Search results for: human and viral genes
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 9311

Search results for: human and viral genes

6461 Meniere's Disease and its Prevalence, Symptoms, Risk Factors and Associated Treatment Solutions for this Disease

Authors: Amirreza Razzaghipour Sorkhab

Abstract:

One of the most common disorders among humans is hearing impairment. This paper provides an evidence base that recovers understanding of Meniere’s disease and highlights the physical and mental health correlates of the disorder. Meniere's disease is more common in the elderly. The term idiopathic endolymphatic hydrops has been attributed to this disease by some in the previous. Meniere’s disease demonstrations a genetic tendency, and a family history is found in 10% of cases, with an autosomal dominant inheritance pattern. The COCH gene may be one of the hereditary factors contributing to Meniere’s disease, and the possibility of a COCH mutation should be considered in patients with Meniere’s disease symptoms. Should be considered Missense mutations in the COCH gene cause the autosomal dominant sensorineural hearing loss and vestibular disorder. Meniere’s disease is a complex, heterogeneous disorder of the inner ear and that is characterized by episodes of vertigo lasting from minutes to hours, fluctuating sensorineural hearing loss, tinnitus, and aural fullness. The existing evidence supports the suggestion that age and sleep disorder are risk factors for Meniere's disease. Many factors have been reported to precipitate the progress of Menier, including endolymphatic hydrops, immunology, viral infection, inheritance, vestibular migraine, and altered intra-labyrinthine fluid dynamics. Although there is currently no treatment that has a proven helpful effect on hearing levels or on the long-term evolution of the disease, however, in the primary stages, the hearing may improve among attacks, but a permanent hearing loss occurs in the majority of cases. Current publications have proposed a role for the intratympanic use of medicine, mostly aminoglycosides, for the control of vertigo. more than 85% of patients with Meniere's disease are helped by either changes in lifestyle and medical treatment or minimally aggressive surgical procedures such as intratympanic steroid therapy, intratympanic gentamicin therapy, and endolymphatic sac surgery. However, unilateral vestibular extirpation methods (intratympanic gentamicin, vestibular nerve section, or labyrinthectomy) are more predictable but invasive approaches to control the vertigo attacks. Medical therapy aimed at reducing endolymph volume, such as low-sodium diet, diuretic use, is the typical initial treatment.

Keywords: meniere's disease, endolymphatic hydrops, hearing loss, vertigo, tinnitus, COCH gene

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6460 An Automated Magnetic Dispersive Solid-Phase Extraction Method for Detection of Cocaine in Human Urine

Authors: Feiyu Yang, Chunfang Ni, Rong Wang, Yun Zou, Wenbin Liu, Chenggong Zhang, Fenjin Sun, Chun Wang

Abstract:

Cocaine is the most frequently used illegal drug globally, with the global annual prevalence of cocaine used ranging from 0.3% to 0.4 % of the adult population aged 15–64 years. Growing consumption trend of abused cocaine and drug crimes are a great concern, therefore urine sample testing has become an important noninvasive sampling whereas cocaine and its metabolites (COCs) are usually present in high concentrations and relatively long detection windows. However, direct analysis of urine samples is not feasible because urine complex medium often causes low sensitivity and selectivity of the determination. On the other hand, presence of low doses of analytes in urine makes an extraction and pretreatment step important before determination. Especially, in gathered taking drug cases, the pretreatment step becomes more tedious and time-consuming. So developing a sensitive, rapid and high-throughput method for detection of COCs in human body is indispensable for law enforcement officers, treatment specialists and health officials. In this work, a new automated magnetic dispersive solid-phase extraction (MDSPE) sampling method followed by high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed for quantitative enrichment of COCs from human urine, using prepared magnetic nanoparticles as absorbants. The nanoparticles were prepared by silanizing magnetic Fe3O4 nanoparticles and modifying them with divinyl benzene and vinyl pyrrolidone, which possesses the ability for specific adsorption of COCs. And this kind of magnetic particle facilitated the pretreatment steps by electromagnetically controlled extraction to achieve full automation. The proposed device significantly improved the sampling preparation efficiency with 32 samples in one batch within 40mins. Optimization of the preparation procedure for the magnetic nanoparticles was explored and the performances of magnetic nanoparticles were characterized by scanning electron microscopy, vibrating sample magnetometer and infrared spectra measurements. Several analytical experimental parameters were studied, including amount of particles, adsorption time, elution solvent, extraction and desorption kinetics, and the verification of the proposed method was accomplished. The limits of detection for the cocaine and cocaine metabolites were 0.09-1.1 ng·mL-1 with recoveries ranging from 75.1 to 105.7%. Compared to traditional sampling method, this method is time-saving and environmentally friendly. It was confirmed that the proposed automated method was a kind of highly effective way for the trace cocaine and cocaine metabolites analyses in human urine.

Keywords: automatic magnetic dispersive solid-phase extraction, cocaine detection, magnetic nanoparticles, urine sample testing

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6459 Introducing the Concept of Sustainable Learning: Redesigning the Social Studies and Citizenship Education Curriculum in the Context of Saudi Arabia

Authors: Aiydh Aljeddani, Fran Martin

Abstract:

Sustainable human development is an essential component of a sustainable economic, social and environmental development. Addressing sustainable learning only through the addition of new teaching methods, or embedding certain approaches, is not sufficient on its own to support the goals of sustainable human development. This research project seeks to explore how the process of redesigning the current principles of curriculum based on the concept of sustainable learning could contribute to preparing a citizen who could later contribute towards sustainable human development. Multiple qualitative methodologies were employed in order to achieve the aim of this study. The main research methods were teachers’ field notes, artefacts, informal interviews (unstructured interview), a passive participant observation, a mini nominal group technique (NGT), a weekly diary, and weekly meeting. The study revealed that the integration of a curriculum for sustainable development, in addition to the use of innovative teaching approaches, highly valued by students and teachers in social studies’ sessions. This was due to the fact that it created a positive atmosphere for interaction and aroused both teachers and students’ interest. The content of the new curriculum also contributed to increasing students’ sense of shared responsibility through involving them in thinking about solutions for some global issues. This was carried out through addressing these issues through the concept of sustainable development and the theory of Thinking Activity in a Social Context (TASC). Students had interacted with sustainable development sessions intellectually and they also practically applied it through designing projects and cut-outs. Ongoing meetings and workshops to develop work between both the researcher and the teachers, and by the teachers themselves, played a vital role in implementing the new curriculum. The participation of teachers in the development of the project through working papers, exchanging experiences and introducing amendments to the students' environment was also critical in the process of implementing the new curriculum. Finally, the concept of sustainable learning can contribute to the learning outcomes much better than the current curriculum and it can better develop the learning objectives in educational institutions.

Keywords: redesigning, social studies and citizenship education curriculum, sustainable learning, thinking activity in a social context

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6458 The Impact of Artificial Intelligence on Medicine Production

Authors: Yasser Ahmed Mahmoud Ali Helal

Abstract:

The use of CAD (Computer Aided Design) technology is ubiquitous in the architecture, engineering and construction (AEC) industry. This has led to its inclusion in the curriculum of architecture schools in Nigeria as an important part of the training module. This article examines the ethical issues involved in implementing CAD (Computer Aided Design) content into the architectural education curriculum. Using existing literature, this study begins with the benefits of integrating CAD into architectural education and the responsibilities of different stakeholders in the implementation process. It also examines issues related to the negative use of information technology and the perceived negative impact of CAD use on design creativity. Using a survey method, data from the architecture department of University was collected to serve as a case study on how the issues raised were being addressed. The article draws conclusions on what ensures successful ethical implementation. Millions of people around the world suffer from hepatitis C, one of the world's deadliest diseases. Interferon (IFN) is treatment options for patients with hepatitis C, but these treatments have their side effects. Our research focused on developing an oral small molecule drug that targets hepatitis C virus (HCV) proteins and has fewer side effects. Our current study aims to develop a drug based on a small molecule antiviral drug specific for the hepatitis C virus (HCV). Drug development using laboratory experiments is not only expensive, but also time-consuming to conduct these experiments. Instead, in this in silicon study, we used computational techniques to propose a specific antiviral drug for the protein domains of found in the hepatitis C virus. This study used homology modeling and abs initio modeling to generate the 3D structure of the proteins, then identifying pockets in the proteins. Acceptable lagans for pocket drugs have been developed using the de novo drug design method. Pocket geometry is taken into account when designing ligands. Among the various lagans generated, a new specific for each of the HCV protein domains has been proposed.

Keywords: drug design, anti-viral drug, in-silicon drug design, hepatitis C virus (HCV) CAD (Computer Aided Design), CAD education, education improvement, small-size contractor automatic pharmacy, PLC, control system, management system, communication

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6457 Characteristics Flakes Product with Dry Residue of Wild Orenago

Authors: Kosutic Milenko, Filipovic Jelena

Abstract:

Cereals constitute the staple food of the human race. In accordance with the modern nutritionist opinions, cereal products, flakes and snack products are the most common foods in the daily diet, such as ready to eat breakfast cereal, flakes, and snacks. Extrusion technology makes it possible to apply different sources of ingredients for the enrichment of cereal-based flakes or snacks products. Substances with strong antioxidant properties such as wild oregano have a positive impact on human health, therefore attracting the attention of scientists, consumers and food industry experts. This paper investigates the effects of simultaneous addition of dry residue of wild oregano (0.5% and 1%), on the physical and colour properties of corn flakes to obtain new products with altered nutritional properties. Post-hoc Tukey’s HSD test at 95% confidence limit showed significant differences between various samples. Addition of dry residue wild oregano positively influenced physical characteristics (decreased bulk density 30.2%, increased expansion rate 44.9%), influenced of decrease hardness 38.1% and work of compression 40.3% also significantly change the color of flakes product. Presented data point that investigated corn flakes is a new product with good physical and sensory properties due to higher level of dry residue of wild oregano.

Keywords: flakes product, wild oregano, phisical properties, colour, sensory properties

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6456 Exploring Simple Sequence Repeats within Conserved microRNA Precursors Identified from Tea Expressed Sequence Tag (EST) Database

Authors: Anjan Hazra, Nirjhar Dasgupta, Chandan Sengupta, Sauren Das

Abstract:

Tea (Camellia sinensis) has received substantial attention from the scientific world time to time, not only for its commercial importance, but also for its demand to the health-conscious people across the world for its extensive use as potential sources of antioxidant supplement. These health-benefit traits primarily rely on some regulatory networks of different metabolic pathways. Development of microsatellite markers from the conserved genomic regions is being worthwhile for studying the genetic diversity of closely related species or self-pollinated species. Although several SSR markers have been reported, in tea the trait-specific Simple Sequence Repeats (SSRs) are yet to be identified, which can be used for marker assisted breeding technique. MicroRNAs are endogenous, noncoding, short RNAs directly involved in regulating gene expressions at the post-transcriptional level. It has been found that diversity in miRNA gene interferes the formation of its characteristic hair pin structure and the subsequent function. In the present study, the precursors of small regulatory RNAs (microRNAs) has been fished out from tea Expressed Sequence Tag (EST) database. Furthermore, the simple sequence repeat motifs within the putative miRNA precursor genes are also identified in order to experimentally validate their existence and function. It is already known that genic-SSR markers are very adept and breeder-friendly source for genetic diversity analysis. So, the potential outcome of this in-silico study would provide some novel clues in understanding the miRNA-triggered polymorphic genic expression controlling specific metabolic pathways, accountable for tea quality.

Keywords: micro RNA, simple sequence repeats, tea quality, trait specific marker

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6455 High Expression Levels and Amplification of rRNA Genes in a Mentally Retarded Child with 13p+: A Familial Case Study

Authors: Irina S. Kolesnikova, Alexander A. Dolskiy, Natalya A. Lemskaya, Yulia V. Maksimova, Asia R. Shorina, Alena S. Telepova, Alexander S. Graphodatsky, Dmitry V. Yudkin

Abstract:

A cytogenetic and molecular genetic study of the family with a male child who had mental retardation and autistic features revealed an abnormal chromosome 13 bearing an enlarged p-arm with amplified ribosomal DNA (rDNA) in a boy and his father. Cytogenetic analysis using standard G-banding and FISH with labeled rDNA probes revealed an abnormal chromosome 13 with an enlarged p-arms due to rDNA amplification in a male child, who had clinically confirmed mental retardation and an autistic behavior. This chromosome is evidently inherited from the father, who has morphologically the same chromosome, but is healthy. The karyotype of the mother was normal. Ag-NOR staining showed brightly stained large whole-p-arm nucleolus organizer regions (NORs) in a child and normal-sized NORs in his father with 13p+-NOR-amount mosaicism. qRT-PCR with specific primers showed highly increased levels of 18S, 28S and 5,8 S ribosomal RNA (rRNA) in the patient’s blood samples compared to a normal healthy control donor. Both patient’s father and mother had no elevated levels of rRNAs expression. Thus, in this case, rRNA level seems to correlate with mental retardation in familial individuals with 13p+. Our findings of rRNA overexpression in a patient with mental retardation and his parents may show a possible link between the karyotype (p-arm enlargement due to rDNA amplification), rDNA functionality (rRNA overexpression), functional changes in the brain and mental retardation. The study is supported by Russian Science Foundation Grant 15-15-10001.

Keywords: mental retardation, ribosomal DNA–rDNA, ribosomal RNA–rRNA, nucleolus organizer region–NOR, chromosome 13

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6454 Real-Time Finger Tracking: Evaluating YOLOv8 and MediaPipe for Enhanced HCI

Authors: Zahra Alipour, Amirreza Moheb Afzali

Abstract:

In the field of human-computer interaction (HCI), hand gestures play a crucial role in facilitating communication by expressing emotions and intentions. The precise tracking of the index finger and the estimation of joint positions are essential for developing effective gesture recognition systems. However, various challenges, such as anatomical variations, occlusions, and environmental influences, hinder optimal functionality. This study investigates the performance of the YOLOv8m model for hand detection using the EgoHands dataset, which comprises diverse hand gesture images captured in various environments. Over three training processes, the model demonstrated significant improvements in precision (from 88.8% to 96.1%) and recall (from 83.5% to 93.5%), achieving a mean average precision (mAP) of 97.3% at an IoU threshold of 0.7. We also compared YOLOv8m with MediaPipe and an integrated YOLOv8 + MediaPipe approach. The combined method outperformed the individual models, achieving an accuracy of 99% and a recall of 99%. These findings underscore the benefits of model integration in enhancing gesture recognition accuracy and localization for real-time applications. The results suggest promising avenues for future research in HCI, particularly in augmented reality and assistive technologies, where improved gesture recognition can significantly enhance user experience.

Keywords: YOLOv8, mediapipe, finger tracking, joint estimation, human-computer interaction (HCI)

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6453 Anticancer Activity of Edible Coprinus Mushroom (Coprinus comatus) on Human Glioblastoma Cell Lines and Interaction with Temozolomide

Authors: Maria Borawska, Patryk Nowakowski, Sylwia K. Naliwajko, Renata Markiewicz-Zukowska, Anna Puscion-Jakubik, Krystyna Gromkowska-Kepka, Justyna Moskwa

Abstract:

Coprinus comatus (O. F. Müll.) Pers.) should not be confused with the common Ink Cap, which contains coprine and can induce coprine poisoning. We study the possibility of applying coprinus mushroom (Coprinus comatus), available in Poland, as food product supporting the treatment of human glioblastoma cells. The U87MG and T98 glioblastoma cell lines were exposed to water (CW) or ethanol 95° (CE) Cantharellus extracts (50-500 μg/ml), with or without temozolomide (TMZ) during 24, 48 or 72 hours. The cell division was examined by the H³-thymidine incorporation. The statistical analysis was performed using Statistica v. 13.0 software. Significant differences were assumed for p < 0.05. We found that both, CW and CE, administrated alone, had inhibitory effect on cell lines growth, but the CE extract had a higher degree of growth inhibition. The anti-tumor effect of TMZ (50 μM) on U87MG was enhanced by mushroom extracts, and the effect was lower to the effect after using Coprinus comatus extracts (CW and CE) alone. A significant decrease (p < 0.05) in pro-MMP2 (82.61 ± 6.3% of control) secretion in U87MG cells was observed after treated with CE (250 μg/ml). We conclude that extracts of Coprinus comatus, edible mushroom, present cytotoxic properties on U87MG and T98 cell lines and may cooperate with TMZ synergistically enhancing its growth inhibiting activity against glioblastoma U87MG cell line.

Keywords: anticancer, glioma, mushroom, temozolomide

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6452 Managing Maritime Security in the Mediterranean Sea: The Roles of the EU in Tackling Irregular Migration

Authors: Shazwanis Shukri

Abstract:

The Mediterranean Sea, at the crossroads of three continents has always been the focus of pan-European and worldwide attention. Over the past decade, the Mediterranean Sea has become a hotbed for irregular migration particularly from the African continent toward the Europe. Among the major transit routes in the Mediterranean Sea include the Strait of Gibraltar, Canary Island and island of Lampedusa. In recent years, Mediterranean Sea has witnessed significant numbers of accidents and shipwrecks involving the irregular migrants and refugees trying to reach Europe via the sea. The shipwrecks and traffickers exploitation of migrants draw most of the attention particularly for the European Union (EU). This incident has been a wakeup call for the EU and become the top political agenda in the EU policy to tackle irregular migration and human smuggling at sea. EU has repeatedly addressed irregular migration as one of the threats the EU and its citizens may be confronted with and therefore immediate measures are crucial to tackle the crisis. In light of this, various initiatives have been adopted by the EU to strengthen external border control and restrict access to irregular migrants, notably through the enforcement of Frontex and Eunavfor Med. This paper analyses current development of counter-migration operations by the EU in response to migration crisis in the Mediterranean Sea. The analysis is threefold. First, this study examines the patterns and trends of irregular migration’s movements from recent perspective. Second, this study concentrates on the evolution of the EU operations that are in place in the Mediterranean Sea, notably by Frontex and Eunavfor Med to curb the influx of irregular migrants to the European countries, including, among others, Greece and Italy. Third, this study investigates the EU approaches to fight against the proliferation of human trafficking networks at sea. This study is essential to determine the roles of the EU in tackling migration crisis and human trafficking in the Mediterranean Sea and the effectiveness of their counter-migration operations to reduce the number of irregular migrants travelling via the sea. Elite interviews and document analysis were used as a methodology in this study. The study discovers that the EU operations have successfully contributed to reduce the numbers of irregular migrant’s arrival to Europe. The study also shows that the operations were effective to disrupt smugglers business models particularly from Libya. This study provides essential understanding about the roles of the EU not limited to tackle the migration crisis and disrupt trafficking networks, but also pledged to prevent further loss of lives at sea.

Keywords: European union, frontex, irregular migration, Mediterranean sea

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6451 Biologically Synthesised Silver Nanoparticles Induces Autophagy and JNK Signaling as a Pro-Survival Response by Abrogating Reactive Oxygen Species Accumulation in Cancer Cells

Authors: Sudeshna Mukherjee, Leena Fageria, R. Venkataramana Dilip, Rajdeep Chowdhury, Jitendra Panwar

Abstract:

Metal nanoparticles in recent years have gained importance in cancer therapy due to their enhanced permeability retention effect. Among various nanomaterials, silver nanoparticles (AgNPs) have received considerable attention due to their unique properties like conductivity, chemical stability, relative lower toxicity and outstanding therapeutic potential, such as anti-inflammatory, antimicrobial and anti-cancerous activities. In this study, we took a greener approach to synthesize silver nanoparticle from fungus and analyze its effects on both epithelial and mesenchymal derived cancer cells. Much research has been done on nanoparticle-induced apoptosis, but little is known about its role in autophagy. In our study, the silver nanoparticles were seen to induce autophagy which was analyzed by studying the expression of several autophagy markers like, LC3B-II and ATG genes. Monodansylcadaverine (MDC) assay also revealed the induction of autophagy upon treatment with AgNPs. Inhibition of autophagy by chloroquine resulted in increased cell death suggesting autophagy as a survival strategy adopted by the cells. In parallel to autophagy induction, silver nanoparticles induced ROS accumulation. Interestingly, autophagy inhibition by chloroquine increased ROS level, resulting in enhanced cell death. We further analyzed MAPK signaling upon AgNP treatment. It was observed that along with autophagy, activation of JNK signaling served as pro-survival while ERK signaling served as a pro-death signal. Our results provide valuable insights into the role of autophagy upon AgNP exposure and provide cues to probabilistic strategies to effectively sensitize cancer cells.

Keywords: autophagy, JNK signalling, reactive oxygen species, silver nanoparticles

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6450 Municipal Employees’ Perceptions of Fairness of Human Resource Management Practices and Employee Organisational Commitment

Authors: Lineo Dzansi

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South African government has been mandated by the Constitution (Act 108 of 1996) to deliver basic services to all who live in it. However, service delivery has always been marred with much criticism and citizens’ dissatisfaction regarding the quality of services rendered to them. This is evidenced by public protests that are common in South Africa lately which they are mostly alleged to link with failure by the government through various municipalities to meet citizens’ service delivery expectations. Municipalities render services through people. People management plays a crucial role in influencing employee and organisational performance and it thus needs to be conducted in a fair and just manner. Literature confirms that there is a relationship between organisational justice perceptions and employee behaviour, and that positive or negative justice perceptions can have an influence on employee attitudes, commitment to their jobs and organisation. The nature of the attachments formed by individuals to their employing organisations depends on the manner in which the organisation treats them. This implies that Municipal employees’ commitment could be linked to fair or unfair perceptions of Human Resource Management practices within their organisations. Unfortunately, the political nature of municipal environment could be a fertile ground for appointments of people based on political affiliation as a reward for political patronage rather than on merit. This paper seeks to investigate the relationship between municipal employees’ perceptions of fairness of Human Resource Management practices and employee commitment from the organisational justice point of view. Research on organisational justice has shown that employees’ organisational justice perceptions link directly with job satisfaction and employee organisational commitment. Quantitative research methods were employed to collect and analyse data from selected managerial and non-managerial municipal employees within selected municipalities in Free State Province of South Africa. Employee commitment has positive relationships with HRM practices at the .05 and .01 levels of significance – indicating that the higher the levels of HRM practices in municipal employees the higher the organisational commitment of employees. Therefore, it is concluded that organisational commitment of municipal employees (EOC) is positively related to their perceptions of fairness of HRM practices (PHF) of municipalities. In other words, fair HRM practices of municipalities promote organisational commitment in municipal employees.

Keywords: organisational Justice, HRM practices, employee organisational commitment, employee attitudes

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6449 Expand Rabies Post-Exposure Prophylaxis to Where It Is Needed the Most

Authors: Henry Wilde, Thiravat Hemachudha

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Human rabies deaths are underreported worldwide at 55,000 annual cases; more than of dengue and Japanese encephalitis. Almost half are children. A recent study from the Philippines of nearly 2,000 rabies deaths revealed that none of had received incomplete or no post exposure prophylaxis. Coming from a canine rabies endemic country, this is not unique. There are two major barriers to reducing human rabies deaths: 1) the large number of unvaccinated dogs and 2) post-exposure prophylaxis (PEP) that is not available, incomplete, not affordable, or not within reach for bite victims travel means. Only the first barrier, inadequate vaccination of dogs, is now being seriously addressed. It is also often not done effectively or sustainably. Rabies PEP has evolved as a complex, prolonged process, usually delegated to centers in larger cities. It is virtually unavailable in villages or small communities where most dog bites occur, victims are poor and usually unable to travel a long distance multiple times to receive PEP. Reseacrh that led to better understanding of the pathophysiology of rabies and immune responses to potent vaccines and immunoglobulin have allowed shortening and making PEP more evidence based. This knowledge needs to be adopted and applied so that PEP can be rendered safely and affordably where needed the most: by village health care workers who have long performed more complex services after appropriate training. Recent research makes this an important and long neglected goal that is now within our means to implement.

Keywords: rabies, post-exposure prophylaxis, availability, immunoglobulin

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6448 NeuroBactrus, a Novel, Highly Effective, and Environmentally Friendly Recombinant Baculovirus Insecticide

Authors: Yeon Ho Je

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A novel recombinant baculovirus, NeuroBactrus, was constructed to develop an improved baculovirus insecticide with additional beneficial properties, such as a higher insecticidal activity and improved recovery, compared to wild-type baculovirus. For the construction of NeuroBactrus, the Bacillus thuringiensis crystal protein gene (here termed cry1-5) was introduced into the Autographa californica nucleopolyhedrovirus (AcMNPV) genome by fusion of the polyhedrin–cry1-5–polyhedrin genes under the control of the polyhedrin promoter. In the opposite direction, an insect-specific neurotoxin gene, AaIT, from Androctonus australis was introduced under the control of an early promoter from Cotesia plutellae bracovirus by fusion of a partial fragment of orf603. The polyhedrin–Cry1-5–polyhedrin fusion protein expressed by the NeuroBactrus was not only occluded into the polyhedra, but it was also activated by treatment with trypsin, resulting in an_65-kDa active toxin. In addition, quantitative PCR revealed that the neurotoxin was expressed from the early phase of infection. NeuroBactrus showed a high level of insecticidal activity against Plutella xylostella larvae and a significant reduction in the median lethal time against Spodoptera exigua larvae compared to those of wild-type AcMNPV. Rerecombinant mutants derived from NeuroBactrus in which AaIT and/or cry1-5 were deleted were generated by serial passages in vitro. Expression of the foreign proteins (B. thuringiensis toxin and AaIT) was continuously reduced during the serial passage of the NeuroBactrus. Moreover, polyhedra collected from S. exigua larvae infected with the serially passaged NeuroBactrus showed insecticidal activity similar to that of wild-type AcMNPV. These results suggested that NeuroBactrus could be recovered to wild-type AcMNPV through serial passaging.

Keywords: baculovirus, insecticide, neurotoxin, neurobactrus

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6447 Assessment of the Entrepreneurial Trends of Agricultural Undergraduates: A Study at Faculty of Agriculture, Eastern University, Sri Lanka

Authors: Tharsinithevy Kirupananthan, Thivahary Geretharan

Abstract:

Since creation of agricultural enterprises going to reflect the micro and macro level development of Sri Lanka, it is vey important to study the entrepreneurial trends of Agricultural Undergraduates. Likert scale scoring method was used to assess the responses of involvement, Role model effect, aware of demands, confidence and willingness. 37.8% were strongly agreed to do full time business. The average score for to do agriculture businesses were between agree and strongly agree. The average scores for role model effects were less than agree. Average score for aware of needs of society was less than agree. 75.7% of them were able to identify the demands of the society. The demands identified were human capital, self sufficient domestic production, safe and nutritional foods. The confidence of having enough skills score was less than agree. 64.1% of them were owned special skills to carry out entrepreneurial activities. Such skills were possession of different human capitals management skills. The willingness responses scores were more than agree. 61.5% of them were discussed their business plan. Their dream plans were development of new food products, Quality planting materials, harmless method of cultivation and floricultural industry. Those were supported by government policies and other related organizations.

Keywords: agricultural undergraduates, entrepreneurial trends, likert scale Sri Lanka

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6446 Roughness Discrimination Using Bioinspired Tactile Sensors

Authors: Zhengkun Yi

Abstract:

Surface texture discrimination using artificial tactile sensors has attracted increasing attentions in the past decade as it can endow technical and robot systems with a key missing ability. However, as a major component of texture, roughness has rarely been explored. This paper presents an approach for tactile surface roughness discrimination, which includes two parts: (1) design and fabrication of a bioinspired artificial fingertip, and (2) tactile signal processing for tactile surface roughness discrimination. The bioinspired fingertip is comprised of two polydimethylsiloxane (PDMS) layers, a polymethyl methacrylate (PMMA) bar, and two perpendicular polyvinylidene difluoride (PVDF) film sensors. This artificial fingertip mimics human fingertips in three aspects: (1) Elastic properties of epidermis and dermis in human skin are replicated by the two PDMS layers with different stiffness, (2) The PMMA bar serves the role analogous to that of a bone, and (3) PVDF film sensors emulate Meissner’s corpuscles in terms of both location and response to the vibratory stimuli. Various extracted features and classification algorithms including support vector machines (SVM) and k-nearest neighbors (kNN) are examined for tactile surface roughness discrimination. Eight standard rough surfaces with roughness values (Ra) of 50 μm, 25 μm, 12.5 μm, 6.3 μm 3.2 μm, 1.6 μm, 0.8 μm, and 0.4 μm are explored. The highest classification accuracy of (82.6 ± 10.8) % can be achieved using solely one PVDF film sensor with kNN (k = 9) classifier and the standard deviation feature.

Keywords: bioinspired fingertip, classifier, feature extraction, roughness discrimination

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6445 Narrative Point of View in Nature Documentary Films: A Study of The Cove (2009), Tale of a Forest (2012), and Before the Flood (2016)

Authors: Sakshi Yadav, Sushila Shekhawat

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This study addresses different types of points of view as seen in nature documentary films with the help of three eco documentaries, and it would be significant in understanding the role of the narrative point of view as a tool for showing and telling in documentaries. Narrative analysis of a film forms an essential aspect of the discourse of scholarship in film studies. Narration is the chain of events occurring in time and space. The notion of narrative provides the idea of coherence and wholeness to the story. There are various components that the narration carries, one of which is the perspective or point of view. The narrator plays the role of a mediator between the film and the audience; thus, his perspective influences the way the audience interprets the film. Feature films have been analyzed through narrative points of view; however, this research intends to conduct it from the angle of a nature documentary film. The study will examine narrative viewpoints unique to nature documentary films using three ecological documentary films-The Cove (2009), Tale of a forest (2012), and Before the flood (2016). This research will apply the framework of narrative theory and will investigate the impact of the different types of narrative points of view, as each portrays the human-nature relationship from a different standpoint, and it will also study the effect that the narrative point of view has on the mode of these eco documentaries.

Keywords: ecodocumentary, narrative, human-nature relationship, point of view

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6444 Realizing the National Disaster Management Policy of Sri Lanka through Public Private Partnerships

Authors: K. W. A. M. Kokila, Matsui Kenichi

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Sri Lanka’s disaster management policy aims to protect lives and developments in disaster affected areas by effectively using resources for disaster risk reduction, emergency management, and community awareness. However, funding for these action programs has posed a serious challenge to the country’s economy. This paper examines the extent to which private-public partnerships (PPPs) can facilitate and expedite disaster management works. In particular, it discusses the results of the questionnaire survey among policymakers, government administrators, NGOs, and private businesses. This questionnaire was conducted in 2017. All respondents were selected based on their experience in PPP projects in the past. The survey focused on clarifying the effectiveness of past PPP projects as well as their efficiency and transparency. The respondents also provided their own opinions and suggestions to improve the future PPP projects in Sri Lanka. The questionnaire was distributed to fifteen persons. The results show that almost all respondents think that PPP projects are beneficial and important for future disaster risk management in Sri Lanka. The respondents, however, showed some reservation about effectiveness and transparency of the PPP process. This paper also discusses the results on the respondents’ perceptions about their capacity regarding human resources and management. This paper, overall, sheds light on technological, financial and human resource management practices in developed countries as well as policy and legislation provisions regarding PPP projects.

Keywords: disaster management, policy, private public partnership, projects

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6443 Nuclear Resistance Movements: Case Study of India

Authors: Shivani Yadav

Abstract:

The paper illustrates dynamics of nuclear resistance movements in India and how peoples’ power rises in response to subversion of justice and suppression of human rights. The need for democratizing nuclear policy runs implicit through the demands of the people protesting against nuclear programmes. The paper analyses the rationale behind developing nuclear energy according to the mainstream development model adopted by the state. Whether the prevalent nuclear discourse includes people’s ambitions and addresses local concerns or not is discussed. Primarily, the nuclear movements across India comprise of two types of actors i.e. the local population as well as the urban interlocutors. The first type of actor is the local population comprising of the people who are residing in the vicinity of the nuclear site and are affected by its construction, presence and operation. They have very immediate concerns against nuclear energy projects but also have an ideological stand against producing nuclear energy. The other types of actors are the urban interlocutors, who are the intellectuals and nuclear activists who have a principled stand against nuclear energy and help to aggregate the aims and goals of the movement on various platforms. The paper focuses on the nuclear resistance movements at five sites in India- Koodankulam (Tamil Nadu), Jaitapur (Maharashtra), Haripur (West Bengal), Mithivirdi (Gujrat) and Gorakhpur (Haryana). The origin, development, role of major actors and mass media coverage of all these movements are discussed in depth. Major observations from the Indian case include: first, nuclear policy discussions in India are confined to elite circles; secondly, concepts like national security and national interest are used to suppress dissent against mainstream policies; and thirdly, India’s energy policies focus on economic concerns while ignoring the human implications of such policies. In conclusion, the paper observes that the anti-nuclear movements question not just the feasibility of nuclear power but also its exclusionary nature when it comes to people’s participation in policy making, endangering the ecology, violation of human rights, etc. The character of these protests is non-violent with an aim to produce more inclusive policy debates and democratic dialogues.

Keywords: anti-nuclear movements, Koodankulam nuclear power plant, non-violent resistance, nuclear resistance movements, social movements

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6442 Examining Individual and Organisational Legal Accountability for Sexual Exploitation Perpetrated by International Humanitarian Workers in Haiti

Authors: Elizabeth Carthy

Abstract:

There is growing recognition that sexual exploitation and abuse (SEA) perpetrated by humanitarian workers is widespread, most recently affirmed by allegations of high-ranking Oxfam officials paying women for sex in post-earthquake Haiti. SEA covers a range of gendered abuses, including rape, sexual assault, and ‘transactional’ or ‘survival’ sex. Holding individuals legally accountable for such behaviors is difficult in all contexts even more so in fragile and conflict-affected settings. Transactional sex, for the purposes of this paper, refers to situations where humanitarian workers exchange aid or assistance for sexual services. This paper explores existing organizational accountability measures relating to transactional sex engaged in by international humanitarian workers through a descriptive and interpretive case study approach-examining the situation in Haiti. It comparatively analyses steps the United Nations has taken to combat this problem. Then it examines the possibility of domestic legal accountability for such conduct in Haiti. Finally, the paper argues that international human rights law can fill in potential gaps in domestic legal frameworks to ensure states hold humanitarian workers and potentially organizations accountable for engaging in and/or perpetuating this gendered abuse of power.

Keywords: gender-based violence, humanitarian action, international human rights law, sexual exploitation

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6441 Methylation Analysis of PHF20L1 and DACT2 Gene Promoters in Women with Breast Cancer

Authors: Marta E. Hernandez-Caballero, Veronica Borgonio-Cuadra, Antonio Miranda-Duarte, Xochitl Rojas-Toledo, Normand Garcia-Hernandez, Maura Cardenas-Garcia, Teresa Abad-Camacho

Abstract:

Breast cancer (BC) is the most common tumor in women over the world. DNA methylation is an epigenetic modification critical in CpG sites, aberrant methylation of CpG islands in promoters is a hallmark of cancer. So, gene expression can be regulated by alterations in DNA methylation. In cell lines DACT2 gene reduces the growth and migration of tumor cells by its participation in the suppression of TGFb/SMAD2/3. PHF20L1 is involved in histone acetylation therefore, it regulates transcription. Our aim was to analyze the methylation status of the DACT2 and PHF20L1 promoter regions in tumoral and healthy mammary tissue from women with BC in different progression states. The study included 77 patients from Centro Medico Nacional La Raza in Mexico City. After identifying a CpG island in DACT2 and PHF20L1 promoters, DNA methylation status was analyzed through sodium bisulfite with subsequent amplification using methylation-specific PCR. Results revealed no changes in methylation status of PHF20L1 and cancer stages (II y III) or in comparison to healthy tissues, it was demethylated. DACT2 promoter methylation was no significant between tumoral stages (II, P = 0.37; III, P = 0.17) or with healthy tissue. Previous data reported DACT2 methylated in nasopharyngeal carcinoma but in this study promoter methylation was not observed. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains, it has been suggested that can stabilize DNMT1 regulating DNA methylation, therefore, was associated with poor prognostic in BC. We found no evidence of methylation in patients and controls in PHF20L1 promoter, so its association with BC may have no direct relation with promoter methylation. More studies including other methylation sites in these genes in BC are necessary.

Keywords: bisulfite conversion, breast cancer, DACT2, DNA methylation, PHF20L1, tumoral status

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6440 The Role of Behavioral Syndromes in Human-Cattle Interactions: A Physiological Approach

Authors: Fruzsina Luca Kézér, Viktor Jurkovich, Ottó Szenci, János Tőzsér, Levente Kovács

Abstract:

Positive interaction between people and animals could have a favorable effect on the welfare and production by reducing stress levels. However, to the repeated contact with humans (e.g. farm staff, veterinarians or herdsmen), animals may respond with escape behavior or avoidance, which both have negative effects on the ease of handling, welfare and may lead to the expression of aggressive behaviors. Rough or aversive handling can impair health and the function of the cardiac autonomic activity due to fear and stress, which also can be determined by certain parameters of heart rate variability (HRV). Although the essential relationships between fear from humans and basal tone of the autonomic nervous system were described by the authors previously, several questions remained unclear in terms of the associations between different coping strategies (behavioral syndromes) of the animals and physiological responsiveness to humans. The main goal of this study was to find out whether human behavior and emotions to the animals have an impact on cardiac function and behavior of animals with different coping styles in response situations. Therefore, in the present study, special (fear, approaching, restraint, novel arena, novel object) tests were performed on healthy, 2-year old heifers (n = 104) differing in coping styles [reactive (passive) vs. proactive (active) coping]. Animals were categorized as reactive or proactive based on the following tests: 1) aggressive behavior at the feeding bunk, 2) avoidance from an approaching person, 3) immobility, and 4) daily activity (number of posture changes). Heart rate, the high frequency (HF) component of HRV as a measure of vagal activity and the ratio between the low frequency (LF) and HF components (LF/HF ratio) as a parameter of sympathetic nervous system activity were calculated for all individual during lying posture (baseline) and for response situations in novel object, novel arena, and unfamiliar person tests (both for 5 min), respectively. The differences between baseline and response were compared between groups. Higher sympathetic (higher heart rates and LF/HF ratios) and lower parasympathetic activity (lower HF) was found for proactive animals in response situations than for reactive (passive) animals either during the novel object, the novel arena and the unfamiliar person test. It suggests that animals with different behavioral traits differ in their immediate autonomic adaptation to novelty and people. Based on our preliminary results, it seems, that the analysis of HRV can help to understand the physiological manifestation of responsiveness to novelty and human presence in dairy cattle with different behavioral syndromes.

Keywords: behavioral syndromes, human-cattle interaction, novel arena test, physiological responsiveness, proactive coping, reactive coping

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6439 The Effective of Training Program Using Neuro- Linguistic Programming (NLP) to Reduce the Test Anxiety through the Use of Biological Feedback

Authors: Mohammed Fakehy, Mohammed Haggag

Abstract:

The problem of test anxiety considered as one of the most important and most complex psychological problems faced by students of King Saud University, where university students in a need to bring their reassurance and psychological comfort, relieves feeling pain and difficulties of the study. Recently, there are programs and science that help human to change, including the science Linguistic Programming this neural science stems from not just the tips of the need to make the effort or continue to work, but provides the keys in which one can be controlled in the internal environment. Even human potential energy is extracted seeking to achieve success and happiness and excellence. Through the work of the researchers as members of the teaching staff at King Saud University and specialists in the field of psychology noticed the suffering of some students of King Saud University, test anxiety. In an attempt by the researchers to mitigate as much as possible of the unity of this concern, students will have a training program in Neuro Linguistic Programming. The main Question of this study is What is the effectiveness of the impact of a training program using NLP to reduce test anxiety by using a biological feedback. Therefore, the results of this study might serve as a good announcement about the usefulness of NLP programs which influence future research to significant effect of NLP on test anxiety.

Keywords: neuro linguistic programming, test anxiety, biological feedback, king saud

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6438 Metabolic Profiling of Populus trichocarpa Family 1 UDP-Glycosyltransferases

Authors: Patricia M. B. Saint-Vincent, Anna Furches, Stephanie Galanie, Erica Teixeira Prates, Piet Jones, Nancy Engle, David Kainer, Wellington Muchero, Daniel Jacobson, Timothy J. Tschaplinski

Abstract:

Uridine diphosphate-glycosyltransferases (UGTs) are enzymes that catalyze sugar transfer to a variety of plant metabolites. UGT substrates, which include plant secondary metabolites involved in lignification, demonstrate new activities and incorporation when glycosylated. Knowledge of UGT function, substrate specificity, and enzyme products is important for plant engineering efforts, especially related to increasing plant biomass through lignification. UGTs in Populus trichocarpa, a biofuel feedstock, and model woody plant, were selected from a pool of gene candidates using rapid prioritization strategies. A functional genomics workflow, consisting of a metabolite genome-wide association study (mGWAS), expression of synthetic codon-optimized genes, and high-throughput biochemical assays with mass spectrometry-based analysis, was developed for determining the substrates and products of previously-uncharacterized enzymes. A total of 40 UGTs from P. trichocarpa were profiled, and the biochemical assay results were compared to predicted mGWAS connections. Assay results confirmed seven of 11 leaf mGWAS associations and demonstrated varying levels of substrate specificity among candidate UGTs. P. trichocarpa UGT substrate processing confirms the role of these newly-characterized enzymes in lignan, flavonoid, and phytohormone metabolism, with potential implications for cell wall biosynthesis, nitrogen uptake, and biotic and abiotic stress responses.

Keywords: Populus, metabolite-gene associations, GWAS, bio feedstocks, glycosyltransferase

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6437 Specialised Financial Institutions and its Role in the Promotion of Small and Medium Enterprises in Kerala, India

Authors: K. V. Venugopalan

Abstract:

Micro, Small and Medium Enterprises (MSMEs) have been accepted as the engine of economic growth and for promoting equitable development. The major advantage of the sector is its employment potential at low capital cost. The labour intensity of the MSME sector is much higher than that of the large enterprises. The MSMEs constitute over 90% of total enterprises in most of the economies and are credited with generating the highest rates of employment growth and account for a major share of industrial production and exports. Kerala is a small state in India with the limited land area with high potential in educated human resources need micro, small and medium enterprises for development. Kerala has the highest Physical Quality of Life Index (PQLI) in India and the highest Human Development Index (HDI) at par with the developed countries SME play an important role in alleviating poverty and contribute significantly towards the growth of developing economies. Financial institutions can play a vital role for the promotion of micro, small and medium enterprises in Kerala. The study entitled “Financial Institutions and its role in the promotion of Small and Medium Enterprises in Kerala “examine the progress of MSME in Kerala and India and also the role of financial institutions and the problems faced by entrepreneurs for getting advances with reference to ‘Kerala Financial Corporation’-an agency set up by the government for promoting small and medium enterprises in the state. This study is based on both secondary and primary data. Primary data for the study was collected from those entrepreneurs who availed advances from financial institutions. The secondary data include the investment made, goods and services provided, the employment generated and the number of units registered in MSME sector for the last 10 years in Kerala. The study concluded that financial institutions providing finance with simple procedures and charging smaller interest rates will increase the number of MSME's and also contribute gross state domestic product and reduce the unemployment problem and poverty in the economy.

Keywords: gross state domestic product, human development index, micro, small and medium enterprises

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6436 Measuring Engagement Equation in Educational Institutes

Authors: Mahfoodh Saleh Al Sabbagh, Venkoba Rao

Abstract:

There is plenty of research, both in academic and consultancy circles, about the importance and benefits of employee engagement and customer engagement and how it gives organization an opportunity to reduce variability and improve performance. Customer engagement is directly related to the engagement level of the organization's employees. It is therefore important to measure both. This research drawing from the work of Human Sigma by Fleming and Asplund, attempts to assess engagement level of customer and employees - the human systems of business - in an educational setup. Student is important to an educational institute and is a customer to be served efficiently and effectively. Considering student as customer and faculty as employees serving them, in–depth interviews were conducted to analyze the relationship between faculty and student engagement in two leading colleges in Oman, one from private sector and another from public sector. The study relied mainly on secondary data sources to understand the concept of engagement. However, the search of secondary sources was extensive to compensate the limited primary data. The results indicate that high faculty engagement is likely to lead to high student engagement. Engaged students were excited about learning, loved the feeling of they being cared as a person by their faculty and advocated the organization to other. The interaction truly represents an opportunity to build emotional connection to the organization. This study could be of interest to organizations interest in building and maintaining engagement with employees and customers.

Keywords: customer engagement, consumer psychology, strategy, educational institutes

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6435 Isolation and Identification of Cytotoxic Compounds from Fruticose Lichen Roccella montagnei, and It’s in Silico Docking Study against CDK-10

Authors: Tripti Mishra, Shipra Shukla, Sanjeev Meena, , Ruchi Singh, Mahesh Pal, D. K. Upreti, Dipak Datta

Abstract:

Roccella montagnei belongs to lichen family Roccelleceae growing luxuriantly along the coastal regions of India. As Roccella has been shown to be bioactive, we prepared methanolic extract and assessed its anticancer potential. The methanolic extract showed significant in vitro cytotoxic activity against four human cancer cell lines such as Colon (DLD-1, SW-620), Breast (MCF-7), Head and Neck (FaDu). This prompted us to isolate bioactive compounds through column chromatography. Two compounds Roccellic acid and Everninic acid have been isolated, out of which Everninic acid is reported for the first time. Both the compounds have been tested for in vitro cytotoxic activity in which Roccellic acid showed strong anticancer activity as compared to the Everninic acid. CDK-10 (Cyclin-dependent kinase) contributes to proliferation of cancer cells, and aberrant activity of these kinases has been reported in a wide variety of human cancers. These kinases, therefore, constitute biomarkers of proliferation and attractive pharmacological targets for the development of anticancer therapeutics. Therefore both the isolated compounds were tested for in silico molecular docking study against CDK-10 isomer enzyme to support the cytotoxic activity.

Keywords: cytotoxic activity, everninic acid, roccellic acid, R. montagnei

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6434 Relatively High Heart-Rate Variability Predicts Greater Survival Chances in Patients with Covid-19

Authors: Yori Gidron, Maartje Mol, Norbert Foudraine, Frits Van Osch, Joop Van Den Bergh, Moshe Farchi, Maud Straus

Abstract:

Background: The worldwide pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-COV2), which began in 2019, also known as Covid-19, has infected over 136 million people and tragically took the lives of over 2.9 million people worldwide. Many of the complications and deaths are predicted by the inflammatory “cytokine storm.” One way to progress in the prevention of death is by finding a predictive and protective factor that inhibits inflammation, on the one hand, and which also increases anti-viral immunity on the other hand. The vagal nerve does precisely both actions. This study examined whether vagal nerve activity, indexed by heart-rate variability (HRV), predicts survival in patients with Covid-19. Method: We performed a pseudo-prospective study, where we retroactively obtained ECGs of 271 Covid-19 patients arriving at a large regional hospital in The Netherlands. HRV was indexed by the standard deviation of the intervals between normal heartbeats (SDNN). We examined patients’ survival at 3 weeks and took into account multiple confounders and known prognostic factors (e.g., age, heart disease, diabetes, hypertension). Results: Patients’ mean age was 68 (range: 25-95) and nearly 22% of the patients had died by 3 weeks. Their mean SDNN (17.47msec) was far below the norm (50msec). Importantly, relatively higher HRV significantly predicted a higher chance of survival, after statistically controlling for patients’ age, cardiac diseases, hypertension and diabetes (relative risk, H.R, and 95% confidence interval (95%CI): H.R = 0.49, 95%CI: 0.26 – 0.95, p < 0.05). However, since HRV declines rapidly with age and since age is a profound predictor in Covid-19, we split the sample by median age (40). Subsequently, we found that higher HRV significantly predicted greater survival in patients older than 70 (H.R = 0.35, 95%CI: 0.16 – 0.78, p = 0.01), but HRV did not predict survival in patients below age 70 years (H.R = 1.11, 95%CI: 0.37 – 3.28, p > 0.05). Conclusions: To the best of our knowledge, this is the first study showing that higher vagal nerve activity, as indexed by HRV, is an independent predictor of higher chances for survival in Covid-19. The results are in line with the protective role of the vagal nerve in diseases and extend this to a severe infectious illness. Studies should replicate these findings and then test in controlled trials whether activating the vagus nerve may prevent mortality in Covid-19.

Keywords: Covid-19, heart-rate Variability, prognosis, survival, vagal nerve

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6433 Human 3D Metastatic Melanoma Models for in vitro Evaluation of Targeted Therapy Efficiency

Authors: Delphine Morales, Florian Lombart, Agathe Truchot, Pauline Maire, Pascale Vigneron, Antoine Galmiche, Catherine Lok, Muriel Vayssade

Abstract:

Targeted therapy molecules are used as a first-line treatment for metastatic melanoma with B-Raf mutation. Nevertheless, these molecules can cause side effects to patients and are efficient on 50 to 60 % of them. Indeed, melanoma cell sensitivity to targeted therapy molecules is dependent on tumor microenvironment (cell-cell and cell-extracellular matrix interactions). To better unravel factors modulating cell sensitivity to B-Raf inhibitor, we have developed and compared several melanoma models: from metastatic melanoma cells cultured as monolayer (2D) to a co-culture in a 3D dermal equivalent. Cell response was studied in different melanoma cell lines such as SK-MEL-28 (mutant B-Raf (V600E), sensitive to Vemurafenib), SK-MEL-3 (mutant B-Raf (V600E), resistant to Vemurafenib) and a primary culture of dermal human fibroblasts (HDFn). Assays have initially been performed in a monolayer cell culture (2D), then a second time on a 3D dermal equivalent (dermal human fibroblasts embedded in a collagen gel). All cell lines were treated with Vemurafenib (a B-Raf inhibitor) for 48 hours at various concentrations. Cell sensitivity to treatment was assessed under various aspects: Cell proliferation (cell counting, EdU incorporation, MTS assay), MAPK signaling pathway analysis (Western-Blotting), Apoptosis (TUNEL), Cytokine release (IL-6, IL-1α, HGF, TGF-β, TNF-α) upon Vemurafenib treatment (ELISA) and histology for 3D models. In 2D configuration, the inhibitory effect of Vemurafenib on cell proliferation was confirmed on SK-MEL-28 cells (IC50=0.5 µM), and not on the SK-MEL-3 cell line. No apoptotic signal was detected in SK-MEL-28-treated cells, suggesting a cytostatic effect of the Vemurafenib rather than a cytotoxic one. The inhibition of SK-MEL-28 cell proliferation upon treatment was correlated with a strong expression decrease of phosphorylated proteins involved in the MAPK pathway (ERK, MEK, and AKT/PKB). Vemurafenib (from 5 µM to 10 µM) also slowed down HDFn proliferation, whatever cell culture configuration (monolayer or 3D dermal equivalent). SK-MEL-28 cells cultured in the dermal equivalent were still sensitive to high Vemurafenib concentrations. To better characterize all cell population impacts (melanoma cells, dermal fibroblasts) on Vemurafenib efficacy, cytokine release is being studied in 2D and 3D models. We have successfully developed and validated a relevant 3D model, mimicking cutaneous metastatic melanoma and tumor microenvironment. This 3D melanoma model will become more complex by adding a third cell population, keratinocytes, allowing us to characterize the epidermis influence on the melanoma cell sensitivity to Vemurafenib. In the long run, the establishment of more relevant 3D melanoma models with patients’ cells might be useful for personalized therapy development. The authors would like to thank the Picardie region and the European Regional Development Fund (ERDF) 2014/2020 for the funding of this work and Oise committee of "La ligue contre le cancer".

Keywords: 3D human skin model, melanoma, tissue engineering, vemurafenib efficiency

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6432 An Evolutionary Perspective on the Role of Extrinsic Noise in Filtering Transcript Variability in Small RNA Regulation in Bacteria

Authors: Rinat Arbel-Goren, Joel Stavans

Abstract:

Cell-to-cell variations in transcript or protein abundance, called noise, may give rise to phenotypic variability between isogenic cells, enhancing the probability of survival under stress conditions. These variations may be introduced by post-transcriptional regulatory processes such as non-coding, small RNAs stoichiometric degradation of target transcripts in bacteria. We study the iron homeostasis network in Escherichia coli, in which the RyhB small RNA regulates the expression of various targets as a model system. Using fluorescence reporter genes to detect protein levels and single-molecule fluorescence in situ hybridization to monitor transcripts levels in individual cells, allows us to compare noise at both transcript and protein levels. The experimental results and computer simulations show that extrinsic noise buffers through a feed-forward loop configuration the increase in variability introduced at the transcript level by iron deprivation, illuminating the important role that extrinsic noise plays during stress. Surprisingly, extrinsic noise also decouples of fluctuations of two different targets, in spite of RyhB being a common upstream factor degrading both. Thus, phenotypic variability increases under stress conditions by the decoupling of target fluctuations in the same cell rather than by increasing the noise of each. We also present preliminary results on the adaptation of cells to prolonged iron deprivation in order to shed light on the evolutionary role of post-transcriptional downregulation by small RNAs.

Keywords: cell-to-cell variability, Escherichia coli, noise, single-molecule fluorescence in situ hybridization (smFISH), transcript

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