Search results for: biological invasion

Authors: Claus Bier, Dennis Binder, Alexander Gruenberger, Dagmar Drobietz, Dietrich Kohlheyer, Anita Loeschcke, Karl Erich Jaeger, Thomas Drepper, Joerg Pietruszka

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Light absorbing chromophoric systems are important optogenetic tools for biotechnical and biophysical investigations. Processes such as fluorescence or photolysis can be triggered by light-absorption of chromophores. These play a central role in life science. Photocaged compounds belong to such chromophoric systems. The photo-labile protecting groups enable them to release biologically active substances with high temporal and spatial resolution. The properties of photocaged compounds are specified by the characteristics of the caging group as well as the characteristics of the linked effector molecule. In our research, we work with different types of photo-labile protecting groups and various effector molecules giving us possible access to a large library of caged compounds. As a function of the caged effector molecule, a nearly limitless number of biological systems can be directed. Our main interest focusses on photocaging carbohydrates (e.g. arabinose) and their derivatives as effector molecules. Based on these resulting photocaged compounds a precisely controlled photoinduced gene expression will give us access to studies of numerous biotechnological and synthetic biological applications. It could be shown, that the regulation of gene expression via light is possible with photocaged carbohydrates achieving a higher-order control over this processes. With the one-step cleavable photocaged carbohydrate, a homogeneous expression was achieved in comparison to free carbohydrates.

Keywords: bacterial gene expression, biotechnology, caged compounds, carbohydrates, optogenetics, photo-removable protecting group

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Authors: Sara Seddiki, Maazouzi Abdelhak

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The main objective of this modest work is to follow the physicochemical and bacteriological evolution of the wastewater from the town of Bechar subjected to purification by filtration according to various local supports, namely Sable and Terrill by reducing nuisances that undergo the receiving environment (Oued Bechar) and therefore make this water source reusable in different areas. The study first made it possible to characterize the urban wastewater of the Bechar wadi, which presents an environmental threat, thus allowing an estimation of the pollutant load, the chemical oxygen demand COD (145 mg / l) and the biological oxygen demand BOD5 (72 mg / l) revealed that these waters are less biodegradable (COD / BOD5 ratio = 0.62), have a fairly high conductivity (2.76 mS/cm), and high levels of mineral matter presented by chlorides and sulphates 390 and 596.1 mg / l respectively, with a pH of 8.1. The characterization of the sand dune (Beni Abbes) shows that quartz (97%) is the most present mineral. The granular analysis allowed us to determine certain parameters like the uniformity coefficient (CU) and the equivalent diameter, and scanning electron microscope (SEM) observations and X-ray analysis were performed. The study of filtered wastewater shows satisfactory and very encouraging treatment results, with complete elimination of total coliforms and streptococci and a good reduction of total aerobic germs in the sand and clay-sand filter. A good yield has been reported in the sand Terrill filter for the reduction of turbidity. The rates of reduction of organic matter in terms of the biological oxygen demand, in chemical oxygen demand recorded, are of the order of 60%. The elimination of sulphates is 40% for the sand filter.

Keywords: urban wastewater, filtration, bacteriological and physicochemical parameters, sand, Terrill, Oued Bechar

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Authors: De Biase Ilaria, Della Rocca Mattia

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Our work aims to highlight the role played by cognitive biases in the reception of environmental information, including scientific communication from expert to the lay public, especially in relationship with environmental data coming from biological and biotechnological recording. Some alternative strategies are suggested in order to maximize public awareness on environmental changes.

Keywords: science communication, environment and psychology, cognitive biases, environmental awareness

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Authors: Stephen Daniel Iduh, Evans Chidi Egwin, Oluwatosin Kudirat Shittu

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The process for the development of reliable and eco-friendly metallic Nanoparticles is an important step in the field of Nanotechnology for biomedical application. To achieve this, use of natural sources like biological systems becomes essential. In the present work, extracellular biosynthesis of gold Nanoparticles using aqueous leave and stembark extracts of K. senegalensis has been attempted. The gold Nanoparticles produced were characterized using High Resolution scanning electron microscopy, Ultra Violet–Visible spectroscopy, zeta-sizer Nano, Energy-Dispersive X-ray (EDAX) Spectroscopy and Fourier Transmission Infrared (FTIR) Spectroscopy. The cytotoxicity of the synthesized gold nanoparticles on MCF-7 cell line was evaluated using MTT assay. The result showed a rapid development of Nano size and shaped particles within 5 minutes of reaction with Surface Plasmon Resonance at 520 and 525nm respectively. An average particle size of 20-90nm was confirmed. The amount of the extracts determines the core size of the AuNPs. The core size of the AuNPs decreases as the amount of extract increases and it causes the shift of Surface Plasmon Resonance band. The FTIR confirms the presence of biomolecules serving as reducing and capping agents on the synthesised gold nanoparticles. The MTT assay shows a significant effect of gold nanoparticles which is concentration dependent. This environment-friendly method of biological gold Nanoparticle synthesis has the potential and can be directly applied in cancer therapy.

Keywords: biosynthesis, gold nanoparticles, characterization, calotropis procera, cytotoxicity

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Authors: Suhair Saleh, Ahmad Aljaberi

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Cationic lipid-mediated delivery of nucleic acids represents an exciting approach for developing therapeutically realistic gene medicines. Elucidation of the molecular and formulation requirements for efficient lipofection is a prerequisite to enhance the biological activity of such delivery systems. To this end, the in vitro lipofection activity of the ionizable asymmetric 1,2-dialkoylamidopropane-based derivatives bearing single primary amine group as the cationic head group was evaluated. The electrostatic interactions of these cationic lipids with plasmid DNA in physiologically relevant medium were investigated by means of gel electrophoresis retardation and Eth-Br quenching assays. The effect of the presence of the helper lipid on these interactions was evaluated. The physicochemical properties of these lipids in terms of bilayer fluidity and extent of ionization were investigated using fluorescence anisotropy and surface potential techniques, respectively. The results showed that only the active lipid, 1,2lmp[5], existed in a liquid crystalline state at physiological temperature. Moreover, the extent of ionization of this lipid in assemblies was significantly higher that it's saturated analogues. Inclusion of the helper lipid DOPE improved the encapsulation and association between 1,2lmp[5] and plasmid DNA, which was reflected by the significant boost of lipofection activity of the 1,2lmp[5]/DOPE formulation as compared to the lipid alone. In conclusion, membrane fluidity and sufficient protonation of ionizable cationic lipid are required for efficient association and encapsulation of plasmid DNA and promoting improved in vitro lipofection activity.

Keywords: cationic lipids, gene delivery, lipofection, membrane fluidity, helper lipids, surface potential

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Authors: Khushbu Priyadarshi, Chandramani Pathak

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Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.

Keywords: drug resistance, gallic acid, PAMAM dendrimer, P-glycoprotein

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Authors: Omoruyi Gold Idemudia, Alexander P. Sadimenko

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The synthesis of N-heterocycles with novel properties, having broad spectrum biological activities that may become alternative medicinal drugs, have been attracting a lot of research attention due to the emergence of medicinal drug’s limitations such as disease resistance and their toxicity effects among others. Acylpyrazolones have been employed as pharmaceuticals as well as analytical reagent and their application as coordination complexes with transition metal ions have been well established. By way of a condensation reaction with amines acylpyrazolone ketones form a more chelating and superior group of compounds known as azomethines. 4-propyl-3-methyl-1-phenyl-2-pyrazolin-5-one was reacted with phenylhydrazine to get a new phenylhydrazone which was further reacted with aqueous solutions of palladium and platinum salts, in an effort towards the discovery of transition metal based synthetic drugs. The compounds were characterized by means of analytical, spectroscopic, thermogravimetric analysis TGA, as well as x-ray crystallography. 4-propyl-3-methyl-1-phenyl-2-pyrazolin-5-one phenylhydrazone crystallizes in a triclinic crystal system with a P-1 (No. 2) space group based on x-ray crystallography. The bidentate ON ligand formed a square planar geometry on coordinating with metal ions based on FTIR, electronic and NMR spectra as well as magnetic moments. Reported compounds showed antibacterial activities against the nominated bacterial isolates using the disc diffusion technique at 20 mg/ml in triplicates. The metal complexes exhibited a better antibacterial activity with platinum complex having an MIC value of 0.63 mg/ml. Similarly, ligand and complexes also showed antioxidant scavenging properties against 2, 2-diphenyl-1-picrylhydrazyl DPPH radical at 0.5mg/ml relative to ascorbic acid (standard drug).

Keywords: acylpyrazolone, antibacterial studies, metal complexes, phenylhydrazone, spectroscopy

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Authors: Zakia Fatima, Liu Lu, Donghao Li

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The analysis of biological fluid metabolites holds significant importance in various areas, such as medical research, food science, and public health. Investigating the levels and distribution of nutrients and their metabolites in biological samples allows researchers and healthcare professionals to determine nutritional status, find hypovitaminosis or hypervitaminosis, and monitor the effectiveness of interventions such as dietary supplementation. Moreover, analysis of nutrient metabolites provides insight into their metabolism, bioavailability, and physiological processes, aiding in the clarification of their health roles. Hence, the exploration of a distinct, efficient, eco-friendly, and simpler methodology is of great importance to evaluate the metabolic content of complex biological samples. In this work, a green and rapid analytical method based on an automated online two-dimensional microscale carbon fiber/activated carbon fiber fractionation system and time-of-flight mass spectrometry (2DμCFs-TOF-MS) was used to evaluate metabolites of urine samples after oral administration of food supplements. The automated 2DμCFs instrument consisted of a microcolumn system with bare carbon fibers and modified carbon fiber coatings. Carbon fibers and modified carbon fibers exhibit different surface characteristics and retain different compounds accordingly. Three kinds of mobile-phase solvents were used to elute the compounds of varied chemical heterogeneities. The 2DμCFs separation system has the ability to effectively separate different compounds based on their polarity and solubility characteristics. No complicated sample preparation method was used prior to analysis, which makes the strategy more eco-friendly, practical, and faster than traditional analysis methods. For optimum analysis results, mobile phase composition, flow rate, and sample diluent were optimized. Water-soluble vitamins, fat-soluble vitamins, and amino acids, as well as 22 vitamin metabolites and 11 vitamin metabolic pathway-related metabolites, were found in urine samples. All water-soluble vitamins except vitamin B12 and vitamin B9 were detected in urine samples. However, no fat-soluble vitamin was detected, and only one metabolite of Vitamin A was found. The comparison with a blank urine sample showed a considerable difference in metabolite content. For example, vitamin metabolites and three related metabolites were not detected in blank urine. The complete single-run screening was carried out in 5.5 minutes with the minimum consumption of toxic organic solvent (0.5 ml). The analytical method was evaluated in terms of greenness, with an analytical greenness (AGREE) score of 0.72. The method’s practicality has been investigated using the Blue Applicability Grade Index (BAGI) tool, obtaining a score of 77. The findings in this work illustrated that the 2DµCFs-TOF-MS approach could emerge as a fast, sustainable, practical, high-throughput, and promising analytical tool for screening and accurate detection of various metabolites, pharmaceuticals, and ingredients in dietary supplements as well as biological fluids.

Keywords: metabolite analysis, sustainability, carbon fibers, urine.

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Authors: Sameen Tariq, Saira Bano, Sayyada Ghufrana Nadeem

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The Ocean, which covers over 70% of the world's surface, is wealthy in biodiversity as well as a rich wellspring of microorganisms with huge potential. The oceanic climate is home to an expansive scope of plants, creatures, and microorganisms. Marine microbial networks, which incorporate microscopic organisms, infections, and different microorganisms, enjoy different benefits in biotechnological processes. Samples were collected from marine environments, including soil and water samples, to cultivate the uncultured marine organisms by using Zobell’s medium, Sabouraud’s dextrose agar, and casein media for this purpose. Following isolation, we conduct microscopy and biochemical tests, including gelatin, starch, glucose, casein, catalase, and carbohydrate hydrolysis for further identification. The results show that more gram-positive and gram-negative bacteria. The isolation process of marine organisms is essential for understanding their ecological roles, unraveling their biological secrets, and harnessing their potential for various applications. Marine organisms exhibit remarkable adaptations to thrive in the diverse and challenging marine environment, offering vast potential for scientific, medical, and industrial applications. The isolation process plays a crucial role in unlocking the secrets of marine organisms, understanding their biological functions, and harnessing their valuable properties. They offer a rich source of bioactive compounds with pharmaceutical potential, including antibiotics, anticancer agents, and novel therapeutics. This study is an attempt to explore the diversity and dynamics related to marine microflora and their role in biofilm formation.

Keywords: marine microorganisms, ecosystem, fungi, biofilm, gram-positive, gram-negative

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Authors: Ingrid R. Avanzi, Marcela dos P. G. Baltazar, Louise H. Gracioso, Luciana J. Gimenes, Bruno Karolski, Elen A. Perpetuo, Claudio Auguto Oller do Nascimento

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In this study was developed a natural process using a biological system for the uptake of Copper and possible removal of copper from wastewater by dead biomass of the strain Burkholderia cenocepacia. Dead and live biomass of Burkholderia cenocepacia was used to analyze the equilibrium and kinetics of copper biosorption by this strain in function of the pH. Living biomass exhibited the highest biosorption capacity of copper, 50 mg g−1, which was achieved within 5 hours of contact, at pH 7.0, temperature of 30°C, and agitation speed of 150 rpm. The dead biomass of Burkholderia cenocepacia may be considered an efficiently bioprocess, being fast and low-cost to production of copper and also a probably nano-adsorbent of this metal ion in wastewater in bioremediation process. In this study was developed a natural process using a biological system for the uptake of Copper and possible removal of copper from wastewater by dead biomass of the strain Burkholderia cenocepacia. Dead and live biomass of Burkholderia cenocepacia was used to analyze the equilibrium and kinetics of copper biosorption by this strain in function of the pH. Living biomass exhibited the highest biosorption capacity of copper, 50 mg g−1, which was achieved within 5 hours of contact, at pH 7.0, temperature of 30°C, and agitation speed of 150 rpm. The dead biomass of Burkholderia cenocepacia may be considered an efficiently bioprocess, being fast and low-cost to production of copper and also a probably nano-adsorbent of this metal ion in wastewater in bioremediation process.

Keywords: biosorption, dead biomass, biotechnology, copper recovery

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Authors: Jamal Radaideh

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Al-Asfar Lake is located about 14 km east of Al-Ahsa and is one of the most important wetland lakes in the Al Ahsa/Eastern Province of Saudi Arabia. Al-Ahsa is may be the largest oasis in the world, having an area of 20,000 hectares, in addition, it is of the largest and oldest agricultural centers in the region. The surplus farm irrigation water beside additional water supplied by treated wastewater from Al-Hofuf sewage station is collected by a drainage network and discharged into Al-Asfar Lake. The lake has good wetlands, sand dunes as well as large expanses of open and shallow water. Salt tolerant vegetation is present in some of the shallow areas around the lake, and huge stands of Phragmites reeds occur around the lake. The lake presents an important habitat for wildlife and birds, something not expected to find in a large desert. Although high evaporation rates in the range of 3250 mm are common, the water remains in the evaporation lakes during all seasons of the year is used to supply cattle with drinking water and for aquifer recharge. Investigations showed that high concentrations of nitrogen (N), phosphorus (P), biological oxygen demand (BOD), chemical oxygen demand (COD) and salinity discharge to Al Asfar Lake from the D2 drain exist. It is expected that the majority of BOD, COD and N originates from wastewater discharge and leachate from surplus irrigation water which also contribute to the majority of P and salinity. The significant content of nutrients and biological oxygen demand reduces available oxygen in the water. The present project aimed to improve the water quality of the lake using constructed wetland trains which will be built around the lake. Phragmites reeds, which already occur around the lake, will be used.

Keywords: Al Asfar lake, constructed wetland, water quality, water treatment

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Authors: Imme Petersen, Wiebke Sick, Regine Kollek

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In systems medicine, high-throughput technologies produce large amounts of data on different biological and pathological processes, including (disturbed) gene expressions, metabolic pathways and signaling. The large volume of data of different types, stored in separate databases and often located at different geographical sites have posed new challenges regarding data handling and processing. Tools based on bioinformatics have been developed to resolve the upcoming problems of systematizing, standardizing and integrating the various data. However, the heterogeneity of data gathered at different levels of biological complexity is still a major challenge in data analysis. To build multilayer disease modules, large and heterogeneous data of disease-related information (e.g., genotype, phenotype, environmental factors) are correlated. Therefore, a great deal of attention in systems medicine has been put on data standardization, primarily to retrieve and combine large, heterogeneous datasets into standardized and incorporated forms and structures. However, this data-centred concept of standardization in systems medicine is contrary to the debate in science and technology studies (STS) on standardization that rather emphasizes the dynamics, contexts and negotiations of standard operating procedures. Based on empirical work on research consortia that explore the molecular profile of diseases to establish systems medical approaches in the clinic in Germany, we trace how standardized data are processed and shaped by bioinformatics tools, how scientists using such data in research perceive such standard operating procedures and which consequences for knowledge production (e.g. modeling) arise from it. Hence, different concepts and meanings of standardization are explored to get a deeper insight into standard operating procedures not only in systems medicine, but also beyond.

Keywords: data, science and technology studies (STS), standardization, systems medicine

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Authors: Pakize Neslihan Taslı, Alev Cumbul, Gul Merve Yalcın, Fikrettin Sahin

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A key experimental trial in the regeneration of large oral and craniofacial defects is the neogenesis of osseous and ligamentous interfacial structures. Currently, oral regenerative medicine strategies are unpredictable for repair of tooth supporting tissues destroyed as a consequence of trauma, chronic infection or surgical resection. A different approach combining the gel-casting method with Hydroxy Apatite HA-based scaffold and different cell lineages as a hybrid system leads to successively mimic the early stage of tooth development, in vitro. HA is widely accepted as a bioactive material for guided bone and tooth regeneration. In this study, it was reported that, HA porous scaffold preparation, characterization and evaluation of structural and chemical properties. HA is the main factor that exists in tooth and it is in harmony with structural, biological, and mechanical characteristics. Here, this study shows mimicking immature tooth at the late bell stage design and construction of HA scaffolds for cell transplantation of human Adipose Stem Cells (hASCs), human Bone Marrow Stem Cells (hBMSCs) and Gingival Epitelial cells for the formation of human tooth dentin-pulp-enamel complexes in vitro. Scaffold characterization was demonstrated by SEM, FTIR and pore size and density measurements. The biological contraction of dental tissues against each other was demonstrated by mRNA gene expressions, histopatologic observations and protein release profile by ELISA tecnique. The tooth shaped constructs with a pore size ranging from 150 to 300 µm arranged by gathering right amounts of materials provide interconnected macro-porous structure. The newly formed tissue like structures that grow and integrate within the HA designed constructs forming tooth cementum like tissue, pulp and bone structures. These findings are important as they emphasize the potential biological effect of the hybrid scaffold system. In conclusion, this in vitro study clearly demonstrates that designed 3D scaffolds shaped as a immature tooth at the late bell stage were essential to form enamel-dentin-pulp interfaces with an appropriate cell and biodegradable material combination. The biomimetic architecture achieved here is providing a promising platform for dental tissue engineering.

Keywords: tooth regeneration, tissue engineering, adipose stem cells, hydroxyapatite tooth engineering, porous scaffold

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Authors: Sarah A. Harris, Fleur L. McIntyre, Paola T. Chivers, Benjamin G. Piggott, Fiona H. Farringdon

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Australian Rules Football (ARF) is an invasion based, contact field sport with over one million participants. The contact nature of the game increases exposure to all injuries, including head trauma. Evidence suggests that both concussion and sub-concussive traumas such as head knocks may damage the brain, in particular the prefrontal cortex. The prefrontal cortex may not reach full maturity until a person is in their early twenties with males taking longer to mature than females. Repeated trauma to the pre-frontal cortex during maturation may lead to negative social, cognitive and emotional effects. It is also during this period that males exhibit high levels of risk taking behaviours. Risk propensity and the incidence of injury is an unexplored area of research. Little research has considered if the level of player’s (especially younger players) risk propensity in everyday life places them at an increased risk of injury. Hence the current study, investigated if a relationship exists between risk propensity and self-reported injuries including diagnosed concussion and head knocks, among male ARF players aged 18 to 31 years. Method: The study was conducted over 22 weeks with one West Australian Football League (WAFL) club during the 2015 competition. Pre-season risk propensity was measured using the 7-item self-report Risk Propensity Scale. Possible scores ranged from 9 to 63, with higher scores indicating higher risk propensity. Players reported their self-perceived injuries (concussion, head knocks, upper body and lower body injuries) fortnightly using the WAFL Injury Report Survey (WIRS). A unique ID code was used to ensure player anonymity, which also enabled linkage of survey responses and injury data tracking over the season. A General Linear Model (GLM) was used to analyse whether there was a relationship between risk propensity score and total number of injuries for each injury type. Results: Seventy one players (N=71) with an age range of 18.40 to 30.48 years and a mean age of 21.92 years (±2.96 years) participated in the study. Player’s mean risk propensity score was 32.73, SD ±8.38. Four hundred and ninety five (495) injuries were reported. The most frequently reported injury was head knocks representing 39.19% of total reported injuries. The GLM identified a significant relationship between risk propensity and head knocks (F=4.17, p=.046). No other injury types were significantly related to risk propensity. Discussion: A positive relationship between risk propensity and head trauma in contact sports (specifically WAFL) was discovered. Assessing player’s risk propensity therefore, may identify those more at risk of head injuries. Potentially leading to greater monitoring and education of these players throughout the season, regarding self-identification of head knocks and symptoms that may indicate trauma to the brain. This is important because many players involved in WAFL are in their late teens or early 20’s hence, may be at greater risk of negative outcomes if they experience repeated head trauma. Continued education and research into the risks associated with head injuries has the potential to improve player well-being.

Keywords: football, head injuries, injury identification, risk

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Authors: Irit Rutman Halili, Tehila Zvulun, Natali Elgabsi, Revaya Cohen, Shlomo Sarig

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Active sand dunes (ASD) may cause significant damage to field crops and livelihood, and therefore, it is necessary to find a treatment that would enhance ADS soil stability. Biological soil crusts (biocrusts) contain microorganisms on the soil surface. Metabolic polysaccharides secreted by biocrust cyanobacteria glue the soil particles into aggregates, thereby stabilizing the soil surface. Filter cake powder (FCP) is a waste by-product in the final stages of the production of sugar from sugarcane, and its disposal causes significant environmental pollution. FCP contains high concentrations of polysaccharides and has recently been shown to be soil stability enhancing agent in ASD. It has been reported that adding FCP to the ASD soil surface by dispersal significantly increases the level of penetration resistance of soil biocrust (PRSB) nine weeks after a single treatment. However, it was not known whether a similar effect could be obtained by administering the FCP in liquid form by means of spraying. It has now been found that spraying a water solution of FCP onto the ASD soil surface significantly increased the level of penetration resistance of soil biocrust (PRSB) three weeks after a single treatment. These results suggest that FCP spraying can be used as a short-term soil stability-enhancing agent for ASD, while administration by dispersal might be more efficient over the long term. Finally, an additional benefit of using FCP as a soil stabilizer, either by dispersal or by spraying, is the reduction in environmental pollution that would otherwise result from the disposal of FCP solid waste.

Keywords: active sand dunes, filter cake powder, biological soil crusts, penetration resistance of soil biocrust

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Authors: Amber C. W. Vandepoele, Michael A. Marciano

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Biological analyses frequently focus on single organisms, however many times, the biological sample consists of more than the target organism; for example, human microbiome research targets bacterial DNA, yet most samples consist largely of human DNA. Therefore, there would be an advantage to removing these contaminating organisms. Conversely, some analyses focus on a single organism but would greatly benefit from the additional information regarding the other organismal components of the sample. Forensic analysis is one such example, wherein most forensic casework, human DNA is targeted; however, it typically exists in complex non-pristine sample substrates such as soil or unclean surfaces. These complex samples are commonly comprised of not just human tissue but also microbial and plant life, where these organisms may help gain more forensically relevant information about a specific location or interaction. This project aims to optimize a ‘phylogenetic’ differential extraction method that will separate mammalian, bacterial and plant cells in a mixed sample. This is accomplished through the use of size exclusion separation, whereby the different cell types are separated through multiple filtrations using 5 μm filters. The components are then lysed via differential enzymatic sensitivities among the cells and extracted with minimal contribution from the preceding component. This extraction method will then allow complex DNA samples to be more easily interpreted through non-targeting sequencing since the data will not be skewed toward the smaller and usually more numerous bacterial DNAs. This research project has demonstrated that this ‘phylogenetic’ differential extraction method successfully separated the epithelial and bacterial cells from each other with minimal cell loss. We will take this one step further, showing that when adding the plant cells into the mixture, they will be separated and extracted from the sample. Research is ongoing, and results are pending.

Keywords: DNA isolation, geolocation, non-human, phylogenetic separation

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Authors: Shuji Fujimoto, Fumiko Kojima, Mika Shigematsu

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Biorisk management (Biosafety/Biosecurity) is required for anyone working in a clinical laboratory (including medical/clinical research laboratories) where infectious agents and potentially hazardous biological materials are examined/stored. Proper education and training based on international standards of biorisk management should be provided not only as a part of laboratory safety program in work place but also as a part of introductory training at educational institutions for continuity and to elevate overall baseline of the biorisk management. We reported results of the pilot study of biorisk management education for graduate students majored in laboratory diagnostics previously. However, postgraduate education is still late in their profession and the participants’ interview also revealed importance and demands of earlier biorisk management education for undergraduates. The aim of this study is to identify the need for biosafety/biosecurity education and training program which is designed for undergraduate students who are entering the profession in clinical microbiology. We modified the previous program to include more basic topics and explanations (risk management, principles of safe clinical lab practices, personal protective equipment, disinfection, disposal of biological substances) and provided incorporating in the routine educational system for faculty of medical sciences in Kyushu University. The results of the pre and post examinations showed that the knowledge of the students on biorisk control had developed effectively as a proof of effectiveness of the program even in the undergraduate students. Our study indicates that administrating the basic biorisk management program in the earlier stage of learning will add positive impact to the understanding of biosafety to the health professional education.

Keywords: biorisk management, biosafety, biosecurity, clinical microbiology, education for undergraduates

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Authors: Ryan Houser

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Introduction: Previous studies have found that public health systems within the United States are inadequately prepared for an act of biological terrorism. As the COVID-19 pandemic continues, few studies have evaluated bioterrorism preparedness of Emergency Medical Services, even in the accelerating environment of biothreats. Methods: This study utilized an Internet-based survey to assess the level of preparedness and willingness to respond to a bioterrorism attack and identify factors that predict preparedness and willingness among Nebraska EMS (Emergency Medical Services ) providers. The survey was available for one month in 2021, during which 190 EMS providers responded to the survey. Results: Only 56.8% of providers were able to recognize an illness or injury as potentially resulting from exposure to a CBRN agent. The provider Clinical Competency levels range from a low of 13.6% (ability to initiate patient care within his/her professional scope of practice and arrange for prompt referral appropriate to the identified condition(s)) to a high of 74% (the ability to respond to an emergency within the emergency management system of his/her practice, institution and community). Only 10% of the respondents are both willing and able to effectively function in a bioterror environment. Discussion: In order to effectively prepare for and respond to a bioterrorist attack, all levels of the healthcare system need to have the clinical skills, knowledge, and abilities necessary to treat patients exposed. Policy changes and increased focus on training and drills are needed to ensure a prepared EMS system which is crucial to a resilient state. EMS entities need to be aware of the extent of their available workforce so that the country can be prepared for the increasing threat of bioterrorism or other novel emerging infectious disease outbreaks. A resilient nation relies on a prepared set of EMS providers who are willing to respond to biological terrorism events.

Keywords: bioterrorism, prehospital, EMS, disaster, emergency, medicine, preparedness, policy

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Authors: Emad Rashwan, M. Mousa, Ayman EL Sabagh, Celaleddin Barutcular

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Flax is a potential winter crop for Egypt that can be grown for both seed and fiber. The study was conducted during two successive winter seasons of 2013/2014, and 2014/2015 in the experimental farm of El-Gemmeiza Agricultural Research Station, Agriculture research Centre, Egypt. The objective of this work was to evaluate the effect of irrigation intervals (25, 35 and 45) on the seed yield and quality of flax cultivars (Sakha1, Giza9 and Giza10). Obtained results indicate that highly significant for all studied traits among irrigation intervals except oil percentage that was not significant in both seasons. Irrigated flax plants every 35 days gave the maximum values for all characters. In contrast, irrigation every 45 days gave the minimum values for all studied characters under this study. In respect to cultivars, significant differences in most yield and quality characters were found. Furthermore, the performance of Sakha1 cultivar was superior in total plant height, main stem diameter, seed index, seed, oil, biological and straw yield /ha as well as fiber length and fiber fineness. Meanwhile, Giza9 and Giza10 cultivars were surpassed in fiber yield/hand fiber percentage, respectively. The interactions between irrigation intervals and flax cultivars were highly significant for total plant height, main stem diameter, seed, oil, biological and straw yields /ha. Based on the results, all flax cultivars recorded the maximum values for major traits were measured under irrigation of flax plants every 35 days.

Keywords: flax, fiber, irrigation intervals, oil, seed yield

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Authors: M. A. Askari, M. A. Nazari, P. Perrier, Y. Payan

Abstract:

Growth and remodeling of biological structures have gained lots of attention over the past decades. Determining the response of the living tissues to the mechanical loads is necessary for a wide range of developing fields such as, designing of prosthetics and optimized surgery operations. It is a well-known fact that biological structures are never stress-free, even when externally unloaded. The exact origin of these residual stresses is not clear, but theoretically growth and remodeling is one of the main sources. Extracting body organs from medical imaging, does not produce any information regarding the existing residual stresses in that organ. The simplest cause of such stresses is the gravity since an organ grows under its influence from its birth. Ignoring such residual stresses might cause erroneous results in numerical simulations. Accounting for residual stresses due to tissue growth can improve the accuracy of mechanical analysis results. In this paper, we have implemented a computational framework based on fixed-point iteration to determine the residual stresses due to growth. Using nonlinear continuum mechanics and the concept of fictitious configuration we find the unknown stress-free reference configuration which is necessary for mechanical analysis. To illustrate the method, we apply it to a finite element model of healthy human face whose geometry has been extracted from medical images. We have computed the distribution of residual stress in facial tissues, which can overcome the effect of gravity and cause that tissues remain firm. Tissue wrinkles caused by aging could be a consequence of decreasing residual stress and not counteracting the gravity. Considering these stresses has important application in maxillofacial surgery. It helps the surgeons to predict the changes after surgical operations and their consequences.

Keywords: growth, soft tissue, residual stress, finite element method

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Authors: Mouna Baassi, Mohamed Moussaoui, Hatim Soufi, Sanchaita RajkhowaI, Ashwani Sharma, Subrata Sinha, Said Belaaouad

Abstract:

Human Immunodeficiency Virus type 1 protease (HIV-1 PR) is one of the most challenging targets of antiretroviral therapy used in the treatment of AIDS-infected people. The performance of protease inhibitors (PIs) is limited by the development of protease mutations that can promote resistance to the treatment. The current study was carried out using statistics and bioinformatics tools. A series of thirty-three compounds with known enzymatic inhibitory activities against HIV-1 protease was used in this paper to build a mathematical model relating the structure to the biological activity. These compounds were designed by software; their descriptors were computed using various tools, such as Gaussian, Chem3D, ChemSketch and MarvinSketch. Computational methods generated the best model based on its statistical parameters. The model’s applicability domain (AD) was elaborated. Furthermore, one compound has been proposed as efficient against HIV-1 protease with comparable biological activity to the existing ones; this drug candidate was evaluated using ADMET properties and Lipinski’s rule. Molecular Docking performed on Wild Type and Mutant Type HIV-1 proteases allowed the investigation of the interaction types displayed between the proteases and the ligands, Darunavir (DRV) and the new drug (ND). Molecular dynamics simulation was also used in order to investigate the complexes’ stability, allowing a comparative study of the performance of both ligands (DRV & ND). Our study suggested that the new molecule showed comparable results to that of Darunavir and may be used for further experimental studies. Our study may also be used as a pipeline to search and design new potential inhibitors of HIV-1 proteases.

Keywords: QSAR, ADMET properties, molecular docking, molecular dynamics simulation.

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Authors: Ahmed Samy Elnoby

Abstract:

Ailanthus altissima native to Asia which belongs to the family Simaroubaceae was subjected to phytochemical screening and biological investigations. Phytochemical screening revealed the presence of carbohydrates, tannins, sterols, flavonoids and traces of saponins. In addition, quantitative determination of phenolics and flavonoid content were performed. The antimicrobial activity of methanolic extract of the leaves was determined against gram-positive, gram-negative bacteria in addition to fungi using a modified Kirby-Bauer disc diffusion method that was compared with standard discs ampicillin which acts as an antibacterial agent and amphotericin B which acts as an antifungal agent. A high potency was observed against gram-positive bacteria mainly staphylococcus aureus, gram-negative bacteria mainly Escherichia coli and showed no potency against fungi mainly Aspergillus flavus and candida albicans. On the other hand, the antioxidant activity of the extract was determined by 1, 1-diphenyl-2- diphenyl-2-picryl-hydrazil (DPPH). A very low potency was shown by using DPPH for the antioxidant effect so IC50 = 0 ug/ml, IC90 =0 ug /ml and remark gave 47.2 % at 100 ug/ml which is very weak. Cytotoxic activity was determined by using MTT assay (3-4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide) against MCF7 (Human Caucasian breast adenocarcinoma) cell line. A moderate potency was shown by using MCF7 cell line for cytotoxic effect so LC50= 90.2 ug/ml, LC90=139.9 ug/ml and the remark gave 55.2% at 100 ug/ml which is of moderate activity so, Ailanthus altissima can be considered to be a promising antimicrobial agent from natural origin.

Keywords: Ailanthus altissima, TLC, HPLC, anti-microbial activity, antifungal activity, antioxidant, cytotoxic activity

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Authors: K. Bencherif, M. Bellifa

Abstract:

The expected climatic changes in North Africa are the increase of both intensity and frequencies of the summer droughts and a reduction in water availability during growing season. The exiting coppices and forest formations in the national park of Tlemcen are dominated by holm oak, zen oak and cork oak. These opened-fragmented structures don’t seem enough strong so to hope durable protection against climate change. According to the observed climatic tendency, the objective is to analyze the climatic context and its evolution taking into account the eventual behaving of the oak species during the next 20-30 years on one side and the landscaped context in relation with the most adequate sylvicultural models to choose and especially in relation with human activities on another side. The study methodology is based on Climatic synthesis and Floristic and spatial analysis. Meteorological data of the decade 1989-2009 are used to characterize the current climate. An another approach, based on dendrochronological analysis of a 120 years sample Aleppo pine stem growing in the park, is used so to analyze the climate evolution during one century. Results on the climate evolution during the 50 years obtained through climatic predictive models are exploited so to predict the climate tendency in the park. Spatially, in each forest unit of the Park, stratified sampling is achieved so to reduce the degree of heterogeneity and to easily delineate different stands using the GPS. Results from precedent study are used to analyze the anthropogenic factor considering the forecasts for the period 2025-2100, the number of warm days with a temperature over 25°C would increase from 30 to 70. The monthly mean temperatures of the maxima’s (M) and the minima’s (m) would pass respectively from 30.5°C to 33°C and from 2.3°C to 4.8°C. With an average drop of 25%, precipitations will be reduced to 411.37 mm. These new data highlight the importance of the risk fire and the water stress witch would affect the vegetation and the regeneration process. Spatial analysis highlights the forest and the agricultural dimensions of the park compared to the urban habitat and bare soils. Maps show both fragmentation state and forest surface regression (50% of total surface). At the level of the park, fires affected already all types of covers creating low structures with various densities. On the silvi cultural plan, Zen oak form in some places pure stands and this invasion must be considered as a natural tendency where Zen oak becomes the structuring specie. Climate-related changes have nothing to do with the real impact that South-Mediterranean forests are undergoing because human constraints they support. Nevertheless, hardwoods stand of oak in the national park of Tlemcen will face up to unexpected climate changes such as changing rainfall regime associated with a lengthening of the period of water stress, to heavy rainfall and/or to sudden cold snaps. Faced with these new conditions, management based on mixed uneven aged high forest method promoting the more dynamic specie could be an appropriate measure.

Keywords: global warming, mediterranean forest, oak shrub-lands, Tlemcen

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Authors: Rakesh Kumar Soni

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Under free living conditions, human biological clocks show a periodicity of 24 hour for numerous physiological, behavioral and biochemical variables. However, this period is not the original period; rather it merely exhibits synchronization with the solar clock. It is, therefore, most important to investigate characteristics of human circadian clock, essentially in shift workers, who normally confront with contrasting social clocks. Aim of the present study was to investigate rest-activity rhythm and to vouch for the best technique for the computation of periods in this rhythm in subjects randomly selected from different groups of shift workers. The rest-activity rhythm was studied in forty-eight shift workers from three different organizations, namely Newspaper Printing Press (NPP), Chhattisgarh State Electricity Board (CSEB) and Raipur Alloys (RA). Shift workers of NPP (N = 20) were working on a permanent night shift schedule (NS; 20:00-04:00). However, in CSEB (N = 14) and RA (N = 14), shift workers were working in a 3-shift system comprising of rotations from night (NS; 22:00-06:00) to afternoon (AS; 14:00-22:00) and to morning shift (MS; 06:00-14:00). Each subject wore an Actiwatch (AW64, Mini Mitter Co. Inc., USA) for 7 and/or 21 consecutive days, only after furnishing a certificate of consent. One-minute epoch length was chosen for the collection of wrist activity data. Period was determined by using Actiware sleep software (Periodogram), Lomb-Scargle Periodogram (LSP) and Spectral analysis software (Spectre). Other statistical techniques, such as ANOVA and Duncan’s multiple-range test were also used whenever required. A statistically significant circadian rhythm in rest-activity, gauged by cosinor, was documented in all shift workers, irrespective of shift work. Results indicate that the efficiency of the technique to determine the period (τ) depended upon the clipping limits of the τs. It appears that the technique of spectre is more reliable.

Keywords: biological clock, rest activity rhythm, spectre, periodogram

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Authors: Thabiso Michael Mokotjomela, Jasper Knight

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Intensification of agricultural food production in sub-Saharan Africa is of paramount importance as a means of increasing the food security of communities that are already experiencing a range of environmental and socio-economic stresses. However, achieving this aim faces several challenges including ongoing climate change, increased resistance of diseases and pests, extreme environmental degradation partly due to biological invasions, land tenure and management practices, socio-economic developments of rural populations, and national population growth. In particular, non-native plant species tend to display greater adaptation capacity to environmental stress than native species that form important food resource base for human beings, thus suggesting a potential for usage to shift accordingly. Based on review of the historical benefits of non-native plant species in food production in sub-Saharan Africa, we propose that use of non-invasive, non-native plant species and/or the genetic modification of native species might be viable options for future agricultural sustainability in this region. Coupled with strategic foresight planning (e.g. use of biological control agents that suppress plant species’ invasions), the consumptive use of already-introduced non-native species might help in containment and control of possible negative environmental impacts of non-native species on native species, ecosystems and biodiversity, and soil fertility and hydrology. Use of non-native species in food production should be accompanied by low cost agroecology practices (e.g. conservation agriculture and agrobiodiversity) that may promote the gradual recovery of natural capital, ecosystem services, and promote conservation of the natural environment as well as enhance food security.

Keywords: food security, invasive species, agroecology, agrobiodiversity, socio-economic stresses

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Authors: Yassets Egaña, Patricio Núñez, Juan C. Rios, Ivan Balic, Alex Manquez, Yarela Flores, Maria C. Gatica, Sergio Diez De Medina, Rocio Tijaro-Rojas

Abstract:

Globally, humans produce millions of tons of waste per year. An important percentage of this waste is plastic, which frequently ends up in landfills and oceans. During the last decades, the greatest plastics production in history have been made, a few amount of this plastic is recycled, the rest ending up as plastic pollution in soils and seas. Plastic pollution is disastrous for the environment, affecting essential species, quality of consumption water, and some economic activities such as tourism, in different parts of the world. Due to its durability and decomposition on micro-plastics, animals and humans are accumulating a variety of plastic components without having clear their effects on human health, economy, and wildlife. In dry regions as the Atacama Desert, up to 95% of the water consumption comes from underground reservoirs, therefore preventing the soil pollution is an urgent need. This contribution focused on isolating, genotyping and optimizing microorganisms that use plastic waste as the only source of food to construct a batch-type bioreactor able to degrade in a faster way the plastic waste before it gets the desert soils and groundwater consumed by people living in this areas. Preliminary results, under laboratory conditions, has shown an improved degradation of polyethylene when three species of bacteria and three of fungi act on a selected plastic material. These microorganisms have been inoculated in dry soils, initially lacking organic matter, under environmental conditions in the laboratory. Our team designed and constructed a prototype using the natural conditions of the region and the best experimental results.

Keywords: biological breakdown, plastic bags, prototype, desert regions

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Authors: Gajanan M. Sonwane

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Nowadays, the entire pharmaceutical industry is facing the challenge of increasing efficiency and innovation. The major hurdles are the growing cost of research and development and a concurrent stagnating number of new chemical entities (NCEs). Hence, the challenge is to select the most druggable targets and to search the equivalent drug-like compounds, which also possess specific pharmacokinetic and toxicological properties that allow them to be developed as drugs. The present research work includes the studies of developing new anticancer heterocycles by using molecular modeling techniques. The heterocycles synthesized through such methodology are much effective as various physicochemical parameters have been already studied and the structure has been optimized for its best fit in the receptor. Hence, on the basis of the literature survey and considering the need to develop newer anticancer agents, new phenazinamine derivatives were designed by subjecting the nucleus to molecular modeling, viz., GQSAR analysis and docking studies. Simultaneously, these designed derivatives were subjected to in silico prediction of biological activity through PASS studies and then in silico toxicity risk assessment studies. In PASS studies, it was found that all the derivatives exhibited a good spectrum of biological activities confirming its anticancer potential. The toxicity risk assessment studies revealed that all the derivatives obey Lipinski’s rule. Amongst these series, compounds 4c, 5b and 6c were found to possess logP and drug-likeness values comparable with the standard Imatinib (used for anticancer activity studies) and also with the standard drug methotrexate (used for antimitotic activity studies). One of the most notable mutations is the threonine to isoleucine mutation at codon 315 (T315I), which is known to be resistant to all currently available TKI. Enzyme assay planned for confirmation of target selective activity.

Keywords: drug design, tyrosine kinases, anticancer, Phenazinamine

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Authors: Paresh Patel, Harshit Patel

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Superficial fungal skin infection is among the most common skin disease. The drug administration through skin has received attention due to several advantages: Avoidance of significant pre-systemic metabolism, drug levels within the therapeutic window, drugs with short biological half-lives, decreased side effects, the non-invasive character, and very high acceptance.

Keywords: transdermal spray, ketoconazole, Eudragit® RLPO, therapeutic window

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Authors: Marziye Hadian, Alireza Jabbari

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Background: The World Health Organization has identified COVID-19 as a public health emergency and is urging governments to stop the virus transmission by adopting appropriate policies. In this regard, authorities have taken different approaches to cut the chain or controlling the spread of the disease. Now, the questions we are facing include what these approaches are? What tools should be used to implement each preventive protocol? In addition, what is the impact of each approach? Objective: The aim of this study was to determine the approaches to biological epidemics and related prevention tools with an emphasis on COVID-19 disease. Data sources: Databases including ISI web of science, PubMed, Scopus, Science Direct, Ovid, and ProQuest were employed for data extraction. Furthermore, authentic sources such as the WHO website, the published reports of relevant countries, as well as the Worldometer website were evaluated for gray studies. The time-frame of the study was from 1 December 2019 to 30 May 2020. Methods: The present study was a systematic study of publications related to the prevention strategies for the COVID-19 disease. The study was carried out based on the PRISMA guidelines and CASP for articles and AACODS for grey literature. Results: The study findings showed that in order to confront the COVID-19 epidemic, in general, there are three approaches of "mitigation", "active control" and "suppression" and four strategies of "quarantine", "isolation", "social distance" and "lockdown" in both individual and social dimensions to deal with epidemics. Selection and implementation of each approach requires specific strategies and has different effects when it comes to controlling and inhibiting the disease. Key finding: One possible approach to control the disease is to change individual behavior and lifestyle. In addition to prevention strategies, use of masks, observance of personal hygiene principles such as regular hand washing and non-contact of contaminated hands with the face, as well as an observance of public health principles such as sneezing and coughing etiquettes, safe extermination of personal protective equipment, must be strictly observed. Have not been included in the category of prevention tools. However, it has a great impact on controlling the epidemic, especially the new coronavirus epidemic. Conclusion: Although the use of different approaches to control and inhibit biological epidemics depends on numerous variables, however, despite these requirements, global experience suggests that some of these approaches are ineffective. The use of previous experiences in the world, along with the current experiences of countries, can be very helpful in choosing the accurate approach for each country in accordance with the characteristics of that country and lead to the reduction of possible costs at the national and international levels.

Keywords: novel corona virus, COVID-19, approaches, prevention tools, prevention strategies

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Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

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