Search results for: gene expression regulation

Authors: Feng-Sheng Wang, Chao-Ting Cheng

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Developing a drug from conception to launch is costly and time-consuming. Computer-aided methods can reduce research costs and accelerate the development process during the early drug discovery and development stages. This study developed a fuzzy multi-objective hierarchical optimization framework for identifying potential anticancer targets in a metabolic model. First, RNA-seq expression data of colorectal cancer samples and their healthy counterparts were used to reconstruct tissue-specific genome-scale metabolic models. The aim of the optimization framework was to identify anticancer targets that lead to cancer cell death and evaluate metabolic flux perturbations in normal cells that have been caused by cancer treatment. Four objectives were established in the optimization framework to evaluate the mortality of cancer cells for treatment and to minimize side effects causing toxicity-induced tumorigenesis on normal cells and smaller metabolic perturbations. Through fuzzy set theory, a multiobjective optimization problem was converted into a trilevel maximizing decision-making (MDM) problem. The applied nested hybrid differential evolution was applied to solve the trilevel MDM problem using two nutrient media to identify anticancer targets in the genome-scale metabolic model of colorectal cancer, respectively. Using Dulbecco’s Modified Eagle Medium (DMEM), the computational results reveal that the identified anticancer targets were mostly involved in cholesterol biosynthesis, pyrimidine and purine metabolisms, glycerophospholipid biosynthetic pathway and sphingolipid pathway. However, using Ham’s medium, the genes involved in cholesterol biosynthesis were unidentifiable. A comparison of the uptake reactions for the DMEM and Ham’s medium revealed that no cholesterol uptake reaction was included in DMEM. Two additional media, i.e., a cholesterol uptake reaction was included in DMEM and excluded in HAM, were respectively used to investigate the relationship of tumor cell growth with nutrient components and anticancer target genes. The genes involved in the cholesterol biosynthesis were also revealed to be determinable if a cholesterol uptake reaction was not induced when the cells were in the culture medium. However, the genes involved in cholesterol biosynthesis became unidentifiable if such a reaction was induced.

Keywords: Cancer metabolism, genome-scale metabolic model, constraint-based model, multilevel optimization, fuzzy optimization, hybrid differential evolution

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Authors: Monika Trzaskowska, Dorota Zielinska, Anna Lepecka, Katarzyna Neffe-Skocinska, Beata Bilska, Marzena Tomaszewska, Danuta Kolozyn-Krajewska

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Microbiological quality and food safety are important food characteristics. Regulation (EU) No 1169/2011 of the European Parliament and of the Council on the provision of food information to consumers introduces the obligation to provide information on the 'use-by' date or the date of minimum durability (DMD). The second term is the date until which the properly stored or transported foodstuff retains its physical, chemical, microbiological and organoleptic properties. The date should be preceded by 'best before'. It is used for durable products, e.g., pasta. In relation to reducing food losses, the question may be asked whether products with the date of minimum durability currently declared retain quality and safety beyond this. The aim of the study was to assess the sensory quality and microbiological safety of selected cereal products, i.e., pasta and millet after DMD. The scope of the study was to determine the markers of microbiological quality, i.e., the total viable count (TVC), the number of bacteria from the Enterobacteriaceae family and the number of yeast and mold (TYMC) on the last day of DMD and after 1 and 3 months of storage. In addition, the presence of Salmonella and Listeria monocytogenes was examined on the last day of DMD. The sensory quality of products was assessed by quantitative descriptive analysis (QDA), the intensity of 14 differentiators and overall quality were defined and determined. In the tested samples of millet and pasta, no pathogenic bacteria Salmonella and Listeria monocytogenes were found. The value of the distinguishing features of selected quality and microbiological safety indicators on the last DMD day was in the range of about 3-1 log cfu/g. This demonstrates the good microbiological quality of the tested food. Comparing the products, a higher number of microorganisms was found in the samples of millet. After 3 months of storage, TVC decreased in millet, while in pasta, it was found to increase in value. In both products, the number of bacteria from the Enterobacretiaceae family decreased. In contrast, the number of TYMCs increased in samples of millet, and in pasta decreased. The intensity of sensory characteristic in the studied period varied. It remained at a similar level or increased. Millet was found to increase the intensity and flavor of 'cooked porridge' 3 months after DMD. Similarly, in the pasta, the smell and taste of 'cooked pasta' was more intense. To sum up, the researched products on the last day of the minimum durability date were characterized by very good microbiological and sensory quality, which was maintained for 3 months after this date. Based on these results, the date of minimum durability of tested products could be extended. The publication was financed on the basis of an agreement with the National Center for Research and Development No. Gospostrateg 1/385753/1/NCBR/2018 for the implementation and financing of the project under the strategic research and development program 'social and economic development of Poland in the conditions of globalizing markets – GOSPOSTRATEG - acronym PROM'.

Keywords: date of minimum durability, food losses, food quality and safety, millet, pasta

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Authors: Supriya Ambawat, Subaran Singh, C. Tara Satyavathi, B. S. Rajpurohit, Ummed Singh, Balraj Singh

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Pearl millet [Pennisetum glaucum (L.) R. Br.] is an important staple food of the arid and semiarid tropical regions of Asia, Africa, and Latin America. It is rightly termed as nutricereal as it has high nutrition value and a good source of carbohydrate, protein, fat, ash, dietary fiber, potassium, magnesium, iron, zinc, etc. Pearl millet has low prolamine fraction and is gluten free which is useful for people having a gluten allergy. It has several health benefits like reduction in blood pressure, thyroid, diabe¬tes, cardiovascular and celiac diseases but its direct consumption as food has significantly declined due to several reasons. Keeping this in view, it is important to reorient the ef¬forts to generate demand through value-addition and quality improvement and create awareness on the nutritional merits of pearl millet. In India, through Indian Council of Agricultural Research-All India Coordinated Research Project on Pearl millet, multilocational coordinated trials for developed hybrids were conducted at various centers. The gene banks of pearl millet contain varieties with high levels of iron and zinc which were used to produce new pearl millet varieties with elevated iron levels bred with the high‐yielding varieties. Thus, using breeding approaches and biochemical analysis, a total of 167 hybrids and 61 varieties were identified and released for cultivation in different agro-ecological zones of the country which also includes some biofortified hybrids rich in Fe and Zn. Further, using several biotechnological interventions such as molecular markers, next-generation sequencing (NGS), association mapping, nested association mapping (NAM), MAGIC populations, genome editing, genotyping by sequencing (GBS), genome wide association studies (GWAS) advancement in millet improvement has become possible by identifying and tagging of genes underlying a trait in the genome. Using DArT markers very high density linkage maps were constructed for pearl millet. Improved HHB67 has been released using marker assisted selection (MAS) strategies, and genomic tools were used to identify Fe-Zn Quantitative Trait Loci (QTL). The draft genome sequence of millet has also opened various ways to explore pearl millet. Further, genomic positions of significantly associated simple sequence repeat (SSR) markers with iron and zinc content in the consensus map is being identified and research is in progress towards mapping QTLs for flour rancidity. The sequence information is being used to explore genes and enzymatic pathways responsible for rancidity of flour. Thus, development and application of several biotechnological approaches along with biofortification can accelerate the genetic gain targets for pearl millet improvement and help improve its quality.

Keywords: Biotechnological approaches, genomic tools, malnutrition, MAS, nutricereal, pearl millet, sequencing.

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Authors: Ilana Singer, Anastasia Lučić, Julie Leclerc

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Introduction: Tics, characterized by repetitive, sudden, non-voluntary motor movements or vocalizations, are prevalent in chronic tic disorder (CT) and Tourette Syndrome (TS). These neurodevelopmental disorders often coexist with various psychiatric conditions, leading to challenges and reduced quality of life. While medication in conjunction with behavioral interventions, such as Habit Reversal Training (HRT), Exposure Response Prevention (ERP), and Comprehensive Behavioral Intervention for Tics (CBIT), has shown efficacy, a significant proportion of patients experience persistent tics. Thus, innovative treatment approaches are necessary to improve therapeutic outcomes, such as mindfulness-based approaches. Nonetheless, the effectiveness of mindfulness-based interventions in the context of CT and TS remains understudied. Objective: The objective of this scoping review is to provide an overview of the current state of research on mindfulness-based interventions for CT and TS, identify knowledge and evidence gaps, discuss the effectiveness of mindfulness-based interventions with other treatment options, and discuss implications for clinical practice and policy development. Method: Using guidelines from Peters (2020) and the PRISMA-ScR, a scoping review was conducted. Multiple electronic databases were searched from inception until June 2023, including MEDLINE, EMBASE, PsychInfo, Global Health, PubMed, Web of Science, and Érudit. Inclusion criteria were applied to select relevant studies, and data extraction was independently performed by two reviewers. Results: Five papers were included in the study. Firstly, we found that mindfulness interventions were found to be effective in reducing anxiety and depression while enhancing overall well-being in individuals with tics. Furthermore, the review highlighted the potential role of mindfulness in enhancing functional connectivity within the Default Mode Network (DMN) as a compensatory function in TS patients. This suggests that mindfulness interventions may complement and support traditional therapeutic approaches, particularly HRT, by positively influencing brain networks associated with tic regulation and control. Conclusion: This scoping review contributes to the understanding of the effectiveness of mindfulness-based interventions in managing CT and TS. By identifying research gaps, this review can guide future investigations and interventions to improve outcomes for individuals with CT or TS. Overall, these findings emphasize the potential benefits of incorporating mindfulness-based interventions as a smaller subset within comprehensive treatment strategies. However, it is essential to acknowledge the limitations of this scoping review, such as the exclusion of a pre-established protocol and the limited number of studies available for inclusion. Further research and clinical exploration are necessary to better understand the specific mechanisms and optimal integration of mindfulness-based interventions with existing behavioral interventions for this population.

Keywords: scoping reviews, Tourette Syndrome, tics, mindfulness-based, therapy, intervention

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Authors: Krithika Lakshmi Sathiya Moorthy

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Introduction: Young people’s attitude towards Intimate partner violence (IPV) is likely to influence their indulgence in or abstinence from IPV in future. We aimed to assess the knowledge and attitudes of college students in a south Indian city regarding IPV, its associated factors and redressal mechanisms. Methods: A convenient sample of 247 students, pursuing medicine and engineering, participated in this analytical cross sectional study. They responded to a self-administered questionnaire developed and pretested for this study. The questionnaire comprises statements from a third person’s perspective and vignettes to reduce social desirability bias. Clearance was obtained from the Institute Ethical Committee of Velammal Medical College Hospital and Research Institute, Madurai, India. Data were entered in Epidata Entry v3.1, Odense, Denmark and analysed using SPSS v20.0. Results: Among 247 students, 116 (47%) were males and 59 (24.9%) hailed from rural areas. About 18% (43) of students believed that IPV was a problem only among females. Almost half of the students had witnessed IPV; at home between their parents (9.7%), other family members (13.4%), in their neighbourhood (13%) or public places (15%). Only 118 (47.8%) were aware that a law was in place in India to address IPV. The perceived risk factors for IPV were alcoholic spouse (78.9%), low income families (53.8%), personality traits (52.2%) and dowry system (51%). A sizeable number of students (38.4%) believed that some amount of physical violence was allowable in a marital relationship while 57.6% even considered IPV as an expression of love. Males as compared to females were more in agreement with negative gender stereotypes such as husband can– ‘threaten wife to ensure welfare of family’ (55% vs. 34%, p < 0.001), ‘spy on wife to check fidelity’ (41% vs. 27%, p < 0.001), ‘financially deprive housewife to punish’ (13% vs. 3.8%, p=0.001) and agreed with the statement that it is ‘duty of wife to comply with demands for sex from the husband’ (9.5% vs 4.6%, p=0.3). About 32% males and 25.6% females foresaw themselves as perpetrators of IPV in future. Conclusion: Knowledge about IPV and the associated risk factors among the study population was satisfactory. However, there was widespread acceptance of negative societal gender stereotypes, more so among males and some degrees of IPV were acceptable between married couples. The study advocates the need to halt the propagation of negative gender stereotypes in the impressionable young minds and the necessity to spread the awareness that no degree of IPV is acceptable. This knowledge is also required to plan the content and choose the appropriate media to effectively communicate the awareness about IPV among young persons.

Keywords: attitude, India, intimate partner violence, knowledge, students

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Authors: Ana L. Gonzalez-Celis, Edgardo Ruiz-Carrillo, Karina Reyes-Jarquin, Margarita Chavez-Becerra

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Since recent decades, the aging has been viewed from a more positive perspective, where is not only about losses and damage, but also about being on a stage where you can enjoy life and live with well-being and quality of life. The challenge to feel better is to find those resources that seniors have. For that reason, psychological well-being has shown interest in the study of the affect and life satisfaction (hedonic well-being), while from a more recent tradition, focus on the development of capabilities and the personal growth, considering both as the main indicators of the quality of life. A resource that can be used in the later age is generativity, which refers to the ability of older people to develop and grow through activities that contribute with the improvement of the context in which they live and participate. In this way the generative interest is understood as a favourable attitude that contribute to the common benefit while strengthening and enriching the social institutions, to ensure continuity between generations and social development. On the other hand, generative behavior, differentiating from generative interest, is the expression of that attitude reflected in activities that make a social contribution and a benefit for generations to come. Hence the purpose of the research was to test if there is an association between the generative behaviour type and the psychological well-being with their dimensions. For this reason 188 Mexican adults from 60 to 94 years old (M = 69.78), 67% women, 33% men, completed two instruments: The Ryff’s Well-Being Scales to measure psychological well-being with 39 items with two dimensions (Hedonic and Eudaimonic well-being), and the Loyola’s Generative Behaviors Scale, grouped in five categories: Knowledge transmitted to the next generation, things to be remember, creativity, be productive, contribution to the community, and responsibility of other people. In addition, the socio-demographic data sheet was tested, and self-reported health status. The results indicated that the psychological well-being and its dimensions were significantly associated with the presence of generative behavior, where the level of well-being was higher when the frequency of some generative behaviour excelled; finding that the behavior with greater psychological well-being (M = 81.04, SD = 8.18) was "things to be remembered"; while with greater hedonic well-being (M = 73.39, SD = 12.19) was the behavior "responsibility of other people"; and with greater Eudaimonic well-being (M = 84.61, SD = 6.63), was the behavior "things to be remembered”. The most important findings highlight the importance of generative behaviors in adulthood, finding empirical evidence that the generativity in the last stage of life is associated with well-being. However, by finding differences in the types of generative behaviors at the level of well-being, is proposed the idea that generativity is not situated as an isolated construct, but needs other contextualized and related constructs that can simultaneously operate at different levels, taking into account the relationship between the environment and the individual, encompassing both the social and psychological dimension.

Keywords: eudaimonic well-being, generativity, hedonic well-being, Mexican elders, psychological well-being

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Authors: Ekatereine Abashidze, Nino Nazarashvili, Dali Gaganidze, Oleg Gorgadze, Mariam Aznarashvili, Eter Gvritishvili

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Potato is one of the leading agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. Potato production is equal to 25,000 ha and its average yield is 20-25 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (Globodera pallida (Stone) Behrens and Globodera rostochiensis (Wollenveber) Behrens) are harmful around the world. PCN is among the most difficult plant pests to control. Cysts protected by a durable wall can survive for over 30 years . Control of PCN (G. pallida and G. rostochiensis) is regulated by Council Directive 2007/33/EE C. There was no legislative regulation of these pests in Georgia before 2016. By Resolution #302 from July 1, 2016, developed within the action plan of the DCFTA (Deep and Comprehensive Free Trade Area) the Government of Georgia established control over potato cyst nematodes. The Agreement about the legal acts approximation to EU legislation concerns the approval of rules of PCN control and research of these pests. Taking into consideration the above mentioned, it is necessary to study PCN (G. pallida and G. rostochiensis) in the potato-growing areas of Georgia. The aim of this research is to conduct survey of potato cyst nematodes (Globodera rostochiensis and G. pallida) in two geographically distinct regions of Georgia - Samtskhe - Javakheti and Svanetii and to identify the species G. Rostochiensis and G. Pallida by the morphological - morphometric and molecular methods. Soil samples were taken in each village, in a zig-zag pattern on the potato fields of the private sector, using the Metlitsky method. Samples were taken also from infested potato plant roots. To extract nematode cysts from soil samples Fanwick can be used according to standard methods by EPPO. Cysts were measured under a stereoscopic microscope (Leica M50). Identification of the nematod species was carried out according to morphological and morphometric characteristics of the cysts and larvae using appropriate protocols EPPO. For molecular identification, a multiplex PCR test was performed by the universal ITS5 and cyst nematodes’ (G. pallida, G. rostochiensis) specific primers. To identify the species of potato cyst nematodes (PCN) in two regions (Samtskhe-Javakheti and Svaneti) were taken 200 samples, among them: 80 samples in Samtskhe-Javakheti region and 120 in Svaneti region. Cysts of Globiodera spp. were revealed in 50 samples obtained from Samtskhe-Javakheti and 80 samples from Svaneti regions. Morphological, morphometric and molecular analysis of two forms of PCN found in investigated regions of Georgia shows that one form of PCN belongs to G. rostoshiensi; the second form is the different species of Globodera sp.t is the subject of future research. Despite the different geographic locations, larvae and cysts of G. rostoshiensi were found in both regions. But cysts and larvae of G. pallida were not reported. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia: Project # FR17_235.

Keywords: cyst nematode, globodera rostochiensis, globodera pallida, morphologic-morphometric measurement

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Authors: Antònio Inês, Davide Silva, Filipa Carvalho, Luís Filipe-Riberiro, Fernando M. Nunes, Luís Abrunhosa, Fernanda Cosme

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The presence of mycotoxins in foodstuff is a matter of concern for food safety. Mycotoxins are toxic secondary metabolites produced by certain molds, being ochratoxin A (OTA) one of the most relevant. Wines can also be contaminated with these toxicants. Several authors have demonstrated the presence of mycotoxins in wine, especially ochratoxin A. Its chemical structure is a dihydro-isocoumarin connected at the 7-carboxy group to a molecule of L-β-phenylalanine via an amide bond. As these toxicants can never be completely removed from the food chain, many countries have defined levels in food in order to attend health concerns. OTA contamination of wines might be a risk to consumer health, thus requiring treatments to achieve acceptable standards for human consumption. The maximum acceptable level of OTA in wines is 2.0 μg/kg according to the Commission regulation No. 1881/2006. Therefore, the aim of this work was to reduce OTA to safer levels using different fining agents, as well as their impact on white wine physicochemical characteristics. To evaluate their efficiency, 11 commercial fining agents (mineral, synthetic, animal and vegetable proteins) were used to get new approaches on OTA removal from white wine. Trials (including a control without addition of a fining agent) were performed in white wine artificially supplemented with OTA (10 µg/L). OTA analyses were performed after wine fining. Wine was centrifuged at 4000 rpm for 10 min and 1 mL of the supernatant was collected and added of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v). Also, the solid fractions obtained after fining, were centrifuged (4000 rpm, 15 min), the resulting supernatant discarded, and the pellet extracted with 1 mL of the above solution and 1 mL of H2O. OTA analysis was performed by HPLC with fluorescence detection. The most effective fining agent in removing OTA (80%) from white wine was a commercial formulation that contains gelatin, bentonite and activated carbon. Removals between 10-30% were obtained with potassium caseinate, yeast cell walls and pea protein. With bentonites, carboxymethylcellulose, polyvinylpolypyrrolidone and chitosan no considerable OTA removal was verified. Following, the effectiveness of seven commercial activated carbons was also evaluated and compared with the commercial formulation that contains gelatin, bentonite and activated carbon. The different activated carbons were applied at the concentration recommended by the manufacturer in order to evaluate their efficiency in reducing OTA levels. Trial and OTA analysis were performed as explained previously. The results showed that in white wine all activated carbons except one reduced 100% of OTA. The commercial formulation that contains gelatin, bentonite and activated carbon reduced only 73% of OTA concentration. These results may provide useful information for winemakers, namely for the selection of the most appropriate oenological product for OTA removal, reducing wine toxicity and simultaneously enhancing food safety and wine quality.

Keywords: wine, ota removal, food safety, fining

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Authors: Hantian Wu, Bo Huang, Yuan Zeng

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Divisions between wealthy and poor, private and public landscapes are propagated by the increasing economic inequality of cities. While these are the spatial reflections of larger social issues and problems, urban design can at least employ spatial techniques that promote more inclusive rather than exclusive, overlapping rather than segregated, interlinked rather than disconnected landscapes. Indeed, the type of edge or border between urban landscapes plays a critical role in the way the environment is perceived. China experiences rapid urbanization, which poses unpredictable environmental challenges. The urban green cover and water body are under changes, which highly relevant to resident wealth and happiness. However, very limited knowledge and data on their rapid changes are available. In this regard, enhancing the monitoring of urban landscape with high-frequency method, evaluating and estimating the impacts of the urban landscape changes, and understating the driving forces of urban landscape changes can be a significant contribution for urban planning and studying. High-resolution remote sensing data has been widely applied to urban management in China. The map of urban land use map for the entire China of 2018 with 10 meters resolution has been published. However, this research focuses on the large-scale and high-resolution remote sensing land use but does not precisely focus on the seasonal change of urban covers. High-resolution remote sensing data has a long-operation cycle (e.g., Landsat 8 required 16 days for the same location), which is unable to satisfy the requirement of monitoring urban-landscape changes. On the other hand, aerial-remote or unmanned aerial vehicle (UAV) sensing are limited by the aviation-regulation and cost was hardly widely applied in the mega-cities. Moreover, those data are limited by the climate and weather conditions (e.g., cloud, fog), and those problems make capturing spatial and temporal dynamics is always a challenge for the remote sensing community. Particularly, during the rainy season, no data are available even for Sentinel Satellite data with 5 days interval. Many natural events and/or human activities drive the changes of urban covers. In this case, enhancing the monitoring of urban landscape with high-frequency method, evaluating and estimating the impacts of the urban landscape changes, and understanding the mechanism of urban landscape changes can be a significant contribution for urban planning and studying. This project aims to use the high spatiotemporal fusion of remote sensing data to create short-cycle, high-resolution remote sensing data sets for exploring the high-frequently urban cover changes. This research will enhance the long-term monitoring applicability of high spatiotemporal fusion of remote sensing data for the urban landscape for optimizing the urban management of landscape border to promoting the inclusive of the urban landscape to all communities.

Keywords: urban land cover changes, remote sensing, high spatiotemporal fusion, urban management

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Authors: Yi Li, Yinan Zheng, Ya Ke, Yungwing Ho

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Motor learning is a process that frequently happens among humans and rodents, which is defined as the changes in the capability to perform a skill that is conformed to have a relatively permanent improvement through practice or experience. There are many ways to learn a behavior, among which is observational learning. Observational learning is the process of learning by watching the behaviors of others, for example, a child imitating parents, learning a new sport by watching the training videos or solving puzzles by watching the solutions. Many research explores observational learning in humans and primates. However, the neuronal mechanism of which, especially observational motor learning, was uncertain. It’s well accepted that mirror neurons are essential in the observational learning process. These neurons fire when the primate performs a goal-directed action and sees someone else demonstrating the same action, which suggests they have high firing activity both completing and watching the behavior. The mirror neurons are assumed to mediate imitation or play a critical and fundamental role in action understanding. They are distributed in many brain areas of primates, i.e., posterior parietal cortex (PPC), premotor cortex (M2), and primary motor cortex (M1) of the macaque brain. However, few researchers report the existence of mirror neurons in rodents. To verify the existence of mirror neurons and the possible role in motor learning in rodents, we performed customised string-pulling behavior combined with multiple behavior analysis methods, photometry, electrophysiology recording, c-fos staining and optogenetics in healthy mice. After five days of training, the demonstrator (demo) mice showed a significantly quicker response and shorter time to reach the string; fast, steady and accurate performance to pull down the string; and more precisely grasping the beads. During three days of observation, the mice showed more facial motions when the demo mice performed behaviors. On the first training day, the observer reduced the number of trials to find and pull the string. However, the time to find beads and pull down string were unchanged in the successful attempts on the first day and other training days, which indicated successful action understanding but failed motor learning through observation in mice. After observation, the post-hoc staining revealed that the c-fos expression was increased in the cognitive-related brain areas (medial prefrontal cortex) and motor cortices (M1, M2). In conclusion, this project indicated that the observation led to a better understanding of behaviors and activated the cognitive and motor-related brain areas, which suggested the possible existence of mirror neurons in these brain areas.

Keywords: observation, motor learning, string-pulling behavior, prefrontal cortex, motor cortex, cognitive

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Authors: Milijana N. Miljkovic, Viktorija M. Dragojevic-Simic, Nemanja K. Rancic, Vesna M. Jacevic, Snezana B. Djordjevic, Momir M. Mikov, Aleksandra M. Kovacevic

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Itraconazole (ITZ) is a weak base and extremely lipophilic compound, with water solubility as a rate-limiting step in its absorption from the gastrointestinal tract. Its absolute bioavailability, about 55%, is maximal when its oral formulation, capsules, are taken immediately after a full meal. Peak plasma concentrations (Cmax) are reached within 2 to 5 hrs after their administration. ITZ undergoes extensive hepatic metabolism by human CYP3A4 isoenzyme and more than 30 different metabolites have been identified. One of the main ones is hydroxyitraconazole (HITZ), in which plasma concentrations are almost twice higher than those of ITZ. Gender differences in drug PK (Pharmacokinetics) have already been recognized, but variations in metabolism are believed to be their major cause. The aim of the study was to investigate the influence of gender on ITZ PK parameters after administration of oral capsule formulation, following 100 mg single dosing in healthy adult volunteers under fed conditions. The single-center, open-label PK study was performed. PK analyses included PK parameters obtained after a single 100 mg dose administration of itraconazole capsules to 48 females and 66 males. Blood samples were collected at pre-dose and up to 72.0 h after administration (1.0, 2.0, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 7.0, 9.0, 12.0, 24.0, 36.0 and 72.0 hrs). The calculated pharmacokinetic parameters, based on the plasma concentrations of itraconazole and hydroxyitraconazole, were Cmax, AUClast, and AUCtot. Plasma concentrations of ITZ and HITZ were determined using a validated liquid chromatographic method with mass spectrometric detection, while pharmacokinetic parameters were estimated using non-compartmental methods. The pharmacokinetic analyses were performed using Kinetica software version 5.0. The mean value of ITZ Cmaxmen was 74.79 ng/ml, and Cmaxwomen was 51.291 ng/ml (independent samples test; p = 0.005). Hydroxyitraconazole had a mean value of Cmaxmen 106.37 ng/ml, and the mean value Cmaxwomen was 70.05 ng/ml. Women had, on average, lower AUClast and Cmax than men. AUClastmen for ITZ was 736.02 ng/mL*h and AUClastwomen was 566.62 ng/mL*h, while AUClastmen for HITZ was 1154.80 was ng/mL*h and AUClastwomen for HITZ was 708.12 ng/mL*h (independent samples test; p = 0.033). The mean values of ITZ AUCtotmen were 884.73 ng/mL*h and AUCtotwomen was 685.10 ng/mL*h. AUCtotmen for HITZ was 1290.41 ng/mL*h, while AUCtotwomen for HIZT was 788.60 ng/mL*h (p < 0.001). The results could point out to lower oral bioavailability of ITZ in women, since values of Cmax, AUClast, and AUCtot of both ITZ and HITZ were significantly lower in women than in men, respectively. The reason may be higher expression and activity of CYP3A4 in women than in men, but there also may be differences in other PK parameters. High variability of both ITZ and HITZ concentrations in both genders confirmed that ITZ is a highly variable drug. Further examinations of its PK are needed to justify strategies for therapeutic drug monitoring in patients treated by this antifungal agent.

Keywords: itraconazole, gender, hydroxyitraconazole, pharmacokinetics

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Authors: Soraia V. M. de Sá, Miguel A. Faria, José O. Fernandes, Sara C. Cunha

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Cereals play a vital role in fulfilling the nutritional needs of children, supplying essential nutrients crucial for their growth and development. However, concerns arise due to children's heightened vulnerability due to their unique physiology, specific dietary requirements, and relatively higher intake in relation to their body weight. This vulnerability exposes them to harmful food contaminants, particularly mycotoxins, prevalent in cereals. Because of the thermal stability of mycotoxins, conventional industrial food processing often falls short of eliminating them. Children, especially those aged 4 months to 12 years, frequently encounter mycotoxins through the consumption of specialized food products, such as instant foods, breakfast cereals, bars, cookie snacks, fruit puree, and various dairy items. A close monitoring of this demographic group's exposure to mycotoxins is essential, as toxins ingestion may weaken children’s immune systems, reduce their resistance to infectious diseases, and potentially lead to cognitive impairments. The severe toxicity of mycotoxins, some of which are classified as carcinogenic, has spurred the establishment and ongoing revision of legislative limits on mycotoxin levels in food and feed globally. While EU Commission Regulation 1881/2006 addresses well-known mycotoxins in processed cereal-based foods and infant foods, the absence of regulations specifically addressing emerging mycotoxins underscores a glaring gap in the regulatory framework, necessitating immediate attention. Emerging mycotoxins have gained mounting scrutiny in recent years due to their pervasive presence in various foodstuffs, notably cereals and cereal-based products. Alarmingly, exposure to multiple mycotoxins is hypothesized to exhibit higher toxicity than isolated effects, raising particular concerns for products primarily aimed at children. This study scrutinizes the presence of 22 mycotoxins of the diverse range of chemical classes in 148 processed cereal-based foods, including 39 breakfast cereals, 25 infant formulas, 27 snacks, 25 cereal bars, and 32 cookies commercially available in Portugal. The analytical approach employed a modified QuEChERS procedure followed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. Given the paucity of information on the risk assessment of children to multiple mycotoxins in cereal and cereal-based products consumed by children of Portugal pioneers the evaluation of this critical aspect. Overall, aflatoxin B1 (AFB1) and aflatoxin G2 (AFG2) emerged as the most prevalent regulated mycotoxins, while enniatin B (ENNB) and sterigmatocystin (STG) were the most frequently detected emerging mycotoxins.

Keywords: cereal-based products, children´s nutrition, food safety, UPLC-MS/MS analysis

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Authors: Beate Englich, Linda Schlittenbauer, Christiane Pfeifer, Isabel Kratochvil, Michael Borte, Gabriele I. Stangl, Martin von Bergen, Thorsten Reemtsma, Irina Lehmann, Kristin M. Junge

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The worldwide production of endocrine disrupting compounds (EDC) has risen dramatically over the last decades, as so has the prevalence for obesity. Many EDCs are believed to contribute to this obesity epidemic, by enhancing adipogenesis or disrupting relevant metabolism. This effect is most tremendous in the early prenatal period when priming effects find a highly vulnerable time window. Therefore, we investigate the impact of parabens on childhood overweight development and adipogenesis in general. Parabens are ester of 4-hydroxy-benzoic acid and part of many cosmetic products or food packing. Therefore, ubiquitous exposure can be found in the westernized world, with exposure already starting during the sensitive prenatal period. We assessed maternal cosmetic product consumption, prenatal paraben exposure and infant BMI z-scores in the prospective German LINA cohort. In detail, maternal urinary concentrations (34 weeks of gestation) of methyl paraben (MeP), ethyl paraben (EtP), n-propyl paraben (PrP) and n-butyl paraben (BuP) were quantified using UPLC-MS/MS. Body weight and height of their children was assessed during annual clinical visits. Further, we investigated the direct influence of those parabens on adipogenesis in-vitro using a human mesenchymal stem cell (MSC) differentiation assay to mimic a prenatal exposure scenario. MSC were exposed to 0.1 – 50 µM paraben during the entire differentiation period. Differentiation outcome was monitored by impedance spectrometry, real-time PCR and triglyceride staining. We found that maternal cosmetic product consumption was highly correlated with urinary paraben concentrations at pregnancy. Further, prenatal paraben exposure was linked to higher BMI Z-scores in children. Our in-vitro analysis revealed that especially the long chained paraben BuP stimulates adipogenesis by increasing the expression of adipocyte specific genes (PPARγ, ADIPOQ, LPL, etc.) and triglyceride storage. Moreover, we found that adiponectin secretion is increased whereas leptin secretion is reduced under BuP exposure in-vitro. Further mechanistic analysis for receptor binding and activation of PPARγ and other key players in adipogenesis are currently in process. We conclude that maternal cosmetic product consumption is linked to prenatal paraben exposure of children and contributes to the development of infant overweight development by triggering key pathways of adipogenesis.

Keywords: adipogenesis, endocrine disruptors, paraben, prenatal exposure

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Authors: Maria Jose Granero Paris, Ana Isabel Jimenez Zarco, Agustin Pablo Alvarez Herranz

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In the industrial sector collaboration with suppliers is key to the development of innovations in the field of processes. Access to resources and expertise that are not available in the business, obtaining a cost advantage, or the reduction of the time needed to carry out innovation are some of the benefits associated with the process. However, the success of this collaborative process is compromised, when from the beginning not clearly rules have been established that govern the relationship. Abundant studies developed in the field of innovation emphasize the strategic importance of the concept of “Governance”. Despite this, there have been few papers that have analyzed how the governance process of the relationship must be designed and managed to ensure the success of the collaboration process. The lack of literature in this area responds to the wide diversity of contexts where collaborative processes to innovate take place. Thus, in sectors such as the car industry there is a strong collaborative tradition between manufacturers and suppliers being part of the value chain. In this case, it is common to establish mechanisms and procedures that fix formal and clear objectives to regulate the relationship, and establishes the rights and obligations of each of the parties involved. By contrast, in other sectors, collaborative relationships to innovate are not a common way of working, particularly when their aim is the development of process improvements. It is in this case, it is when the lack of mechanisms to establish and regulate the behavior of those involved, can give rise to conflicts, and the failure of the cooperative relationship. Because of this the present paper analyzes the similarities and differences in the processes of governance in collaboration with suppliers in the European aerospace industry With these ideas in mind, we present research is twofold: Understand the importance of governance as a key element of the success of the collaboration in the development of product and process innovations, Establish the mechanisms and procedures to ensure the proper management of the processes of collaboration. Following the methodology of the case study, we analyze the way in which manufacturers and suppliers cooperate in the development of new products and processes in two industries with different levels of technological intensity and collaborative tradition: the automotive and aerospace. The identification of those elements playing a key role to establish a successful governance and relationship management and the compression of the mechanisms of regulation and control in place at the automotive sector can be use to propose solutions to some of the conflicts that currently arise in aerospace industry. The paper concludes by analyzing the strategic implications for the aerospace industry entails the adoption of some of the practices traditionally used in other industrial sectors. Finally, it is important to highlight that in this paper are presented the first results of a research project currently in progress describing a model of governance that explains the way to manage outsourced services to suppliers in the European aerospace industry, through the analysis of companies in the sector located in Germany, France and Spain.

Keywords: supplier collaboration, supplier relationship governance, innovation management, product innovation, process innovation

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

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

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

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Authors: Rayanne Branco Dos Santos Lima, Lorena Pinheiro Barbosa, Kamila Ferreira Lima, Victor Manuel Tegoma Ruiz, Monyka Brito Lima Dos Santos, Maria Wendiane Gueiros Gaspar, Luzia Camila Coelho Ferreira, Leandro Cardozo Dos Santos Brito, Deyse Maria Alves Rocha

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Introduction: The lack of regulation of the baby's sleep-wake pattern in the first years of life affects the health of thousands of women. Maternal sleep deprivation can trigger or aggravate psychosomatic problems such as depression, anxiety and stress that can directly influence maternal safety, with consequences for the baby's and mother's sleep. Such conditions can affect the family's quality of life and child development. Objective: To correlate maternal security with maternal state anxiety scores and the mother's and baby's total sleep time. Method: Cross-sectional study carried out with 96 mothers of babies aged 10 to 24 months, accompanied by nursing professionals linked to a Federal University in Northeast Brazil. Study variables were maternal security, maternal state anxiety scores, infant latency and sleep time, and total nocturnal sleep time of mother and infant. Maternal safety was calculated using a four-point Likert scale (1=not at all safe, 2=somewhat safe, 3=very safe, 4=completely safe). Maternal anxiety was measured by State-Trait Anxiety Inventory, state-anxiety subscale whose scores vary from 20 to 80 points, and the higher the score, the higher the anxiety levels. Scores below 33 are considered mild; from 33 to 49, moderate and above 49, high. As for the total nocturnal sleep time, values between 7-9 hours of sleep were considered adequate for mothers, and values between 9-12 hours for the baby, according to the guidelines of the National Sleep Foundation. For the sleep latency time, a time equal to or less than 20 min was considered adequate. It is noteworthy that the latency time and the time of night sleep of the mother and the baby were obtained by the mother's subjective report. To correlate the data, Spearman's correlation was used in the statistical package R version 3.6.3. Results: 96 women and babies participated, aged 22 to 38 years (mean 30.8) and 10 to 24 months (mean 14.7), respectively. The average of maternal security was 2.89 (unsafe); Mean maternal state anxiety scores were 43.75 (moderate anxiety). The babies' average sleep latency time was 39.6 min (>20 min). The mean sleep times of the mother and baby were, respectively, 6h and 42min and 8h and 19min, both less than the recommended nocturnal sleep time. Maternal security was positively correlated with maternal state anxiety scores (rh=266, p=0.009) and negatively correlated with infant sleep latency (rh= -0.30. P=0.003). Baby sleep time was positively correlated with maternal sleep time. (rh 0.46, p<0.001). Conclusion: The more secure the mothers considered themselves, the higher the anxiety scores and the shorter the baby's sleep latency. Also, the longer the baby sleeps, the longer the mother sleeps. Thus, interventions are needed to promote the quality and efficiency of sleep for both mother and baby.

Keywords: sleep, anxiety, infant, mother-child relations

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Authors: Hortènsia Curell, Ana Fernández-Montraveta

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This paper presents the creation, development, and analysis of a corpus designed to study the presence of attitude markers and author’s stance in research articles in two different areas of linguistics (theoretical linguistics and sociolinguistics). These two disciplines are expected to behave differently in this respect, given the disparity in their discursive conventions. Attitude markers in this work are understood as the linguistic elements (adjectives, nouns and verbs) used to convey the writer's stance towards the content presented in the article, and are crucial in understanding writer-reader interaction and the writer's position. These attitude markers are divided into three broad classes: assessment, significance, and emotion. In addition to them, we also consider first-person singular and plural pronouns and possessives, modal verbs, and passive constructions, which are other linguistic elements expressing the author’s stance. The corpus, Corpus of Attitude in Academic Writing (CAAW), comprises a collection of 21 articles, collected from six journals indexed in JCR. These articles were originally written in English by a single native-speaker author from the UK or USA and were published between 2022 and 2023. The total number of words in the corpus is approximately 222,400, with 106,422 from theoretical linguistics (Lingua, Linguistic Inquiry and Journal of Linguistics) and 116,022 from sociolinguistics journals (International Journal of the Sociology of Language, Language in Society and Journal of Sociolinguistics). Together with the corpus, we present the tool created for the creation and storage of the corpus, along with a tool for automatic annotation. The steps followed in the compilation of the corpus are as follows. First, the articles were selected according to the parameters explained above. Second, they were downloaded and converted to txt format. Finally, examples, direct quotes, section titles and references were eliminated, since they do not involve the author’s stance. The resulting texts were the input for the annotation of the linguistic features related to stance. As for the annotation, two articles (one from each subdiscipline) were annotated manually by the two researchers. An existing list was used as a baseline, and other attitude markers were identified, together with the other elements mentioned above. Once a consensus was reached, the rest of articles were annotated automatically using the tool created for this purpose. The annotated corpus will serve as a resource for scholars working in discourse analysis (both in linguistics and communication) and related fields, since it offers new insights into the expression of attitude. The tools created for the compilation and annotation of the corpus will be useful to study author’s attitude and stance in articles from any academic discipline: new data can be uploaded and the list of markers can be enlarged. Finally, the tool can be expanded to other languages, which will allow cross-linguistic studies of author’s stance.

Keywords: academic writing, attitude, corpus, english

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Authors: B. Ratchaneepun, N.S. Bardell

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It is recognized that people are the main drivers in almost all the processes that affect airworthiness assurance. This is especially true in the area of aircraft maintenance, which is an essential part of continuing airworthiness. This work investigates what impact English language proficiency, the intersection of the military and Thai cultures, and the lack of initial and continuing human factors training have on the work performance of maintenance personnel in the Royal Thai Air Force (RTAF). A quantitative research method based on a cross-sectional survey was used to gather data about these three key aspects of “people” in a military airworthiness environment. 30 questions were developed addressing the crucial topics of English language proficiency, impact of culture, and human factors training. The officers and the non-commissioned officers (NCOs) who work for the Aeronautical Engineering Divisions in the RTAF comprised the survey participants. The survey data were analysed to support various hypotheses by using a t-test method. English competency in the RTAF is very important since all of the service manuals for Thai military aircraft are written in English. Without such competency, it is difficult for maintenance staff to perform tasks and correctly interpret the relevant maintenance manual instructions; any misunderstandings could lead to potential accidents. The survey results showed that the officers appreciated the importance of this more than the NCOs, who are the people actually doing the hands-on maintenance work. Military culture focuses on the success of a given mission, and leverages the power distance between the lower and higher ranks. In Thai society, a power distance also exists between younger and older citizens. In the RTAF, such a combination tends to inhibit a just reporting culture and hence hinders safety. The survey results confirmed this, showing that the older people and higher ranks involved with RTAF aircraft maintenance believe that the workplace has a positive safety culture and climate, whereas the younger people and lower ranks think the opposite. The final area of consideration concerned human factors training and non-technical skills training. The survey revealed that those participants who had previously attended such courses appreciated its value and were aware of its benefits in daily life. However, currently there is no regulation in the RTAF to mandate recurrent training to maintain such knowledge and skills. The findings from this work suggest that the people involved in assuring the continuing airworthiness of the RTAF would benefit from: (i) more rigorous requirements and standards in the recruitment, initial training and continuation training regarding English competence; (ii) the development of a strong safety culture that exploits the uniqueness of both the military culture and the Thai culture; and (iii) providing more initial and recurrent training in human factors and non-technical skills.

Keywords: aircraft maintenance, continuing airworthiness, military culture, people, Royal Thai Air Force

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Authors: Marta Kwiatkowska, Paweł Jarosiewicz, Bożena Bukowska

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Glyphosate (N-phosphonomethylglycine) is a non-selected broad spectrum ingredient in the herbicide (Roundup) used for over 35 years for the protection of agricultural and horticultural crops. Glyphosate was believed to be environmentally friendly but recently, a large body of evidence has revealed that glyphosate can negatively affect on environment and humans. It has been found that glyphosate is present in the soil and groundwater. It can also enter human body which results in its occurrence in blood in low concentrations of 73.6 ± 28.2 ng/ml. Research conducted for potential genotoxicity and cytotoxicity can be an important element in determining the toxic effect of glyphosate. Due to regulation of European Parliament 1107/2009 it is important to assess genotoxicity and cytotoxicity not only for the parent substance but also its impurities, which are formed at different stages of production of major substance – glyphosate. Moreover verifying, which of these compounds are more toxic is required. Understanding of the molecular pathways of action is extremely important in the context of the environmental risk assessment. In 2002, the European Union has decided that glyphosate is not genotoxic. Unfortunately, recently performed studies around the world achieved results which contest decision taken by the committee of the European Union. World Health Organization (WHO) in March 2015 has decided to change the classification of glyphosate to category 2A, which means that the compound is considered to "probably carcinogenic to humans". This category relates to compounds for which there is limited evidence of carcinogenicity to humans and sufficient evidence of carcinogenicity on experimental animals. That is why we have investigated genotoxicity and cytotoxicity effects of the most commonly used pesticide: glyphosate and its impurities: N-(phosphonomethyl)iminodiacetic acid (PMIDA) and bis-(phosphonomethyl)amine on human peripheral blood mononuclear cells (PBMCs), mostly lymphocytes. DNA damage (analysis of DNA strand-breaks) using the single cell gel electrophoresis (comet assay) and ATP level were assessed. Cells were incubated with glyphosate and its impurities: PMIDA and bis-(phosphonomethyl)amine at concentrations from 0.01 to 10 mM for 24 hours. Evaluating genotoxicity using the comet assay showed a concentration-dependent increase in DNA damage for all compounds studied. ATP level was decreased to zero as a result of using the highest concentration of two investigated impurities, like bis-(phosphonomethyl)amine and PMIDA. Changes were observed using the highest concentration at which a person can be exposed as a result of acute intoxication. Our survey leads to a conclusion that the investigated compounds exhibited genotoxic and cytotoxic potential but only in high concentrations, to which people are not exposed environmentally. Acknowledgments: This work was supported by the Polish National Science Centre (Contract-2013/11/N/NZ7/00371), MSc Marta Kwiatkowska, project manager.

Keywords: cell viability, DNA damage, glyphosate, impurities, peripheral blood mononuclear cells

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Authors: Adeiza Matthew, Oluwadamilola Abubakar

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Wood energy, also known as wood fuel, is a renewable energy source that is derived from woody biomass, which is organic matter that is harvested from forests, woodlands, and other lands. Woody biomass includes trees, branches, twigs, and other woody debris that can be used as fuel. Wood energy can be classified based on its sources, such as trees outside forests, residues from wood harvesting and conversion, and energy plantations. There are several policy frameworks that support the use of wood energy, including participatory forest management and agroforestry. These policies aim to promote the sustainable use of woody biomass as a source of energy while also protecting forests and wildlife habitats. There are several options for using wood as a fuel, including central heating systems, pellet-based systems, wood chip-based systems, log boilers, fireplaces, and stoves. Each of these options has its own benefits and drawbacks, and the most appropriate option will depend on factors such as the availability of woody biomass, the heating needs of the household or facility, and the local climate. In order to use wood as a fuel, it must be harvested and stored properly. Hardwood or softwood can be used as fuel, and the heating value of firewood depends on the species of tree and the degree of moisture content. Proper harvesting and storage of wood can help to minimize environmental impacts and improve wildlife habitats. The use of wood energy has several environmental impacts, including the release of greenhouse gases during combustion and the potential for air pollution from combustion by-products. However, wood energy can also have positive environmental impacts, such as the sequestration of carbon in trees and the reduction of reliance on fossil fuels. The regulation and legislation of wood energy vary by country and region, and there is an ongoing debate about the potential use of wood energy in renewable energy technologies. Wood energy is a renewable energy source that can be used to generate electricity, heat, and transportation fuels. Woody biomass is abundant and widely available, making it a potentially significant source of energy for many countries. The use of wood energy can create local economic and employment opportunities, particularly in rural areas. Wood energy can be used to reduce reliance on fossil fuels and reduce greenhouse gas emissions. Properly managed forests can provide a sustained supply of woody biomass for energy, helping to reduce the risk of deforestation and habitat loss. Wood energy can be produced using a variety of technologies, including direct combustion, co-firing with fossil fuels, and the production of biofuels. The environmental impacts of wood energy can be minimized through the use of best practices in harvesting, transportation, and processing. Wood energy is regulated and legislated at the national and international levels, and there are various standards and certification systems in place to promote sustainable practices. Wood energy has the potential to play a significant role in the transition to a low-carbon economy and the achievement of climate change mitigation goals.

Keywords: biomass, timber, charcoal, firewood

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Authors: M. G. Shilina

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The interstate economic interaction phenomenon is complex. ‘Economic integration’, as one of its types, can be explored through the prism of international law, the theories of the world economy, politics and international relations. The most objective study of the phenomenon requires a comprehensive multifactoral approach. In new geopolitical realities, the problems of coexistence and possible interconnection of various mechanisms of interstate economic interaction are actively discussed. Currently, the Eurasian continent states support the direction to economic integration. At the same time, the existing international economic law fragmentation in Eurasia is seen as the important problem. The Eurasian space is characterized by a various types of interstate relations: international agreements (multilateral and bilateral), and a large number of cooperation formats (from discussion platforms to organizations aimed at deep integration). For their harmonization, it is necessary to have a clear vision to the phased international economic relations regulation options. In the conditions of rapid development of international economic relations, the modeling (including prognostic) can be optimally used as the main scientific method for presenting the phenomenon. On the basis of this method, it is possible to form the current situation vision and the best options for further action. In order to determine the most objective version of the integration development, the combination of several approaches were used. The normative legal approach- the descriptive method of legal modeling- was taken as the basis for the analysis. A set of legal methods was supplemented by the international relations science prognostic methods. The key elements of the model are the international economic organizations and states' associations existing in the Eurasian space (the Eurasian Economic Union (EAEU), the European Union (EU), the Shanghai Cooperation Organization (SCO), Chinese project ‘One belt-one road’ (OBOR), the Commonwealth of Independent States (CIS), BRICS, etc.). A general term for the elements of the model is proposed - the interstate interaction mechanisms (IIM). The aim of building a model of current and future Eurasian economic integration is to show optimal options for joint economic development of the states and IIMs. The long-term goal of this development is the new economic and political space, so-called the ‘Great Eurasian Community’. The process of achievement this long-term goal consists of successive steps. Modeling the integration architecture and dividing the interaction into stages led us to the following conclusion: the SCO is able to transform Eurasia into a single economic space. Gradual implementation of the complex phased model, in which the SCO+ plays a key role, will allow building an effective economic integration for all its participants, to create an economically strong community. The model can have practical value for politicians, lawyers, economists and other participants involved in the economic integration process. A clear, systematic structure can serve as a basis for further governmental action.

Keywords: economic integration, The Eurasian Economic Union, The European Union, The Shanghai Cooperation Organization, The Silk Road Economic Belt

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Authors: Avadh Bihari

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Corporate social responsibility (CSR) has become a global parameter to define corporates' ethical and responsible behaviour. It was a voluntary practice in India till 2013, driven by various guidelines, which has become a mandate since 2014 under the Companies Act, 2013. This has compelled the corporates to redesign their CSR strategies by bringing in structures, planning, accountability, and transparency in their processes with a mandate to 'comply or explain'. Based on the author's M.Phil. dissertation, this paper presents the changes in organisational practices and institutional mechanisms of third-sector organisations (TSOs) with the theoretical frameworks of institutionalism and co-optation. It became an interesting case as India is the only country to have a law on CSR, which is not only mandating the reporting but the spending too. The space of CSR in India is changing rapidly and affecting multiple institutions, in the context of the changing roles of the state, market, and TSOs. Several factors such as stringent regulation on foreign funding, mandatory CSR pushing corporates to look out for NGOs, and dependency of Indian NGOs on CSR funds have come to the fore almost simultaneously, which made it an important area of study. Further, the paper aims at addressing the gap in the literature on the effects of mandated CSR on the functioning of TSOs through the empirical and theoretical findings of this study. The author had adopted an interpretivist position in this study to explore changes in organisational practices from the participants' experiences. Data were collected through in-depth interviews with five corporate officials, eleven officials from six TSOs, and two academicians, located at Mumbai and Delhi, India. The findings of this study show the legislation has institutionalised CSR, and TSOs get co-opted in the process of implementing mandated CSR. Seventy percent of the corporates implement their CSR projects through TSOs in India; this has affected the organisational practices of TSOs to a large extent. They are compelled to recruit expert workforce, create new departments for monitoring & evaluation, communications, and adopt management practices of project implementation from corporates. These are attempts to institutionalise the TSOs so that they can produce calculated results as demanded by corporates. In this process, TSOs get co-opted in a struggle to secure funds and lose their autonomy. The normative, coercive, and mimetic isomorphisms of institutionalism come into play as corporates are mandated to take up CSR, thereby influencing the organisational practices of TSOs. These results suggest that corporates and TSOs require an understanding of each other's work culture to develop mutual respect and work towards the goal of sustainable development of the communities. Further, TSOs need to retain their autonomy and understanding of ground realities without which they become an extension of the corporate-funder. For a successful CSR project, engagement beyond funding is required from corporate, through their involvement and not interference. CSR-led community development can be structured by management practices to an extent, but cannot overshadow the knowledge and experience of TSOs.

Keywords: corporate social responsibility, institutionalism, organisational practices, third-sector organisations

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Authors: B. K. Manjunatha Goud, Aruna Chanu Oinam

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The undergraduate medical curriculum is oriented towards training the students to undertake the responsibilities of a physician. During the training period, adequate emphasis is placed on inculcating logical and scientific habits of thought; clarity of expression and independence of judgment; and ability to collect and analyze information and to correlate them. At Ras Al Khaimah Medical and Health Sciences University (RAKMHSU), Biochemistry a basic medical science subject is taught in the 1st year of 5 years medical course with vertical interdisciplinary interaction with all subjects, which needs to be taught and learned adequately by the students to be related to clinical case or clinical problem in medicine and future diagnostics so that they can practice confidently and skillfully in the community. Based on these facts study was done to know the extent of usage of library resources by the students and the impact of study materials on their preparation for examination. It was a comparative cross sectional study included 100 and 80 1st and 2nd-year students who had successfully completed Biochemistry course. The purpose of the study was explained to all students [participants]. Information was collected on a pre-designed, pre-tested and self-administered questionnaire. The questionnaire was validated by the senior faculties and pre tested on students who were not involved in the study. The study results showed that 80.30% and 93.15% of 1st and 2nd year students have the clear idea of course outline given in course handout or study guide. We also found a statistically significant number of students agreed that they were benefited from the practical session and writing notes in the class hour. A high percentage of students [50% and 62.02%] disagreed that that reading only the handouts is enough for their examination as compared to other students. The study also showed that only 35% and 41% of students visited the library on daily basis for the learning process, around 65% of students were using lecture notes and text books as a tool for learning and to understand the subject and 45% and 53% of students used the library resources (recommended text books) compared to online sources before the examinations. The results presented here show that students perceived that e-learning resources like power point presentations along with text book reading using SQ4R technique had made a positive impact on various aspects of their learning in Biochemistry. The use of library by students has overall positive impact on learning process especially in medical field enhances the outcome, and medical students are better equipped to treat the patient. But it’s also true that use of library use has been in decline which will impact the knowledge aspects and outcome. In conclusion, a student has to be taught how to use the library as learning tool apart from lecture handouts.

Keywords: medical education, learning resources, study guide, biochemistry

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Authors: Steven G. Sclan

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Background: Since December 2019, the world has been afflicted by the spread of the Severe Acute Respiratory Syndrome-Corona Virus-2 (SARS-CoV-2), which is responsible for the condition referred to as Covid-19. The illness has had a cataclysmic impact on the political, social, economic, and overall well-being of the population of the entire globe. While Covid-19 has had a substantial universal fatality impact, it may have an even greater effect on the socioeconomic, medical well-being, and healthcare planning for remaining societies. Significance: As these numbers illustrate, many more persons survive the infection than die from it, and many of those patients have noted ongoing, persistent symptoms after successfully enduring the acute phase of the illness. Recognition and understanding of these symptoms are crucial for developing and arranging efficacious models of care for all patients (whether or not having been hospitalized) surviving acute covid illness and plagued by post-acute symptoms. Furthermore, regarding Covid infection in children (< 18 y/o), although it may be that Covid “+” children are not major vectors of infective transmission, it now appears that many more children than initially thought are carrying the virus without accompanying obvious symptomatic expression. It seems reasonable to wonder whether viral effects occur in children – those children who are Covid “+” and now asymptomatic – and if, over time, they might also experience similar symptoms. An even more significant question is whether Covid “+” asymptomatic children might manifest increased multiple health problems as they grow – i.e., developmental complications (e.g., physical/medical, metabolic, neurobehavioral, etc.) – in comparison to children who had been consistently Covid “ - ” during the pandemic. Topics Addressed and Theoretical Importance: This review is important because of the description of both quantitative and qualitative methods for clinical and biomedical research. Topics reviewed will consider the importance of well-designed, comprehensive (i.e., quantitative and qualitative methods) longitudinal studies of Post Covid-19 symptoms in both adults and children. Also reviewed will be general characteristics of longitudinal studies and a presentation of a model for a proposed study. Also discussed will be the benefit of longitudinal studies for the development of efficacious interventions and for the establishment of cogent, practical, and efficacious community healthcare service planning for post-acute covid patients. Conclusion: Results of multi-dimensional, longitudinal studies will have important theoretical implications. These studies will help to improve our understanding of the pathophysiology of long COVID and will aid in the identification of potential targets for treatment. Such studies can also provide valuable insights into the long-term impact of COVID-19 on public health and socioeconomics.

Keywords: COVID-19, post-COVID-19, long COVID, longitudinal research, quantitative research, qualitative research

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Authors: N. Manjunatha, K. T. Rangswamy, N. Nagaraju, H. A. Prameela, P. Rudraswamy, M. Krishnareddy

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The identification of virus in cowpea especially potyviruses is confusing. Even though there are several studies on viruses causing diseases in cowpea, difficult to distinguish based on symptoms and serological detection. The differentiation of potyviruses considering as a constraint, the present study is initiated for molecular characterization, host plant resistance and epidemiology of the BCMV infecting cowpea. The etiological agent causing cowpea mosaic was identified as Bean Common Mosaic Virus (BCMV) on the basis of RT-PCR and electron microscopy. An approximately 750bp PCR product corresponding to coat protein (CP) region of the virus and the presence of long flexuous filamentous particles measuring about 952 nm in size typical to genus potyvirus were observed under electron microscope. The characterized virus isolate genome had 10054 nucleotides, excluding the 3’ terminal poly (A) tail. Comparison of polyprotein of the virus with other potyviruses showed similar genome organization with 9 cleavage sites resulted in 10 functional proteins. The pairwise sequence comparison of individual genes, P1 showed most divergent, but CP gene was less divergent at nucleotide and amino acid level. A phylogenetic tree constructed based on multiple sequence alignments of the polyprotein nucleotide and amino acid sequences of cowpea BCMV and potyviruses showed virus is closely related to BCMV-HB. Whereas, Soybean variant of china (KJ807806) and NL1 isolate (AY112735) showed 93.8 % (5’UTR) and 94.9 % (3’UTR) homology respectively with other BCMV isolates. This virus transmitted to different leguminous plant species and produced systemic symptoms under greenhouse conditions. Out of 100 cowpea genotypes screened, three genotypes viz., IC 8966, V 5 and IC 202806 showed immune reaction in both field and greenhouse conditions. Single marker analysis (SMA) was revealed out of 4 SSR markers linked to BCMV resistance, M135 marker explains 28.2 % of phenotypic variation (R2) and Polymorphic information content (PIC) value of these markers was ranged from 0.23 to 0.37. The correlation and regression analysis showed rainfall, and minimum temperature had significant negative impact and strong relationship with aphid population, whereas weak correlation was observed with disease incidence. Path coefficient analysis revealed most of the weather parameters exerted their indirect contributions to the aphid population and disease incidence except minimum temperature. This study helps to identify specific gaps in knowledge for researchers who may wish to further analyse the science behind complex interactions between vector-virus and host in relation to the environment. The resistant genotypes identified are could be effectively used in resistance breeding programme.

Keywords: cowpea, epidemiology, genotypes, virus

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Authors: Arkadiusz Gut, Robert Mirski

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The nativist-constructivist debate constitutes a considerable part of current research on mindreading. Peter Carruthers and his colleagues are known for their nativist position in the debate and take issue with constructivist views proposed by other researchers, with Henry Wellman, Alison Gopnik, and Ian Apperly at the forefront. More specifically, Carruthers together with Evan Westra propose a nativistic explanation of Theory of Mind Scale study results that Wellman et al. see as supporting constructivism. While allowing for development of the innate mindreading system, Westra and Carruthers base their argumentation essentially on a competence-performance gap, claiming that cross-cultural differences in Theory of Mind Scale progression as well as discrepancies between infants’ and toddlers’ results on verbal and non-verbal false-belief tasks are fully explainable in terms of acquisition of other, pragmatic, cognitive developments, which are said to allow for an expression of the innately present Theory of Mind understanding. The goal of the present paper is to bring together arguments against the view offered by Westra and Carruthers. It will be shown that even though Carruthers et al.’s interpretation has not been directly controlled for in Wellman et al.’s experiments, there are serious reasons to dismiss such nativistic views which Carruthers et al. advance. The present paper discusses the following issues that undermine Carruthers et al.’s nativistic conception: (1) The concept of innateness is argued to be developmentally inaccurate; it has been dropped in many biological sciences altogether and many developmental psychologists advocate for doing the same in cognitive psychology. Reality of development is a complex interaction of changing elements that is belied by the simplistic notion of ‘the innate.’ (2) The purported innate mindreading conceptual system posited by Carruthers ascribes adult-like understanding to infants, ignoring the difference between first- and second-order understanding, between what can be called ‘presentation’ and ‘representation.’ (3) Advances in neurobiology speak strongly against any inborn conceptual knowledge; neocortex, where conceptual knowledge finds its correlates, is said to be largely equipotential at birth. (4) Carruthers et al.’s interpretations are excessively charitable; they extend results of studies done with 15-month-olds to conclusions about innateness, whereas in reality at that age there has been plenty of time for construction of the skill. (5) Looking-time experiment paradigm used in non-verbal false belief tasks that provide the main support for Carruthers’ argumentation has been criticized on methodological grounds. In the light of the presented arguments, nativism in theory of mind research is concluded to be an untenable position.

Keywords: development, false belief, mindreading, nativism, theory of mind

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Authors: Gleb Fomin, Kairat Tabynov, Nurkeldy Turebekov, Dinara Turegeldiyeva, Rinat Islamov

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The level of major cytokines in the blood of patients with COVID-19 varies greatly depending on age, gender, duration and severity of infection, and comorbidity. There are two clinically significant cytokines, IL-6 and TNF-α, which increase in levels in patients with severe COVID-19. However, in a model of COVID-19 in hamsters, TNF-α levels are unchanged or reduced, while the expression of other cytokines reflects the profile of cytokines found in patients’ plasma. The aim of our study was to evaluate the relationship between the level of cytokines in the blood, lungs, and lung damage in the model of the Syrian hamster (Mesocricetus auratus) infected with the SARS-CoV-2 strain. The study used outbred female and male Syrian hamsters (n=36, 4 groups) weighing 80-110 g and 5 months old (protocol IACUC, #4, 09/22/2020). Animals were infected intranasally with the hCoV-19/Kazakhstan/KazNAU-NSCEDI-481/2020 strain and euthanized at 3 d.p.i. The level of cytokines IL-6, TNF-α, IFN-α, and IFN-γ was determined by ELISA MyBioSourse (USA) for hamsters. Lung samples were subjected to histological processing. The presence of pathological changes in histological preparations was assessed on a 3-point scale. The work was carried out in the ABSL-3 laboratory. The data were analyzed in GraphPad Prism 6.00 (GraphPad Software, La Jolla, California, USA). The work was supported by the MES RK grant (AP09259865). In the blood, the level of TNF-α increased in males (p=0.0012) and IFN-γ in males and females (p=0.0001). On the contrary, IFN-α production decreased (p=0.0006). Only TNF-α level increased in lung tissues (p=0.0011). Correlation analysis showed a negative relationship between the level of IL-6 in the blood and lung damage in males (r -0.71, p=0.0001) and females (r-0.57, p=0.025). On the contrary, in males, the level of IL-6 in the lungs and score is positively correlated (r 0.80, p=0.01). The level of IFN-γ in the blood (r -0.64, p=0.035) and lungs (r-0.72, p=0.017) in males has a negative correlation with lung damage. No links were found for TNF-α and IFN-α. The study showed a positive association between lung injury and tissue levels of IL-6 in male hamsters. It is known that in humans, high concentrations of IL-6 in the lungs are associated with suppression of cellular immunity and, as a result, with an increase in the severity of COVID-19. TNF-α and IFN-γ play a key role in the pathogenesis of COVID-19 in hamsters. However, the mechanisms of their activity require more detailed study. IFN-α plays a lesser role in direct lung injury in a Syrian hamster model. We have shown the significance of tissue IL-6 and IFN-γ as predictors of the severity of lung damage in COVID-19 in the Syrian hamster model. Changes in the level of cytokines in the blood may not always reflect pathological processes in the lungs with COVID-19.

Keywords: syrian hamster, COVID-19, cytokines, biological model

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Authors: Ratna Sharma, Kaveri Dutta

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Assam (India), the gate way to the Northeast India is mainly known for its exquisite silks, the art and craft. The state has a rich collection of traditional jewellery which is unique and exclusive to the state. These jewelleries hold a special place in the heart of the Assamese women. Similarly handloom industry of Assam is basically silk oriented. Among the wild silk, Eri silk fabric has remained as “the poor man’s silk” but it is closely attached to the assamese society, dress for it's warm quality. In view of the changing market trends, fashion and consumer demands, Silk is emerging as a fashion fabric both in India and abroad. In case of Eri silk fabric it has limited use in clothing and accessories. Hence the restructured and redesigned traditional jewellery motifs of Assam (India) over Eri silk products will have greater potential in reviving the decline of art, generate revenue, self employment towards craftsmen and also recognition of the art. The information incorporated in the paper is primary and the data have been collected by purposive sampling method. This work of art was expressed on Eri silk fabric in the form of traditional hand embroidery as it is closely connected with the era of the individual in history of mankind and reflects the personal expression of an entity. For this study selected traditional motifs of Assamese ornaments was used. Some of the popular traditional Assamese jewellery include earrings with exquisite Lokaparo, Keru, Thuriya, Jangphai, etc. An array of necklaces including Golpata, Satsori, Jon biri, Bena, Gejera, Dhol biri, Doog doogi, Biri Moni, Mukuta Moni, Poalmoni, Silikha Moni and Magardana and diversified rings including Senpata, Horinsakua, Jethinejia, bakharpata and others. Selected two motifs each from necklace, earring and finger ring designs. Selected motifs were further developed into 3 categories- the border, the main motif and all over butta followed by placement of developed patterns on products. Products developed were stoles, scarf’s, purses, brooch pins, skirts for women and ties, handkerchief, jackets for men. The developed products were surveyed by selected respondents. From the present study it can be observed that the embellished traditional jewellery motifs resulted in fresh and colourful pattern on developed Eri silk products. Moreover the motifs which were gradually fading among the community itself showed a very good recognition towards art. The embroidered Eri silk fabric also created a huge change in a positive way among craftsman.

Keywords: Art and craft of Assam, eri silk, hand embroidery, traditional Assamese jewellery motifs

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Authors: Jong H. Kim, Kathleen L. Chan, Luisa W. Cheng

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Control of fungal pathogens, such as foodborne mycotoxin producers, is problematic as effective antimycotic agents are often very limited. Mycotoxin contamination significantly interferes with the safe production of foods or crops worldwide. Moreover, expansion of fungal resistance to commercial drugs or fungicides is a global human health concern. Therefore, there is a persistent need to enhance the efficacy of commercial antimycotic agents or to develop new intervention strategies. Disruption of the cellular antioxidant system should be an effective method for pathogen control. Such disruption can be achieved with safe, redox-active compounds. Natural phenolic derivatives are potent redox cyclers that inhibit fungal growth through destabilization of the cellular antioxidant system. The goal of this study is to identify novel, redox-active compounds that disrupt the fungal antioxidant system. The identified compounds could also function as sensitizing agents to conventional antimycotics (i.e., chemosensitization) to improve antifungal efficacy. Various benzo derivatives were tested against fungal pathogens. Gene deletion mutants of the yeast Saccharomyces cerevisiae were used as model systems for identifying molecular targets of benzo analogs. The efficacy of identified compounds as potent antifungal agents or as chemosensitizing agents to commercial drugs or fungicides was examined with methods outlined by the Clinical Laboratory Standards Institute or the European Committee on Antimicrobial Susceptibility Testing. Selected benzo derivatives possessed potent antifungal or antimycotoxigenic activity. Molecular analyses by using S. cerevisiae mutants indicated antifungal activity of benzo derivatives was through disruption of cellular antioxidant or cell wall integrity system. Certain benzo analogs screened overcame tolerance of Aspergillus signaling mutants, namely mitogen-activated protein kinase mutants, to fludioxonil fungicide. Synergistic antifungal chemosensitization greatly lowered minimum inhibitory or fungicidal concentrations of test compounds, including inhibitors of mitochondrial respiration. Of note, salicylaldehyde is a potent antimycotic volatile that has some practical application as a fumigant. Altogether, benzo derivatives targeting cellular antioxidant system of fungi (along with cell wall integrity system) effectively suppress fungal growth. Candidate compounds possess the antifungal, antimycotoxigenic or chemosensitizing capacity to augment the efficacy of commercial antifungals. Therefore, chemogenetic approaches can lead to the development of novel antifungal intervention strategies, which enhance the efficacy of established microbe intervention practices and overcome drug/fungicide resistance. Chemosensitization further reduces costs and alleviates negative side effects associated with current antifungal treatments.

Keywords: antifungals, antioxidant system, benzo derivatives, chemosensitization

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Authors: Hooriya Sadr Dadras, Shiva Seyed Erfani

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Speaking skills merit meticulous attention both on the side of the learners and the teachers. In particular, accuracy is a critical component to guarantee the messages to be conveyed through conversation because a wrongful change may adversely alter the content and purpose of the talk. Different types of tasks have served teachers to meet numerous educational objectives. Besides, negotiation of meaning and the use of different strategies have been areas of concern in socio-cultural theories of SLA. Negotiation of meaning is among the conversational processes which have a crucial role in facilitating the understanding and expression of meaning in a given second language. Conversational strategies are used during interaction when there is a breakdown in communication that leads to the interlocutor attempting to remedy the gap through talk. Therefore, this study was an attempt to investigate if there was any significant difference between the effect of reasoning gap tasks and information gap tasks on the frequency of conversational strategies used in negotiation of meaning in classrooms on one hand, and on the accuracy in speaking of Iranian intermediate EFL learners on the other. After a pilot study to check the practicality of the treatments, at the outset of the main study, the Preliminary English Test was administered to ensure the homogeneity of 87 out of 107 participants who attended the intact classes of a 15 session term in one control and two experimental groups. Also, speaking sections of PET were used as pretest and posttest to examine their speaking accuracy. The tests were recorded and transcribed to estimate the percentage of the number of the clauses with no grammatical errors in the total produced clauses to measure the speaking accuracy. In all groups, the grammatical points of accuracy were instructed and the use of conversational strategies was practiced. Then, different kinds of reasoning gap tasks (matchmaking, deciding on the course of action, and working out a time table) and information gap tasks (restoring an incomplete chart, spot the differences, arranging sentences into stories, and guessing game) were manipulated in experimental groups during treatment sessions, and the students were required to practice conversational strategies when doing speaking tasks. The conversations throughout the terms were recorded and transcribed to count the frequency of the conversational strategies used in all groups. The results of statistical analysis demonstrated that applying both the reasoning gap tasks and information gap tasks significantly affected the frequency of conversational strategies through negotiation. In the face of the improvements, the reasoning gap tasks had a more significant impact on encouraging the negotiation of meaning and increasing the number of conversational frequencies every session. The findings also indicated both task types could help learners significantly improve their speaking accuracy. Here, applying the reasoning gap tasks was more effective than the information gap tasks in improving the level of learners’ speaking accuracy.

Keywords: accuracy in speaking, conversational strategies, information gap tasks, reasoning gap tasks

Procedia PDF Downloads 290