Search results for: toehold mediated DNA switch

Authors: Mary H Moen, Lauren H. Mandel

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Creating a sense of community in online learning is important for student engagement and success. The integration of discussion forums within online learning environments presents an opportunity to explore how this computer mediated communications format can cultivate a sense of community among students in accelerated master’s degree programs. This research has two aims, to delve into the ways instructors utilize this communications technology to create community and to understand the feelings and experiences of graduate students participating in these forums in regard to its effectiveness in community building. This study is a two-phase approach encompassing qualitative and quantitative methodologies. The data will be collected at an online accelerated Master of Library and Information Studies program at a public university in the northeast of the United States. Phase 1 is a content analysis of the syllabi from all courses taught in the 2023 calendar year, which explores the format and rules governing discussion forum assignments. Four to six individual interviews of department faculty and part time faculty will also be conducted to illuminate their perceptions of the successes and challenges of their discussion forum activities. Phase 2 will be an online survey administered to students in the program during the 2023 calendar year. Quantitative data will be collected for statistical analysis, and short answer responses will be analyzed for themes. The survey is adapted from the Classroom Community Scale Short-Form (CSS-SF), which measures students' self-reported responses on their feelings of connectedness and learning. The prompts will contextualize the items from their experience in discussion forums during the program. Short answer responses on the challenges and successes of using discussion forums will be analyzed to gauge student perceptions and experiences using this type of communication technology in education. This research study is in progress. The authors anticipate that the findings will provide a comprehensive understanding of the varied approaches instructors use in discussion forums for community-building purposes in an accelerated MLIS program. They predict that the more varied, flexible, and consistent student uses of discussion forums are, the greater the sense of community students will report. Additionally, students’ and instructors’ perceptions and experiences within these forums will shed light on the successes and challenges faced, thereby offering valuable recommendations for enhancing online learning environments. The findings are significant because they can contribute actionable insights for instructors, educational institutions, and curriculum designers aiming to optimize the use of discussion forums in online accelerated graduate programs, ultimately fostering a richer and more engaging learning experience for students.

Keywords: accelerated online learning, discussion forums, LIS programs, sense of community, g

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Authors: Barbara Behre, Verolien Cauberghe, Dieneke Van de Sompel

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Sustainable consumption is on the rise, yet it has still not entered the mainstream in several industries, such as the fashion industry. In online fashion contexts, sustainability cues have been used to signal the sustainable benefits of certain garments to promote sustainable consumption. These sustainable cues may focus on the ecological or social dimension of sustainability. Since sustainability, in general, relates to distant, abstract benefits, the current study aims to examine if and how psychological distance may mediate the effects of exposure to different sustainability cues on consumption outcomes. Following the framework of Construal Level Theory of Psychological Distance, reduced psychological distance renders the construal level more concrete, which may influence attitudes and subsequent behavior in situations like fashion shopping. Most studies investigated sustainability as a composite, failing to differentiate between ecological and societal aspects of sustainability. The few studies examining sustainability more in detail uncovered that environmental sustainability is rather perceived in abstract cognitive construal, whereas social sustainability is linked to concrete construal. However, the construal level affiliation of the sustainability dimensions likely is not universally applicable to different domains and stages of consumption, which further suggest a need to clarify the relationships between environmental and social sustainability dimensions and the construal level of psychological distance within fashion brand consumption. While psychological distance and construal level have been examined in the context of sustainability, these studies yielded mixed results. The inconsistent findings of past studies might be due to the context-dependence of psychological distance as inducing construal differently in diverse situations. Especially in a hedonic consumption context like online fashion shopping, the role of visual processing of information could determine behavioural outcomes as linked to situational construal. Given the influence of the mode of processing on psychological distance and construal level, the current study examines the moderating role of verbal versus non-verbal presentation of the sustainability cues. In a 3 (environmental sustainability vs. social sustainability vs. control) x 2 (non-verbal message vs. verbal message) between subjects experiment, the present study thus examines how consumers evaluate sustainable brands in online shopping contexts in terms of psychological distance and construal level, as well as the impact on brand attitudes and buying intentions. The results among 246 participants verify the differential impact of the sustainability dimensions on fashion brand purchase intent as mediated by construal level and perceived psychological distance. The ecological sustainability cue is perceived as more concrete, which might be explained by consumer bias induced by the predominance of pro-environmental sustainability messages. The verbal versus non-verbal presentation of the sustainability cue neither had a significant influence on distance perceptions and construal level nor on buying intentions. This study offers valuable contributions to the sustainable consumption literature, as well as a theoretical basis for construal-level framing as applied in sustainable fashion branding.

Keywords: construal level theory, environmental vs social sustainability, online fashion shopping, sustainable fashion

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Authors: Al Sayed Al Soudy, Valeria Maselli, Gianluca Polese, Anna Di Cosmo

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Cephalopods are considered as a model organism with a rich behavioral repertoire. Sophisticated behaviors were widely studied and described in different species such as Octopus vulgaris, who has evolved the largest and more complex nervous system among invertebrates. In O. vulgaris, cognitive abilities in problem-solving tasks and learning abilities are associated with long-term memory and spatial memory, mediated by highly developed sensory organs. They are equipped with sophisticated eyes, able to discriminate colors even with a single photoreceptor type, vestibular system, ‘lateral line analogue’, primitive ‘hearing’ system and olfactory organs. They can recognize chemical cues either through direct contact with odors sources using suckers or by distance through the olfactory organs. Cephalopods are able to detect widespread waterborne molecules by the olfactory organs. However, many volatile odorant molecules are insoluble or have a very low solubility in water, and must be perceived by direct contact. O. vulgaris, equipped with many chemosensory neurons located in their suckers, exhibits a peculiar behavior that can be provocatively described as 'smell by touch'. The aim of this study is to establish the priority given to chemical vs. visual perception in food choice. Materials and methods: Three different types of food (anchovies, clams, and mussels) were used, and all sessions were recorded with a digital camera. During the acclimatization period, Octopuses were exposed to the three types of food to test their natural food preferences. Later, to verify if food preference is maintained, food was provided in transparent screw-jars with pierced lids to allow both visual and chemical recognition of the food inside. Subsequently, we tested alternatively octopuses with food in sealed transparent screw-jars and food in blind screw-jars with pierced lids. As a control, we used blind sealed jars with the same lid color to verify a random choice among food types. Results and discussion: During the acclimatization period, O. vulgaris shows a higher preference for anchovies (60%) followed by clams (30%), then mussels (10%). After acclimatization, using the transparent and pierced screw jars octopus’s food choices resulted in 50-50 between anchovies and clams, avoiding mussels. Later, guided by just visual sense, with transparent but not pierced jars, their food preferences resulted in 100% anchovies. With pierced but not transparent jars their food preference resulted in 100% anchovies as first food choice, the clams as a second food choice result (33.3%). With no possibility to select food, neither by vision nor by chemoreception, the results were 20% anchovies, 20% clams, and 60% mussels. We conclude that O. vulgaris uses both chemical and visual senses in an integrative way in food choice, but if we exclude one of them, it appears clear that its food preference relies on chemical sense more than on visual perception.

Keywords: food choice, Octopus vulgaris, olfaction, sensory organs, visual sense

Procedia PDF Downloads 193

Authors: Pui-sze Yeung, Connie Suk-han Ho, David Wai-ock Chan, Kevin Kien-hoa Chung

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Background: Recent findings have shown that transcription skills play a unique and significant role in Chinese word reading and spelling (i.e. word dictation), and written composition development. The interrelationships among component skills of transcription, word reading, word spelling, and written composition in Chinese have rarely been examined in the literature. Is the contribution of component skills of transcription to Chinese written composition mediated by word level skills (i.e., word reading and spelling)? Methods: The participants in the study were 249 Chinese children in Grade 1, Grade 3, and Grade 5 in Hong Kong. They were administered measures of general reasoning ability, orthographic knowledge, stroke sequence knowledge, word spelling, handwriting fluency, word reading, and Chinese narrative writing. Orthographic knowledge- orthographic knowledge was assessed by a task modeled after the lexical decision subtest of the Hong Kong Test of Specific Learning Difficulties in Reading and Writing (HKT-SpLD). Stroke sequence knowledge: The participants’ performance in producing legitimate stroke sequences was measured by a stroke sequence knowledge task. Handwriting fluency- Handwriting fluency was assessed by a task modeled after the Chinese Handwriting Speed Test. Word spelling: The stimuli of the word spelling task consist of fourteen two-character Chinese words. Word reading: The stimuli of the word reading task consist of 120 two-character Chinese words. Written composition: A narrative writing task was used to assess the participants’ text writing skills. Results: Analysis of covariance results showed that there were significant between-grade differences in the performance of word reading, word spelling, handwriting fluency, and written composition. Preliminary hierarchical multiple regression analysis results showed that orthographic knowledge, word spelling, and handwriting fluency were unique predictors of Chinese written composition even after controlling for age, IQ, and word reading. The interaction effects between grade and each of these three skills (orthographic knowledge, word spelling, and handwriting fluency) were not significant. Path analysis results showed that orthographic knowledge contributed to written composition both directly and indirectly through word spelling, while handwriting fluency contributed to written composition directly and indirectly through both word reading and spelling. Stroke sequence knowledge only contributed to written composition indirectly through word spelling. Conclusions: Preliminary hierarchical regression results were consistent with previous findings about the significant role of transcription skills in Chinese word reading, spelling and written composition development. The fact that orthographic knowledge contributed both directly and indirectly to written composition through word reading and spelling may reflect the impact of the script-sound-meaning convergence of Chinese characters on the composing process. The significant contribution of word spelling and handwriting fluency to Chinese written composition across elementary grades highlighted the difficulty in attaining automaticity of transcription skills in Chinese, which limits the working memory resources available for other composing processes.

Keywords: orthographic knowledge, transcription skills, word reading, writing

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Authors: Bill D. Geis

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Suicide wrongful death forensic cases are the fastest rising tort in mental health law. It is estimated that suicide-related cases have accounted for 15% of U.S. malpractice claims since 2006. Most suicide-related personal injury claims fall into the legal category of “wrongful death.” Though mental health experts may be called on to address a range of forensic questions in wrongful death cases, the central consultation that most experts provide is about the negligence element—specifically, the issue of whether the clinician met the clinical standard of care in assessing, treating, and managing the deceased person’s mental health care. Standards of care, varying from U.S. state to state, are broad and address what a reasonable clinician might do in a similar circumstance. This fact leaves the issue of the suicide standard of care, in each case, up to forensic experts to put forth a reasoned estimate of what the standard of care should have been in the specific case under litigation. Because the general state guidelines for standard of care are broad, forensic experts are readily retained to provide scientific and clinical opinions about whether or not a clinician met the standard of care in their suicide assessment, treatment, and management of the case. In the past and in much of current practice, the assessment of suicide has centered on the elicitation of verbalized suicide ideation. Research in recent years, however, has indicated that the majority of persons who end their lives do not say they are suicidal at their last medical or psychiatric contact. Near-term risk assessment—that goes beyond verbalized suicide ideation—is needed. Our previous research employed structural equation modeling to predict lethal suicide risk--eight negative thought patterns (feeling like a burden on others, hopelessness, self-hatred, etc.) mediated by nine transdiagnostic clinical factors (mental torment, insomnia, substance abuse, PTSD intrusions, etc.) were combined to predict acute lethal suicide risk. This structural equation model, the Lethal Suicide Risk Pattern (LSRP), Acute model, had excellent goodness-of-fit [χ2(df) = 94.25(47)***, CFI = .98, RMSEA = .05, .90CI = .03-.06, p(RMSEA = .05) = .63. AIC = 340.25, ***p < .001.]. A further SEQ analysis was completed for this paper, adding a measure of Acute Suicide Ideation to the previous SEQ. Acceptable prediction model fit was no longer achieved [χ2(df) = 3.571, CFI > .953, RMSEA = .075, .90% CI = .065-.085, AIC = 529.550].This finding suggests that, in this additional study, immediate verbalized suicide ideation information was unhelpful in the assessment of lethal risk. The LSRP and other dynamic, near-term risk models (such as the Acute Suicide Affective Disorder Model and the Suicide Crisis Syndrome Model)—going beyond elicited suicide ideation—need to be incorporated into current clinical suicide assessment training. Without this training, the standard of care for suicide assessment is out of sync with current research—an emerging dilemma for the forensic evaluation of suicide wrongful death cases.

Keywords: forensic evaluation, standard of care, suicide, suicide assessment, wrongful death

Procedia PDF Downloads 43

Authors: Caroline Lavratti, Pedro Dal Lago, Gustavo Reinaldo, Gilson Dorneles, Andreia Bard, Laira Fuhr, Daniela Pochmann, Alessandra Peres, Luciane Wagner, Viviane Elsner

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Approximately 1% of the world's population is affected by schizophrenia (SZ), a chronic and debilitating neurodevelopmental disorder. Among possible factors, reduced levels of Brain-derived neurotrophic factor (BDNF) has been recognized in physiopathogenesis and course of SZ. In this context, peripheral BDNF levels have been used as a biomarker in several clinical studies, since this neurotrophin is able to cross the blood-brain barrier in a bi-directional manner and seems to present a strong correlation with the central nervous system fluid levels. The patients with SZ usually adopts a sedentary lifestyle, which has been partly associated with the increase in obesity incidence rates, metabolic syndrome, type 2 diabetes and coronary heart disease. On the other hand, exercise, a non-invasive and low cost intervention, has been considered an important additional therapeutic option for this population, promoting benefits to physical and mental health. To our knowledge, few studies have been pointed out that the positive effects of exercise in SZ patients are mediated, at least in part, to enhanced levels of BDNF after training. However, these studies are focused on evaluating the effect of single bouts of exercise of chronic interventions, data concerning the short- and long-term exercise outcomes on BDNF are scarce. Therefore, this study aimed to evaluate the effect of a concurrent exercise protocol (CEP) on plasma BDNF levels of SZ patients in different time-points. Material and Methods: This study was approved by the Research Ethics Committee of the Centro Universitário Metodista do IPA (no 1.243.680/2015). The participants (n=15) were subbmited to the CEP during 90 days, 3 times a week for 60 minutes each session. In order to evaluate the short and long-term effects of exercise, blood samples were collected pre, 30, 60 and 90 days after the intervention began. Plasma BDNF levels were determined with the ELISA method, from Sigma-Aldrich commercial kit (catalog number RAB0026) according to manufacturer's instructions. Results: A remarkable increase on plasma BDNF levels at 90 days after training compared to baseline (p=0.006) and 30 days (p=0.007) values were observed. Conclusion: Our data are in agreement with several studies that show significant enhancement on BDNF levels in response to different exercise protocols in SZ individuals. We might suggest that BDNF upregulation after training in SZ patients acts in a dose-dependent manner, being more pronounced in response to chronic exposure. Acknowledgments: This work was supported by Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS)/Brazil.

Keywords: exercise, BDNF, schizophrenia, time-points

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Authors: Sofia Mariani, Irene Leo

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The objective of the present study is to define the mediating role of attitudes that objectify and devalue women (hostile sexism, benevolent sexism, and sexual objectification of women) in the indirect correlation between exposure to televised sports and acceptance of rape myths. A second goal is to contribute to research on the topic by defining the role of mediators in exposure to different types of sports, following the traditional gender classification of sports. Data collection was carried out by means of an online questionnaire, measuring television sport exposure, sport type, hostile sexism, benevolent sexism, and sexual objectification of women. Data analysis was carried out using IBM SPSS software. The model used was created using Ordinary Least Squares (OLS) regression path analysis. The predictor variable in the model was television sports exposure, the outcome was rape myths acceptance, and the mediators were (1) hostile sexism, (2) benevolent sexism, and (3) sexual objectification of women. Correlation analyses were carried out dividing by sport type and controlling for the participants’ gender. As seen in existing literature, television sports exposure was found to be indirectly and positively related to rape myth acceptance through the mediating role of: (1) hostile sexism, (2) benevolent sexism, and (3) sexual objectification of women. The type of sport watched influenced the role of the mediators: hostile sexism was found to be the common mediator to all sports type, exposure to traditionally considered feminine or neutral sports showed the additional mediation effect of sexual objectification of women. In line with existing literature, controlling for gender showed that the only significant mediators were hostile sexism for male participants and benevolent sexism for female participants. Given the prevalence of men among the viewers of traditionally considered masculine sports, the correlation between television sports exposure and rape myth acceptance through the mediation of hostile sexism is likely due to the gender of the participants. However, this does not apply to the viewers of traditionally considered feminine and neutral sports, as this group is balanced in terms of gender and shows a unique mediation: the correlation between television sports exposure and rape myth acceptance is mediated by both hostile sexism and sexual objectification. Given that hostile sexism is defined as hostility towards women who oppose or fail to conform to traditional gender roles, these findings confirm that sport is perceived as a non-traditional activity for women. Additionally, these results imply that the portrayal of women in traditionally considered feminine and neutral sports - which are defined as such because of their aesthetic characteristics - may have a strong component of sexual objectification of women. The present research contributes to defining the association between sports exposure and rape myth acceptance through the mediation effects of sexist attitudes and sexual objectification of women. The results of this study have practical implications, such as supporting the feminine sports teams who ask for more practical and less revealing uniforms, more similar to their male colleagues and therefore less objectifying.

Keywords: television exposure, sport, rape myths, objectification, sexism

Procedia PDF Downloads 69

Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

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Authors: Olfat Mohamed Hussien Yousef

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Tamoxifen (TM) is a synthetic non-steroidal antiestrogen. It is one of the most effective drugs for treatment of estrogen-dependent cancer by binding to estrogen receptors, suppressing of epithelial proliferation and as a chemotherapeutic agent. Recently, more attention has been paid to the protective effects of natural antioxidants against toxicities induced by anti-cancer drugs involving free radical-mediated oxidative stress and tissue injury. Vitamin C is a potent antioxidant that has the ability to scavenge factors causing free radical formation in animals receiving tamoxifen. The present study aims at pinpointing the TM-induced histopathological and ultrastructural changes in the kidneys and to assess the possible chemoprotective role of vitamin C against such TM-induced microscopic changes. Thirty adult male CD-1 mice, 25-30 g in weight and 3 months old, were divided into three groups. The first group served as control. The second group received the therapeutic dose of TM at daily oral dose of 40 mg/kg body weight for 28 days. The third group received the therapeutic dose of vitamin C at a daily dose of 500 mg/kg body weight simultaneously with the therapeutic dose of TM used in group two for 28 days. Animals were sacrificed and kidney samples were obtained and processed for histological and ultrastructural examination. Histological changes induced by TM included damage of the renal corpuscles including obliteration of the subcapsular space, congestion of the glomerular blood capillaries, segmental mesangial cell proliferation with matrix expansion, capsular adhesions with the glomerular tuft especially at the urinary pole of the corpuscles. Moreover, some proximal and distal tubules suffered various degrees of degeneration in some lining cells. Haemorrhage and inflammatory cell infiltration were also observed in the intertubular spaces. Ultrastructural observations revealed damage of the parietal epithelium of Bowman’s capsule, fusion and destruction of the foot processes of podocytes and great increase of mesangial cells and mesangial matrix. The cells of the proximal convoluted tubules displayed marked destruction of the microvilli constituting the brush borders and degeneration of the mitochondria; besides, abundant lysosomes, numerous vacuoles and pyknotic nuclei were observed. The distal convoluted tubules displayed marked distruction of both the basal infolding and the mitochondria in some areas. Histological and ultrastructural results revealed that treatment of male mice with TM simultaneously with vitamin C led to apparent repair of the injured renal tissue. This might suggest that vitamin C (an antioxidant agent) can minimize the toxic effects of TM (an antiestrogen).

Keywords: tamoxifen, vitamin c, mammalian kidney, histology, ultrastructure

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Authors: Benjamin Osei Tutu, Sharon Annison

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On 21^st November, 2016, an outbreak of foodborne illness occurred after a buffet lunch served during a stakeholders’ consultation meeting held in Accra. An investigation was conducted to characterise the affected people, determine the etiologic food, the source of contamination and the etiologic agent and to implement appropriate public health measures to prevent future occurrences. A retrospective cohort study was conducted via telephone interviews, using a structured questionnaire developed from the buffet menu. A case was defined as any person suffering from symptoms of foodborne illness e.g. diarrhoea and/or abdominal cramps after eating food served during the stakeholder consultation meeting in Accra on 21^st November, 2016. The exposure status of all the members of the cohort was assessed by taking the food history of each respondent during the telephone interview. The data obtained was analysed using Epi Info 7. An environmental risk assessment was conducted to ascertain the source of the food contamination. Risks of foodborne infection from the foods eaten were determined using attack rates and odds ratios. Data was obtained from 54 people who consumed food served during the stakeholders’ meeting. Out of this population, 44 people reported with symptoms of food poisoning representing 81.45% (overall attack rate). The peak incubation period was seven hours with a minimum and maximum incubation periods of four and 17 hours, respectively. The commonly reported symptoms were diarrhoea (97.73%, 43/44), vomiting (84.09%, 37/44) and abdominal cramps (75.00%, 33/44). From the incubation period, duration of illness and the symptoms, toxin-mediated food poisoning was suspected. The environmental risk assessment of the implicated catering facility indicated a lack of time/temperature control, inadequate knowledge on food safety among workers and sanitation issues. Limited number of food samples was received for microbiological analysis. Multivariate analysis indicated that illness was significantly associated with the consumption of the snacks served (OR 14.78, P < 0.001). No stool and blood or samples of etiologic food were available for organism isolation; however, the suspected etiologic agent was Staphylococcus aureus or Clostridium perfringens. The outbreak could probably be due to the consumption of unwholesome snack (tuna sandwich or chicken. The contamination and/or growth of the etiologic agent in the snack may be due to the breakdown in cleanliness, time/temperature control and good food handling practices. Training of food handlers in basic food hygiene and safety is recommended.

Keywords: Accra, buffet, conference, C. perfringens, cohort study, food poisoning, gastroenteritis, office workers, Staphylococcus aureus

Procedia PDF Downloads 198

Authors: Syed Benazir Firdaus, Debosree Ghosh, Aindrila Chattyopadhyay, Kuladip Jana, Debasish Bandyopadhyay

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Gastro-toxic effect of piroxicam, a classical non-steroidal anti-inflammatory drug (NSAID), has restricted its use in arthritis and similar diseases. The present study aims to find if a combination of melatonin and Murraya koenigii leaf extract therapy can protect against piroxicam induced ulcerative damage in rats. For this study, rats were divided into four groups namely control group where rats were orally administered distilled water, only combination treated group, piroxicam treated group and combination pre-administered piroxicam treated group. Each group of rats consisted of six animals. Melatonin at a dose of 20mg/kg body weight and antioxidant rich Murraya koenigii leaf extract at a dose of 50 mg /kg body weight were successively administered at 30 minutes interval one hour before oral administration of piroxicam at a dose of 30 mg/kg body weight to Wistar rats in the combination pre-administered piroxicam treated group. The rats of the animal group which was only combination treated were administered both the drugs respectively without piroxicam treatment whereas the piroxicam treated animal group was administered only piroxicam at 30mg/kg body weight without any pre-treatment with the combination. Macroscopic examination along with histo-pathological study of gastric tissue using haemotoxylin-eosin staining and alcian blue dye staining showed protection of the gastric mucosa in the combination pre-administered piroxicam treated group. Determination of adherent mucus content biochemically and collagen content through Image J analysis of picro-sirius stained sections of rat gastric tissue also revealed protective effects of the combination in piroxicam mediated toxicity. Gelatinolytic activity of piroxicam was significantly reduced by pre-administration of the drugs which was well exhibited by the gelatin zymography study of the rat gastric tissue. Mean ulcer index determined from macroscopic study of rat stomach reduced to a minimum (0±0.00; Mean ± Standard error of mean and number of animals in the group=6) indicating the absence of ulcer spots on pre-treatment of rats with the combination. Gastro-friendly prostaglandin (PGE2) which otherwise gets depleted on piroxicam treatment was also well protected when the combination was pre-administered in the rats prior to piroxicam treatment. The requirement of the individual drugs in low doses in this combinatorial therapeutic approach will possibly minimize the cost of therapy as well as it will eliminate the possibility of any pro-oxidant side effects on the use of high doses of antioxidants. Beneficial activity of this combination therapy in the rat model raises the possibility that similar protective actions might be also observed if it is adopted by patients consuming NSAIDs like piroxicam. However, the introduction of any such therapeutic approach is subject to future studies in human.

Keywords: gastro-protective action, melatonin, Murraya koenigii leaf extract, piroxicam

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Authors: Sakshi Piplani, Ajit Kumar

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Valproic acid (VPA) is the first synthetic therapeutic agent used to treat epileptic disorders, which account for affecting nearly 1% world population. Teratogenicity caused by VPA has prompted the search for next generation drug with better efficacy and lower side effects. Recent studies have posed HDAC8 as direct target of VPA that causes the teratogenic effect in foetus. We have employed molecular dynamics (MD) and docking simulations to understand the binding mode of VPA and their analogues onto HDAC8. A total of twenty 3D-structures of human HDAC8 isoforms were selected using BLAST-P search against PDB. Multiple sequence alignment was carried out using ClustalW and PDB-3F07 having least missing and mutated regions was selected for study. The missing residues of loop region were constructed using MODELLER and energy was minimized. A set of 216 structural analogues (>90% identity) of VPA were obtained from Pubchem and ZINC database and their energy was optimized with Chemsketch software using 3-D CHARMM-type force field. Four major neurotransmitters (GABAt, SSADH, α-KGDH, GAD) involved in anticonvulsant activity were docked with VPA and its analogues. Out of 216 analogues, 75 were selected on the basis of lower binding energy and inhibition constant as compared to VPA, thus predicted to have anti-convulsant activity. Selected hHDAC8 structure was then subjected to MD Simulation using licenced version YASARA with AMBER99SB force field. The structure was solvated in rectangular box of TIP3P. The simulation was carried out with periodic boundary conditions and electrostatic interactions and treated with Particle mesh Ewald algorithm. pH of system was set to 7.4, temperature 323K and pressure 1atm respectively. Simulation snapshots were stored every 25ps. The MD simulation was carried out for 20ns and pdb file of HDAC8 structure was saved every 2ns. The structures were analysed using castP and UCSF Chimera and most stabilized structure (20ns) was used for docking study. Molecular docking of 75 selected VPA-analogues with PDB-3F07 was performed using AUTODOCK4.2.6. Lamarckian Genetic Algorithm was used to generate conformations of docked ligand and structure. The docking study revealed that VPA and its analogues have more affinity towards ‘hydrophobic active site channel’, due to its hydrophobic properties and allows VPA and their analogues to take part in van der Waal interactions with TYR24, HIS42, VAL41, TYR20, SER138, TRP137 while TRP137 and SER138 showed hydrogen bonding interaction with VPA-analogues. 14 analogues showed better binding affinity than VPA. ADMET SAR server was used to predict the ADMET properties of selected VPA analogues for predicting their druggability. On the basis of ADMET screening, 09 molecules were selected and are being used for in-vivo evaluation using Danio rerio model.

Keywords: HDAC8, docking, molecular dynamics simulation, valproic acid

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Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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Protein apoferritin seems to be a very promising structure for use as a nanocarrier. It is prepared from intracellular ferritin protein naturally found in most organisms. The role of ferritin proteins is to store and transport ferrous ions. Apoferritin is a hollow protein cage without ferrous ions that can be prepared from ferritin by reduction with thioglycolic acid or dithionite. The structure of apoferritin is composed of 24 protein subunits, creating a sphere with 12 nm in diameter. The inner cavity has a diameter of 8 nm. The drug encapsulation process is based on the response of apoferritin structure to the pH changes of surrounding solution. In low pH, apoferritin is disassembled into individual subunits and its structure is “opened”. It can then be mixed with any desired cytotoxic drug and after adjustment of pH back to neutral the subunits are reconnected again and the drug is encapsulated within the apoferritin particles. Excess drug molecules can be removed by dialysis. The receptors for apoferritin, SCARA5 and TfR1 can be found in the membrane of both healthy and cancer cells. To enhance the specific targeting of apoferritin nanocarrier, it is possible to modify its surface with targeting moieties, such as antibodies. To ensure sterically correct complex, we used a a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (ApoDox) was coated either with gold nanoparticles (ApoDox-Nano) or gold (III) chloride hydrate reduced with sodium borohydride (ApoDox-HAu). The applied amount of gold in form of gold (III) chloride hydrate was 10 times higher than in the case of gold nanoparticles. However, after removal of the excess unbound ions by electrophoretic separation, the concentration of gold on the surface of apoferritin was only 6 times higher for ApoDox-HAu in comparison with ApoDox-Nano. Moreover, the reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties (excitation maximum at 480 nm with emission maximum at 600 nm) and thus its biological activity. Fluorescent properties of ApoDox-Nano were similar to the unmodified ApoDox, therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, we used ELISA-like method with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, we applied ApoDox without targeting antibodies and ApoDox-Nano modified with targeting antibodies (human IgG antibodies). The amount of unmodified ApoDox on antigen after incubation and subsequent rinsing with water was 5 times lower than in the case of ApoDox-Nano modified with targeting antibodies. The modification of non-gold ApoDox with antibodies caused no change in its targeting properties. It can therefore be concluded that the demonstrated procedure allows us to create nanocarrier with enhanced targeting properties, suitable for nanomedicine.

Keywords: apoferritin, doxorubicin, nanocarrier, targeting antibodies

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Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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Authors: Shahira M. Ezzat, Marwa I. Ezzat, Mona M. Okba, Esther T. Menze, Ashraf B. Abdel-Naim, Shahnas O. Mohamed

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Background: In order to decrease the burden of the high cost of synthetic drugs, it is important to focus on phytopharmaceuticals. The aim of our study was to search for the mechanism of ginger (Zingiber officinale Roscoe) anti-inflammatory potential and to correlate it to its biophytochemicals. Methods: Various extracts viz. water, 50%, 70%, 80%, and 90% ethanol were prepared from ginger rhizomes. Fractionation of the aqueous extract (AE) was accomplished using Diaion HP-20. In vitro anti-inflammatory activity of the different extracts and isolated compounds was evaluated by protein denaturation inhibition, membrane stabilization, protease inhibition, and anti-lipoxygenase assays. In vivo anti-inflammatory activity of AE was estimated by assessment of rat paw oedema after carrageenan injection. Prostaglandin E2 (PGE2), certain inflammation markers (TNF-α, IL-6, IL-1α, IL-1β, INFr, MCP-1MIP, RANTES, and Nox) levels and MPO activity in the paw edema exudates were measured. Total antioxidant capacity (TAC) was also determined. Histopathological alterations of paw tissues were scored. Results: All the tested extracts showed significant (p < 0.1) anti-inflammatory activities. The highest percentage of heat induced albumin denaturation (66%) was exhibited by the 50% ethanol (250 μg/ml). The 70 and 90% ethanol extracts (500 μg/ml) were more potent as membrane stabilizers (34.5 and 37%, respectively) than diclofenac (33%). The 80 and 90% ethanol extracts (500 μg/ml) showed maximum protease inhibition (56%). The strongest anti-lipoxygenase activity was observed for the AE. It showed more significant lipoxygenase inhibition activity than that of diclofenac (58% and 52%, respectively) at the same concentration (125 μg/ml). Fractionation of AE yielded four main fractions (Fr I-IV) which showed significant in vitro anti-inflammatory. Purification of Fr-III and IV led to the isolation of 6-poradol (G1), 6-shogaol (G2); methyl 6- gingerol (G3), 5-gingerol (G4), 6-gingerol (G5), 8-gingerol (G6), 10-gingerol (G7), and 1-dehydro-6-gingerol (G8). G2 (62.5 ug/ml), G1 (250 ug/ml), and G8 (250 ug/ml) exhibited potent anti-inflammatory activity in all studied assays, while G4 and G5 exhibited moderate activity. In vivo administration of AE ameliorated rat paw oedema in a dose-dependent manner. AE (at 200 mg/kg) showed significant reduction (60%) of PGE2 production. The AE at different doses (at 25-200 mg/kg) showed significant reduction in inflammatory markers except for IL-1α. AE (at 25 mg/kg) is superior to indomethacin in reduction of IL-1β. Treatment of animals with the AE (100, 200 mg/kg) or indomethacin (10 mg/kg) showed significant reduction in TNF-α, IL-6, MCP-1, and RANTES levels, and MPO activity by about (31, 57 and 32% ) (65, 60 and 57%) (27, 41 and 28%) (23, 32 and 23%) (66, 67 and 67%) respectively. AE at 100 and 200 mg/kg was equipotent to indomethacin in reduction of NOₓ level and in increasing the TAC. Histopathological examination revealed very few inflammatory cells infiltration and oedema after administration of AE (200 mg/kg) prior to carrageenan. Conclusion: Ginger anti-inflammatory activity is mediated by inhibiting macrophage and neutrophils activation as well as negatively affecting monocyte and leukocyte migration. Moreover, it produced dose-dependent decrease in pro-inflammatory cytokines and chemokines and replenished the total antioxidant capacity. We strongly recommend future investigations of ginger in the potential signal transduction pathways.

Keywords: anti-lipoxygenase activity, inflammatory markers, 1-dehydro-6-gingerol, 6-shogaol

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Authors: Kirti Hira, A. Sajeli Begum, S. Mahibalan, Poorna Chandra Rao

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Introduction: Cancer is one of the leading causes of death worldwide. Although existing therapy effectively kills cancer cells, they do affect normal growing cells leading to many undesirable side effects. Hence there is need to develop effective as well as safe drug molecules to combat cancer, which is possible through phyto-research. The currently available plant-derived blockbuster drugs are the example for this. In view of this, an investigation was done to identify cytotoxic lead molecules from Hedyotis umbellata (Family Rubiaceae), a widely distributed weed in India. Materials and Methods: The methanolic extract of the whole plant of H. umbellata (MHU), prepared through Soxhlet extraction method was further fractionated with diethyl ether and n-butanol, successively. MHU, ether fraction (EMHU) and butanol fraction (BMHU) were lyophilized and were tested for the cytotoxic effect using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay against non-small cell lung cancer (NSCLC) A549 cell lines. The potentially active EMHU was subjected to chromatographic purification using normal-phase silica columns, in order to isolate the responsible bioactive compounds. The isolated pure compounds were tested for their cytotoxic effect by MTT assay against A549 cells. Compound-3, which was found to be most active, was characterized using IR, 1H- and 13C-NMR and MS analysis. The study was further extended to decipher the mechanism of action of cytotoxicity of compound-3 against A549 cells through various in vitro cellular models. Cell cycle analysis was done using flow cytometry following PI (Propidium Iodide) staining. Protein analysis was done using Western blot technique. Results: Among MHU, EMHU, and BMHU, the non-polar fraction EMHU demonstrated a significant dose-dependent cytotoxic effect with IC50 of 67.7μg/ml. Chromatography of EMHU yielded seven compounds. MTT assay of isolated compounds explored compound-3 as potentially active one, which inhibited the growth of A549 cells with IC50value of 14.2μM. Further, compound-3 was identified as cedrelopsin, a coumarin derivative having molecular weight of 260. Results of in vitro mechanistic studies explained that cedrelopsin induced cell cycle arrest at G2/M phase and down-regulated the expression of G2/M regulatory proteins such as cyclin B1, cdc2, and cdc25C, dose dependently. This is the first report that explores the cytotoxic mechanism of cedrelopsin. Conclusion: Thus a potential small lead molecule, cedrelopsin isolated from H. umbellata, showing antiproliferative effect mediated by G2/M arrest in A549 cells was discovered. The effect of cedrelopsin against other cancer cell lines followed by in vivo studies can be performed in future to develop a new drug candidate.

Keywords: A549, cedrelopsin, G2/M phase, Hedyotis umbellata

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Authors: Felice Elefant, Akanksha Bhatnaghar, Keegan Krick, Elizabeth Heller

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Context: The severity of Alzheimer’s Disease (AD) progression involves an interplay of genetics, age, and environmental factors orchestrated by histone acetyltransferase (HAT) mediated neuroepigenetic mechanisms. While disruption of Tip60 HAT action in neural gene control is implicated in AD, alternative mechanisms underlying Tip60 function remain unexplored. Altered RNA splicing has recently been highlighted as a widespread hallmark in the AD transcriptome that is implicated in the disease. Research Aim: The aim of this study was to identify a novel RNA binding/splicing function for Tip60 in human hippocampus and impaired in brains from AD fly models and AD patients. Methodology/Analysis: The authors used RNA immunoprecipitation using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. To identify Tip60’s RNA targets, they performed genome sequencing (DNB-SequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Findings: The authors' transcriptomic analysis of RNA bound to Tip60 by Tip60-RNA immunoprecipitation (RIP) revealed Tip60 RNA targets enriched for critical neuronal processes implicated in AD. Remarkably, 79% of Tip60’s RNA targets overlap with its chromatin gene targets, supporting a model by which Tip60 orchestrates bi-level transcriptional regulation at both the chromatin and RNA level, a function unprecedented for any HAT to date. Since RNA splicing occurs co-transcriptionally and splicing defects are implicated in AD, the authors investigated whether Tip60-RNA targeting modulates splicing decisions and if this function is altered in AD. Replicate multivariate analysis of transcript splicing (rMATS) analysis of RNA-Seq data sets from wild-type and AD fly brains revealed a multitude of mammalian-like AS defects. Strikingly, over half of these altered RNAs were bonafide Tip60-RNA targets enriched for in the AD-gene curated database, with some AS alterations prevented against by increasing Tip60 in fly brain. Importantly, human orthologs of several Tip60-modulated spliced genes in Drosophila are well characterized aberrantly spliced genes in human AD brains, implicating disruption of Tip60’s splicing function in AD pathogenesis. Theoretical Importance: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology. Data Collection: The authors collected data from RNA immunoprecipitation experiments using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. They also performed genome sequencing (DNBSequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Questions: The question addressed by this study was whether Tip60 has a novel RNA binding/splicing function in human hippocampus and whether this function is impaired in brains from AD fly models and AD patients. Conclusions: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology.

Keywords: Alzheimer's disease, cognition, aging, neuroepigenetics

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Authors: Gary J. Pickering, Sarah Lucas, Catherine E. Klodnicki, Nicole J. Gaudette

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Two taste pheno types that are of interest in the study of habitual diet-related risk factors and disease are 6-n-propylthiouracil (PROP) responsiveness and thermal tasting. Individuals differ considerable in how intensely they experience the bitterness of PROP, which is partially explained by three major single nucleotide polymorphisms associated with the TAS2R38 gene. Importantly, this variable responsiveness is a useful proxy for general taste responsiveness, and links to diet-related disease risk, including body mass index, in some studies. Thermal tasting - a newly discovered taste phenotype independent of PROP responsiveness - refers to the capacity of many individuals to perceive phantom tastes in response to lingual thermal stimulation, and is linked with TRPM5 channels. Thermal tasters (TTs) also experience oral sensations more intensely than thermal non-tasters (TnTs), and this was shown to associate with differences in self-reported food preferences in a previous survey from our lab. Here we report on two related studies, where we sought to determine whether PROP responsiveness and thermal tasting would associate with perceptual differences in the oral sensations elicited by sampled energy-dense foods, and whether in turn this would influence liking. We hypothesized that hyper-tasters (thermal tasters and individuals who experience PROP intensely) would (a) rate sweet and high-fat foods more intensely than hypo-tasters, and (b) would differ from hypo-tasters in liking scores. (Liking has been proposed recently as a more accurate measure of actual food consumption). In Study 1, a range of energy-dense foods and beverages, including table cream and chocolate, was assessed by 25 TTs and 19 TnTs. Ratings of oral sensation intensity and overall liking were obtained using gVAS and gDOL scales, respectively. TTs and TnTs did not differ significantly in intensity ratings for most stimuli (ANOVA). In a 2nd study, 44 female participants sampled 22 foods and beverages, assessing them for intensity of oral sensations (gVAS) and overall liking (9-point hedonic scale). TTs (n=23) rated their overall liking of creaminess and milk products lower than did TnTs (n=21), and liked milk chocolate less. PROP responsiveness was negatively correlated with liking of food and beverages belonging to the sweet or sensory food grouping. No other differences in intensity or liking scores between hyper- and hypo-tasters were found. Taken overall, our results are somewhat unexpected, lending only modest support to the hypothesis that these taste phenotypes associate with energy-dense food liking and consumption through differences in the oral sensations they elicit. Reasons for this lack of concordance with expectations and some prior literature are discussed, and suggestions for future research are advanced.

Keywords: taste phenotypes, sensory evaluation, PROP, thermal tasting, diet-related health risk

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Authors: Asma Naim, Warda Mushtaq

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Staphylococcus species are the most versatile and adaptive organism. They are widespread and naturally found on the skin, mucosa and nose in humans. Among these, Staphylococcus aureus is the most important species. These organisms act as opportunistic pathogens and can infect various organs of the host, causing minor skin infection to severe toxin mediated diseases, and life threatening nosocomial infections. Staphylococcus aureus has acquired resistance against β-lactam antibiotics by the production of β-lactamase, and Methicillin-Resistant Staphylococcus aureus (MRSA) strains have also been reported with increasing frequency. MRSA strains have been associated with nosocomial as well as community acquired infections. Medicinal plants have enormous potential as antimicrobial substances and have been used in traditional medicine. Search for medicinally valuable plants with antimicrobial activity is being emphasized due to increasing antibiotic resistance in bacteria. In the present study, the antibacterial potential of Rosmarinus officinalis (Rosemary) and Murraya koenigii (curry leaves) was evaluated. These are common household herbs used in food as enhancer of flavor and aroma. The crude aqueous infusion, decoction and ethanolic extracts of curry leaves and rosemary and essential oil of rosemary were investigated in the present study for antibacterial activity against multi-drug resistant Staphylococcus strains using well diffusion method. In the present study, 60 Multi-drug resistant clinical isolates of S. aureus (43) and Coagulase Negative Staphylococci (CoNS) (17) were screened against different concentrations of crude extracts of Rosmarinus officinalis and Murraya koenigii. Out of these 60 isolates, 43 were sensitive to the aqueous infusion of rosemary; 23 to aqueous decoction and 58 to ethanolic extract whereas, 24 isolates were sensitive to the essential oil. In the case of the curry leaves, no antibacterial activity was observed in aqueous infusion and decoction while only 14 isolates were sensitive to the ethanolic extract. The aqueous infusion of rosemary (50% concentration) exhibited a zone of inhibition of 21(±5.69) mm. against CoNS and 17(±4.77) mm. against S. aureus, the zone of inhibition of 50% concentration of aqueous decoction of rosemary was also larger against CoNS 17(±5.78) mm. then S. aureus 13(±6.91) mm. and the 50% concentrated ethanolic extract showed almost similar zone of inhibition in S. aureus 22(±3.61) mm. and CoNS 21(±7.64) mm. whereas, the essential oil of rosemary showed greater zone of inhibition against S. aureus i.e., 16(±4.67) mm. while CoNS showed 15(±6.94) mm. These results show that ethanolic extract of rosemary has significant antibacterial activity. Aqueous infusion and decoction of curry leaves revealed no significant antibacterial potential against all Staphylococcal species and ethanolic extract also showed only a weak response. Staphylococcus strains were susceptible to crude extracts and essential oil of rosemary in a dose depend manner, where the aqueous infusion showed highest zone of inhibition and ethanolic extract also demonstrated antistaphylococcal activity. These results demonstrate that rosemary possesses antistaphylococcal activity.

Keywords: antibacterial activity, curry leaves, multidrug resistant, rosemary, S. aureus

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Authors: Mikko Salminen, Niklas Ravaja

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Emotional intelligence (EI) consists of skills for monitoring own emotions and emotions of others, skills for discriminating different emotions, and skills for using this information in thinking and actions. EI enhances, for example, work outcomes and organizational climate. We suggest that the role and manifestations of EI should also be studied in real leadership situations, especially during the emotional, social interaction. Leadership is essentially a process to influence others for reaching a certain goal. This influencing happens by managerial processes and computer-mediated communication (e.g. e-mail) but also by face-to-face, where facial expressions have a significant role in conveying emotional information. Persons with high EI are typically perceived more positively, and they have better social skills. We hypothesize, that during social interaction high EI enhances the ability to detect other’s emotional state and controlling own emotional expressions. We suggest, that emotionally intelligent leader’s experience less stress during social leadership situations, since they have better skills in dealing with the related emotional work. Thus the high-EI leaders would be more able to enjoy these situations, but also be more efficient in choosing appropriate expressions for building constructive dialogue. We suggest, that emotionally intelligent leaders show more positive emotional expressions than low-EI leaders. To study these hypotheses we observed performance review discussions of 40 leaders (24 female) with 78 (45 female) of their followers. Each leader held a discussion with two followers. Psychophysiological methods were chosen because they provide objective and continuous data from the whole duration of the discussions. We recorded sweating of the hands (electrodermal activation) by electrodes placed to the fingers of the non-dominant hand to assess the stress-related physiological arousal of the leaders. In addition, facial electromyography was recorded from cheek (zygomaticus major, activated during e.g. smiling) and periocular (orbicularis oculi, activated during smiling) muscles using electrode pairs placed on the left side of the face. Leader’s trait EI was measured with a 360 questionnaire, filled by each leader’s followers, peers, managers and by themselves. High-EI leaders had less sweating of the hands (p = .007) than the low-EI leaders. It is thus suggested that the high-EI leaders experienced less physiological stress during the discussions. Also, high scores in the factor “Using of emotions” were related to more facial muscle activation indicating positive emotional expressions (cheek muscle: p = .048; periocular muscle: p = .076, almost statistically significant). The results imply that emotionally intelligent managers are positively relaxed during s social leadership situations such as a performance review discussion. The current study also highlights the importance of EI in face-to-face social interaction, given the central role facial expressions have in interaction situations. The study also offers new insight to the biological basis of trait EI. It is suggested that the identification, forming, and intelligently using of facial expressions are skills that could be trained during leadership development courses.

Keywords: emotional intelligence, leadership, performance review discussion, psychophysiology, social interaction

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Authors: Carla Teixeira, Diana Azevedo, Jonatas Bessa, Maria Guilam

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The flipped classroom is a blended learning modality that combines face-to-face and virtual activities of self-learning, mediated by digital information and communication technologies, which reverses traditional teaching approaches and presents, as a presupposition, the previous study of contents by students. In the following face-to-face activities, the contents are discussed, producing active learning. This work aims to describe the systematization process of the use of flipped classrooms as a method to develop complementary national activities in PROFSAÚDE, a professional master's program in the area of public health, offered as a distance learning course, in the network, in Brazil. The complementary national activities were organized with the objective of strengthening and qualifying students´ learning process. The network gathers twenty-two public institutions of higher education in the country. Its national coordination conducted a survey to detect complementary educational needs, supposed to improve the formative process and align important content sums for the program nationally. The activities were organized both asynchronously, making study materials available in Google classrooms, and synchronously in a tele presential way, organized on virtual platforms to reach the largest number of students in the country. The asynchronous activities allowed each student to study at their own pace and the synchronous activities were intended for deepening and reflecting on the themes. The national team identified some professors' areas of expertise, who were contacted for the production of audiovisual content such as video classes and podcasts, guidance for supporting bibliographic materials and also to conduct synchronous activities together with the technical team. The contents posted in the virtual classroom were organized by modules and made available before the synchronous meeting; these modules, in turn, contain “pills of experience” that correspond to reports of teachers' experiences in relation to the different themes. In addition, activity was proposed, with questions aimed to expose doubts about the contents and a learning challenge, as a practical exercise. Synchronous activities are built with different invited teachers, based on the participants 'discussions, and are the forum where teachers can answer students' questions, providing feedback on the learning process. At the end of each complementary activity, an evaluation questionnaire is available. The responses analyses show that this institutional network experience, as pedagogical innovation, provides important tools to support teaching and research due to its potential in the participatory construction of learning, optimization of resources, the democratization of knowledge and sharing and strengthening of practical experiences on the network. One of its relevant aspects was the thematic diversity addressed through this method.

Keywords: active learning, flipped classroom, network education experience, pedagogic 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: Anastasia V. Kovaleva, Alena A. Ryabova, Vladimir N. Kasatkin

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Background: The most simple form of entrainment to a sensory (typically auditory) rhythmic stimulus involves perceiving and synchronizing movements with an isochronous beat with one level of periodicity, such as that produced by a metronome. Children with pediatric cancer usually treated with chemo- and radiotherapy. Because of such treatment, psychologists and health professionals declare cognitive and motor abilities decline in cancer patients. The purpose of our study was to measure working memory characteristics with association with audio-motor synchronization tasks, also involved some memory resources, in children with different degrees of central nervous system lesions: posterior fossa tumors, acute lymphoblastic leukemia, and healthy controls. Methods: Our sample consisted of three groups of children: children treated for posterior fossa tumors (PFT-group, n=42, mean age 12.23), children treated for acute lymphoblastic leukemia (ALL-group, n=11, mean age 11.57) and neurologically healthy children (control group, n=36, mean age 11.67). Participants were tested for working memory characteristics with Cambridge Neuropsychological Test Automated Battery (CANTAB). Pattern recognition memory (PRM) and spatial working memory (SWM) tests were applied. Outcome measures of PRM test include the number and percentage of correct trials and latency (speed of participant’s response), and measures of SWM include errors, strategy, and latency. In the synchronization tests, the instruction was to tap out a regular beat (40, 60, 90 and 120 beats per minute) in synchrony with the rhythmic sequences that were played. This meant that for the sequences with an isochronous beat, participants were required to tap into every auditory event. Variations of inter-tap-intervals and deviations of children’s taps from the metronome were assessed. Results: Analysis of variance revealed the significant effect of group (ALL, PFT and control) on such parameters as short-term PRM, SWM strategy and errors. Healthy controls demonstrated more correctly retained elements, better working memory strategy, compared to cancer patients. Interestingly that ALL patients chose the bad strategy, but committed significantly less errors in SWM test then PFT and controls did. As to rhythmic ability, significant associations of working memory were found out only with 40 bpm rhythm: the less variable were inter-tap-intervals of the child, the more elements in memory he/she could retain. The ability to audio-motor synchronization may be related to working memory processes mediated by the prefrontal cortex whereby each sensory event is actively retrieved and monitored during rhythmic sequencing. Conclusion: Our results suggest that working memory, tested with appropriate cognitive methods, is associated with the ability to synchronize movements with rhythmic sounds, especially in sub-second intervals (40 per minute).

Keywords: acute lymphoblastic leukemia (ALL), audio-motor synchronization, posterior fossa tumor, working memory

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Authors: Stephen Kyakulumbye, Vivian Kobusingye

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Generally, World over, COVID-19 has had adverse effects on all sectors but with more debilitating effects on the education sector. After reactive lockdowns, education institutions that could continue teaching and learning had to go a distance mediated by digital technological tools. In Uganda, the Ministry of Education thereby issued COVID-19 Online Distance E-learning (ODeL) emergent guidelines. Despite such guidelines, academic institutions in Uganda and similar developing contexts with academically constrained resource environments were caught off-guard and ill-prepared to transform from face-to-face learning to online distance learning mode. Most academic institutions that migrated spontaneously did so with no deliberate tools, systems, strategies, or software to cause active, meaningful, and engaging learning for students. By experience, most of these academic institutions shifted to Zoom and WhatsApp and instead conducted online teaching in real-time than blended synchronous and asynchronous tools. This paper provides students’ experiences while blending synchronous and asynchronous content-creating and learning tools within a technological resource-constrained environment to navigate in such a challenging Uganda context. These conceptual case-based findings, using experience from Uganda Christian University (UCU), point at the design of learning activities with two certain characteristics, the enhancement of synchronous learning technologies with asynchronous ones to mitigate the challenge of system breakdown, passive learning to active learning, and enhances the types of presence (social, cognitive and facilitatory). The paper, both empirical and experiential in nature, uses online experiences from third-year students in Bachelor of Business Administration student lectured using asynchronous text, audio, and video created with Open Broadcaster Studio software and compressed with Handbrake, all open-source software to mitigate disk space and bandwidth usage challenges. The synchronous online engagements with students were a blend of zoom or BigBlueButton, to ensure that students had an alternative just in case one failed due to excessive real-time traffic. Generally, students report that compared to their previous face-to-face lectures, the pre-recorded lectures via Youtube provided them an opportunity to reflect on content in a self-paced manner, which later on enabled them to engage actively during the live zoom and/or BigBlueButton real-time discussions and presentations. The major recommendation is that lecturers and teachers in a resource-constrained environment with limited digital resources like the internet and digital devices should harness this approach to offer students access to learning content in a self-paced manner and thereby enabling reflective active learning through reflective and high-order thinking.

Keywords: synchronous learning, asynchronous learning, active learning, reflective learning, resource-constrained environment

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Authors: Tristan R. Fraser, Christopher D. Steadman, Christopher J. Boos

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Myopericarditis is an inflammation syndrome characterised by clinical diagnostic criteria for pericarditis, such as chest pain, combined with evidence of myocardial involvement, such as elevation of biomarkers of myocardial damage, e.g., troponins. It can rarely be a complication of therapeutics used for dysregulated immune-mediated diseases such as inflammatory bowel disease (IBD), for example, mesalazine. The infrequency of mesalazine-induced myopericarditis adds to the challenge in its recognition. Rapid diagnosis and the early introduction of treatment are crucial. This case report follows a 24-year-old professional footballer with a past medical history of ulcerative colitis, recently started on mesalazine for disease control. Three weeks after mesalazine was initiated, he was admitted with fever, shortness of breath, and chest pain worse whilst supine and on deep inspiration, as well as elevated venous blood cardiac troponin T level (cTnT, 288ng/L; normal: <13ng/L). Myocarditis was confirmed on initial inpatient cardiac MRI, revealing the presence of florid myocarditis with preserved left ventricular systolic function and an ejection fraction of 67%. This was a longitudinal case study following the progress of a single individual with myopericarditis over four acute hospital admissions over nine weeks, with admissions ranging from two to five days. Parameters examined included clinical signs and symptoms, serum troponin, transthoracic echocardiogram, and cardiac MRI. Serial measurements of cardiac function, including cardiac MRI and transthoracic echocardiogram, showed progressive deterioration of cardiac function whilst mesalazine was continued. Prior to cessation of mesalazine, transthoracic echocardiography revealed a small global pericardial effusion of < 1cm and worsening left ventricular systolic function with an ejection fraction of 45%. After recognition of mesalazine as a potential cause and consequent cessation of the drug, symptoms resolved, with cardiac MRI performed as an outpatient showing resolution of myocardial oedema. The patient plans to make a return to competitive sport. Patients suffering from myopericarditis are advised to refrain from competitive sport for at least six months in order to reduce the risk of cardiac remodelling and sudden cardiac death. Additional considerations must be taken in individuals for whom competitive sport is an essential component of their livelihood, such as professional athletes. Myopericarditis is an uncommon, however potentially serious medical condition with a wide variety of aetiologies, including viral, autoimmune, and drug-related causes. Management is mainly supportive and relies on prompt recognition and removal of the aetiological process. Mesalazine-induced myopericarditis is a rare condition; as such increasing awareness of mesalazine as a precipitant of myopericarditis is vital for optimising the management of these patients.

Keywords: myopericarditis, mesalazine, inflammatory bowel disease, professional athlete

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Authors: Faris Mohsin Alabeedi

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Pemphigus Vulgaris (PV) is a life-threatening autoimmune blistering disease characterized by the presence of autoantibodies directed against the epidermis's surface proteins. There are two forms of PV, mucocutaneous and mucosal-dominant PV. Disruption of the cell junctions is a hallmark of PV due to the autoantibodies targeting the desmosomal cadherins, desmoglein-3 (Dsg3) and desmoglein-1, leading to acantholysis in the skin and mucous membrane. Although the pathogenesis of PV is known, the detailed molecular events remain not fully understood. Our recent study has shown that both the PV sera and pathogenic anti-Dsg3 antibody AK23 can induce ROS and cause oxidative stress in cultured keratinocytes. In line with our finding, other independent studies also demonstrate oxidative stress in PV. Since Nrf2 plays a crucial role in cellular anti-oxidative stress response, we hypothesize that the expression of Nrf2 may alter in PV. Thus, treatment of cells with PV sera or AK23 may cause changes in Nrf2 expression and distribution. The purpose of this study was to examine the effect of AK23 and PV sera on Nrf2 in a normal human keratinocyte cell line, such as NTERT cells. Both a time-course and dose-dependent experiments with AK23, alongside the matched isotype control IgG, were performed in keratinocyte cultures and analysed by immunofluorescence for Nrf2 and Dsg3. Additionally, the same approach was conducted with the sera from PV patients and healthy individuals that served as a control in this study. All the fluorescent images were analysed using ImageJ software. Each experiment was repeated twice. In general, variations were observed throughout this study. In the dose-response experiments, although enhanced Dsg3 expression was consistently detected in AK23 treated cells, the expression of Nrf2 showed no consistent findings between the experiments, although changes in its expression were noticeable in cells treated with AK23. In the time-course study, a trend with induction of Nrf2 over time was shown in control cells treated with mouse isotype IgG. Treatment with AK23 showed a reduction of Nrf2 in a time-dependent manner, especially at the 24-hour time point. However, the earlier time points, such as 2 hours and 6 hours with AK23 treatments, detected somewhat variations. Finally, PV sera caused a decrease of Dsg3, but on the other hand, variations were observed in Nrf2 expression in PV sera treated cells. In general, PV sera seemed to cause a reduction of Nrf2 in the majority of PV sera treated samples. In addition, more pronounced cytoplasmic expression of Nrf2 has been observed in PV sera treated cells than those treated with AK23, suggesting that polyclonal and monoclonal IgG might induce a different effect on Nrf2 expression and distribution. Further experimental studies are crucial to obtain a more coincide global view of Nrf2-mediated gene regulation. In particular, Pemphigus Voulgaris studies assessing how the Nrf2-dependent network changes from a physiological to a pathological condition can provide insight into disease mechanisms and perhaps initiate further treatment approaches.

Keywords: pemphigus vulgaris, monoclonal antibody against desmoglein-3, Nrf2 oxidative stress, keratinocyte cultures

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Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

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Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

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Authors: Supriya Majumder, Pabitra Banik

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Fates of Arsenic (As) on the soil-plant environment belong to the critical emerging issue, which in turn to appraises the threatening implications of a human health risk — assessing the dynamics of As in soil solid components are likely to impose its potential availability towards plant uptake. In the present context, we introduced an improved Sequential Extraction Procedure (SEP) questioning to identify solid-phase speciation of As in paddy soil under variable soil environmental conditions during two consecutive seasons of rice cultivation practices. We coupled gradients of water management practices with the addition of fertilizer amendments to assess the changes in a partition of As through a field experimental study during monsoon and post-monsoon season using two rice cultivars. Water management regimes were varied based on the methods of cultivation of rice by Conventional (waterlogged) vis-a-vis System of Rice Intensification-SRI (saturated). Fertilizer amendment through the nutrient treatment of absolute control, NPK-RD, NPK-RD + Calcium silicate, NPK-RD + Ferrous sulfate, Farmyard manure (FYM), FYM + Calcium silicate, FYM + Ferrous sulfate, Vermicompost (VC), VC + Calcium silicate, VC + Ferrous sulfate were selected to construct the study. After harvest, soil samples were sequentially extracted to estimate partition of As among the different fractions such as: exchangeable (F1), specifically sorbed (F2), As bound to amorphous Fe oxides (F3), crystalline Fe oxides (F4), organic matter (F5) and residual phase (F6). Results showed that the major proportions of As were found in F3, F4 and F6, whereas F1 exhibited the lowest proportion of total soil As. Among the nutrient treatment mediated changes on As fractions, the application of organic manure and ferrous sulfate were significantly found to restrict the release of As from exchangeable phase. Meanwhile, conventional practice produced much higher release of As from F1 as compared to SRI, which may substantially increase the environmental risk. In contrast, SRI practice was found to retain a significantly higher proportion of As in F2, F3, and F4 phase resulting restricted mobilization of As. This was critically reflected towards rice grain As bioavailability where the reduction in grain As concentration of 33% and 55% in SRI concerning conventional treatment (p <0.05) during monsoon and post-monsoon season respectively. Also, prediction assay for rice grain As bioavailability based on the linear regression model was performed. Results demonstrated that rice grain As concentration was positively correlated with As concentration in F1 and negatively correlated with F2, F3, and F4 with a satisfactory level of variation being explained (p <0.001). Finally, we conclude that F1, F2, F3 and F4 are the major soil. As fractions critically may govern the potential availability of As in soil and suggest that rice cultivation with the SRI treatment is particularly at less risk of As availability in soil. Such exhaustive information may be useful for adopting certain management practices for rice grown in contaminated soil concerning to the environmental issues in particular.

Keywords: arsenic, fractionation, paddy soil, potential availability

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Authors: Georgios Samaras

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New media such as Facebook, YouTube and Twitter introduced the world to a new era of instant communication. An era where online interactions could replace a lot of offline actions. Technology can create a mediated environment in which participants can communicate (one-to-one, one-to-many, and many-to-many) both synchronously and asynchronously and participate in reciprocal message exchanges. Currently, social networks are attracting similar academic attention to that of the internet after its mainstream implementation into public life. Websites and platforms are seen as the forefront of a new political change. There is a significant backdrop of previous methodologies employed to research the effects of social networks. New approaches are being developed to be able to adapt to the growth of social networks and the invention of new platforms. Golden Dawn was the first openly neo-Nazi party post World War II to win seats in the parliament of a European country. Its racist rhetoric and violent tactics on social networks were rewarded by their supporters, who in the face of Golden Dawn’s leaders saw a ‘new dawn’ in Greek politics. Mainstream media banned its leaders and members of the party indefinitely after Ilias Kasidiaris attacked Liana Kanelli, a member of the Greek Communist Party, on live television. This media ban was seen as a treasonous move by a significant percentage of voters, who believed that the system was desperately trying to censor Golden Dawn to favor mainstream parties. The shocking attack on live television received international coverage and while European countries were condemning this newly emerged neo-Nazi rhetoric, almost 7 percent of the Greek population rewarded Golden Dawn with 18 seats in the Greek parliament. Many seem to think that Golden Dawn mobilised its voters online and this approach played a significant role in spreading their message and appealing to wider audiences. No strict online censorship existed back in 2012 and although Golden Dawn was openly used neo-Nazi symbolism, it was allowed to use social networks without serious restrictions until 2017. This paper used qualitative methods to investigate Golden Dawn’s rise in social networks from 2012 to 2019. The focus of the content analysis was set on three social networking platforms: Facebook, Twitter and YouTube, while the existence of Golden Dawn’s website, which was used as a news sharing hub, was also taken into account. The content analysis included text and visual analyses that sampled content from their social networking pages to translate their political messaging through an ideological lens focused on extreme-right populism. The absence of hate speech regulations on social network platforms in 2012 allowed the free expression of those heavily ultranationalist and populist views, as they were employed by Golden Dawn in the Greek political scene. On YouTube, Facebook and Twitter, the influence of their rhetoric was particularly strong. Official channels and MPs profiles were investigated to explore the messaging in-depth and understand its ideological elements.

Keywords: populism, far-right, social media, Greece, golden dawn

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Authors: Neelam Taneja, Abhishek Mewara, Ajay Kumar

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Multidrug resistant shigellae have emerged as a therapeutic challenge in India. At our 2000 bed tertiary care referral centre in Chandigarh, North India, which caters to a large population of 7 neighboring states, antibiotic resistance in Shigella is being constantly monitored. Shigellae are isolated from 3 to 5% of all stool samples. In 1990 nalidixic acid was the drug of choice as 82%, and 63% of shigellae were resistant to ampicillin and cotrimoxazole respectively. Nalidixic acid resistance emerged in 1992 and rapidly increased from 6% during 1994-98 to 86% by the turn of 21st century. In the 1990s, the WHO recommended ciprofloxacin as the drug of choice for empiric treatment of shigellosis in view of the existing high level resistance to agents like chloramphenicol, ampicillin, cotrimoxazole and nalidixic acid. First resistance to ciprofloxacin in S. flexneri at our centre appeared in 2000 and rapidly rose to 46% in 2007 (MIC>4mg/L). In between we had an outbreak of ciprofloxacin resistant S.dysenteriae serotype 1 in 2003. Therapeutic failures with ciprofloxacin occurred with both ciprofloxacin-resistant S. dysenteriae and ciprofloxacin-resistant S. flexneri. The severity of illness was more with ciprofloxacin-resistant strains. Till 2000, elsewhere in the world ciprofloxacin resistance in S. flexneri was sporadic and uncommon, though resistance to co-trimoxazole and ampicillin was common and in some areas resistance to nalidixic acid had also emerged. Fluoroquinolones due to extensive use and misuse for many other illnesses in our region are thus no longer the preferred group of drugs for managing shigellosis in India. WHO presently recommends ceftriaxone and azithromycin as alternative drugs to fluoroquinolone-resistant shigellae, however, overreliance on this group of drugs also seems to soon become questionable considering the emerging cephalosporin-resistant shigellae. We found 15.1% of S. flexneri isolates collected over a period of 9 years (2000-2009) resistant to at least one of the third-generation cephalosporins (ceftriaxone/cefotaxime). The first isolate showing ceftriaxone resistance was obtained in 2001, and we have observed an increase in number of isolates resistant to third generation cephalosporins in S. flexneri 2005 onwards. This situation has now become a therapeutic challenge in our region. The MIC values for Shigella isolates revealed a worrisome rise for ceftriaxone (MIC90:12 mg/L) and cefepime (MIC90:8 mg/L). MIC values for S. dysenteriae remained below 1 mg/L for ceftriaxone, however for cefepime, the MIC90 has raised to 4 mg/L. These infections caused by ceftriaxone-resistant S. flexneri isolates were successfully treated by azithromycin at our center. Most worrisome development in the present has been the emergence of DSA(Decreased susceptibility to azithromycin) which surfaced in 2001 and has increased from 4.3% till 2011 to 34% thereafter. We suspect plasmid-mediated resistance as we detected qnrS1-positive Shigella for the first time from the Indian subcontinent in 2 strains from 2010, indicating a relatively new appearance of this PMQR determinant among Shigella in India. This calls for a continuous and strong surveillance of antibiotic resistance across the country. The prevention of shigellosis by developing cost-effective vaccines is desirable as it will substantially reduce the morbidity associated with diarrhoea in the country

Keywords: Shigella, antimicrobial, resistance, India

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