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Authors: Junaid Jibran Jawed, Prasanta Saini, Subrata Majumdar

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Background: Leishmania donovani infection causes severe host immune-suppression through the modulation of pathogen recognition receptors. Apart from TLRs (Toll Like Receptor), recent studies focus on the important contribution of NLR (NOD-Like Receptor) family member NOD1 and NOD2 as these receptors are capable of triggering host innate immunity. The aim of this study was to decipher the role of NOD1/NOD2 receptors during experimental visceral leishmaniasis (VL) and the important link between host failure and parasite evasion strategy. Method: The status of NOD1 and NOD2 receptors were analysed in uninfected and infected cells through western blotting and RT-PCR. The active contributions of these receptors in reducing parasite burden were confirmed by siRNA mediated silencing, and over-expression studies and the parasite numbers were calculated through microscopic examination of the Giemsa-stained slides. In-vivo studies were done by using non-toxic dose of Mw (Mycobacterium indicus pranii), Ara-LAM(Arabinoasylated lipoarabinomannan) along with MDP (Muramyl dipeptide) administration. Result: Leishmania donovani infection of the macrophages reduced the expression of NOD2 receptors whereas NOD1 remain unaffected. MDP, a NOD2-ligand, treatment during over-expression of NOD2, reduced the parasite burden effectively which was associated with increased pro-inflammatory cytokine generation and NO production. In experimental mouse model, Ara-LAM treatment increased the expression of NOD2 and in combination with MDP it showed active therapeutic potential against VL and found to be more effective than Mw which was already reported to be involved in NOD2 modulation. Conclusion: This work explores the essential contribution of NOD2 during experimental VL and mechanistic understanding of Ara-LAM + MDP combination therapy to work against this disease and highlighted NOD2 as an essential therapeutic target.

Keywords: Ara-LAM (Arabinoacylated Lipoarabinomannan), NOD2 (nucleotide binding oligomerization receptor 2), MDP (muramyl di peptide), visceral Leishmaniasis

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Authors: Nayab Kanwal, Noor Us Saher

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The management and utilization of food resources is becoming a big issue due to rapid urbanization, wastage and non-sustainable use of food, especially in developing countries. Therefore, the use of seafood as alternative sources is strongly promoted worldwide. Marine pollution strongly affects marine organisms, which ultimately decreases their export quality. The monitoring of contamination in marine organisms is a good indicator of the environmental quality as well as seafood quality. Monitoring the accumulation of chemical elements within various tissues of organisms has become a useful tool to survey current or chronic levels of heavy metal exposure within an environment. In this perspective, this study was carried out to compare the previous and current levels (Year 2012 and 2014) of heavy metals (Cd, Pb, Cr, Cu and Zn) in crabs marketed in Karachi and to estimate the toxicological risk associated with their intake. The accumulation of metals in marine organisms, both essential (Cu and Zn) and toxic (Pb, Cd and Cr), natural and anthropogenic, is an actual food safety issue. Significant (p>0.05) variations in metal concentrations were found in all crab species between the two years, with most of the metals showing high accumulation in 2012. For toxicological risk assessment, EWI (Estimated weekly intake), Target Hazard quotient (THQ) and cancer risk (CR) were also assessed and high EWI, Non- cancer risk (THQ < 1) showed that there is no serious threat associated with the consumption of shellfish species on Karachi coast. The Cancer risk showed the highest risk from Cd and Pb pollution if consumed in excess. We summarize key environmental health research on health effects associated with exposure to contaminated seafood. It could be concluded that considering the Pakistan coast, these edible species may be sensitive and vulnerable to the adverse effects of environmental contaminants; more attention should be paid to the Pb and Cd metal bioaccumulation and to toxicological risks to seafood and consumers.

Keywords: cancer risk, edible crabs, heavy metals pollution, risk assessment

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Authors: Raudone Lina, Raudonis Raimondas, Gaivelyte Kristina, Pukalskas Audrius, Janulis Valdimaras, Viskelis Pranas

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Sorbus L. species are widely distributed in the Northern hemisphere and have been used for medicinal purposes in various traditional medicine systems and as food ingredients. Various Sorbus L. raw materials, fruits, leaves, inflorescences, barks, possess diuretic, anti-inflammatory, hypoglycemic, anti-diarrheal and vasoprotective activities. Phenolics, to whom main pharmacological activities are attributed, are compounds of interest due to their notable antioxidant activity. The aim of this study was to determine the antioxidant profiles of fruits and leaves of selected Sorbus L. species (S. anglica, S. aria f. latifolia, S. arranensis, S. aucuparia, S. austriaca, S. caucasica, S. commixta, S. discolor, S. gracilis, S. hostii, S. semi-incisa, S. tianschanica) and to identify the phenolic compounds with potent contribution to antioxidant activity. Twenty two constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers (UPLC–QTOF–MS). Reducing activity of individual constituents was determined using high performance liquid chromatography (HPLC) coupled to post-column FRAP assay. Signicantly greatest trolox equivalent values corresponding up to 45% of contribution to antioxidant activity were assessed for neochlorogenic and chlorogenic acids, which were determined as markers of antioxidant activity in samples of leaves and fruits. Characteristic patterns of antioxidant profiles obtained using HPLC post-column FRAP assay significantly depend on specific Sorbus L. species and raw materials and are suitable for equivalency research of Sorbus L. fruits and leaves. Selecting species and target plant organs with richest phenolic composition and strongly expressed antioxidant power is the first step in further research of standardized extracts.

Keywords: FRAP, antioxidant, phenolic, Sorbus L., chlorogenic acid, neochlorogenic acid

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Authors: Abdelkrim Khadir, Sina Kavalakatt, Preethi Cherian, Ali Tiss

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Soluble epoxide hydrolase (sEH) is an emerging therapeutic target in several chronic states that have inflammation as a common underlying cause such as immunometabolic diseases. Indeed, sEH is known to play a pro-inflammatory role by metabolizing anti-inflammatory, epoxyeicosatrienoic acids (EETs) to pro-inflammatory diols. Recently, it was shown sEH to be linked to diet and microbiota interaction in rat models of obesity. Nevertheless, the functional contribution of sEH and its anti-inflammatory substrates EETs in obesity remain poorly understood. In the current study, we compared the expression pattern of sEH between lean and obese nondiabetic human subjects using subcutaneous adipose tissue (SAT) and peripheral blood mononuclear cells (PBMCs). Using RT-PCR, western blot and immunofluorescence confocal microscopy, we show here that the level of sEH mRNA and protein to be significantly increased in obese subjects with concomitant increase in endoplasmic reticulum (ER) stress components (GRP78 and ATF6α) and inflammatory markers (TNF-α, IL-6) when compared to lean controls. The observation that sEH was overexpressed in obese subjects’ prompt us to investigate whether physical exercise could reduce its expression. In this study, we report here 3-months supervised physical exercise significantly attenuated the expression of sEH in both the SAT and PBMCs, with a parallel decrease in the expression of ER stress markers along with attenuated inflammatory response. On the other hand, homocysteine, a sulfur containing amino acid deriving from the essential amino acid methionine was shown to be directly associated with insulin resistance. When 3T3-L1 preadipocytes cells were treated with homocysteine our results show increased sEH levels along with ER stress markers. Collectively, our data suggest that sEH upregulation is strongly linked to ER stress in adiposity and that physical exercise modulates its expression. This gives further evidence that exercise might be useful as a strategy for managing obesity and preventing its associated complications.

Keywords: obesity, adipose tissue, epoxide hydrolase, ER stress

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Authors: Thomas Weber, Nina Strobel, Thomas Kohne, Eberhard Abele

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In context of the energy transition in Germany, the importance of so-called virtual power plants in the energy supply continues to increase. The progressive dismantling of the large power plants and the ongoing construction of many new decentralized plants result in great potential for optimization through synergies between the individual plants. These potentials can be exploited by mathematical optimization algorithms to calculate the optimal application planning of decentralized power and heat generators and storage systems. This also includes linear or linear mixed integer optimization. In this paper, procedures for reducing the number of decision variables to be calculated are explained and validated. On the one hand, this includes combining n similar installation types into one aggregated unit. This aggregated unit is described by the same constraints and target function terms as a single plant. This reduces the number of decision variables per time step and the complexity of the problem to be solved by a factor of n. The exact operating mode of the individual plants can then be calculated in a second optimization in such a way that the output of the individual plants corresponds to the calculated output of the aggregated unit. Another way to reduce the number of decision variables in an optimization problem is to reduce the number of time steps to be calculated. This is useful if a high temporal resolution is not necessary for all time steps. For example, the volatility or the forecast quality of environmental parameters may justify a high or low temporal resolution of the optimization. Both approaches are examined for the resulting calculation time as well as for optimality. Several optimization models for virtual power plants (combined heat and power plants, heat storage, power storage, gas turbine) with different numbers of plants are used as a reference for the investigation of both processes with regard to calculation duration and optimality.

Keywords: CHP, Energy 4.0, energy storage, MILP, optimization, virtual power plant

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Authors: Theresa Belema Chris-Biriowu

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The change in trends in relation to gender representation in beer brand commercials was the thrust of the study. The study investigated how beer brand commercials reflect societal realities in their portrayals of gender roles within the span of a decade. The major objective of the study was to find out how gender was contextualized in selected beer brand commercials that both air on Nigerian television and stream on YouTube. The study was anchored on the muted group theory. The population of the study was in two streams: the total number of beer beverages that are produced by the eleven breweries in Nigeria and the registered advertising agencies in Lagos, Nigeria. The sample size was also two-pronged: the purposive selection of beer brands that have their commercials on television and YouTube and the purposive selection of an ad agency that has produced running commercials for beer brands within the period between 2010s and 2020s. They adopted visual framing analysis and narrative analysis research techniques. The study qualitatively analyzed the contents of beer brand commercials and conducted an interview with the management of the ad agency for data collection. The data was presented in images and words. The findings showed that females are underrepresented and misrepresented in the beer brand commercials and that the beer brands are not producing commercials that adequately reflect the realities of present times. It was also found that very little has changed in the ad industry between the periods studied, and commercial screenplays are not written with a specific aim to either target the female demographics or give them equal opportunities to thrive in the beer economy. The study concluded that the gender gap in beer commercials subsists and translates to gender discrimination, especially since it is established that females are also stakeholders in the beer economy. The study recommends that beer brands should produce commercials that appeal to their audience irrespective of gender, reflect contemporary realities, and give all genders equal opportunities to thrive in the increasingly competitive industry.

Keywords: beer brands, commercials, gender representation, visuals, television, YouTube

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Authors: Salim H. Al Salmani, Kevin Tansey, Mohammed S. Ozigis

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The study of the growth of built-up areas and settlement expansion is a major exercise that city managers seek to undertake to establish previous and current developmental trends. This is to ensure that there is an equal match of settlement expansion needs to the appropriate levels of services and infrastructure required. This research aims at demonstrating the potential of satellite image processing technique, harnessing the utility of single feature probability-object based image analysis technique in assessing the urban growth dynamics of the Muscat Governorate in Oman for the period 1990, 2002 and 2013. This need is fueled by the continuous expansion of the Muscat Governorate beyond predicted levels of infrastructural provision. Landsat Images of the years 1990, 2002 and 2013 were downloaded and preprocessed to forestall appropriate radiometric and geometric standards. A novel approach of probability filtering of the target feature segment was implemented to derive the spatial extent of the final Built-Up Area of the Muscat governorate for the three years period. This however proved to be a useful technique as high accuracy assessment results of 55%, 70%, and 71% were recorded for the Urban Landcover of 1990, 2002 and 2013 respectively. Furthermore, the Normalized Differential Built – Up Index for the various images were derived and used to consolidate the results of the SFP-OBIA through a linear regression model and visual comparison. The result obtained showed various hotspots where urbanization have sporadically taken place. Specifically, settlement in the districts (Wilayat) of AL-Amarat, Muscat, and Qurayyat experienced tremendous change between 1990 and 2002, while the districts (Wilayat) of AL-Seeb, Bawshar, and Muttrah experienced more sporadic changes between 2002 and 2013.

Keywords: urban growth, single feature probability, object based image analysis, landcover change

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Authors: Anshar Ajatasatru

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The decline of coal prices over past years have been significantly increasing the awareness of effective mining operation. A viable step must be undertaken in becoming more cost competitive while striving for best mining practice especially at Melak Coal Mine in East Kalimantan, Indonesia. This paper aims to show how effective dozer push measurement method can be implemented as it is controlled by contract rate on the unit basis of USD ($) per bcm. The method emerges from an idea of daily dozer push activity that continually shifts the overburden until final target design by mine planning. Volume calculation is then performed by calculating volume of each time overburden is removed within determined distance using cut and fill method from a high precision GNSS system which is applied into dozer as a guidance to ensure the optimum result of overburden removal. Accumulation of daily to weekly dozer push volume is found 95 bcm which is multiplied by average sell rate of $ 0,95, thus the amount monthly revenue is $ 90,25. Furthermore, the payment mechanism is then based on push distance and push grade. The push distance interval will determine the rates that vary from $ 0,9 - $ 2,69 per bcm and are influenced by certain push slope grade from -25% until +25%. The amount payable rates for dozer push operation shall be specifically following currency adjustment and is to be added to the monthly overburden volume claim, therefore, the sell rate of overburden volume per bcm may fluctuate depends on the real time exchange rate of Jakarta Interbank Spot Dollar Rate (JISDOR). The result indicates that dozer push measurement can be one of the surface mining alternative since it has enabled to refine method of work, operating cost and productivity improvement apart from exposing risk of low rented equipment performance. In addition, payment mechanism of contract rate by dozer push operation scheduling will ultimately deliver clients by almost 45% cost reduction in the form of low and consistent cost.

Keywords: contract rate, cut-fill method, dozer push, overburden volume

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Authors: Havva Nur Tarakci, Bahar Urhan Torun

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The transformation that is seen in every area of life by technology, especially internet technology changes the structure of political communications too. Internet, which is at the top of new communication technologies, affects political communications with its structure in a way that no traditional communication tools ever have and enables interaction and the channel between receiver and sender, and it becomes one of the most effective tools preferred among the political communication applications. This state as a result of technological convergence makes Internet an unobtainable place for political communication campaigns. Political communications, which means every kind of communication strategies that political parties called 'actors of political communications' use with the aim of messaging their opinions and party programmes to their present and potential voters who are a target group for them, is a type of communication that is frequently used also among social media tools at the present day. The electorate consisting of different structures is informed, directed, and managed by social media tools. Political parties easily reach their electorate by these tools without any limitations of both time and place and also are able to take the opinions and reactions of their electorate by the element of interaction that is a feature of social media. In this context, Facebook, which is a place that political parties use in social media at most, is a communication network including in our daily life since 2004. As it is one of the most popular social networks today, it is among the most-visited websites in the global scale. In this way, the research is based on the question, “How do the political parties use Facebook at the campaigns, which they conduct during the election periods, for informing their voters?” and it aims at clarifying the Facebook using practices of the political parties. In direction of this objective the official Facebook accounts of the four political parties (JDP–AKParti, PDP–BDP, RPP-CHP, NMP-MHP), which reach their voters by social media besides other communication tools, are treated, and a frame for the politics of Turkey is formed. The time of examination is constricted with totally two weeks, one week before the mayoral elections and one week after the mayoral elections, when it is supposed that the political parties use their Facebook accounts in full swing. As a research method, the method of content analysis is preferred, and the texts and the visual elements that are gotten are interpreted based on this analysis.

Keywords: Facebook, political communications, social media, electrorate

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Authors: Singh Divya, Rohan Singh, Anjana Pandey

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Reviewers' Comments: The study provides valuable insights into the anti-pneumonia properties of flavonoids against LytB. Authors could further validate findings through in vitro studies and consider exploring combination therapies for enhanced efficacy Response: Thankyou for your valuable comments. This study has been conducted further via experimental validation of the in-silico findings. The study uses Streptococcus pneumoniae D39 strain and examine the anti-pneumonia effect of kaempferol, quercetin and apigenin at various concentrations ranging from 9ug/ml to 200ug/ml. From results, it can be concluded that the kaempferol has shown the highest cytotoxic effect (72.1% of inhibition) against S. pneumoniae at concentration of 40ug/ml compare to apigenin and quercetin. The treatment of S. pneumoniae with concoction of kaempferol, quercetin and apigenin has also been performed, it is noted that conc. of 200ug/ml was most effect in achieving 75% inhibition. As S. pneumoniae D39 is a virulent encapsulated strain, the capsule interferes with the uptake of large size drug formulation. For instance, S. pneumoniae D39 with kaempferol and gold nano urchin (GNU) formulation, but the large size of GNU has resulted in reduced cytotoxic effect of kaempferol (27%). To achieve near 100% cytotoxic effect on the MDR S. pneumoniae D39 strain, the study will target the development of kaempferol-engineered gold nano-urchin’ conjugates, where gold nanocrystal will be of small size (less than or equal to 5nm) and decorated with hydroxyl, sulfhydryl, carboxyl, amine and groups. This approach is expected to enhance the anti-pneumonia effect of kaempferol (polyhydroxylated flavonoid). The study will also examine the interactive study among lung epithelial cell line (A549), kaempferol-engineered gold nano urchins, and S. pneumoniae for exploring the colonization, invasion, and biofilm formation of S. pneumoniae on A549 cells resembling the upper respiratory surface of humans.

Keywords: streptococcus pneumoniae, β-endo-Acetylglucosaminidase, apigenin, quercetin kaempferol, molecular dynamic simulation, interactome study and GROMACS

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Authors: Jodie Bradshaw, Rebecca Dooley, Habib Kadiri, Holly Bird, Aaliyah Felix-West, Udit Mahalingam, Ella Thomson

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The sharp rise of civil orders has led to an expansion of police powers, particularly in the realm of stop and search activities. The broad scope and objectives of these civil orders –addressing issues as varied as public safety, crime prevention, and counter-terrorism – has led to ‘mission-creep’, whereby orders were being imposed in a wider range of contexts than initially intended. The ever-widening purview of civil orders in practice necessitates proactive measures by law enforcement which often rely heavily on the utilisation of stop and search, leading to an expansion of stop and search practices and the regulation of public space. Civil liberties organisations, criminal justice and legal practitioners, activist groups, and researchers have argued that civil orders dilute and undermine foundational legal principles, pose a threat to our basic rights and freedoms, facilitate dangerous criminal justice net-widening, and disproportionately target young, working-class people of colour. Many of the provisions in these orders are potentially incompatible with the right to liberty and security. The conditions of an order (whether negative restrictions or positive obligations) tend to be extremely easy to breach –and in some cases, almost impossible for the person subject to the order not to breach. When the conditions of an order are breached, the result is criminal punishment – often in the form of imprisonment. This paper argues that civil orders set people up to fail, sending them down a path towards incarceration and the ultimate deprivation of liberty. The proclaimed intentions underpinning these civil orders – to tackle purportedly ‘undesirable’ behaviour (which in and of itself is not a crime) committed by ‘undesirable’ people – paves the way for justifying violent and racially disproportionate policing practices.

Keywords: civil orders, policing, stop and search, crime, civil liberties, criminal punishment, anti-social behaviour

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Authors: Ahmed Stifi, Sascha Gentes

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The contract in its simplest definition is an agreement involving parties with a number of documents which may be as little as a marriage contract involving two parties or as big as a contract of construction and operation of a nuclear power plant involving companies and stakeholders with hundreds or even thousands of documents. All parties in the construction industry, not only the contract experts, agree that the success of a project is linked primarily to the form of contract regulating the relationship between stakeholders of the project. Therefore it is essential for the construction industry to study, analyze and improve its contracts forms continuously. However, it should be mentioned that different contract forms are developed to suit the construction evolution in term of its machinery, materials and construction process. There exist some similarities in some clauses and variations in many of these forms depending upon the type of project, the kind of clients and more importantly the laws and regulations governing the transaction in the country where the project is carried out. This paper will discuss the most important forms of construction contracts starting from national level, intended to the contract form in Germany and moving on to the international level introducing FIDIC contracts and its different forms, some newly developed contracts forms namely the integrated form of agreement, the new engineering contract and the project alliance agreement. The result of the study shows that many of the contract’s paragraphs are similar and the main difference comes in the approach of the relationship between the parties. Is it based on co-operation and mutual trust, or in some cases a load of responsibility for a particular party which increases the problems and disputes that affects the success of the project negatively. Thus we can say that the form of the contract, that plays an essential role in the approach of the project management, which is ultimately the key factor for the success of the project. So we advise to use a form of contract, which enhance the mutual trust between the project parties, contribute to support the cooperation between them, distribute responsibility and risks on an equitable basis and build on the principle “win-win". In additional to the conventional role of the contract it should integrate all parties into one team to achieve the target value of the project.

Keywords: contract, FIDIC, integrated form of agreement, new engineering contract, project alliance agreemen

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Authors: Petra Borilova Linhartova, Michaela Ruckova, Sabina Sevcikova, Natalie Mlcuchova, Jan Bohm, Katerina Zukalova, Monika Vlachova, Jiri Dolina, Lumir Kunovsky, Radek Kroupa, Zdenek Pavlovsky, Zdenek Danek, Tereza Deissova, Lydie Izakovicova Holla, Ondrej Slaby, Zdenek Kala

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Esophageal adenocarcinoma (EAC) is a highly aggressive malignancy that frequently develops from Barrett's esophagus (BE), a premalignant pathologic change occurring in the lower end of the esophagus. Specific microRNAs (miRNAs), small non-coding RNAs that function as posttranscriptional regulators of gene expression, were repeatedly proved to play key roles in the pathogenesis of these diseases. This pilot study aimed to analyze four selected miRNAs in esophageal tissues from healthy controls (HC) and patients with reflux esophagitis (RE)/BE/EAC, as well as to compare expression at the site of Barrett's mucosa/adenocarcinoma and healthy esophageal tissue outside the area of the main pathology in patients with BE/EAC. In this pilot study, 22 individuals (3 HC, 8 RE, 5 BE, 6 EAC) were included and endoscopically examined. RNA was isolated from the fresh-frozen esophageal tissue (stored in the RNAlater™ Stabilization Solution −70°C) using the AllPrep DNA/RNA/miRNA Universal Kit. Subsequent RT-qPCR analysis was performed using selected TaqMan MicroRNA Assays for miR-21, miR-34a, miR-196a, miR-196b, and endogenous control (RNU44). While the expression of miR-21 in the esophageal tissue with the main pathology was decreased in BE and EAC patients in comparison to the group of HC and RE patients (p=0.01), the expression of miR-196a was increased in the BE and EAC patients (p<0.01). Correlations between those miRNAs expression in tissue and severity of diagnosis were observed (p<0.05). In addition, miR-196a was significantly more expressed at the site with the main pathology than in paired adjacent esophageal tissue in BE and EAC patients (p<0.01). In conclusion, our pilot results showed that miR-196a, which regulates the proliferation, invasion, and migration (and was previously associated with esophageal squamous cell carcinoma and marked as a potential therapeutic target), could be a diagnostic tissue biomarker for BE and EAC as well.

Keywords: microRNA, barrett´s esophagus, esophageal adenocarcinoma, biomarker

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Authors: Hwang Ahreum, Kang Taejoon, Kim Bongsoo

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Nanosensors for high sensitive detection of diseases have been widely studied to improve the quality of life. Here, we suggest robust nano-plasmonic diagnostic sensor using cysteine tagged protein G (Cys3-protein G) and ultraflat, ultraclean and single-crystalline Au nanoplates. Protein G formed on an ultraflat Au surface provides ideal background for dense and uniform immobilization of antibodies. The Au is highly stable in diverse biochemical environment and can immobilize antibodies easily through Au-S bonding, having been widely used for various biosensing applications. Especially, atomically smooth single-crystalline Au nanomaterials synthesized using chemical vapor transport (CVT) method are very suitable to fabricate reproducible sensitive sensors. As the C-reactive protein (CRP) is a nonspecific biomarker of inflammation and infection, it can be used as a predictive or prognostic marker for various cardiovascular diseases. Cys3-protein G immobilized uniformly on the Au nanoplate enable CRP antibody (anti-CRP) to be ordered in a correct orientation, making their binding capacity be maximized for CRP detection. Immobilization condition for the Cys3-protein G and anti-CRP on the Au nanoplate is optimized visually by AFM analysis. Au nanoparticle - Au nanoplate (NPs-on-Au nanoplate) assembly fabricated from sandwich immunoassay for CRP can reduce zero-signal extremely caused by nonspecific bindings, providing a distinct surface-enhanced Raman scattering (SERS) enhancement still in 10-18 M of CRP concentration. Moreover, the NP-on-Au nanoplate sensor shows an excellent selectivity against non-target proteins with high concentration. In addition, comparing with control experiments employing a Au film fabricated by e-beam assisted deposition and linker molecule, we validate clearly contribution of the Au nanoplate for the attomolar sensitive detection of CRP. We expect that the devised platform employing the complex of single-crystalline Au nanoplates and Cys3-protein G can be applied for detection of many other cancer biomarkers.

Keywords: Au nanoplate, biomarker, diagnostic sensor, protein G, SERS

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Authors: Noor Abdelhamid, Donovan Nelson, Cara Prosser

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The architectural design process could be mapped out as a dialogue between designer and user that is constructed across multiple phases with the overarching goal of aligning design outcomes with user needs. Traditionally, this dialogue is bounded within a preliminary phase of determining factors that will direct the design intent, and a completion phase, of handing off the project to the client. Pre- and post-occupancy evaluations (P/POE’s) could provide an alternative process by extending this dialogue on both ends of the design process. The purpose of this research is to study the influence of systematic pre-occupancy data collection in achieving design goals by conducting post-occupancy evaluations of two case studies. In the context of this study, systematic pre-occupancy data collection is defined as the preliminary documentation of the existing conditions that helps portray stakeholders’ needs. When implemented, pre-occupancy occurs during the early phases of the architectural design process, utilizing the information to shape the design intent. Investigative POE’s are performed on two case studies with distinct early design approaches to understand how the current space is impacting user needs, establish design outcomes, and inform future strategies. The first case study underwent systematic pre-occupancy data collection and synthesis, while the other represents the traditional, uncoordinated practice of informally collecting data during an early design phase. POE’s target the dynamics between the building and its occupants by studying how spaces are serving the needs of the users. Data collection for this study consists of user surveys, audiovisual materials, and observations during regular site visits. Mixed methods of qualitative and quantitative analyses are synthesized to identify patterns in the data. The paper concludes by positioning value on both sides of the architectural design process: the integration of systematic pre-occupancy methods in the early phases and the reinforcement of a continued dialogue between building and design team after building completion.

Keywords: architecture, design process, pre-occupancy data, post-occupancy evaluation

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Authors: Tamunotonye Mayowa Braide, Isaac Oluwatayo

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This study investigates multidimensional poverty, and its correlates among rural households in Sekhukhune and Capricorn District municipalities (SDM & CDM) in Limpopo Province, South Africa. Primary data were collected from 407 rural households selected through purposive and simple random sampling techniques. Analytical techniques employed include descriptive statistics, principal component analysis (PCA), and the Alkire Foster (A-F) methodology. The results of the descriptive statistics showed there are more females (66%) than males (34%) in rural areas of Limpopo Province, with about 45% of them having secondary school education as the highest educational level attained and only about 3% do not have formal education. In the analysis of deprivation, eight dimensions of deprivation, constructed from 21 variables, were identified using the PCA. These dimensions include type and condition of dwelling water and sanitation, educational attainment and income, type of fuel for cooking and heating, access to clothing and cell phone, assets and fuel for light, health condition, crowding, and child health. In identifying the poor with poverty cut-off (0.13) of all indicators, about 75.9% of the rural households are deprived in 25% of the total dimensions, with the adjusted headcount ratio (M0) being 0.19. Multidimensional poverty estimates showed higher estimates of poor rural households with 71%, compared to 29%, which fall below the income poverty line. The study conducted poverty decomposition, using sub-groups within the area by examining regions and household characteristics. In SDM, there are more multidimensionally poor households than in CDM. The water and sanitation dimension is the largest contributor to the multidimensional poverty index (MPI) in rural areas of Limpopo Province. The findings can, therefore, assist in better design of welfare policy and target poverty alleviation programs and as well help in efficient resource allocation at the provincial and local municipality levels.

Keywords: Alkire-Foster methodology, Limpopo province, multidimensional poverty, principal component analysis, South Africa

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Authors: Mohit Wadhawan, Sushma Rathaur

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Lymphatic filariasis, also called elephantiasis is a tropical disease afflicting over 120 million people in 81 countries worldwide. Existing anti filarial drugs are effective against the larval stages of filarial parasites which call for an urgent need of drugs which are macrofilaricidal. Identification of molecular targets crucial for survival of filarial parasites is a prerequisite for drug designing. Prolyl oligopeptidase (POP) is one such crucial enzyme involved in the maturation and degradation of neuropeptides and peptide hormones. We have identified this peptidase in the bovine filarial parasite, Setaria cervi. Effect of inhibition of POP on the proteome profile of filarial parasite has been discussed in this study. Filarial parasites were exposed to Z-pro-prolinal (ZPP), a specific POP inhibitor for 8 h and the motility and viability of the parasites was observed. It significantly reduced the motility and viability of the parasites. To study the proteome profile, the cytosolic, endoplasmic reticulum (ER) and mitochondrial extracts of the adult female parasites were subjected to 2-dimensional electrophoresis. As analyzed by the PD-Quest software, the ZPP caused the alteration in the different subcellular proteins, and the significantly altered proteins were identified using MALDI-MS/MS spectrometry. The major proteins identified were found to play important role in diverse biological functions like signaling, redox regulation, energy metabolism, stress response, and cytoskeleton formation. Moreover, we found upregulation in the calcium binding proteins such as calreticulin, calponin, and calpain-6 suggesting that POP inhibition regulates calcium release. This relates to earlier reports that POP plays non-catalytic role in inositol 1,4,5-trisphosphate (IP3) signaling inducing release of calcium from ER. Taken together, the data demonstrated that inhibition of prolyl oligopeptidase alter the overall proteome signifying its role in survival of the filarial parasites. Thus this study provides a basis for the use of POP as a chemotherapeutic target for the treatment of lymphatic filariasis.

Keywords: lymphatic filariasis, setaria cervi, prolyl oligopeptidase, proteomics

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Authors: Farheen Tabassum, Shoab Ahmed Khan

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The brutality of attacks on networks and decisive infrastructures are on the climb over recent years and appears to continue to do so. Distributed Denial of service attack is the most prevalent and easy attack on the availability of a service due to the easy availability of large botnet computers at cheap price and the general lack of protection against these attacks. Application layer DDoS attack is DDoS attack that is targeted on wed server, application server or database server. These types of attacks are much more sophisticated and challenging as they get around most conventional network security devices because attack traffic often impersonate normal traffic and cannot be recognized by network layer anomalies. Conventional techniques of single-hosted security systems are becoming gradually less effective in the face of such complicated and synchronized multi-front attacks. In order to protect from such attacks and intrusion, corporation among all network devices is essential. To overcome this issue, a collaborative intrusion detection system (CIDS) is proposed in which multiple network devices share valuable information to identify attacks, as a single device might not be capable to sense any malevolent action on its own. So it helps us to take decision after analyzing the information collected from different sources. This novel attack detection technique helps to detect seemingly benign packets that target the availability of the critical infrastructure, and the proposed solution methodology shall enable the incident response teams to detect and react to DDoS attacks at the earliest stage to ensure that the uptime of the service remain unaffected. Experimental evaluation shows that the proposed collaborative detection approach is much more effective and efficient than the previous approaches.

Keywords: Distributed Denial-of-Service (DDoS), Collaborative Intrusion Detection System (CIDS), Slowloris, OSSIM (Open Source Security Information Management tool), OSSEC HIDS

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Authors: Denise Schrama, Claudia Raposo, Pedro Rodrigues

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Background: Food allergies are conducted by a hypersensitive response of the immune system. These allergies are a global concern for the public health. Consumption of fish is increasing worldwide as it is a healthy meat with high nutritional value. Unfortunately, fish can cause adverse immune-mediate reactions, affecting part of the population with higher incidence in children. β-parvalbumin, a small, highly conserved stable, calcium or magnesium binding muscle protein is the main fish allergen. In fish-allergic patients, cross-reactivity between different fish species exist due to recognition of highly identical protein regions. Enolases, aldolases, or fish gelatin are other identified fish allergens in some fish species. With no available cure for fish allergies, clinical management is only based on an avoidance diet aiming at the total exclusion of offending food. Methods: Mediterranean fish (S. aurata and D. labrax) were fed specifically designed diets, enriched in components that target the expression or inactivation of parvalbumin (creatine and EDTA, respectively). After 90 days fish were sampled and biological tissues were excised. Proteomics was used to access fish allergens characterization and expression in muscle while IgE assays to confirm the lower allergenic potential are conducted in patients with history of fish allergies. Fish welfare and quality of flesh were established with biochemical, texture and sensorial analysis. Results: Fish welfare shows no major impact between diets. In case of creatine supplementation in D. labrax proteomic analysis show a slight decrease in parvalbumin expression. No accumulation of this compound was found in muscle. For EDTA supplementation in S. aurata IgE assay show a slight decrease in allergenicity when using sera of fish allergic patients. Conclusion: Supplementation with these two compounds seems to change slightly the allergenicity of the two mean Mediterranean species.

Keywords: fish allergies, fish nutrition, proteomics, aquaculture

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Authors: Saraswati Suna

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The outbreak of the COVID-19 epidemic and the subsequent lockdown have significantly impacted India's political, structural, and economic systems and a rising gap between the rich and the disadvantaged, upper and lower caste. For Dalit women, such forms of subjugation were followed by socioeconomic uncertainty due to the pandemic's economic shutdown and labour oppressions. Dalit women have been the victims of the most oppression among the nation's underprivileged groups. Dalit women undergo systemic oppression at the hands of the state, caste, class, gender, and religious hegemons historically. Dalit women hold a subordinate position within the gender to their male counterparts and caste to their upper-caste counterparts. This paper examines how Brahminical patriarchy and state power severely affected Dalit/Adivasi women during COVID-19 in Odisha, India. In order to understand caste-based sexual violence, a total of five cases have been analysed from newspapers. Findings revealed that Covid-19 appears to have a significant physical, psychological, and economic impact on Dalit women. The intention of sexual harassment and rape perpetrated by upper caste men is to maintain power and patriarchal culture in society. Dalit women are economically, socially, and culturally marginalised, which effectively exacerbates the sense of impunity by perpetrators of violence against Dalit women. This issue requires special attention to end atrocities against Dalit women. Dalit women become the target of rape, sexual assault, and murder. Sexual violence against Dalit women cannot be fully explained without linkage to caste, gender, and power. Dominant caste comes through caste privilege-socio-economic and politically; these factors contribute to sexual violence against Dalit women. The findings revealed that state police manipulate sexual violence, and in so doing, they create and deny access to both services to get justice. This article has argued that understanding Brahminical culture and the legal impacts of state police on Dalit women's identity requires a nuanced analysis.

Keywords: COVID-19, dalit women, sexual violence, brahminical patriarchy, power

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Authors: Małgorzata Tomczyńska, Joanna Saluk-Bijak

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Background: Graves’ disease is a frequent organ-specific autoimmune thyroid disease, which characterized by the presence of different kind autoantibodies, that, in most cases, act as agonists of the thyrotropin receptor, leading to hyperthyroidism. Role of platelets and lymphocytes can be modulated in the pathophysiology of thyroid autoimmune diseases. Interference in the physiology of platelets can lead to enhanced activity of these cells. Activated platelets can bind to circulating lymphocytes and to affect lymphocyte adhesion. Platelets and lymphocytes can regulate mutual functions. Therefore, the activation of T lymphocytes, as well as blood platelets, is associated with the development of inflammation and oxidative stress within the target tissue. The present study was performed to investigate a platelet-lymphocyte relation by assessing the degree of their mutual aggregation in whole blood of patients with Graves’ disease. Also, the purpose of this study was to examine the impact of platelet interaction on lymphocyte migration capacity. Methods: 30 patients with Graves’ disease were recruited in the study. The matched 30 healthy subjects were served as the control group. Immunophenotyping of lymphocytes was carried out by flow cytometry method. A CytoSelect™ Cell Migration Assay Kit was used to evaluate lymphocyte migration and adhesion to blood platelets. Visual assessment of lymphocyte-platelet aggregate morphology was done using confocal microscope after magnetic cell isolation by Miltenyi Biotec. Results: The migration and functional responses of lymphocytes to blood platelets were greater in the group of Graves’ disease patients compared with healthy controls. The group of Graves’ disease patients exhibited a reduced T lymphocyte and a higher B cell count compared with controls. Based on microscopic analysis, more platelet-lymphocyte aggregates were found in patients than in control. Conclusions: Studies have shown that in Graves' disease, lymphocytes show increased platelet affinity, more strongly migrating toward them, and forming mutual cellular conglomerates. This may be due to the increased activation of blood platelets in this disease.

Keywords: blood platelets, cell migration, Graves’ disease, lymphocytes, lymphocyte-platelet aggregates

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Authors: Alexandra P. Alexandropoulou, Andreas Fousteris, Eleni Didaskalou, Dimitrios A. Georgakellos

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As urbanization is becoming a major stress factor not only for the urban environment but also for the wellbeing of city dwellers, incorporating the issues of urban health in strategic city planning and policy-making has never been more relevant. The impact of urbanization can vary from low to severe and relates to all non-communicable diseases caused by the different functions of cities. Air pollution, noise pollution, water and soil pollution, availability of open green spaces, and urban heat island are the major factors that can compromise citizens' health. Urban health describes the effects of the social environment, the physical environment, and the availability and accessibility to health and social services. To assess the quality of urban wellbeing, all urban characteristics that might have an effect on citizens' health must be considered, evaluated, and introduced in integrated local planning. A series of indices and indicators can be used to better describe these effects and set the target values in policy making. Local strategic planning is one of the most valuable development tools a local city administration can possess; thus, it has become mandatory under Greek law for all municipalities. It involves a two-stage procedure; the first aims to collect, analyse and evaluate data on the current situation of the city (administrative data, population data, environmental data, social data, swot analysis), while the second aims to introduce a policy vision described and supported by distinct (nevertheless integrated) actions, plans and measures to be implemented with the aim of city development and citizen wellbeing. In this procedure, the element of health is often neglected or under-evaluated. A relative survey was conducted among all Greek local authorities in order to shed light on the current situation. Evidence shows that the rate of incorporation of health in strategic planning is lacking behind. The survey also highlights key hindrances and concerns raised by local officials and suggests a path for the way forward.

Keywords: urban health, strategic planning, local authorities, integrated development

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Authors: Armando A. Rodríguez, Emilio Salceda, Anoland Garateix, André J. Zaharenko, Steve Peigneur, Omar López, Tirso Pons, Michael Richardson, Maylín Díaz, Yasnay Hernández, Ludger Ständker, Jan Tytgat, Enrique Soto

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Acid-sensing ion channels are cation (Na+) channels activated by a pH drop. These proteins belong to the ENaC/degenerin superfamily of sodium channels. ASICs are involved in sensory perception, synaptic plasticity, learning, memory formation, cell migration and proliferation, nociception, and neurodegenerative disorders, among other processes; therefore those molecules that specifically target these channels are of growing pharmacological and biomedical interest. Sea anemones produce a large variety of ion channels peptide toxins; however, those acting on ligand-gated ion channels, such as Glu-gated, Ach-gated ion channels, and acid-sensing ion channels (ASICs), remain barely explored. The peptide PhcrTx1 is the first compound characterized from the sea anemone Phymanthus crucifer, and it constitutes a novel ASIC inhibitor. This peptide was purified by chromatographic techniques and pharmacologically characterized on acid-sensing ion channels of mammalian neurons using patch-clamp techniques. PhcrTx1 inhibited ASIC currents with an IC50 of 100 nM. Edman degradation yielded a sequence of 32 amino acids residues, with a molecular mass of 3477 Da by MALDI-TOF. No similarity to known sea anemone peptides was found in protein databases. The computational analysis of Cys-pattern and secondary structure arrangement suggested that this is a structurally ICK (Inhibitor Cystine Knot)-type peptide, a scaffold that had not been found in sea anemones but in other venomous organisms. These results show that PhcrTx1 represents the first member of a new structural group of sea anemones toxins acting on ASICs. Also, this peptide constitutes a novel template for the development of drugs against pathologies related to ASICs function.

Keywords: animal toxin, inhibitor cystine knot, ion channel, sea anemone

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Authors: Gezahegn Gizaw

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The FDRE Tourism Training Institute organizes forums for debates, best practices exchange and focus group discussions to forge a sustainable and growing tourism sector while minimizing negative impacts on the environment, communities, and cultures. This study aimed at applying empirical research method to identify and quantify relative importance of success factors and individual engagement indicators that were identified in these forums. Response to the 12-question survey was collected from a total of 437 respondents in academic training institutes (212), business executive and employee (204) and non-academic government offices (21). Overall, capacity building was perceived as the most important driver of success for stakeholder engagement. Business executive and employee category rated capacity building as the most important driver of success (53%), followed by decision-making process (27%) and community participation (20%). Among educators and students, both capacity building and decision-making process were perceived as the most important factors (40% of respondents), whereas community participation was perceived as the most important success factor only by 20% of respondents. Individual engagement score in capacity building, decision-making process and community participation showed highest variability by educational level of participants (variance of 3.4% - 5.2%, p<0.001). Individual engagement score in capacity building was highly correlated to perceived benefit of training on improved efficiency, job security, higher customer satisfaction and self-esteem. On the other hand, individual engagement score in decision making process was highly correlated to its perceived benefit on lowering business costs, improving ability to meet the needs of a target market, job security, self-esteem and more teamwork. The study provides a set of recommendations that help educators, business executives and policy makers to maximize the individual and synergetic effect of training, decision making process on sustainability and growth of the tourism sector in Ethiopia.

Keywords: engagement score, driver of success, capacity building, tourism

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Authors: Merfat Algethami, Anton Blencowe, Bryce Feltis, Stephen Best, Moshi Geso

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Nano-materials with high atomic number atoms have been demonstrated to enhance the effective radiation dose and thus potentially could improve therapeutic efficacy in radiotherapy. The optimal nanoparticulate agents require high X-ray absorption coefficients, low toxicity, and should be cost effective. The focus of our research is the development of a nanoparticle therapeutic agent that can be used in radiotherapy to provide optimal enhancement of the radiation effects on the target. In this study, we used bismuth (Bi) nanoparticles coated with starch and bismuth sulphide nanoparticles (Bi2S3) coated with polyvinylpyrrolidone (PVP). These NPs are of low toxicity and are one of the least expensive heavy metal-based nanoparticles. The aims of this study were to synthesise Bi2S3 and Bi NPs, and examine their cytotoxicity to human lung adenocarcinoma epithelial cells (A549). The dose enhancing effects of NPs on A549 cells were examined at both KV and MV energies. The preliminary results revealed that bismuth based nanoparticles show increased radio-sensitisation of cells, displaying dose enhancement with KV X-ray energies and to a lesser degree for the MV energies. We also observed that Bi NPs generated a greater dose enhancement effect than Bi2S3 NPs in irradiated A549 cells. The maximum Dose Enhancement Factor (DEF) was obtained at lower energy KV range when cells treated with Bi NPs (1.5) compared to the DEF of 1.2 when cells treated with Bi2S3NPs. Less radiation dose enhancement was observed when using high energy MV beam with higher DEF value of Bi NPs treatment (1.26) as compared to 1.06 DEF value with Bi2S3 NPs. The greater dose enhancement was achieved at KV energy range, due the effect of the photoelectric effect which is the dominant process of interaction of X-ray. The cytotoxic effect of Bi NPs on enhancing the X-ray dose was higher due to the higher amount of elemental Bismuth present in Bi NPs compared to Bi2S3 NPs. The results suggest that Bismuth based NPs can be considered as valuable dose enhancing agents when used in clinical applications.

Keywords: A549 lung cancer cells, Bi2S3 nanoparticles, dose enhancement effect, radio-sensitising agents

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Authors: Abbas Ali Husseini, Ali Mohammad Yazdani, Fatemeh Ghadiri, Alper Şişman

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We developed a microchip device that uses surface acoustic waves for rapid lysis of low level of cell samples. The device incorporates sharp-edge glass microparticles for improved performance. We optimized the lysis conditions for high efficiency and evaluated the device's feasibility for point-of-care applications. The microchip contains a 13-finger pair interdigital transducer with a 30-degree focused angle. It generates high-intensity acoustic beams that converge 6 mm away. The microchip operates at a frequency of 16 MHz, exciting Rayleigh waves with a 250 µm wavelength on the LiNbO3 substrate. Cell lysis occurs when Candida albicans cells and glass particles are placed within the focal area. The high-intensity surface acoustic waves induce centrifugal forces on the cells and glass particles, resulting in cell lysis through lateral forces from the sharp-edge glass particles. We conducted 42 pilot cell lysis experiments to optimize the surface acoustic wave-induced streaming. We varied electrical power, droplet volume, glass particle size, concentration, and lysis time. A regression machine-learning model determined the impact of each parameter on lysis efficiency. Based on these findings, we predicted optimal conditions: electrical signal of 2.5 W, sample volume of 20 µl, glass particle size below 10 µm, concentration of 0.2 µg, and a 5-minute lysis period. Downstream analysis successfully amplified a DNA target fragment directly from the lysate. The study presents an efficient microchip-based cell lysis method employing acoustic streaming and microparticle collisions within microdroplets. Integration of a surface acoustic wave-based lysis chip with an isothermal amplification method enables swift point-of-care applications.

Keywords: cell lysis, surface acoustic wave, micro-glass particle, droplet

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Authors: Davod Ahmadi, Narges Ebadi, Ethan Wang, Hugo Melgar-Quiñonez

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Compared to the other Least Developed Countries (LDCs), major countries in sub-Saharan Africa (SSA) have limited access to the clean water. People in this region, and more specifically females, suffer from acute water scarcity problems. They are compelled to spend too much of their time bringing water for domestic use like drinking and washing. Apart from domestic use, water through affecting agriculture and livestock contributes to the food security status of people in vulnerable regions like SSA. Livestock needs water to grow, and agriculture requires enormous quantities of water for irrigation. The main objective of this study is to explore the association between access to water and individuals’ food security status. Data from 2017 Gallup World Poll (GWP) for SSA were analyzed (n=35,000). The target population in GWP is the entire civilian, non-institutionalized, aged 15 and older population. All samples selection is probability based and nationally representative. The Gallup surveys an average of 1,000 samples of individuals per country. Three questions related to water (i.e., water quality, availability of water for crops and availability of water for livestock) were used as the exposure variables. Food Insecurity Experience Scale (FIES) was used as the outcome variable. FIES measures individuals’ food security status, and it is composed of eight questions with simple dichotomous responses (1=Yes and 0=No). Different statistical analyses such as descriptive, crosstabs and binary logistic regression, form the basis of this study. Results from descriptive analyses showed that more than 50% of the respondents had no access to enough water for crops and livestock. More than 85% of respondents were categorized as “food insecure”. Findings from cross-tabulation analyses showed that food security status was significantly associated with water quality (0.135; P=0.000), water for crops (0.106; P=0.000) and water for livestock (0.112; P=0.000). In regression analyses, the probability of being food insecure increased among people who expressed no satisfaction with water quality (OR=1.884 (OR=1.768-2.008)), not enough water for crops (OR=1.721 (1.616-1.834)) and not enough water for livestock (OR=1.706 (1.819)). In conclusion, it should note that water access affects food security status in SSA.

Keywords: water access, agriculture, livestock, FIES

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Authors: R. Fardid, R. Coppes

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Introduction: In mediastinal radiotherapy and to a lesser extend also in total-body irradiation (TBI) radiation exposure may lead to development of cardiac diseases. Radiation-induced heart disease is dose-dependent and it is characterized by a loss of cardiac function, associated with progressive heart cells degeneration. We aimed to determine the in-vivo radiation effects on fibronectin, ColaA1, ColaA2, galectin and TGFb1 gene expression levels in left ventricle heart tissues of rats after irradiation. Material and method: Four non-treatment adult Wistar rats as control group (group A) were selected. In group B, 4 adult Wistar rats irradiated to 20 Gy single dose of 150 Mev proton beam locally in heart only. In heart plus lung irradiate group (group C) 4 adult rats was irradiated by 50% of lung laterally plus heart radiation that mentioned in before group. At 8 weeks after radiation animals sacrificed and left ventricle heart dropped in liquid nitrogen for RNA extraction by Absolutely RNA® Miniprep Kit (Stratagen, Cat no. 400800). cDNA was synthesized using M-MLV reverse transcriptase (Life Technologies, Cat no. 28025-013). We used Bio-Rad machine (Bio Rad iQ5 Real Time PCR) for QPCR testing by relative standard curve method. Results: We found that gene expression of fibronectin in group C significantly increased compared to control group, but it was not showed significant change in group B compared to group A. The levels of gene expressions of Cola1 and Cola2 in mRNA did not show any significant changes between normal and radiation groups. Changes of expression of galectin target significantly increased only in group C compared to group A. TGFb1 expressions in group C more than group B showed significant enhancement compared to group A. Conclusion: In summary we can say that 20 Gy of proton exposure of heart tissue may lead to detectable damages in heart cells and may distribute function of them as a component of heart tissue structure in molecular level.

Keywords: gene expression, heart damage, proton irradiation, radiotherapy

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Authors: Umar Saeed

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Peptidyl-prolyl cis/trans isomerases Par14 and Par17 in humans play crucial roles in diverse cellular processes, including protein folding, chromatin remodeling, DNA binding, ribosome biogenesis, and cell cycle progression. However, the effects of Par14 and Par17 on viral replication have been explored to a limited extent. We first time discovered their influential roles in promoting Hepatitis B Virus replication. In this study, we observed that in the presence of HBx, either Par14 or Par17 could upregulate HBV replication. However, in the absence of HBx, neither Par14 nor Par17 had any effect on replication. Their mechanism of action involves binding to specific motifs within HBc and HBx proteins. Notably, they target the conserved 133Arg-Pro134 (RP) motif of HBc and the 19RP20-28RP29 motifs of HBx. This interaction is fundamental for the stability of HBx, core particles, and HBc. Par14 and Par17 exhibit versatility by binding both outside and inside core particles, thereby facilitating core particle assembly through their participation in HBc dimer-dimer interactions. NAGE and immunoblotting analyses unveiled the binding of Par14/Par17 to core particles. Co-immunoprecipitation experiments further demonstrated the interaction of Par14/Par17 with core particle assembly-defective and dimer-positive HBc-Y132A. It's essential to emphasize that R133 is the key residue in the HBc RP motif that governs their interaction with Par14/Par17. Chromatin immunoprecipitation conducted on HBV-infected cells elucidated the participation of residues S19 and E46/D74 in Par14 and S44 and E71/D99 in Par17 in the recruitment of 133RP134 motif-containing HBc into cccDNA. Depleting PIN4 in liver cell lines results in a significant reduction in cccDNA levels, pgRNA, sgRNAs, HBc, core particle assembly, and HBV DNA synthesis. Notably, parvulin inhibitors like juglone and PiB have proven to be effective in substantially reducing HBV replication. These inhibitors weaken the interaction between HBV core particles and Par14/Par17, underscoring the dynamic nature of this interaction. It's also worth noting that specific Par14/Par17 inhibitors hold promise as potential therapeutic options for chronic hepatitis B.

Keywords: Par14Par17, HBx, HBc, cccDNA, HBV

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Authors: Fei Sun, Meilan Xu, Jianghui Zhu, Maria Stefanie Dwiyanti, Cheolwoo Park, Fanjiang Kong, Baohui Liu, Tetsuya Yamada, Jun Abe

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Reduced or lack of sensitivity to long daylengths is a key character for soybean, a short-day crop, to adapt to higher latitudinal environments. However, the photoperiod-insensitivity often results in a reduction of the duration of vegetative growth and final yield. To overcome this limitation, a photoperiod insensitive line (RIL16) was developed in this study that delayed flowering from the recombinant inbred population derived from a cross between a photoperiod-insensitive cultivar AGS292 and a late-flowering Thai cultivar K3. Expression analyses under SD and LD conditions revealed that the expression levels of FLOWERING LOCUS T (FT) orthologues, FT2a and FT5a, were lowered in RIL16 relative to AGS292, although the expression of E1, a soybean-specific suppressor for FTs, was inhibited in both conditions. A soybean orthologue of TARGET OF EAT1 (TOE1), another suppressor of FT, showed an upregulated expression in RIL16, which appeared to reflect a lower expression of miR172a. Our data suggest that the delayed flowering of RIL16 most likely is controlled by genes involved in an age-dependent pathway in flowering. The QTL analysis based on 1,125 SNPs obtained from Restriction Site Associated DNA Sequencing revealed two major QTLs for flowering dates in Chromosome 16 and two minor QTLs in Chromosome 4, all of which accounted for 55% and 48% of the whole variations observed in natural day length and artificially-induced long day length conditions, respectively. The intervals of the major QTLs harbored FT2a and FT5a, respectively, on the basis of annotated genes in the Williams 82 reference genome. Sequencing analysis further revealed a nonsynonymous mutation in FT2a and an SNP in the 3′ UTR region of FT5a. A further study may elucidate a detailed mechanism underlying the QTL for late flowering. The alleles from K3 at the two QTLs can be used singly or in combination to retain an appropriate duration of vegetative growth to maximize the final yield of photoperiod-insensitive soybeans.

Keywords: FT genes, miR72a, photoperiod-insensitive, soybean flowering

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