the proposed research to enhance the effectiveness of their maximum automation capabilities and expenditure of resources to reduce the recommended system analysis of the methodological concept-based approach, as modern science and technology major particularity of one, it will allow all family structures to be identified for the same work stages and procedures, which makes it possible to exclude subjectivity and addresses the problem of the optimal direction. It discussed the methodology of the project and to establish a major step forward in the construction trades and practical assistance to engineers, supervisors, and technical experts in the construction of the settlement of the problem.

Keywords: building, reinforced concrete, expertise, stone structures

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Authors: Shariful Islam, Sharun Akter Khushbu, S. M. Shaqib, Shahriar Sultan Ramit

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In the construction sector, ensuring worker safety is of the utmost significance. In this study, a deep learning-based technique is presented for identifying safety gear worn by construction workers, such as helmets, goggles, jackets, gloves, and footwear. The suggested method precisely locates these safety items by using the YOLO v7 (You Only Look Once) object detection algorithm. The dataset utilized in this work consists of labeled images split into training, testing and validation sets. Each image has bounding box labels that indicate where the safety equipment is located within the image. The model is trained to identify and categorize the safety equipment based on the labeled dataset through an iterative training approach. We used custom dataset to train this model. Our trained model performed admirably well, with good precision, recall, and F1-score for safety equipment recognition. Also, the model's evaluation produced encouraging results, with a [email protected] score of 87.7%. The model performs effectively, making it possible to quickly identify safety equipment violations on building sites. A thorough evaluation of the outcomes reveals the model's advantages and points up potential areas for development. By offering an automatic and trustworthy method for safety equipment detection, this research contributes to the fields of computer vision and workplace safety. The proposed deep learning-based approach will increase safety compliance and reduce the risk of accidents in the construction industry.

Keywords: deep learning, safety equipment detection, YOLOv7, computer vision, workplace safety

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Authors: Ana-Katarina Nikich

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Background: The kinesiotaping method is often used in physiotherapy and rehabilitation. The purpose of this study was to evaluate the effectiveness of taping in the rehabilitation process of patients. Materials and methods: The study involved 90 male and female patients (the average age was 40-50 years) with various conditions requiring rehabilitation, such as injuries of the musculoskeletal system, sports injuries and other ailments. All patients were divided into two groups: experimental (n=40) and control (n=50). Both groups received 20 days of standard rehabilitation. In the experimental group, kinesiotaping methods were used, taking into account the individual characteristics of each patient. The control group performed regular exercises and physical therapy, but without using kinesiotape. During the study, physical parameters were monitored, interviews were conducted and the conditions of patients from both groups were compared. Results and discussion: The use of the kinesiotaping method in the rehabilitation process led to a significant improvement in physical parameters and pain reduction in patients. Significant improvement (p <0.005) was observed in all evaluated parameters among the patients of the experimental group. The control group also showed sufficient improvement (p <0.005), but the percentage of the experimental group was higher. As a result of the observation, the patients of the experimental group showed faster and more complete rehabilitation compared to the control group. The use of the kinesiotaping method allows to reduce the load on the damaged areas, improve blood circulation and lymphatic drainage, as well as increase stability and coordination of movements. Conclusions: Kinesiotaping as one of the modern therapeutic methods has shown its effectiveness in the rehabilitation process, contributing to the optimal recovery of patients with various conditions requiring rehabilitation. The use of tapes should be included in a comprehensive rehabilitation program to achieve the best results and reduce recovery time.

Keywords: kinesiotaping, rehabilitation, therapy, pain

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Authors: Nipun Ansal

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People guard their beliefs and opinions with their lives. Beliefs that they’ve formed over a period of time, and can go to any lengths to defy, desist from, resist and negate any outward stimulus that has the potential to shake them. Cognitive dissonance is term used to describe it in theory. And every human being, in order to defend himself from cognitive dissonance applies 4 rings of defense viz. Selective Exposure, Selective Perception, Selective Attention, and Selective Retention. This paper is a discursive analysis on how the onslaught of social media, complete with its intrusive weaponry, has amorphized the external ring of defense: the selective exposure. The stimulus-response model of communication is one of the most inherent model that encompasses communication behaviours of children and elderly, individual and masses, humans and animals alike. The paper deliberates on how information bombardment through the uncontrollable channels of the social media, Facebook and Twitter in particular, have dismantled our outer sphere of exposure, leading users online to a state of constant dissonance, and thus feeding impulsive action-taking. It applies case study method citing an example to corroborate how knowledge generation has given in to the information overload and the effect it has on decision making. With stimulus increasing in number of encounters, opinion formation precedes knowledge because of the increased demand of participation and decrease in time for the information to permeate from the outer sphere of exposure to the sphere of retention, which of course, is through perception and attention. This paper discusses the challenge posed by this fleeting, stimulus rich, peer-dominated media on the traditional models of communication and meaning-generation.

Keywords: communication, discretion, exposure, social media, stimulus

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Authors: Walid Tawfik

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The interaction of high intensity ultrashort laser pulses with plasma generates many significant applications, including soft x-ray lasers, time-resolved laser induced plasma spectroscopy LIPS, and laser-driven accelerators. The development in producing of femtosecond down to ten femtosecond optical pulses has facilitates scientists with a vital tool in a variety of ultrashort phenomena, such as high field physics, femtochemistry and high harmonic generation HHG. In this research, we generate a two-octave-wide ultrashort supercontinuum pulses with an optical spectrum extending from 3.5 eV (ultraviolet) to 1.3 eV (near-infrared) using a capillary fiber filled with neon gas. These pulses are formed according to nonlinear self-phase modulation in the neon gas as a nonlinear medium. The investigations of the created pulses were made using spectral phase interferometry for direct electric-field reconstruction (SPIDER). A complete description of the output pulses was considered. The observed characterization of the produced pulses includes the beam profile, the pulse width, and the spectral bandwidth. After reaching optimization conditions, the intensity of the reconstructed pulse autocorrelation function was applied for the shorts pulse duration to achieve transform limited ultrashort pulses with durations below 6-fs energies up to 600μJ. Moreover, the effect of neon pressure variation on the pulse width was examined. The nonlinear self-phase modulation realized to be increased with the pressure of the neon gas. The observed results may lead to an advanced method to control and monitor ultrashort transit interaction in femtochemistry.

Keywords: supercontinuum, ultrafast, SPIDER, ultra-broadband

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Authors: Sriram Venkatesh, V. Murali Mohan, T. V. Karthikeyan

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The main advantage of multi-directionally reinforced composites is the freedom to orient selected fibre types and hence derives the benefits of varying fibre volume fractions and there by accommodate the design loads of the final structure of composites. This technology provides the means to produce tailored composites with desired properties. Due to the high level of fibre integrity with through thickness reinforcement those composites are expected to exhibit superior load bearing characteristics with capability to carry load even after noticeable and apparent fracture. However a survey of published literature indicates inadequacy in the design and test data base for the complete characterization of the multidirectional composites. In this paper the research objective is focused on the development and testing of 4-D orthogonal composites with different preform configurations and resin systems. A preform is the skeleton 4D reinforced composite other than the matrix. In 4-D preforms fibre bundles are oriented in three directions at 1200 with respect to each other and they are on orthogonal plane with the fibre in 4th direction. This paper addresses the various types of 4-D composite manufacturing processes and the mechanical test methods followed for the material characterization. A composite analysis is also made, experiments on course and fine woven preforms are conducted and the findings of test results are discussed in this paper. The interpretations of the test results reveal several useful and interesting features. This should pave the way for more widespread use of the perform configurations for allied applications.

Keywords: multi-directionally reinforced composites, 4-D orthogonal preform, course weave, fine weave, fibre bundle spools, unit cell, fibre architecture, fibre volume fraction, fibre distribution

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Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

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Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

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Authors: Giorgio Visentin, Alexei A. Buchachenko

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Recent proposals to use Yb dimer as an optical clock and as a sensor for non-Newtonian gravity imply the knowledge of its interaction potential. Here, the ground-state Born-Oppenheimer Yb₂ potential energy curve is represented by a semi-analytical function, consisting of short- and long-range contributions. For the former, the systematic ab initio all-electron exact 2-component scalar-relativistic CCSD(T) calculations are carried out. Special care is taken to saturate diffuse basis set component with the atom- and bond-centered primitives and reach the complete basis set limit through n = D, T, Q sequence of the correlation-consistent polarized n-zeta basis sets. Similar approaches are used to the long-range dipole and quadrupole dispersion terms by implementing the CCSD(3) polarization propagator method for dynamic polarizabilities. Dispersion coefficients are then computed through Casimir-Polder integration. The semiclassical constraint on the number of the bound vibrational levels known for the ¹⁷⁴Yb isotope is used to scale the potential function. The scaling, based on the most accurate ab initio results, bounds the interaction energy of two Yb atoms within the narrow 734 ± 4 cm⁻¹ range, in reasonable agreement with the previous ab initio-based estimations. The resulting potentials can be used as the reference for more sophisticated models that go beyond the Born-Oppenheimer approximation and provide the means of their uncertainty estimations. The work is supported by Russian Science Foundation grant # 17-13-01466.

Keywords: ab initio coupled cluster methods, interaction potential, semi-analytical function, ytterbium dimer

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Authors: Robert G. Nini

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Soil consolidation is one of the biggest problem facing engineers. The consolidation process has an important role in settlement analysis for the embankments and footings resting on clayey soils. The settlement amount is related to the compression and the swelling indexes of the soil. Because the predominant upper soil layer in Lebanon is consisting mainly of clay, this layer is a real challenge for structural and highway engineering. To determine the effect of load and drainage on the engineering consolidation characteristics of Lebanese soil, a full experimental and synthesis study was conducted on different soil samples collected from many locations. This study consists of two parts. During the first part which is an experimental one, the Proctor test and the consolidation test were performed on the collected soil samples. After it, the identifications soil tests as hydrometer, specific gravity and Atterberg limits are done. The consolidation test which is the main test in this research is done by loading the soil for some days then an unloading cycle was applied. It takes two weeks to complete a typical consolidation test. Because of these reasons, during the second part of our research which is based on the analysis of the experiments results, some correlations were found between the main consolidation parameters as compression and swelling indexes with the other soil parameters easy to calculate. The results show that the compression and swelling indexes of Lebanese clays may be roughly estimated using a model involving one or two variables in the form of the natural void ratio and the Atterberg limits. These correlations have increasing importance for site engineers, and the proposed model also seems to be applicable to a wide range of clays worldwide.

Keywords: atterberg limits, clay, compression and swelling indexes, settlement, soil consolidation

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Authors: Salama A. Ouf, Amera A. El-Adly, Abdelaleam H. Mohamed

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Dermatophytosis is the infection of keratinized tissues such as hair, nail and the stratum corneum of the skin by dermatophytic fungi. Infection is generally cutaneous and restricted to the non-living cornified layers because of the inability of the fungi to penetrate the deeper tissues or organs of immunocompetent hosts. In Saudi Arabia, Onychomycosis is the most frequent infection (40.3%), followed by tinea capitis (21.9%), tinea pedis (16%), tinea cruris (15.1%), and tinea corporis (6.7%). Several azole compounds have been tried to control dermatophytic infection, however, the azole-containing medicines may interfere with the activity of hepatic microsomal enzymes, sex and thyroid hormones, and testosterone biosynthesis. In this research, antibody-conjugated nanoparticles stimulated by cold plasma and laser were evaluated in vitro against some dermatophytes isolated from the common types of tinea. Different types of nanomaterials were tested but silver nanoparticles (AgNPs) were proved to be most effective against the dermatophytes under test. The use of cold plasma coupled with antibody-conjugated nano-particles has severe impact on dermatophytes where the inhibition of growth, spore germination keratinase activity was more than 88% in the case of Trichophyton rubrum, T. violaceum, Microsprum canis and M. gypseum. Complete inhibition of growth for all dermatophytes was brought about by the interaction of conjugated nanoparticles, with cold plasma and laser treatment. The in vivo test with inoculated guinea pigs achieved promising results where the recovery from the infection reached 95% in the case of M. canis –inoculated pigs treated with AgNPs pretreated with cold plasma and laser.

Keywords: cold plasma, dermatophytes, laser, silver nanoparticles

Procedia PDF Downloads 364

Authors: A. Kherraf, S. Ouarrak, L. Azzouzi, R. Habbal

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Introduction: Preserved LVEF heart failure is an important cause of mortality and morbidity in hypertensive patients. A strong correlation between impaired diastolic function and longitudinal systolic dysfunction. could have several explanations, first, the diastole is an energy dependent process, especially during its first phase, it also includes active systolic components during the phase of iso volumetric relaxation, in addition, the impairment of the intrinsic myocytic function is part of hypertensive pathology as evidenced by recent studies. METHODS AND MATERIALS: This work consists of performing in a series of 333 hypertensive patients (aged 25 to 75 years) a complete echocardiographic study, including LVEF by Simpson biplane method, the calculation of the indexed left ventricular mass, the analysis of the diastolic function, and finally, the study of the longitudinal deformation of the LV by the technique of speckletracking (calculation of the GLS). Patients with secondary hypertension, leaky or stenosing valve disease, arrhythmia, and a history of coronary insufficiency were excluded from this study. RESULTS: Of the 333 hypertensive patients, 225 patients (67.5%) had impaired diastolic function, of which 60 patients (18%) had high filling pressures. 49.39% had echocardigraphic HVG, Almost all of these patients (60 patients) had low GLS. There is a statistically very significant relationship between lower GLS and increased left ventricular filling pressures in hypertensive patients. These results suggest that increased filling pressures are closely associated with atrioventricular interaction in patients with hypertension, with a strong correlation with impairment of longitudinal systolic function and diastolic function CONCLUSION: Overall, a linear relationship is established between increased left ventricular mass, diastolic dysfunction, and longitudinal LV systolic dysfunction

Keywords: hypertension, diastolic function, left ventricle, heart failure

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Authors: Mohsin Raza, Rizwan Latif, Syed Adnan Qasim, Imran Shafi

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High torque low-speed diesel engine has a wide range of industrial and commercial applications. In literature, it’s found that lot of work has been done for the high-speed diesel engine and research on High Torque low-speed is rare. The fuel injection plays a key role in the efficiency of engine and reduction in exhaust emission. The fuel breakup plays a critical role in air-fuel mixture and spray combustion. The current study explains numerically an important phenomenon in spray combustion which is deformation and breakup of liquid drops in compression ignition internal combustion engine. The secondary breakup and its influence on spray and characteristics of compressed gas in-cylinder have been calculated by using simulation software in the backdrop of high torque low-speed diesel like conditions. The secondary spray breakup is modeled with KH - RT instabilities. The continuous field is described by turbulence model and dynamics of the dispersed droplet is modeled by Lagrangian tracking scheme. The results by using KH - RT model are compared against other default methods in OpenFOAM and published experimental data from research and implemented in CFD (Computational Fluid Dynamics). These numerical simulation, done in OpenFoam and Matlab, results are analyzed for the complete 720- degree 4 stroke engine cycle at a low engine speed, for favorable agreement to be achieved. Results thus obtained will be analyzed for better evaporation in near nozzle region. The proposed analyses will further help in better engine efficiency, low emission and improved fuel economy.

Keywords: diesel fuel, KH-RT, Lagrangian , Open FOAM, secondary breakup

Procedia PDF Downloads 262

Authors: F. Zouai, F. Z. Benabid, S. Bouhelal, D. Benachour

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Reinforced plastics or nanocomposites have attracted considerable attention in scientific and industrial fields because a very small amount of clay can significantly improve the properties of the polymer. The polymeric matrices used in this work are two saturated polyesters, i.e., polyethylene terephthalate (PET) and polyethylene naphthalate (PEN). The success of processing compatible blends, based on poly(ethylene terephthalate) (PET)/poly(ethylene naphthalene) (PEN)/clay nanocomposites in one step by reactive melt extrusion is described. Untreated clay was first purified and functionalized ‘in situ’ with a compound based on an organic peroxide/ sulfur mixture and (tetramethylthiuram disulfide) as the activator for sulfur. The PET and PEN materials were first separately mixed in the molten state with functionalized clay. The PET/4 wt% clay and PEN/7.5 wt% clay compositions showed total exfoliation. These compositions, denoted nPET and nPEN, respectively, were used to prepare new n(PET/PEN) nanoblends in the same mixing batch. The n(PET/PEN) nanoblends were compared to neat PET/PEN blends. The blends and nanocomposites were characterized using various techniques. Microstructural and nanostructural properties were investigated. Fourier transform infrared spectroscopy (FTIR) results showed that the exfoliation of tetrahedral clay nanolayers is complete, and the octahedral structure totally disappears. It was shown that total exfoliation, confirmed by wide-angle X-ray scattering (WAXS) measurements, contributes to the enhancement of impact strength and tensile modulus. In addition, WAXS results indicated that all samples are amorphous. The differential scanning calorimetry (DSC) study indicated the occurrence of one glass transition temperature Tg, one crystallization temperature Tc and one melting temperature Tm for every composition.

Keywords: exfoliation, DRX, DSC, montmorillonite, nanocomposites, PEN, PET, plastograph, reactive melt-mixing

Procedia PDF Downloads 322

Authors: Fahsai Pawapootanont, Sasithon Yuwakosol

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The objectives of this research study are as follows: 1) Study the information-seeking behavior of followers of influencers on Facebook in making hotel booking decisions and 2) Study the characteristics of travel influencers that affect their followers' hotel booking decisions. The Data was collected by interviewing 35 key informants, consisting of 25 Thai tourists who were followers of travel influencers and 10 travel influencers, as well as collecting data using online questionnaires from a sample of 400 Thai tourists and using statistical data analysis: percentage, standard deviation, mean, T-Test and One-Way Analysis of Variance: ANOVA. The results of the influence of travel influencers on Facebook on hotel booking decisions in Thailand revealed the following: People in different age groups have different information-seeking behaviours. Depends on experience and aptitude in using technology. The sample group did not seek information from only one source. There is also a search for information from various places in order to get comparative information and the most truthful information to make decisions. In addition, travel influencers should be those who present honest, clear, and complete content. And present services honestly. In addition to the characteristics of travel influencers affecting hotel booking decisions, Presentation formats and platforms also affect hotel booking decisions. But it must be designed and presented to suit the behavior of the group of people we want. As for the influence of travel influencers, it can be concluded that The influence of travel influencers can influence their followers' interests and hotel booking decisions. However, it was found that there are other factors that followers of travel influencers on Facebook will factor into their decision to book a hotel, such as Whether the hotel's comfort meets your needs or not; location, price, and promotions also play an important role in deciding to book a hotel.

Keywords: influencer, travel, facebook, hotel booking decisions, Thailand

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Authors: Aussara Panya, Nunghathai Sawasdee, Mutita Junking, Chatchawan Srisawat, Kiattawee Choowongkomon, Pa-Thai Yenchitsomanus

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Dengue virus (DENV) infection is a global public health problem with approximately 100 million infected cases a year. Presently, there is no approved vaccine or effective drug available; therefore, the development of anti-DENV drug is urgently needed. The clinical reports revealing the positive association between the disease severity and viral titer has been reported previously suggesting that the anti-DENV drug therapy can possibly ameliorate the disease severity. Although several anti-DENV agents showed inhibitory activities against DENV infection, to date none of them accomplishes clinical use in the patients. The surface envelope (E) protein of DENV is critical for the viral entry step, which includes attachment and membrane fusion; thus, the blocking of envelope protein is an attractive strategy for anti-DENV drug development. To search the safe anti-DENV agent, this study aimed to search for novel peptide inhibitors to counter DENV infection through the targeting of E protein using a structure-based in silico design. Two selected strategies has been used including to identify the peptide inhibitor which interfere the membrane fusion process whereby the hydrophobic pocket on the E protein was the target, the destabilization of virion structure organization through the disruption of the interaction between the envelope and membrane proteins, respectively. The molecular docking technique has been used in the first strategy to search for the peptide inhibitors that specifically bind to the hydrophobic pocket. The second strategy, the peptide inhibitor has been designed to mimic the ectodomain portion of membrane protein to disrupt the protein-protein interaction. The designed peptides were tested for the effects on cell viability to measure the toxic to peptide to the cells and their inhibitory assay to inhibit the DENV infection in Vero cells. Furthermore, their antiviral effects on viral replication, intracellular protein level and viral production have been observed by using the qPCR, cell-based flavivirus immunodetection and immunofluorescence assay. None of tested peptides showed the significant effect on cell viability. The small peptide inhibitors achieved from molecular docking, Glu-Phe (EF), effectively inhibited DENV infection in cell culture system. Its most potential effect was observed for DENV2 with a half maximal inhibition concentration (IC50) of 96 μM, but it partially inhibited other serotypes. Treatment of EF at 200 µM on infected cells also significantly reduced the viral genome and protein to 83.47% and 84.15%, respectively, corresponding to the reduction of infected cell numbers. An additional approach was carried out by using peptide mimicking membrane (M) protein, namely MLH40. Treatment of MLH40 caused the reduction of foci formation in four individual DENV serotype (DENV1-4) with IC50 of 24-31 μM. Further characterization suggested that the MLH40 specifically blocked viral attachment to host membrane, and treatment with 100 μM could diminish 80% of viral attachment. In summary, targeting the hydrophobic pocket and M-binding site on the E protein by using the peptide inhibitors could inhibit DENV infection. The results provide proof of-concept for the development of antiviral therapeutic peptide inhibitors to counter DENV infection through the use of a structure-based design targeting conserved viral protein.

Keywords: dengue virus, dengue virus infection, drug design, peptide inhibitor

Procedia PDF Downloads 351