Search results for: genomic regions of differences

Authors: Yun Yu Stephanie Law

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Sociometer Theory is well tested among young adults in western population, however, limited research is found for other age groups, like adolescent and middle-adulthood in Asia population. Thus, one of the main purposes of this study is to verify the validity of Sociometer Theory in different age groups among Asian. To be specific, we hypothesized that an increase in one’s perceived social rejection is associated to a decrease in his/her state self-esteem among all age groups in Asian population. And we expected that this association can be found among all age groups including adolescent, young adults and middle-adults group in our first study. In this way, we can verify the validity of Sociometer Theory across different age groups as well as its significance in Asian population. Furthermore, those participants who received rejection about ‘mate-role’ would also receive some negative feedbacks regarding their current/future capacity of being a good mate. Results suggested that participants’ state self-esteem sensitivity for mating-capacity rejection is higher when comparing to that of friend-capacity rejection, i.e. greater drop in state self-esteem when receiving mating-capacity feedbacks then receiving friend-capacity feedbacks. These results, however, is just applicable on young adults. Thus, the main purpose of study two would be testing the state self-esteem sensitivity towards social rejection in different domains among three age groups. We hypothesized that group differences would be found for three age groups regarding state self-esteem sensitivity. Research question 1: perceived social rejection is associated to decrease in state self-esteem, is applicable among different age groups in Asia population. Research question 2: there are significant group differences for three age groups regarding state self-esteem sensitivity. Methods: 300 subjects are divided into three age groups, adolescents group, young adult group and middle-adult group, with 100 subjects in each group. Two questionnaires were used in testing this fundamental concept. Subjects were then asked to rate themselves on questionnaire in measuring their current state self-esteem in order to obtain the baseline measurements for later comparison. In order to avoid demand characteristics from subjects, other unrelated tasks like word matching were also given after the first test. Results: A positive correlation between scores in questionnaire 1 and questionnaire 2 among all age groups. Conclusion: State self-esteem decrease to both imagined social rejection (study1) and experienced social rejection (study2). Moreover, level of decrease in state self-esteem vary when receiving different domains of social rejection. Implications: a better understanding of self-esteem development for various age group might bring insights for education systems and policies for teaching approaches and learning methods among different age groups.

Keywords: state self-esteem, social rejection, stage theory, self-feelings

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Authors: Pejman Taghibeikzadehbadr

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Muscle contraction stimulates a transient change of myogenic factors, partly related to the mode of contractions. Here, we assessed the response of Insulin-like growth factor 1Ea (IGF-1Ea), Insulin-like growth factor 1Eb (IGF-1Eb), Insulin-like growth factor 1Ec (IGF-1Ec), Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α-1), Peroxisome proliferator-activated receptor gamma coactivator 4-alpha (PGC1α-4), and myostatin to the eccentric Vs the concentric contraction in human skeletal muscle. Ten healthy males were performed an acute eccentric and concentric exercise bout (n = 5 per group). For each contraction type, participants performed 12 sets of 10 repetitions knee extension by the dominant leg. Baseline and post-exercise muscle biopsy were taken 4 weeks before and immediately after experimental sessions from Vastus Lateralis muscle. Genes expression was measured by real-time PCR technique. There was a significant increase in PGC1α-1, PGC1α-4, IGF-1Ea and, IGF-1Eb mRNA after concentric contraction (p ≤ 0.05), while the PGC1α-4 and IGF-1Ec significantly increased after eccentric contraction (p ≤ 0.05). It is intriguing to highlight that; no significant differences between groups were evident for changes in any variables following exercise bouts (p ≥ 0.05). Our results found that concentric and eccentric contractions presented different responses in PGC1α-1, IGF-1Ea, IGF-1Eb, and IGF-1Ec mRNA. However, a similar significant increase in mRNA content was observed in PGC1α-4. Further, no apparent differences could be found between the response of genes to eccentric and concentric contraction.

Keywords: eccentric contraction, concentric contraction, gene expression, PGC-1 alpha, IGF-1 Myostatin

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Authors: Theresa Campbell, Camille Bowen-Forbes, William Aalbersberg

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Differences in the sensory properties of two subtly distinct varieties of Rubus rosifolius lead to the examination of their anthocyanin, essential oil and polyphenol profiles. In both cases, notable differences were identified. Pelargonidin-3-rhutinoside (17.2 mg/100 g FW) and Cyanidin-3-glucoside (66.2 mg/100g FW) proved to be the dominant anthocyanins in the red and wine red varieties respectively. Linalool and terpineol were the major constituents of the essential oil from the red variety; however, those of the wine red variety are unidentified. In regard to phenolic compounds, caffeic acid and quercetin were in a higher concentration in the red variety (1.85 and 0.73 mg/100g FW respectively, compared to 1.22 and 0.34 mg/100g FW respectively in the wine red fruits); while ellagic acid and ferulic acid were of a higher concentration in the wine red variety (0.92 and 0.84mg/100g FW respectively, compared to 0.15 and 0.48 mg/100g FW respectively in the red variety). The methanol extract of both fruit varieties showed great antioxidant activity. Analysis of the antimicrobial activity of the fruit extracts against the growth of drug resistant pathogens revealed that they are active against methicillin resistant S. aureus (MRSA), rifampicin resistant S. aureus (RRSA), wild-type S. aureus (WTSA) and vancomycin-resistant Enterococcus faecium (VREF). Activity was also reported against several food-borne pathogens including two strains of E. coli, L. monocytogenes and Enterobacter aerogenes. The cytotoxicity of the various extracts was assessed and the essential oil extracts exhibited superior activity. The phenolic composition and biological activity of the fruits indicate that their consumption is beneficial to health and also that their incorporation into functional foods and nutraceuticals should be considered.

Keywords: phytochemicals, antimicrobial, cytotoxic, Rubus rosifolius

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Authors: Ayman El Sabagh, Celaleddin Barutçular, Hirofumi Saneoka

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Drought is one of the most serious problems posing a grave threat to cereals production including maize. Maize improvement in drought-stress tolerance poses a great challenge as the global need for food and bio-enegry increases. Thus, the current study was planned to explore the variations and determine the performance of target traits of maize hybrids at grain growth stage under drought conditions during 2014 under Adana, Mediterranean climate conditions, Turkey. Maize hybrids (Sancia, Indaco, 71May69, Aaccel, Calgary, 70May82, 72May80) were evaluated under (irrigated and water stress). Results revealed that, grain yield and yield traits had a negative effects because of water stress conditions compared with the normal irrigation. As well as, based on the result under normal irrigation, the maximum biological yield and harvest index were recorded. According to the differences among hybrids were found that, significant differences were observed among hybrids with respect to yield and yield traits under current research. Based on the results, grain weight had more effect on grain yield than grain number during grain filling growth stage under water stress conditions. In this concern, according to low drought susceptibility index (less grain yield losses), the hybrid (Indaco) was more stable in grain number and grain weight. Consequently, it may be concluded that this hybrid would be recommended for use in the future breeding programs for production of drought tolerant hybrids.

Keywords: drought susceptibility index, grain growth, grain yield, maize, water stress

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Authors: Drew Commandeur, Ryan Brodie, Sandra Hundza, Marc Klimstra

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The amplitude of raw electromyography (EMG) is affected by recording conditions and often requires normalization to make meaningful comparisons. Bioelectric methods normalize with an EMG signal recorded during a standardized task or from the experimental protocol itself, while biomechanical methods often involve measurements with an additional sensor such as a force transducer. Common bioelectric normalization techniques for treadmill walking include maximum voluntary isometric contraction (MVIC), dynamic EMG peak (EMGPeak) or dynamic EMG mean (EMGMean). There are several concerns with using MVICs to normalize EMG, including poor reliability and potential discomfort. A limitation of bioelectric normalization techniques is that they could result in a misrepresentation of the absolute magnitude of force generated by the muscle and impact the interpretation of EMG between functionally disparate groups. Additionally, methods that normalize to EMG recorded during the task may eliminate some real inter-individual variability due to biological variation. This study compared biomechanical and bioelectric EMG normalization techniques during treadmill walking to assess the impact of the normalization method on the functional interpretation of EMG data. For the biomechanical method, we normalized EMG to a target torque (EMGTS) and the bioelectric methods used were normalization to the mean and peak of the signal during the walking task (EMGMean and EMGPeak). The effect of normalization on muscle activation pattern, EMG amplitude, and inter-individual variability were compared between disparate cohorts of OLD (76.6 yrs N=11) and YOUNG (26.6 yrs N=11) adults. Participants walked on a treadmill at a self-selected pace while EMG was recorded from the right lower limb. EMG data from the soleus (SOL), medial gastrocnemius (MG), tibialis anterior (TA), vastus lateralis (VL), and biceps femoris (BF) were phase averaged into 16 bins (phases) representing the gait cycle with bins 1-10 associated with right stance and bins 11-16 with right swing. Pearson’s correlations showed that activation patterns across the gait cycle were similar between all methods, ranging from r =0.86 to r=1.00 with p<0.05. This indicates that each method can characterize the muscle activation pattern during walking. Repeated measures ANOVA showed a main effect for age in MG for EMGPeak but no other main effects were observed. Interactions between age*phase of EMG amplitude between YOUNG and OLD with each method resulted in different statistical interpretation between methods. EMGTS normalization characterized the fewest differences (four phases across all 5 muscles) while EMGMean (11 phases) and EMGPeak (19 phases) showed considerably more differences between cohorts. The second notable finding was that coefficient of variation, the representation of inter-individual variability, was greatest for EMGTS and lowest for EMGMean while EMGPeak was slightly higher than EMGMean for all muscles. This finding supports our expectation that EMGTS normalization would retain inter-individual variability which may be desirable, however, it also suggests that even when large differences are expected, a larger sample size may be required to observe the differences. Our findings clearly indicate that interpretation of EMG is highly dependent on the normalization method used, and it is essential to consider the strengths and limitations of each method when drawing conclusions.

Keywords: electromyography, EMG normalization, functional EMG, older adults

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Authors: Abu Salim Mustafa

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Rv3873 is a relatively large size protein (371 amino acids in length) and its gene is located in the immunodominant genomic region of difference (RD)1 that is present in the genome of Mycobacterium tuberculosis but deleted from the genomes of all the vaccine strains of Bacillus Calmette Guerin (BCG) and most other mycobacteria. However, when tested for cellular immune responses using peripheral blood mononuclear cells from tuberculosis patients and BCG-vaccinated healthy subjects, this protein was found to be a major stimulator of cell mediated immune responses in both groups of subjects. In order to further identify the sequence of immunodominant epitopes and explore their Human Leukocyte Antigen (HLA)-restriction for epitope recognition, 24 peptides (25-mers overlapping with the neighboring peptides by 10 residues) covering the sequence of Rv3873 were synthesized chemically using fluorenylmethyloxycarbonyl chemistry and tested in cell mediated immune responses. The results of these experiments helped in the identification of an immunodominant peptide P9 that was recognized by people expressing varying HLA-DR types. Furthermore, it was also predicted to be a promiscuous binder with multiple epitopes for binding to HLA-DR, HLA-DP and HLA-DQ alleles of HLA-class II molecules that present antigens to T helper cells, and to HLA-class I molecules that present antigens to T cytotoxic cells. In addition, the evaluation of peptide P9 using an immunogenicity predictor server yielded a high score (0.94), which indicated a greater probability of this peptide to elicit a protective cellular immune response. In conclusion, P9, a peptide with multiple epitopes and ability to bind several HLA class I and class II molecules for presentation to cells of the cellular immune response, may be useful as a peptide-based vaccine against tuberculosis.

Keywords: mycobacterium tuberculosis, PPE68, peptides, vaccine

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Authors: Sukru Merey, Caglar Sinayuc

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Gas hydrate deposits which are found in deep ocean sediments and in permafrost regions are supposed to be a fossil fuel reserve for the future. The Black Sea is also considered rich in terms of gas hydrates. It abundantly contains gas hydrates as methane (CH₄~80 to 99.9%) source. In this study, by using the literature, seismic and other data of the Black Sea such as salinity, porosity of the sediments, common gas type, temperature distribution and pressure gradient, the optimum gas production method for the Black Sea gas hydrates was selected as mainly depressurization method. Numerical simulations were run to analyze gas production from gas hydrate deposited in turbidites in the Black Sea by depressurization.

Keywords: CH4 hydrate, Black Sea hydrates, gas hydrate experiments, HydrateResSim

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Authors: Maciej Sobczak, Julita Pietrzak, Tomasz Płoszaj, Agnieszka Robaszkiewicz

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Brg1, a member of SWI/SNF complex, plays a role in chromatin remodeling, therefore, regulates expression of many genes. Brg1 is an ATPase of SWI/SNF complex, thus its activity requires ATP. Through its bromodomain recognizes acetylated histone residues and evicts them, thus promoting transcriptionally active state of chromatin. One of the enzymes that is responsible for acetylation of histone residues is Ep300. It was previously shown in the literature that cooperation of Brg1 and Ep300 occurs at the promoter regions that have binding sites for E2F-family transcription factors as well as CpG islands. According to literature, approximately 20% of human cancer possess mutation in Brg1 or any other crucial SWI/SNF subunit. That phenomenon makes Brg1-Ep300 a very promising target for anti-cancer therapy. Therefore in our study, we investigated if physical interaction between Brg1 and Ep300 exists and what impact those two proteins have on key for breast cancer cells processes such as DNA damage repair and cell proliferation. Bioinformatical analysis pointed out, that genes involved in cell proliferation and DNA damage repair are overexpressed in MCF7 and MDA-MB-231 cells. Moreover, promoter regions of these genes are highly acetylated, which suggests high transcriptional activity of those sites. Notably, many of those gene possess within their promoters an E2F, Brg1 motives, as well as CpG islands and acetylated histones. Our data show that Brg1 physically interacts with Ep300, and together they regulate expression of genes involved in DNA damage repair and cell proliferation. Upon inhibiting Brg1 or Ep300, expression of vital for cancer cell survival genes such as CDK2/4, BRCA1/2, PCNA, and XRCC1 is decreased in MDA-MB-231 and MCF7 cells. Moreover, inhibition or silencing of either Brg1 or Ep300 leads to cell cycle arrest in G1. After inhibition of BRG1 or Ep300 on tested gene promoters, the repressor complex including Rb, HDAC1, and EZH2 is formed, which inhibits gene expression. These results highlight potentially significant target for targeted anticancer therapy to be introduced as a supportive therapy.

Keywords: brg1, ep300, breast cancer, epigenetics

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Authors: Athicha Cherdpunt, Patompong Satapornpong

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The medical cannabis is made up of components also known as cannabinoids, which consists of two ingredients which are Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD). Interestingly, the Cannabinoid can be used for many treatments such as chemotherapy, including nausea and vomiting, cachexia, anorexia nervosa, spinal cord injury and disease, epilepsy, pain, and many others. However, the adverse drug reactions (ADRs) of THC can cause sedation, anxiety, dizziness, appetite stimulation and impairments in driving and cognitive function. Furthermore, genetic polymorphisms of CYP2C9, CYP2C19 and CYP3A4 influenced the THC metabolism and might be a cause of ADRs. Particularly, CYP2C19*17 allele increases gene transcription and therefore results in ultra-rapid metabolizer phenotype (UM). The aim of this study, is to investigate the frequency of CYP2C19*17 alleles in Thai patients who have been treated with medical cannabis. We prospectively enrolled 60 Thai patients who were treated with medical cannabis and clinical data from College of Pharmacy, Rangsit University. DNA of each patient was isolated from EDTA blood, using the Genomic DNA Mini Kit. CYP2C19*17 genotyping was conducted using the real time-PCR ViiA7 (ABI, Foster City, CA, USA). 30 patients with medical cannabis-induced ADRs group, 20 (67%) were female, and 10 (33%) were male, with an age range of 30-69 years. On the other hand, 30 patients without medical cannabis-induced ADRs (control group) consist of 17 (57%) female and 13 (43%) male. The most ADRs for medical cannabis treatment in the case group were dry mouth and dry throat (77%), tachycardia (70%), nausea (30%) and arrhythmia(10%). Accordingly, the case group carried CYP2C19*1/*1 (normal metabolizer) approximately 93%, while 7% patients carrying CYP2C19*1/*17 (ultra rapid metabolizers) exhibited in this group. Meanwhile, we found 90% of CYP2C19*1/*1 and 10% of CYP2C19*1/*17 in control group. In this study, we identified the frequency of CYP2C19*17 allele in Thai population which will support the pharmacogenetics biomarkers for screening and avoid ADRs of medical cannabis treatment.

Keywords: CYP2C19, allele frequency, ultra rapid metabolizer, medical cannabis

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Authors: Ömer Faruk Ada, Türker Baş, M. Yaman Öztek

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In this study, we aim to investigate the skills to be owned by procurement professionals in order to fulfill their developing and changing role completely. Market conditions, competitive pressure, and high financial costs make it more important than ever for organizations to be able to use resources more efficiently. Research shows that procurement expenses consist more than 50 % of the operating expenses. With increasing profit impact of procurement, reviewing the position of the procurement function within the organization has become inevitable. This study is significant as it indicates the necessary skills that procurement professionals must have to keep in step with the transformation of procurement units from transaction oriented to value chain oriented. In this study, the transformation of procurement is investigated from the perspective of procurement professionals and we aim to answer following research questions: • How do procurement professionals perceive their role within the organization? • How has their role changed and what challenges have they had to face? • What portfolio of skills do they believe will enable them to fulfill their role effectively? Literature review consists of the first part of the study by investigating the changing role of procurement from different perspectives. In the second part, we present the results of the in-depth interviews with 15 procurement professionals and we used descriptive analysis as a methodology. In the light of these results, we classified procurement skills under operational, tactical and strategic levels and Procurement Skills Framework has been developed. This study shows the differences in the perception of purchasing by professionals and the organizations. The differences in the perception are considered as an important barrier beyond the procurement transformation. Although having the necessary skills has a significant effect for procurement professionals to fulfill their role completely and keep in step with the transformation of the procurement function, It is not the only factor and the degree of high-level management and organizational support has also a direct impact during this transformation.

Keywords: procuement skills, procurement transformation, strategic procurement, value chain

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Authors: Reza Farahmand Rad, Farnad Imani, Azadeh Emami, Reza Salehi, Ali Reza Ghavamy, Ali Nima Shariat

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Background: One of the methods of pain control after pediatric surgical procedures is regional techniques, including caudal block, despite their limitations. Objectives: In this study, the pain score and complications of caudal tramadol were evaluated in pediatrics following lower abdom- inal surgery. Methods: In this study, 46 children aged 3 to 10 years were allocated into two equal groups (R and TR) for performing caudal anal- gesia after lower abdominal surgery. The injectate contained 0.2% ropivacaine 1 mL/kg in the R group (control group) and tramadol (2 mg/kg) and ropivacaine in the TR group. The pain score, duration of pain relief, amount of paracetamol consumption, hemody- namic alterations, and possible complications at specific times (1, 2, and 6 hours) were evaluated in both groups. Results: No considerable difference was observed in the pain score between the groups in the first and second hours (P > 0.05). However, in the sixth hour, the TR group had a significantly lower pain score than the R group (P < 0.05). Compared to the R group, the TR group had a longer period of analgesia and lower consumption of analgesic drugs (P < 0.05). Heart rate and blood pressure differences were not significant between the two groups (P > 0.05). Similarly, the duration of operation and recovery time were not remarkably different between the two groups (P > 0.05). Complications had no apparent differences between these two groups, as well (P > 0.05). Conclusions: In this study, the addition of tramadol to caudal ropivacaine in pediatric lower abdominal surgery promoted pain relief without complications.

Keywords: tramadol, ropivacaine, caudal block, pediatric, lower abdominal surgery, postoperative pain

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Authors: Shiori Ishida, Hiromi Okuno, Hisato Igarashi, Akemi Yamazaki, Hiroko Takahashi

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The purpose of this study was to clarify the differences and similarities regarding the social support of fathers and mothers towards considering increased assistance for the paternity of children with developmental disabilities. Written questionnaires were completed by fathers (n=85) and mothers (n=101) of children using rehabilitation facilities between infancy and 5 years of age. The survey contained multiple-choice questions on four categories: information support (6 items), emotional support (7 items), evaluation support (3 items), and daily living support (3 items). Regarding information support, fathers answered ‘spouse’ as the provider in over 50% of cases for all 6 items, which was significantly different compared with mothers (all p < 0.001). For emotional support, fathers were significantly more likely to get support from the workplace (p < 0.001) and from spouse (p < 0.001). The ‘evaluation support’ did not have significant differences for fathers in all the items, but the most frequent support providers were ‘spouses’. ‘Daily living support’ was significantly different from fathers in the workplace (p < 0.000) in terms of make allowances for work and duties. Thus, it appeared that fathers had fewer social support sources as compared with mothers and limited non-spouse support. The understanding of developmental disabilities, acquisition of methods of rehabilitation, and sources of support might have been inadequately addressed among fathers, which could be a hindrance to the involvement of fathers in the rearing of children with developmental disabilities. On the other hand, we also observed that some fathers were involved in the care of developmentally troubled children while providing mental support for their spouse, cooperating with housework, and adjusting their work life. However, the results on the external and social backgrounds of fathers indicated a necessity for greater empowerment and peer support to improve the paternal care of children with developmental disabilities in the family survey.

Keywords: children with developmental disabilities, family support, father, social support

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Authors: Thulane Paepae, Tumisang Seodigeng

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This paper demonstrates the use of a method of synthesizing process flowsheets using a graphical tool called the GH-plot and in particular, to look at how it can be used to compare the reactions of a combined simultaneous process with regard to their thermodynamics. The technique uses fundamental thermodynamic principles to allow the mass, energy and work balances locate the attainable region for chemical processes in a reactor. This provides guidance on what design decisions would be best suited to developing new processes that are more effective and make lower demands on raw material and energy usage.

Keywords: attainable regions, dimethyl ether, optimal reaction network, GH Space

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Authors: Stephanie Lehmann, Esther Ruf, Sabina Misoch

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Background: Due to demographic change and shortage of skilled nursing staff, assistive robots are built to support older adults at home and nursing staff in care institutions. When assistive robots facilitate tasks that are usually performed by humans, user acceptance is essential. Even though they are an important aspect of acceptance, emotions towards different assistive robots and different situations of robot-use have so far not been examined in detail. The appearance of assistive robots can trigger emotions that affect their acceptance. Acceptance of robots is assumed to be greater when they look more human-like; however, too much human similarity can be counterproductive. Regarding different groups, it is assumed that older adults have a more negative attitude towards robots than younger adults. Within the framework of a simulated robot study, the aim was to investigate emotions of older adults compared to students towards robots with different appearances and in different situations and so contribute to a deeper view of the emotions influencing acceptance. Methods: In a questionnaire study, vignettes were used to assess emotions toward robots in different situations and of different appearance. The vignettes were composed of two situations (service and care) shown by video and four pictures of robots varying in human similarity (machine-like to android). The combination of the vignettes was randomly distributed to the participants. One hundred forty-two older adults and 35 bachelor students of nursing participated. They filled out a questionnaire that surveyed 30 positive and 30 negative emotions. For each group, older adults and students, a sum score of “positive emotions” and a sum score of “negative emotions” was calculated. Mean value, standard deviation, or n for sample size and % for frequencies, according to the scale level, were calculated. For differences in the scores of positive and negative emotions for different situations, t-tests were calculated. Results: Overall, older adults reported significantly more positive emotions than students towards robots in general. Students reported significantly more negative emotions than older adults. Regarding the two different situations, the results were similar for the care situation, with older adults reporting more positive emotions than students and less negative emotions than students. In the service situation, older adults reported significantly more positive emotions; negative emotions did not differ significantly from the students. Regarding the appearance of the robot, there were no significant differences in emotions reported towards the machine-like, the mechanical-human-like and the human-like appearance. Regarding the android robot, students reported significantly more negative emotions than older adults. Conclusion: There were differences in the emotions reported by older adults compared to students. Older adults reported more positive emotions, and students reported more negative emotions towards robots in different situations and with different appearances. It can be assumed that older adults have a different attitude towards the use of robots than younger people, especially young adults in the health sector. Therefore, the use of robots in the service or care sector should not be rejected rashly based on the attitudes of younger persons, without considering the attitudes of older adults equally.

Keywords: emotions, robots, seniors, young adults

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Authors: Chih-Yang Liao

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Baseball is usually played outdoors in the warmest months of the year. Therefore, baseball players are susceptible to the influence of the hot environment. It has been shown that hitting performance is increased in games played in warm weather, compared to in cold weather, in Major League Baseball. Intermittent cooling during sporting events can prevent the risk of hyperthermia and increase endurance performance. However, the effects of cooling during baseball games played in a hot environment are unclear. This study adopted a cross-over design. Ten Division I collegiate male baseball players in Taiwan volunteered to participate in this study. Each player played two simulated baseball games, with one day in between. Five of the players received intermittent cooling during the first simulated game, while the other five players received intermittent cooling during the second simulated game. The participants were covered in neck and forehand regions for 6 min with towels that were soaked in icy salt water 3 to 4 times during the games. The participants received the cooling treatment in the dugout when they were not on the field for defense or hitting. During the 2 simulated games, the temperature was 31.1-34.1°C and humidity was 58.2-61.8%, with no difference between the two games. Ratings of perceived exertion, thermal sensation, tympanic and forehead skin temperature immediately after each defensive half-inning and after cooling treatments were recorded. Ratings of perceived exertion were measured using the Borg 10-point scale. The thermal sensation was measured with a 6-point scale. The tympanic and skin temperature was measured with infrared thermometers. The data were analyzed with a two-way analysis of variance with repeated measurement. The results showed that intermitted cooling significantly reduced ratings of perceived exertion and thermal sensation. Forehead skin temperature was also significantly decreased after cooling treatments. However, the tympanic temperature was not significantly different between the two trials. In conclusion, intermittent cooling in the neck and forehead regions was effective in alleviating the perceived exertion and heat sensation. However, this cooling intervention did not affect the core temperature. Whether intermittent cooling has any impact on hitting or pitching performance in baseball players warrants further investigation.

Keywords: baseball, cooling, ratings of perceived exertion, thermal sensation

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Authors: Sarah Almuhayya, Andrew Mcbain, Gavin Humphreys

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Background. The microbiome of the upper respiratory tract (URT) has received less research attention than other body sites. This study aims to investigate the microbial ecology of the human URT with a focus on the antagonism between the corynebacteria and staphylococci. Methods. Mucosal swabs were collected from the anterior nares and nasal turbinates of 20 healthy adult subjects. Genomic DNA amplification targeting the (V4) of the 16Sr RNA gene was conducted and analyzed using QIIME. Nasal swab isolates were cultured and identified using near full-length sequencing of the 16S rRNA gene. Isolates identified as corynebacteria or staphylococci were typed using (rep-PCR). Antagonism was determined using an agar-based inhibition assay. Results. Four major bacterial phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) were identified from all volunteers. The typing of cultured staphylococci and corynebacteria suggested that intra-individual strain diversity was limited. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between staphylococci and corynebacteria. Despite the apparent antagonism between these genera, it was limited when investigated on agar. Of 1000 pairwise interactions, observable zones of inhibition were only reported between a single strain of C.pseudodiphtheriticum and S.aureus. Imaging under EM revealed this effect to be bactericidal with clear lytic effects on staphylococcal cell morphology. Conclusion. Nasal microbiota is complex, but culturable staphylococci and corynebacteria were limited in terms of clone type. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between these genera suggesting an antagonism or competition between these taxonomic groups.

Keywords: nasal, microbiota, S.aureus, microbioal interaction

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Authors: Yinghua Huang, Tsu-Hong Yen

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The latest advance in leadership research has revealed that leadership is co-created through the combined action of leading and following. The role of followers is as important as leaders in the leadership process. However, the previous leadership studies often conceptualize leadership as a leader-centric process, while the role of followers is largely neglected in the literature. Until recently, followership studies receives more attention because the character and behavior of followers are as vital as the leader during the leadership process. Yet, there is a dearth of followership research in the context of tourism and hospitality industries. Therefore, this study seeks to fill in the gap of knowledge and investigate the followership styles in the U.S. hospitality workforce. In particular, the objectives of this study are to identify popular followership practices among hospitality employees and evaluate hospitality employees' followership styles using Kelley’s followership typology framework. This study also compared the generational differences in followership styles among hospitality employees. According to the U.S. Bureau of Labor Statistics, the workforce in the lodging and foodservice sectors consists of around 12% baby boomers, 29% Gen Xs, 23% Gen Ys, and 36% Gen Zs in 2019. The diversity of workforce demographics in the U.S. hospitality industry calls for more attention to understand the generational differences in followership styles and organizational performance. This study conducted an in-depth interview and a questionnaire survey to collect both qualitative and quantitative data. A snowball sampling method was used to recruit participants working in the hospitality industry in the San Francisco Bay Area, California, USA. A total of 120 hospitality employees participated in this study, including 22 baby boomers, 32 Gen Xs, 30 Gen Ys, and 36 Gen Zs. 45% of the participants were males, and 55% were female. The findings of this study identified good followership practices across the multi-generational participants. For example, a Gen Y participant said that 'followership involves learning and molding oneself after another person usually an expert in an area of interest. I think of followership as personal and professional development. I learn and get better by hands-on training and experience'. A Gen X participant said that 'I can excel by not being fearful of taking on unfamiliar tasks and accepting challenges.' Furthermore, this study identified five typologies of Kelley’s followership model among the participants: 45% exemplary followers, 13% pragmatist followers, 2% alienated followers, 18% passive followers, and 23% conformist followers. The generational differences in followership styles were also identified. The findings of this study contribute to the hospitality human resource literature by identifying the multi-generational perspectives of followership styles among hospitality employees. The findings provide valuable insights for hospitality leaders to understand their followers better. Hospitality leaders were suggested to adjust their leadership style and communication strategies based on employees' different followership styles.

Keywords: followership, hospitality workforce, generational diversity, Kelley’s followership topology

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Authors: Emmanuel Kamal Aziz Saba

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Aim of the work: This study proposed to assess the role of the median versus ulnar medial thenar motor (MTM) recording in supporting the diagnosis of carpal tunnel syndrome (CTS). Patients and methods: The present study included 130 hands (70 CTS and 60 controls). Clinical examination was done for all patients. The following tests were done (using surface electrodes recording) for patients and control: (1) sensory nerve conduction studies: median nerve, ulnar nerve and median versus ulnar digit four sensory study; (2) motor nerve conduction studies: median nerve, ulnar nerve, median (second lumbrical) versus ulnar (interosseous) (2-LINT) motor study and median versus ulnar (MTM) study. Results: The tests with higher sensitivity in diagnosing CTS were median versus ulnar (2-LINT) motor latency difference (87.1%), median versus ulnar (MTM) motor latency difference (80%) and median versus ulnar digit four sensory latency differences (91.4%). There was no statistically significant difference between median versus ulnar (MTM) motor latency difference with both median versus ulnar (2-LINT) motor latency difference and median versus ulnar digit four sensory latency difference (P > 0.05) as regards the confirmation of CTS. Conclusions: Median versus ulnar (MTM) motor latency difference has high sensitivity and specificity for the diagnosis of CTS as for both median versus ulnar (2-LINT) motor latency difference and median versus ulnar digit four sensory latency differences. It can be considered a useful neurophysiological test to be used in combination with another median versus ulnar comparative tests for confirming the diagnosis of CTS beside other well-known electrophysiological tests.

Keywords: carpal tunnel syndrome, medial thenar motor, median nerve, ulnar nerve

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Authors: T. N. Girish, B. E. Pradeep, Ravi Parkash

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Insects from tropical wet or dry seasons are likely to cope starvation stress through seasonal phenotypic plasticity in energy metabolites. Accordingly, we analyzed such plastic changes in Zaprionus indianus flies reared under wet or dry season-specific conditions; and also after adult acclimation at 32℃ for 1 to 6 days; and to low (40% RH) or high (70% RH) humidity. Both thermal or humidity acclimation revealed significant accumulation of body lipids for wet season flies but low humidity acclimation did not change the level of body lipids in dry season flies. Developmental and adult acclimation showed sex specific differences i.e., starvation resistance and body lipids were higher in the males of dry season but in females of wet season. We found seasonal and sex specific differences in the relative level for body lipids at death; and in the rates of accumulation or utilization of energy metabolites (body lipids, carbohydrates and proteins). Body lipids constitute the preferred energy source under starvation for flies of both the seasons. However, utilization of carbohydrates (~20% to 30%) and proteins (~20% to 25%) was evident only in dry season flies. Higher starvation resistance after thermal or humidity acclimation is achieved by increased accumulation of lipids. Adult acclimation of wet or dry season flies revealed plastic changes in mean daily fecundity despite reduction in fecundity under starvation. Thus, thermal or humidity induced plastic responses in body lipids support starvation resistance under wet or dry seasons.

Keywords: heat or humidity acclimation, plastic changes in body lipids and starvation resistance, tropical drosophilid, Wet- or Dry seasons, Zaprionus indianus

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Authors: Aya M. H. Eissa, Ayman H. A. Mahmoud

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When it comes to energy efficient thermal regulation system, natural systems do not only offer an inspirational source of innovative strategies but also sustainable and even regenerative ones. Using biomimetic design an energy efficient thermal regulation system can be developed. Although, conventional design process methods achieved fairly efficient systems, they still had limitations which can be overcome by using parametric design software. Accordingly, the main objective of this study is to apply and assess the efficiency of heat regulation strategies inspired from termite mounds in residential buildings’ thermal regulation system. Parametric design software is used to pave the way for further and more complex biomimetic design studies and implementations. A hot arid region is selected due to the deficiency of research in this climatic region. First, the analysis phase in which the stimuli, affecting, and the parameters, to be optimized, are set mimicking the natural system. Then, based on climatic data and using parametric design software Grasshopper, building form and openings height and areas are altered till settling on an optimized solution. Finally, an assessment of the efficiency of the optimized system, in comparison with a conventional system, is determined by firstly, indoors airflow and indoors temperature, by Ansys Fluent (CFD) simulation. Secondly by and total solar radiation falling on the building envelope, which was calculated using Ladybug, Grasshopper plugin. The results show an increase in the average indoor airflow speed from 0.5m/s to 1.5 m/s. Also, a slight decrease in temperature was noticed. And finally, the total radiation was decreased by 4%. In conclusion, despite the fact that applying a single bio-inspired heat regulation strategy might not be enough to achieve an optimum system, the concluded system is more energy efficient than the conventional ones as it aids achieving indoors comfort through passive techniques. Thus demonstrating the potential of parametric design software in biomimetic design.

Keywords: biomimicry, heat regulation systems, hot arid regions, parametric design, thermal comfort

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Authors: Timma Reddy, Srikonda Ramesh

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India has been traditionally vulnerable to natural disasters such as Floods, droughts, cyclones, earthquakes and landslides. It has become a recurrent phenomenon as observed in last five decades. The survey indicates that about 60% of the landmass is prone to earthquakes of various intensities; over 40 million hectares is prone to floods; about 8% of the total area is prone to cyclones and 68% of the area is susceptible to drought. Climate change is likely to be perceived through experience of extreme weather events. There is growing societal concern about climate change, given the potential impacts of associated natural hazards such as cyclones, flooding, earthquakes, landslides etc, hence it is essential and crucial to strengthening our settlements to respond to such calamities. So, the research paper focus is to analyze the effective planning strategy/mechanism to integrate disaster mitigation measures in coastal regions in general and Capital Region of Andhra Pradesh in particular. The basic hypothesis is to govern the appropriate special planning considerations would facilitate to have organized way of protective life and properties from natural disasters. And further to integrate the infrastructure planning with conscious direction would provide an effective mitigations measures. It has been planned and analyzed to Vijayawada city with conscious land use planning with reference to space syntax trajectory in accordance to required social infrastructure such as health facilities, institution areas and recreational and other open spaces. It has been identified that the geographically ideal location with reference to the population densities based on GIS tools the properness strategies can be effectively integrated to protect the life and to save the properties by means of reducing the damage/impact of natural disasters in general earth quake/cyclones or floods in particularly.

Keywords: modular, trajectories, social infrastructure, evidence based syntax, drills and equipments, GIS, geographical micro zoning, high resolution satellite image

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Authors: Sohan Sengupta, Arnab Pramanik, Abhrajyoti Ghosh, Maitree Bhattacharyya

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Newly emerged phyto-pathogens and multi drug resistance have been threating the world for last few decades. Actinomycetes, the most endowed group of microorganisms isolated from unexplored regions of the world may be the ultimate solution to these problems. Thus the aim of this study was to isolate several bioactive actinomycetes strains capable of producing antimicrobial secondary metabolite from Sundarbans, the only mangrove tiger land of the world. Fifty four actinomycetes were isolated and analyzed for antimicrobial activity against fifteen test organisms including three phytopathogens. Nine morphologically distinct and biologically active isolates were subjected to polyphasic identification study. 16s rDNA sequencing indicated eight isolates to reveal maximum similarity to the genus streptomyces, whereas one isolate presented only 93.57% similarity with Streptomyces albogriseolus NRRL B-1305T. Seventy-one carbon sources and twenty-three chemical sources utilization assay revealed their metabolic relatedness. Among these nine isolates three specific strains were found to have notably higher degree of antimicrobial potential effective in a broader range including phyto-pathogenic fungus. PCR base whole genome screen for PKS and NRPS genes, confirmed the occurrence of bio-synthetic gene cluster in some of the isolates for novel antibiotic production. Finally the strain SMS_SU21, which showed antimicrobial activity with MIC value of 0.05 mg ml-1and antioxidant activity with IC50 value of 0.242±0.33 mg ml-1 was detected to be the most potential one. True prospective of this strain was evaluated utilizing GC-MS and the bioactive compound responsible for antimicrobial activity was purified and characterized. Rare bioactive actinomycetes were isolated from unexplored heritage site. Diversity of the biosynthetic gene cluster for antimicrobial compound production has also been evaluated. Antimicrobial compound SU21-C has been identified and purified which is active against a broad range of pathogens.

Keywords: actinomycetes, sundarbans, antimicrobial, pks nrps, phyto-pathogens, GC-MS

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Authors: Defne Uz

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Horizontal Tail of helicopter is exposed to repeated oscillatory loading generated by aerodynamic and inertial loads, and bending moments depending on operating conditions and maneuvers of the helicopter. In order to ensure that maximum stress levels do not exceed certain fatigue limit of the material and to prevent damage, a numerical analysis approach can be utilized through the Finite Element Method. Therefore, in this paper, fatigue analysis of the Horizontal Tail model is studied numerically to predict high-cycle and low-cycle fatigue life related to defined loading. The analysis estimates the stress field at stress concentration regions such as around fastener holes where the maximum principal stresses are considered for each load case. Critical element identification of the main load carrying structural components of the model with rivet holes is performed as a post-process since critical regions with high-stress values are used as an input for fatigue life calculation. Once the maximum stress is obtained at the critical element and the related mean and alternating components, it is compared with the endurance limit by applying Soderberg approach. The constant life straight line provides the limit for several combinations of mean and alternating stresses. The life calculation based on S-N (Stress-Number of Cycles) curve is also applied with fully reversed loading to determine the number of cycles corresponds to the oscillatory stress with zero means. The results determine the appropriateness of the design of the model for its fatigue strength and the number of cycles that the model can withstand for the calculated stress. The effect of correctly determining the critical rivet holes is investigated by analyzing stresses at different structural parts in the model. In the case of low life prediction, alternative design solutions are developed, and flight hours can be estimated for the fatigue safe operation of the model.

Keywords: fatigue analysis, finite element method, helicopter horizontal tail, life prediction, stress concentration

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Authors: Li Lin, Hein Roelfsema

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To achieve aggressive expansion goals, many Chinese companies are seeking resources and market around the world. To an increasing extent, Chinese enterprises recognized the Netherlands as their gateway to Europe Market. Yet, large cultural gaps (e.g. individualism/collectivism, power distance) may influence expat leaders’ influencing process, in turn affect intercultural teamwork. Lessons and suggestions from Chinese expat leaders could provide profound knowledge for managerial practice and future research. The current research focuses on the cultural difference between China and the Netherlands, along with leadership tactics for coping and handling differences occurring in the international business work. Exclusive 47 in-depth interviews with Chinese expat leaders were conducted. Within each interview, respondents were asked what were the main issues when working with Dutch employees, and what they believed as the keys to successful leadership in Dutch-Chinese cross-cultural workplaces. Consistent with previous research, the findings highlight the need to consider the cultural context within which leadership adapts. In addition, the findings indicated the importance of recognizing and applying the cultural advantages from which leadership originates. The results address observation ability as a crucial key for Chinese managers to lead Dutch/international teams. Moreover, setting a common goal help a leader to overcome the challenges due to cultural differences. Based on the analysis, we develop a process model to illustrate the dynamic mechanisms. Our study contributes to the better understanding of transference of management practices, and has important practical implications for managing Dutch employees.

Keywords: Chinese managers, Dutch employees, leadership, interviews

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Authors: Sydney Burnett

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The history of the relationship between elder abuse and its tendency to go underreported is rooted in the oppressive nature of ageism and victimization. Approximately 8% of the world's population was aged sixty or over in 1950, whereas, in 2020, the number more than doubled to 16.9%. By 2050, that number is expected to reach 22%. Although difficult for individuals of any age to feel completely supported in society, this task proves especially difficult for the elderly demographic. And as the elderly population continues to grow, the systemic abuse and neglect that this group encounters, and thus its underreported nature, multiply at a similar rate. Although a recent increase in awareness has initiated stronger efforts towards addressing the meager resources, processes, and personnel present to manage elder abuse, both reported and unreported, the destructive complexities of ageism and victimization persist. Examining the byproducts of the rapidly growing elderly demographic in China, the United States, and Israel, in cohesion with the inherent challenges in the context of terminology, definition, and typologies of elder abuse should provide insight into the pernicious influences of elder abuse that contribute to the non-identification and non-recognition of elder maltreatment present in these three countries in different stages of development. This investigation aims to understand the intricacy of elder abuse and its correlation to a lack of acknowledgment as well as its consequences in society by exploring the variation between China, the United States, and Israel's attitudes surrounding the subject. Furthermore, the systemic abuse and neglect embedded in global ageism can be revealed by the differences between the three countries' approaches to reporting elder abuse.

Keywords: elder abuse, ageism, mistreatment, underreported

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Authors: O. Fatemi

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Considering growing building construction industry in developing countries and sustainable development concept, as well as the importance of taking care of the future generations, codifying buildings scoring system based on environmental criteria, has always been a subject for discussion. The existing systems cannot be used for all the regions due to several reasons, including but not limited to variety in regional variables. In this article, the most important common LEED (Leadership in Energy and Environmental Design) and BREEAM (Building Research Establishment Environmental Assessment Method) common and Global environmental scoring systems, used in UK, USA, and Japan, respectively, have been discussed and compared with a special focus on CASBEE (Comprehensive Assessment System for Built Environment Efficiency), to credit assigning field (weighing and scores systems) as well as sustainable development criteria in each system. Then, converging and distinct fields of the foregoing systems are examined considering National Iranian Building Code. Furthermore, the common credits in the said systems not mentioned in National Iranian Building Code have been identified. These credits, which are generally included in well-known fundamental principles in sustainable development, may be considered as offered options for the Iranian building environmental scoring system. It is suggested that one of the globally and commonly accepted systems is chosen considering national priorities in order to offer an effective method for buildings environmental scoring, and then, a part of credits is added and/or removed, or a certain credit score is changed, and eventually, a new scoring system with a new title is developed for the country. Evidently, building construction industry highly affects the environment, economy, efficiency, and health of the relevant occupants. Considering the growing trend of cities and construction, achieving building scoring systems based on environmental criteria has always been a matter of discussion. The existing systems cannot be used for all the regions due to several reasons, including but not limited to variety in regional variables.

Keywords: scoring system, sustainability assessment, water efficiency, national Iranian building code

Procedia PDF Downloads 172

Authors: Alice Robba, Renaud Bouchet, Celine Barchasz, Jean-Francois Colin, Erik Elkaim, Kristina Kvashnina, Gavin Vaughan, Matjaz Kavcic, Fannie Alloin

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With their high theoretical energy density (~2600 Wh.kg-1), lithium/sulfur (Li/S) batteries are highly promising, but these systems are still poorly understood due to the complex mechanisms/equilibria involved. Replacing S8 by Li2S as the active material allows the use of safer negative electrodes, like silicon, instead of lithium metal. S8 and Li2S have different conductivity and solubility properties, resulting in a profoundly changed activation process during the first cycle. Particularly, during the first charge a high polarization and a lack of reproducibility between tests are observed. Differences observed between raw Li2S material (micron-sized) and that electrochemically produced in a battery (nano-sized) may indicate that the electrochemical process depends on the particle size. Then the major focus of the presented work is to deepen the understanding of the Li2S material charge mechanism, and more precisely to characterize the effect of the initial Li2S particle size both on the mechanism and the electrode preparation process. To do so, Li2S nanoparticles were synthetized according to two ways: a liquid path synthesis and a dissolution in ethanol, allowing Li2S nanoparticles/carbon composites to be made. Preliminary chemical and electrochemical tests show that starting with Li2S nanoparticles could effectively suppress the high initial polarization but also influence the electrode slurry preparation. Indeed, it has been shown that classical formulation process - a slurry composed of Polyvinylidone Fluoride polymer dissolved in N-methyle-2-pyrrolidone - cannot be used with Li2S nanoparticles. This reveals a complete different Li2S material behavior regarding polymers and organic solvents when going at the nanometric scale. Then the coupling between two operando characterizations such as X-Ray Diffraction (XRD) and X-Ray Absorption and Emission Spectroscopy (XAS/XES) have been carried out in order to interpret the poorly understood first charge. This study discloses that initial particle size of the active material has a great impact on the working mechanism and particularly on the different equilibria involved during the first charge of the Li2S based Li-ion batteries. These results explain the electrochemical differences and particularly the polarization differences observed during the first charge between micrometric and nanometric Li2S-based electrodes. Finally, this work could lead to a better active material design and so to more efficient Li2S-based batteries.

Keywords: Li-ion/Sulfur batteries, Li2S nanoparticles effect, Operando characterizations, working mechanism

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Authors: Milica Ž. Karadžić, Lidija R. Jevrić, Sanja Podunavac-Kuzmanović, Strahinja Z. Kovačević, Anamarija I. Mandić, Katarina Penov-Gaši, Andrea R. Nikolić, Aleksandar M. Oklješa

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This study considered selection of the in silico molecular descriptors and the models for newly synthesized steroid derivatives description and their characterization using chemometric techniques. Multiple linear regression (MLR) models were established and gave the best molecular descriptors for quantitative structure-retention relationship (QSRR) modeling of the retention of the investigated molecules. MLR models were without multicollinearity among the selected molecular descriptors according to the variance inflation factor (VIF) values. Used molecular descriptors were ranked using generalized pair correlation method (GPCM). In this method, the significant difference between independent variables can be noticed regardless almost equal correlation between dependent variable. Generated MLR models were statistically and cross-validated and the best models were kept. Models were ranked using sum of ranking differences (SRD) method. According to this method, the most consistent QSRR model can be found and similarity or dissimilarity between the models could be noticed. In this study, SRD was performed using average values of experimentally observed data as a golden standard. Chemometric analysis was conducted in order to characterize newly synthesized steroid derivatives for further investigation regarding their potential biological activity and further synthesis. This article is based upon work from COST Action (CM1105), supported by COST (European Cooperation in Science and Technology).

Keywords: generalized pair correlation method, molecular descriptors, regression analysis, steroids, sum of ranking differences

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Authors: Zita Fomukong Andam

Abstract:

Civil disobedience according to John Rawls (1971) is a public nonviolent and conscientious breach of laws undertaken with the aim of bringing about a change in government laws and policies. Thus individuals who engage themselves in such an act are aware and ready to accept the consequences of their actions. Cameroon more precisely the Northwest and the Southwest region which are the English part are considered as one of the societies facing this act of civil disobedience. It has been a tormenting issue in the country affecting its economy and the tourism sector. This is because these regions known as one of the best touristic sites of the country is not more considered as a destination to be visited by tourist because of its insecurities. Many commercial buildings have been burning down, leaving many young Cameroonians jobless. Education has been hindered, and youths are forced to relocate to nearby cities in order to continue their education. This crisis has created a lot of insecurity throughout the regions thus youths now have one common interest to travel abroad either to seek refuge or to continue their education and even search for jobs. The purpose of this research is to assess the issue of civil disobedience, trying to understand why it is affected only by a specific region in a country while the others are doing fine. A deep research discourse was conducted with randomly selected individuals aging between 15 to 40 years living both in the destination and abroad. Survey questionnaires and interviews were carried out as a method to collect data. The results show that this crisis has impacted the local residents psychologically and has injected a lot of fears into tourists and they are no more willing to visit the destination. In addition, it has brought a negative impact on the county’s economy since tourism is considered as the key sector in a country’s economy. On the other hand, the results showed that many local residents have remained jobless, others have lost family members, and the daily routine life has been affected. Understanding these results, the national government and international bodies might be able to propose possible and efficient solutions in order to attain stability and security in this region.

Keywords: civil disobedience, economic impact, local residents, tourist

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Authors: Mohammad K. Parvez, Mohammed S. Al-Dosari

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Hepatitis E virus (HEV) is an emerging RNA virus that causes acute and chronic liver disease with a global mortality rate of about 2%. Despite milestone developments in understanding of HEV biology, there is still lack of a robust culture system or animal model. Therefore, in a novel approach, two recombinant-baculoviruses (vBac-ORF2 and vBac-ORF3) that could overexpress HEV ORF2 (structural/capsid) and ORF3 (nonstructural/regulatory) proteins, respectively were constructed. The established HEV-SAR55 (genotype 1) replicon that contained GFP gene, in place of ORF2/ORF3 sequences was in vitro transcribed, and GFP production in RNA transfected S10-3 cells was scored by FACS. Enhanced infectivity, if any, of nascent virions produced by exogenously-supplied ORF2 and viral RNA by co-expression of ORF3 was tested on naïve HepG2 cells. Co-transduction with vBac-ORF2/vBac-ORF3 (108 pfu/microL) produced high amounts of native ORF2/ORF3 in approximately 60% of S10-3 cells, determined by immunofluorescence microscopy and Western analysis. FACS analysis showed about 9% GFP positivity of S10-3 cells on day6 post-transfection (i.e, day5 post-transduction). Further, FACS scoring indicated that lysates from S10-3 cultures receiving the RNA plus vBac-ORF2 were capable of producing HEV particles with about 4% infectivity in HepG2 cells. However, lysates of cultures co-transduced with vBac-ORF3, were found to further enhance virion infectivity by approximately 17%. This supported a previously proposed role of ORF3 as a minor-structural protein in HEV virion assembly and infectivity. In conclusion, the present model for efficient genomic RNA packaging and production of infectious virions could be a valuable tool to study various aspects of HEV molecular biology, in vitro.

Keywords: chronic liver disease, hepatitis E virus, ORF2, ORF3, replicon

Procedia PDF Downloads 250